Companies Developing Modified Plant Varieties as Improved Biofuel Feedstocks (Part 3)

The following are brief profiles of companies that are developing engineered or modified plant varieties for the production of biofuels, that were listed in Part 1 of this blog entry. The profiles are presented in alphabetical order by company name, with 10 companies profiled here following the initial 10 profiles in Part 2 of this entry. These profiles have been adapted or excerpted from company websites and/or other publicly available information, and I don’t assume any liability for the accuracy, comprehensiveness or use of the information. 

FuturaGene PLC (the FGN Group) is a leader in plant genetic research and development for global biofuel, forestry and agricultural markets. The Group was formed by the merger of FGN PLC with CBD Technologies and has integrated the abiotic stress platform with the yield enhancement and processability technology from these entities into a powerful new package for the enhancement of biofuel, forestry and agricultural crops. The FGN Group has strategic agreements with leading commercial companies in the forestry and agricultural sectors around the world and continues to actively partner its technologies. The Group has a strong and growing intellectual property portfolio that addresses environmental stresses in plants such as salinity, drought, cold, and heat and protects its developments on the modification of plant cell walls to enhance plant growth rates, yield and processability of plant fiber. FuturaGene technologies, by allowing plants to grow on marginal lands and to produce more usable fiber for industry, are a major contributor to ensuring food security and enhancing the sustainability of agriculture, forest product and biofuel production. 

FuturaGene’s biofuel program is focused on renewable feedstocks with enhanced yields and processability and which are environmentally sustainable. FuturaGene technology can be used to create biofuel feedstock with increased biomass, shorter crop cycles and improved cellulose accessibility and processability for use in biofuel. FuturaGene’s technology is suitable for a large range of cellulosic biomass crops such as switchgrass, Miscanthus, hybrid poplar and willow which are being developed as biofuel crops. The research approach involves manipulation of genes such as cbd and cel1, which encode cell wall polysaccharide modifying enzymes, so as to change the composition of the plant cell wall. These polysaccharides dissolve in preprocessing of cellulosic biomass, allowing enhanced penetration of  chemicals and enzymes to the cellulose and hemicellulose fractions, thus increasing the efficiency of deconstruction for bioethanol production. 

Infinite Enzymes is generating an improved system of producing commercially available enzymes for converting cellulosic biomass to ethanol and other biobased products using a plant biotechnology platform for manufacturing. The technology utilizes the transgenic maize production system—producing enzymes in the embryo, or germ, of the corn seed. The starch in the endosperm is available to be converted to ethanol, allowing dual utilization of the crop that is being grown for ethanol and additionally the enzymes—enabling very inexpensive enzyme production.  The method combines a propriety transgenic plant-based production system and an integrated agribusiness logistics model which complements the current corn-to-ethanol industry. 

The global demand for biofuels is increasing rapidly and 2nd generation processes using cellulose from trees, corn stalks, rice straw and other sources will be the preferred feedstock. The Infinite Enzymes’ products provide the necessary ingredients to convert crops into fuels and other valuable chemicals. Infinite Enzymes is also involved in producing other enzymes in transgenic plants, as well as commercializing other biobased products technologies.

The primary expression system results in the production of enzymes in transgenic maize kernels. Maize seed is an excellent system in which to produce proteins for industrial applications. Seed provides a stable environment for the protein—over millennia the seed has evolved mechanisms to stably store proteins for later use by the germinating embryo. Processing costs can be kept to a minimum in a commodity crop like maize, allowing inexpensive recovery of various fractions of the grain. One of the major advantages of using the embryo (germ) instead of the endosperm to produce industrial proteins, particularly cellulases, is that the endosperm starch can be separated from the embryo and converted into ethanol. Thus, if the major use of the crop is for ethanol anyway, the production of the germ is a by-product, making the enzyme in the germ virtually free. The oil can be used in food/feed or biodiesel applications, creating another co-product. 

The first target products are cellulases for converting plant biomass into sugars for fermentation into ethanol or other biobased products. The cellulases are the most expensive technology still required for the biomass-to-ethanol process. The transgenic corn seed system is the most cost-effective and volume-permissive system available. Each of the required cellulases for biomass conversion are produced as single activities—endo and exo-cellulases, with high accumulation and genetic stability.  Infinite Enzymes received several permits from the USDA’s Biotechnology Regulatory Services in 2008 and 2009 for field tests in Illinois, Arkansas and Puerto Rico of transgenic corn expressing a cellulase. 

Kaiima is  a  next  generation  seed  and  breeding  company.    The company uses its CGM™ technology to develop new non-GMO crops with dramatically improved productivity and improved land and water-use efficiencies. Use of these new high-performance varieties and thoughtful agricultural practices can enable farmers all over the world to generate more food and more energy while sustaining land and water resources. 

Kaiima’s CGM™ (Clean Genomic Multiplication) technology is a non-transgenic biotechnology platform developed in 2002 that induces clean polyploidy in plants (i.e., multiplying the number of chromosomes found in the plant). It includes a proprietary set of protocols and methods that direct the active chemicals used in the genome-multiplication process away from the sensitive DNA, which stays unharmed, unlike past methods for inducing polyploidy, thus keeping the plant fertile and genetically stable. The company claims that its technology provides advantages including higher plant yield, greater biomass accumulation, enhanced photosynthesis and other features. 

Kaiima is using CGM™ and other advanced and proprietary genomic-based breeding technologies to develop high-yielding energy crops for the production of biodiesel, bioethanol, and biomass energy. The biodiesel strategy involves breeding castor varieties that can yield up to 10 tons of seeds (or 5 tons of oil) per hectare per year compared to the global average of between 1-1.5 tons of seeds. The company expects that these high yields, will make fuel from castor we economically competitive with the price of petroleum.  The company is also collaborating with the prominent rapeseed program at the Anhui Academy of Agriculture in China to increase the yield of their market-leading rapeseed by several tens of percents, for use as a feedstock for biodiesel. Kaiima has also started its own research programs to develop high-yielding varieties of sugarcane for ethanol and eucalyptus for biomass energy production. 

Medicago, Inc. is a biotechnology company focused on developing highly effective and affordable vaccines based on proprietary manufacturing technologies using transient gene expression in plants.  In October 2009, the company announced that it had been awarded a proof of concept contract by the U.S. Army Research, Development and Engineering Command laboratory specifically the Edgewood Chemical Biological Center (“ECBC”) Research & Technology Directorate, to work with ECBC to investigate the affordable production of industrial enzymes in the field of biofuels. This new project builds on Medicago’s proprietary plant-based manufacturing platform and its potential for applications beyond the biological drug market. The company believes that the high cost of enzymes is a major hurdle in the production of biofuels using biomass and that its manufacturing platform could be suitable for the production of affordable enzymes. 

Mendel Biotechnology, Inc. is an innovative biotechnology company serving large agricultural companies with new genetic and chemical solutions and developing novels seeds to serve the bioenergy industry.   Mendel Biotechnology’s vision is that the company’s knowledge about regulation of plant gene and pathway function will enable accelerated improvement in plant varieties, and the delivery of associated services, to meet global agricultural and energy production needs. 

Mendel is focusing on a class of genes encoding transcription factors, which act as master regulators of gene networks. Over a period of approximately five years, Mendel scientists identified essentially all of the transcription factor genes from a model plant species (Arabidopsis thaliana) and systematically analyzed the function of each of the encoded proteins by producing experimental plants that had increased or decreased amounts of the target protein, and assaying for traits such as  abiotic stress tolerance, disease resistance, and metabolic composition. These initial efforts have resulted in a large number of novel discoveries about the function of key transcription factors, their molecular mode of action, and the local genetic networks that they regulate, and this knowledge has enabled Mendel to develop both genetic and chemical approaches to deliver valuable traits to target crops. 

Mendel entered into a strategic long-term collaboration with BP in May 2007 for the development of its BioEnergy Seeds and Feedstocks business. The focus of the collaboration is the development and commercialization of seed products, both conventional and biotech varieties, for dedicated energy crops such as Miscanthus and switchgrass, to serve the emerging 2nd generation biofuels industry both in the United States and abroad. Under this arrangement, Mendel will own the technology developed through the collaboration, and will own and operate the seed and feedstock business, while BP receives royalties on seed sales. 

Mendel’s BioEnergy Seeds (MBS) division is developing new varieties of very productive, non-food energy grasses to enable the delivery of large scale supplies of high value biomass feedstocks produced on marginal and under-utilized lands in a highly sustainable manner. Mendel’s product portfolio includes the largest research and development program in the world focused on the most promising perennial grass species, Miscanthus.  C4 perennial grasses, and, particularly Miscanthus, can collect and store that energy more efficiently than any other system yet devised. Domestication of commercial Miscanthus varieties in the US, combined with continued advances in fuel conversion technologies is an integral component of the country’s renewable energy solution. Mendel is applying its validated trait technology and advanced breeding techniques to develop superior, proprietary Miscanthus varieties and other energy crop products. Additional species in development include high-biomass sorghum through Mendel’s collaboration with MMR Genetics and Richardson Seeds and “Miscanes”. 

Metabolix, Inc. is developing and commercializing sustainable biological solutions for the world’s needs in plastics, chemicals, and energy with unequaled environmental benefits. Known for its Mirel brand of bioplastics, produced in genetically modified microorganisms and plants, Metabolix is also developing proprietary technology that can produce energy from switchgrass, oilseeds, sugarcane and other crops. 

Metabolix has developed considerable expertise in genetic pathway engineering in switchgrass. Switchgrass is a commercially and ecologically attractive, non-food energy crop that is indigenous to North America and is generally considered to be a leading candidate for cellulose-derived production of ethanol and other biofuels. In 2001, Metabolix was awarded a $15 million Industries of the Future cost-shared grant from the U.S. Department of Energy to help fund the development of a biomass biorefinery based on switchgrass. The goal of this five-year program was to produce bioplastic in green tissue plants and, after polymer extraction, use the residual plant biomass for energy generation. The combined polymer and energy production from a biomass biorefinery would reduce the nation’s dependence on foreign oil imports for use in materials feedstock and processing. 

Metabolix also has a collaboration with the Australian Cooperative Research Centre for genetic pathway engineering in sugarcane for co-production of products In February 2008, Metabolix established a strategic research collaboration with the Donald Danforth Plant Science Center to develop an advanced industrial oilseed crop for co-production of bioplastics along with energy. 

Rahan Meristem is an Israeli company with more than 30 years experience in plant propagation and biotechnology, and in the laboratory production of tissue culture plants. In January 2010, the company was reported to have announced its plans to engage in developing protocols for the mass propagation and genetic transformation of castor beans and Jatropha to produce biodiesel. Genetic engineering  can be used to increase plant oil yield, enhance the oxidative stability of the oil, render the plants resistant to biotic and abiotic stress factors, and control plant height for ease of mechanical harvesting. The company has reportedly isolated several genes from unicellular algae that confer resistance to drought and salinity, which it said are expected to greatly improve the efficacy of the culture of biodiesel crops in low rainfall climates. The company’s scientific director was quoted as saying that their main goal is to develop and bring to market transgenic Jatropha and castor bean clones that confer resistance to salinity and drought. 

SG Biofuels is a plant oil company specializing in the development of Jatropha as a low cost, sustainably produced oil that can be used for a variety of bio-based materials including biodiesel and feedstock substitutes for the petrochemical and jet fuel industries. SG Biofuels brings together a world-class leadership team of executive management, energy, biotechnology and agribusiness veterans with a proven track record developing sustainable, for-profit projects in Latin America. The executive team and advisory board include a broad base of successful leaders in technology, energy, government, petrochemical, biotechnology and agriculture. 

SG Biofuels has established a Jatropha Genetic Resource Center (GRC) to further accelerate profitable, large-scale production of Jatropha as a low-cost, sustainable source of feedstock for biofuel. With research sites in San Diego and several Latin American countries, SG Biofuels says that it possesses the largest, most genetically diverse library of Jatropha genetic material in the world. The GRC enables the company’s efforts to drive genetic improvements that will enhance yield, improve agronomic practices and broaden the effective growing range of this promising subtropical crop. This germplasm foundation in combination with modern biotechnological advances and practices is providing the platform for significant improvements in this renewable fuel crop. The company’s scientific team has already identified many strains with promising characteristics and SG Biofuels has begun evaluating thousands of diverse accessions of Jatropha obtained from a range of geographical and climatic conditions. Through additional genetic improvements and breeding, a range of opportunities exist to improve Jatropha’s oil yield and develop improved strains, including those that can further enhance production in colder climates of the United States and other nations. 

In January 2010,  SG Biofuels announced a strategic alliance with Life Technologies Corporation, a leading supplier of products for the life science research market, to advance the development of Jatropha as a sustainable biofuel. The alliance brings together SG Biofuels’ Genetic Resource Center with the advanced biotechnology and synthetic biology tools of Life Technologies. The partnership will initially include sequencing the Jatropha curcas genome, allowing for the rapid introduction of new traits targeted toward increasing the yield of the oil-producing plant. Life Technologies will also become a strategic partner in SG Biofuels. 

In February 2010, SG Biofuels introduced JMax 100, the world’s first elite Jatropha cultivar, providing growers and plantation developers with access to the highest yielding and most profitable Jatropha. The company says that JMax 100 increases the profitability of Jatropha to greater than $400 per acre — more than 300 percent above existing commercial varieties. This equates to more than 350 gallons per acre at $1.39 per gallon, enabling the large-scale growth of the nation’s renewable fuel industry and development opportunities for community farmers, plantation developers and renewable energy investors. 

Syngenta is a world-leading, multinational agribusiness company. Its two main commercial areas  are the sale of seeds and crop protection products. The company was formed in 2000 by the merger of the agricultural businesses of Novartis and AstraZeneca, but its roots extend back 250 years to the earliest industrial predecessor companies of these founder companies. Syngenta has now grown into one of the world’s leading companies with more than 25,000 employees in over 90 countries. 

Syngenta’s major activity in biofuels is a line of transgenic corn expressing an engineered thermostable amylase for enhanced ethanol production. This corn variety, which the company refers to as its “output trait corn amylase”, is sold under the brand name Enogen. Originally known by the internal product name “Corn [Maize] event 3272”, this line was developed using recombinant DNA technology to introduce into corn the amy797E gene and the pmi marker gene. The amy797E gene is derived from alpha-amylase genes from three hyperthermophilic microorganisms of the archaeal order Thermococcales, and it encodes a thermostable AMY797E alpha-amylase enzyme which catalyses the hydrolysis of starch by cleaving the internal alpha-1,4-glucosidic bonds into dextrins, maltose and glucose. The pmi gene from Escherichia coli encodes the phosphomannose isomerase (PMI) enzyme, which allows the plant to utilize mannose as a carbon source. This corn variety has been extensively field tested throughout the world, and has been approved for commercial sale in several countries including Canada (but as of this writing not yet in the U.S., where USDA has been considering approval for the past 2-3 years and where some public opposition has emerged). The company says that use of this corn variety will offer ethanol producers a significant cost advantage of between 8 and 15 cents per gallon. 

Targeted Growth, Inc. (TGI) is a crop biotechnology company focused on developing products with enhanced yield and improved quality for the agriculture and energy industries. Founded in 1998, TGI has developed a technology portfolio based on the principle that regulating cell cycle processes can directly and significantly enhance plant yields.  Over the past ten years, TGI has applied modern breeding and biotechnology techniques to a variety of commercial and emerging crops to produce significant yield increases, already validated over multiple years and in large-scale field trials.  TGI has incorporated modern molecular breeding techniques with targeted genetic adaptations and biotechnology for a comprehensive approach to optimizing the characteristics of plants for use in both food and fuel crops.  TGI’s current development programs include enhancements to corn, soybean, canola, rice, wheat, and Camelina, for purposes including providing improved raw materials while reducing input costs; optimizing agricultural crops for biofuel endproducts, and recapturing non-arable and underutilized land for energy solutions.  

The company is using conventional breeding and biotechnology to enhance the suitability of select energy crops for use as fuels, by improving sugar, starch and oil profiles, as well as characteristics of cell wall formation. For ethanol production TGI is focused on corn and sorghum; for biodiesel, soybean, canola, and Camelina. TGI has field tested engineered Camelina and soybean varieties at numerous locations in the U.S. and Canada for the past several years. 

D. Glass Associates, Inc. is a consulting company specializing in several fields of biotechnology. David Glass, Ph.D. is a veteran of nearly thirty years in the biotech industry, with expertise in patents, technology licensing, industrial biotechnology regulatory affairs, and market and technology assessments. This blog provides back-up and expanded content to complement a presentation Dr. Glass made at the EUEC 2010 conference on February 2, 2010 entitled “Prospects for the Use of Genetic Engineering in Biofuel Production.” The slides from that presentation are available at www.slideshare.net/djglass99.

Companies Developing Modified Plant Varieties as Improved Biofuel Feedstocks (Part 2)

The following are brief profiles of the companies that are developing engineered or modified plant varieties for the production of biofuels, that were listed in Part 1 of this blog entry. The profiles are presented in alphabetical order by company name, with 10 companies profiled here and an additional 10 profiles to follow in Part 3 of this entry. These profiles have been adapted or excerpted from company websites and/or other publicly available information, and I don’t assume any liability for the accuracy, comprehensiveness or use of the information. 

Agragen, LLC is a biotechnology company based in Cincinnati, Ohio, with offices in Helsinki, Finland, and Grand Forks, North Dakota that genetically modifies Camelina sativa to increase agronomically desirable traits, based on breeding expertise and access to germplasm. Camelina is a low input alternative plant for oil used in downstream biofuel production, while also producing a high quality meal comparable to soybean meal. Agragen is reported to have a patent portfolio that includes nine major Camelina-based patents, four patents pending, and numerous additional intellectual property licenses related to the improvement of Camelina. 

In June 2009, Agragen entered into a partnership with Great Plains Oil and Exploration (which calls itself  “The Camelina Company”), aimed at increasing Camelina’s oil content, viability in expanded locations, and resistance to disease, weeds and pests. In August 2009, Great Plains announced that Agragen had filed for patent protection on a novel method designed to increase the tolerance of Camelina to Group 2 herbicides. Agragen’s science team has reportedly introduced specific modifications that increase Camelina’s tolerance to Group 2 herbicides by more than 300-fold in laboratory testing.  This unique approach is designed to make Camelina more tolerant for planting in areas where residual Group 2 herbicides in the soil limit farmer’s cropping options.  The introduction of an herbicide-tolerant Camelina will potentially open up the crop to millions of additional acres of rotational land. The resulting intellectual property will belong to Great Plains, and will expand Great Plains’ germplasm collection, which already comprises the majority of the world’s Camelina germplasm for commercial use. 

In November 2009, Agragen announced the acquisition of intellectual property rights that will increase oil content in Camelina. Through an agreement with the University of Alberta and Agriculture and Agri-Food Canada (AAFC), Agragen will use its patented and patent-pending technology to introduce into Camelina a gene encoding a key enzyme in oil synthesis. Further testing will be needed to demonstrate that the introduction of the gene encoding this enzyme will result in elevation of oil production in Camelina. However, researchers with the University of Alberta and AAFC have already demonstrated in other systems that the introduction of this enzyme results in significant elevation of oil content. In fact, it is expected that the enhancement could increase oil content by as much as five percent. Once ready for commercialization, Agragen plans to transfer the resulting intellectual property to Great Plains. 

In 2005, Agragen proposed to use transgenic flax to make albumin and a recombinant form of omega-3 fatty acids from flax grown in North Dakota. Like several other proposals in the mid-2000s to use transgenic plants to produce pharmaceutical products, this project drew controversy and it is not clear if it ever proceeded to fruition. 

Agrisoma Biosciences, Inc. uses a proprietary chromosome engineering technology (“Engineered Trait Loci”) to create novel bio-energy crops engineered for maximum production of optimized oils for use in renewable fuels. The company’s goal is to create high-yielding, optimized oilseed crops for production of biodiesel. Engineered crops such as Brassica and Jatropha are expected to provide high quality feedstock at reduced costs. 

Agrisoma’s ETL system is used to engineer new chromosome structures (“Engineered Trait Loci”) that carry new combinations of traits within an optimal genetic environment, providing consistency, performance and stability when transformed into a host species.  Unlike common methods that randomly insert traits into the chromosomes of a plant, Agrisoma’s ETL system is a precision technology that introduces traits into a specific, selected chromosome to provide performance and stability advantages.  The ETL system has produced multiple trait combinations that have gone from initial design to the field in under 2 years, demonstrating the speed and efficiency of the ETL technology.  The technology has been successfully applied to a number of major global crops including major food crops and new, emerging industrial crops such as Brassica and Jatropha oilseeds, both highly productive and sustainable bio-energy crops.  

In 2009, Agrisoma conducted its first field tests in Canada, under the regulations of the Canadian Food Inspection Agency, of engineered Brassica and soybean, improved in oil quality, content and seed size.  Agrisoma applied its proprietary ETL process to produce a new soybean line that carries an independent mini-chromosome comprising multiple new regions for efficient gene stacking in soybean. The company says that this variety is the first engineered crop variety based on a minichromosome to be taken to the field and assessed under field conditions. The maturation of the ETL varieties in the field duplicates that of control varieties, showing an identical rate of seed development and pod fill. The company has also been testing engineered oilseed Brassica varieties, have shown significant improvement in oil quality, content and overall seed size. The company says that these trials  have shown that ETL engineered crops can hold up under periods of stresses, particularly during a  year in which the lack of moisture hit many crop production regions very hard The company says that, in regions affected by low moisture, selections of ETL engineered lines showed good flowering and seed set. In regions where moisture has not been a problem, the ETL crops are performing as anticipated. The Brassica field trials took place in Manitoba and Saskatchewan. 

Agrivida, Inc. is an agricultural biotechnology company developing varieties of switchgrass, sugarcane, sorghum, corn, and other energy crops for the production of chemicals, fuels, and bioproducts. By enabling the production of cheap sugars from this non-food, cellulosic biomass, the company expects that its energy crops and processing technology can reduce costs by over 30% for these industrial biotechnology products. 

The key to Agrivida’s technology is the introduction of genetic modifications that allow the plants to produce inactive enzymes within the plant cell wall. After harvest, the enzymes are activated on demand to degrade the cellulose into sugars for downstream production at costs the company says are comparable to $50/bbl petroleum. Because Agrivida’s energy crop varieties produce their own enzymes, the cellulosic biomass can be broken down with less severe pretreatments and without addition of exogenous enzymes. 

In November 2009, Agrivida received two separate awards for the continued development of energy crops expressing cell wall-degrading enzymes. Funding from the United States Department of Agriculture, which could amount to almost $2 million in total, is part of the Biomass Research and Development Initiative, while a $4.6 million award from the Department of Energy is from the Advanced Research Projects Agency – Energy. In awarding the grant, ARPA-E stated that projects like this will lower the cost of biofuels and chemicals and “will help establish a sustainable market for non-food biomass resources to bolster the development of biorefinery jobs and commerce and create carbon neutral transportation fuels”. 

ArborGen is the global leader in improving trees through advanced genetics, and is dedicated to improving the sustainability and productivity of purpose grown working forests, providing more wood on less land while preserving native habitats. Global demand for wood-based products, including renewable energy, is predicted to grow significantly into the foreseeable future. To meet this growing demand for biofuels, ArborGen is researching and developing purpose grown trees to produce cellulosic ethanol as an environmentally friendly and renewable alternative to fossil fuels. The trees ArborGen is targeting, particularly U.S. plantation hardwoods, are ideally suited to produce cellulose-based ethanol from commercial working forests. Among the potential bioenergy products the company is developing are cold tolerant eucalyptus; short rotation hardwoods; and short rotation pine varieties. One of the fastest growing hardwoods, Eucalyptus, along with other hardwoods, has the potential to provide a ready source of biomass for biofuels as well as high quality wood for pulp and paper. ArborGen has conducted numerous field tests of engineered trees, and has obtained a number of recent USDA permits for field testing of Eucalyptus hybrids with altered lignin biosynthesis. 

CanaVialis S.A is a Brazilian company whose mission is to develop and provide sugarcane varieties to growers as well as services, products and expert advice that enable the achievement of the highest degree of productivity in their sugarcane fields. Since its creation in 2003, CanaVialis has applied the best available knowledge, professionals and technologies to the Brazilian sugarcane industry, and the company hopes to be the best and largest developer and supplier of genetic solutions in sugarcane through the use of state-of-the-art technological processes and high-quality employees. 

The company utilizes traditional breeding technologies as well as cutting-edge biotechnology. In a partnership with Alellyx (a Brazilian plant biotech and genomics company that is affiliated with the Monsanto Company), CanaVialis has invested in the improvement of sugarcane with the use of genetic engineering. This partnership has undertaken projects focused on using genetic modification to develop superior sugarcane varieties.  In May 2007, CanaVialis and Alellyx signed an agreement with Monsanto which will allow CanaVialis to make technologies developed by Monsanto available to the sugar and alcohol business sector. The initial products resulting from this agreement will be insect attack resistant and herbicide-tolerant varieties. These technologies will allow CanaVialis customers to increase the productivity of cane fields and decrease production costs. 

Ceres, Inc. develops & markets low-carbon, non-food grasses for advanced biofuels and biopower. The company’s energy crops can provide more fuel or electricity, new opportunities for growers and a cleaner environment. Using advanced plant breeding and biotechnology, Ceres is developing dedicated energy crops as raw materials for a new generation of biofuels made from plant stems, stalks and leaves. This will enable the large-scale production of biofuels by increasing yields, lowering costs, facilitating processing and refining, and even helping reduce greenhouse gases. The company’s first products, high-yielding switchgrass cultivars and high-biomass sorghum hybrids, are now available under the Blade Energy Crops brand. Other crops in the pipeline include sweet sorghum, Miscanthus and energycane. 

Using marker-assisted breeding and other technology platforms, the company plans to introduce a succession of enhancements, including further increases in biomass yield and other agronomic and compositional improvements. For sugar-based biofuel production, Ceres will introduce sweet sorghum hybrids (also called sweet-stem sorghum) to complement or expand feedstock supplies in sugarcane-growing regions. This includes Gulf Coast states in the United States and other sub-tropical or tropical growing environments. Miscanthus is a tall Asian perennial grass that grows unusually well in temperate climates. It has been used as an energy crop across Europe for two decades. Ceres is using multiple technology platforms to improve this crop, which can produce high yields of biomass. The company’s goal is to expand the scale and range where farmers can grow this crop. 

In November 2009, Ceres announced that it plans to expand an advanced trait development project to increase biomass yields of several energy grasses by as much as 40% in coming years, while simultaneously decreasing the use of inputs such as nitrogen fertilizers. The project, which was selected by the U.S. Department of Energy from among 3,700 renewable energy proposals, will be funded in part by a $5 million research grant from the Advanced Research Projects Agency – Energy. The three-year project was expected to begin in late 2009. Ceres researchers will test its advanced traits in a variety of energy grasses such as switchgrass, sorghum and Miscanthus. Productivity and inputs requirements, such as fertilizer, will be evaluated as well as expected improvements to carbon and nitrogen cycles. 

Chromatin, Inc. has developed patented mini-chromosome technologies that enable the development of new seed products and the delivery of multiple genetic traits. The company expects that consumers, growers, seed companies, and processors will derive greater value from crop plants through the application of Chromatin’s technologies. These benefits are expected to include more efficient and faster product development, greater product differentiation, and creation of novel products. 

Chromatin initially developed its gene stacking technology for applications in traditional crops (corn, soy, cotton, canola), forming partnerships with industry leaders that have market validated trait technologies. Chromatin is now entering the bioenergy feedstock market, where there are significant opportunities to create and capture value using the company’s transformation technology. Its first feedstock products are targeting crops such as switchgrass, Miscanthus, sorghum and sugarcane where the addition of traits can improve crop and sugar yield and allow digestion of cellulosic fiber. Chromatin is also using this synthetic biology technology to develop scalable and competitive solutions for the North American cellulosic biofuels market. 

Edenspace Systems Corporation is a plant biotechnology company that is developing innovative new crops to lower the cost of renewable fuels. The company is also a commercial leader in the use of living plants to restore and protect the environment (phytoremediation). Responding to the market need for lower-cost ways to produce cellulosic biofuels such as ethanol and butanol, Edenspace is developing Energy Corn™ and other enhanced energy crops. To support commercial sales and licensing of its energy crops, the Company is implementing a five-part market strategy: 

  • Bioengineer corn so that the stover can be processed in existing or planned corn grain ethanol production facilities.
  • Apply the technology platform developed for Energy Corn™ to other crops such as sorghum, switchgrass, sugarcane and trees.
  • Sell proprietary Edenspace seed to farmers as well as license traits to seed companies for incorporation in their own proprietary crop seed.
  • Develop energy crops under cooperative agreements with government agencies and industry leaders, and maintain key research and development links with universities and other centers of excellence for ongoing plant research relating to biofuels applications.
  • Develop and expand a strong intellectual property estate

The Energy Corn™ line of products are expected to deliver significant benefits to various customers. For the farmer, these varieties will have a greater yield per acre, increasing ethanol yield and increasing the farmer’s profits, while also enabling farmers to enter new markets. These products are also expected to benefit the biorefinery by leading to lower enzyme costs and therefore lower preprocessing costs. Finally, benefits to consumers will derive from lower fuel costs and other environmental benefits of biofuel crops. 

Edenspace received permits from the USDA in 2009 to conduct field trials of transgenic corn and poplar that had been engineered for improved digestibility.  In November 2008, Edenspace announced the commercial availability of a non-transgenic, high-yielding forage sorghum that it says is an excellent feedstock for production of cellulosic biofuels such as ethanol and butanol. The new feedstock was selected from more than 100 sorghum varieties screened in 2008 for biomass yield and low processing cost. 

Evogene Ltd. is an agricultural biotechnology company geared toward developing improved plants for the agriculture and biofuel industries through the use of plant genomics. The company was founded in 2002 as a plant-focused spin-off of Compugen, aimed at utilizing predictive discovery capabilities in the agricultural biotechnology field. The company’s mission is to be a world leader in delivering improved plant traits to the agbiotech industry through the use of a continuously improving proprietary platform combining state-of-the-art computational genomics, molecular biology and advanced breeding methods. The company combines state-of-the-art computational gene discovery technologies, molecular biology, high-through-put assays, classical and advanced breeding methods, and uses these methods to discover and develop genes for the improvement of plant traits, and improved high oil yielding plant varieties for biofuels production. Evogene holds pending IP rights to over 300 novel genes and more than 40 promoters validated in plants relating to key plant traits, such as yield under normal and various environmental stress conditions (such as drought), fertilizer utilization and more. 

Evogene has currently two complementary ongoing projects aimed at developing improved oil yielding crops suitable for biodiesel production: 

  • Increased yield (oil yield) of canola and soybean. Evogene has discovered a set of proprietary candidate genes that have high potential to increase economic oil yield and are currently undergoing validation in model plant systems. 
  • Develop new crops (feedstocks) for biodiesel. In September 2007, Evogene formed a joint venture with OrFuel, a fully owned US subsidiary of Ormat Industries, that was aimed at combining industry needs with plant development capabilities to produce an optimized feedstock which is tailored for the biodiesel industry. The collaboration, which has recently been terminated, was focused on using advanced breeding techniques on oil yielding crops for the development of non edible commercial crops for biodiesel, suitable for growth in non arable lands under harsh condition and that require low inputs.  Evogene is continuing the project on its own.

 In February 2010, Evogene announced the establishment of field trials in Texas, USA and northeastern Brazil, for the evaluation of its proprietary castor bean lines. These castor bean lines, designed for higher yield and growth on semi-arid land, are being developed by Evogene to serve as a sustainable and cost efficient second-generation feedstock for biofuel production. Having recently completed two years of field trials under semi-arid conditions in Israel, the purpose of the field trials in Texas and Brazil is to evaluate the lines when grown in the two main target locations for future commercial growth, and further develop them to local conditions. The field trials will be operated in collaboration with Texas AgriLife Research, part of the Texas A&M System in the U.S., and South Cone Agriculture in Brazil.  These field tests are a continuation of the project under the terminated OrFuel collaboration.

Farmacule BioIndustries Pty Ltd. is developing molecular farming technology to cost effectively mass produce high-value industrial and therapeutic proteins and biofuels. The company’s “In-Plant Activation technology” (INPACT) allows Farmacule to insert a molecular switch into the genome of a plant, to control the way proteins are produced, ultimately allowing large scale high value protein production. The goal is to use the INPACT molecular switch to create engineered plants that synthesize desired proteins in targeted areas such as leaves, roots and seeds. The technology also allows Farmacule to turn the production of genes on and off in plants when required – either in response to specific external or internal influences. The INPACT technology was initially developed by a team of molecular biologists under Prof. James Dale’s lead at the Queensland University of Technology, from which INPACT has licensed the technology. 

Farmacule’s goal is to select the most appropriate crop species to use as an efficient bioreactor in a commercial molecular farming operation. The initial focus has been on finding crops which can be genetically enhanced, harvested with ease and processed with minimum complications, while placing particular importance on the safety and containment of gene flow to other species. In this way, the company feels that any risks arising from the release of genetically modified organisms into the environment can be eliminated using the INPACT technology. In particular, INPACT enables the creation of male sterility in plants and therefore has commercial application in the production of hybrid seed and preventing the release of modified genetic material into neighboring crops. Farmacule’s efforts have also concentrated on ‘safe’ plants such as tobacco, bananas and sugarcane, which provide several advantages over other potential plant bioreactors. 

In October 2007, Farmacule entered into a partnership with Syngenta, the world’s largest agribusiness company, the Queensland University of Technology (QUT) and qutbluebox, to establish the Syngenta Centre for Sugarcane Biofuel Development which will develop cellulosic ethanol and biofuels derived from sugarcane. 

D. Glass Associates, Inc. is a consulting company specializing in several fields of biotechnology. David Glass, Ph.D. is a veteran of nearly thirty years in the biotech industry, with expertise in patents, technology licensing, industrial biotechnology regulatory affairs, and market and technology assessments. This blog provides back-up and expanded content to complement a presentation Dr. Glass made at the EUEC 2010 conference on February 2, 2010 entitled “Prospects for the Use of Genetic Engineering in Biofuel Production.” The slides from that presentation are available at www.slideshare.net/djglass99.

Companies Developing Modified Plant Varieties as Improved Biofuel Feedstocks (Part 1)

These next entries of the blog will discuss the companies that are using biotechnology to create improved varieties of the plant species that are, or are proposed to be, used as feedstocks for creation of ethanol or other biofuels. As described in an earlier entry of the blog, the focus of much of these efforts is to use recombinant DNA genetic engineering techniques to introduce into plants certain genes encoding enzymes that are needed for the breakdown of the complex carbohydrates found in cellulosic feedstocks, with the goal of cutting processing costs by having these enzymes inherently present in the biomass used for fuel production. Other technical strategies involving genetically engineered (“transgenic”) plants are also being pursued, for example to increase plant growth rates or possibly to engineer into these species the same sorts of insect or herbicide resistance that are now common in food crops. There are also several companies planning to use genetically modified plants as the production platform to manufacture industrial enzymes for use in preprocessing feedstocks for cellulosic fuel manufacture. However, some of the companies profiled in this installment of the blog are not creating transgenic plants per se, but are using other advanced biotechnology techniques to make traditional plant breeding activities more predictable and more efficient. 

Among the notable things about the companies listed below is how many of them have entered the biofuel arena from other sectors of the plant biotechnology industry. Included here are agbiotech or plant genomics companies (e.g. Chromatin, Mendel Biotechnology), seed companies like Syngenta, the forestry biotechnology leader Arborgen, a company originally dedicated to phytoremediation (Edenspace), and at least two companies using plants as the manufacturing platform for high-value commercial products (Medicago, whose main business in producing vaccines in plants, and Metabolix, a leader in bio-based production of precursors used to create biodegradable plastics). Only a relative handful of the companies profiled were started specifically to address biofuel markets, as opposed to the microbial and algal sectors, where most of the small-company players were founded as biofuel companies. However, this can be seen as a positive development for this sector of the biofuels industry, in that these “diversifying” companies bring considerable expertise in plant biotechnology, plant breeding, and the commercial aspects of the seed business, which will no doubt be beneficial as companies begin to commercialize the specialized “energy crops” they are developing.  

It is harder to predict the chances for success of the companies in this sector of the industry, since in a way they are creating a brand-new market. Current biomass-based production of ethanol or other fuels typically relies on plant species already grown and harvested for food or other purposes, or in some cases uses cellulosic waste products. The business model that companies can develop and sell plant varieties (or seeds) to be grown in a dedicated manner solely for use in fuel production is one that is just beginning to be utilized, by the small number of companies that have already introduced classically-bred, nontransgenic plant lines tailored for biofuel use (for example, high-yielding forage sorghum varieties developed by Edenspace, new Camelina varieties introduced by Great Plains Oil and Exploration,  and Jatropha lines introduced by SG Biofuels). And some of these companies are developing plant species that have never been used and sold in commercial agriculture to any significant extent (e.g. switchgrass, Miscanthus).  

So commercial success may require addressing and solving issues that have not previously been faced in the biofuel industry. For the most part, these are issues that have been faced in the agbiotech industry with some success, including the need to convince farmers to buy and grow plant varieties that, once harvested, would need to be segregated from other crops to be sold to entirely different buyers at higher value than the typical commodity crop. This point is critical, because profit margins in the seed business are typically small, and commercial success will likely depend on these companies’ abilities to sell seed for these varieties at a premium, in the expectation that the grower will recoup his/her costs in the greater selling price the energy crop will provide. Another challenge that some companies may face is how to adequately protect intellectual property. In commercial agriculture today, seed for many important crop plants is sold as hybrids, and because hybrids don’t breed true, farmers cannot save seeds from one year’s crop to plant the next year, and they instead must go back to the seed company to buy seed each year. For those crops not typically sold as hybrids, such as soybeans, farmers have traditionally been able to save seed from one year’s crop to plant the next year, and this has led to legal battles between such “seed-saving” farmers and the seed companies seeking to protect their intellectual property in transgenic plant varieties (and their ability to get annual sales from farmers for such varieties). Because so many of the projected “energy crops” are species that have not historically been sold as commercial crops, it is likely that they would not be sold as hybrid seed and therefore likely that similar issues may arise in this industry sector.       

Finally, timelines for plant biotechnology tend to be longer than those in microbial or even algal biotech, due to the longer life cycle of the organism, the ordinary need for years of field testing, and (in commercial agriculture) the frequent need for repeated cycles of cross-breeding to ensure that introduced traits are expressed in the desired genetic background. Many of the companies here have already begun field-testing their new varieties, and in one case, that of Syngenta, approvals are in hand to commercialize a transgenic variety in some countries of the world. But the products of most of these companies are still several years away, creating the challenge for the smaller companies to obtain and conserve sufficient capital to stay in business long enough to survive until products can be introduced.  

Whether or not the efforts to develop transgenic or other modified plant varieties as energy crops proves successful, it is undoubtedly a good thing that research programs pursued by companies like these are shifting attention away from “traditional” biofuel species like corn and sugarcane towards nonconventional non-food species like switchgrass, Camelina, Jatropha and woody plants. Most observers would agree with the need for the biofuel industry to increasingly look towards fast-growing, non-food species as the source of biomass for biofuel production, and significant germplasm improvements in these species can be made even without advanced biotechnology. This need is evidenced by the existence of government programs such as the joint USDA/DOE Biomass Research and Development Initiative, one segment of which is devoted to funding innovative approaches to development of new feedstocks for biofuel production, and which has awarded grants to many of the companies profiled here. The broadened focus that these companies, as well as academic research labs around the world, bring to these nontraditional crops will pay off in the long term, regardless of the success of any one company.  

The following are the companies that are using biotechnology to create new plant varieties as improved feedstocks for biofuel production. Profiles of these companies will follow in upcoming installments of the blog over the next several days.  

Conventional Feedstocks
(Note that many of these companies are also developing newer feedstocks)

  • Agrivida: corn
  • ArborGen: purpose-grown trees
  • CanaVialis S.A.: sugarcane
  • Edenspace Systems: corn
  • FuturaGene: hybrid poplar and willow
  • Targeted Growth: corn

Newer Feedstocks

  • Agragen: Camelina
  • Agrisoma: Brassica and Jatropha
  • Agrivida: switchgrass, sugarcane, sorghum, others
  • Ceres: non-food grasses
  • Chromatin: switchgrass, Miscanthus, sorghum and sugarcane
  • Edenspace Systems: switchgrass
  • Evogene: canola, soybean, others
  • Farmacule BioIndustries: sugarcane, tobacco
  • FuturaGene: switchgrass, Miscanthus
  • Kaiima: castor beans (non-GMO)
  • Mendel Biotechnology: grasses, others
  • Metabolix: switchgrass, oil crops, others
  • Rahan Meristem: Jatropha, castor beans
  • SG Biofuels: Jatropha
  • Targeted Growth: Camelina, canola, others

Enzyme Manufacture in Plants

  • Infinite Enzymes (transgenic plants)
  • Medicago (transient expression)
  • Syngenta (transgenic plants)

D. Glass Associates, Inc. is a consulting company specializing in several fields of biotechnology. David Glass, Ph.D. is a veteran of nearly thirty years in the biotech industry, with expertise in patents, technology licensing, industrial biotechnology regulatory affairs, and market and technology assessments. This blog provides back-up and expanded content to complement a presentation Dr. Glass made at the EUEC 2010 conference on February 2, 2010 entitled “Prospects for the Use of Genetic Engineering in Biofuel Production.” The slides from that presentation are available at www.slideshare.net/djglass99.

Companies Reported to be Developing Genetically Modified Algae for Biofuels

The algae biofuels industry has received more publicity and media attention in the last 12-18 months than any other sector of the biofuels industry. Recent developments, more on the business side than the technology side, have brought algae into the spotlight, and this has brought the usual amounts of hype and hope (on the one hand) and criticism and skepticism (on the other). It is far beyond the scope of this blog to try to exhaustively summarize everything that has been said or written about the potential for algal biofuels, but I will try to put some of these issues into context and assess the chances for success of biodiesel, jet fuels, ethanol and other transportation fuels produced using algae, particularly those made with algal strains enhanced using modern biotechnology. 

Because algae are naturally capable of producing large amounts of fatty acids, and can be grown on nonarable land (without competition with food crops) using only sunlight and water for growth, this category of organism has long been considered to have potential for producing transportation fuels, particularly biodiesel, which is produced by the chemical transesterification of fatty acids. Algae drew the attention of the U.S. Department of Energy in the wake of the energy crisis of the late 1970s, and from 1978 to 1996, the DOE’s Office of Fuels Development funded a program to develop renewable transportation fuels from algae. The main focus of the program, known as the Aquatic Species Program, was the production of biodiesel from high lipid-content algae grown in ponds, in particular utilizing waste CO2 from coal fired power plants. Although this program supported significant scientific advances in algal biology and in the design and operation of open ponds for large-scale algal growth, the project concluded that the costs of producing biodiesel using algae remained too high for this route to be a competitive source of transportation fuel. As stated in the 1998 DOE close-out publication for this program: 

“The factors that most influence cost are biological, and not engineering-related. These analyses point to the need for highly productive organisms capable of near-theoretical levels of conversion of sunlight to biomass. Even with aggressive assumptions about biological productivity, we project costs for biodiesel which are two times higher than current petroleum diesel fuel costs”.

Because of these and other technical hurdles, very little progress was seen towards increased large-scale commercial use of algae for biodiesel or other fuels, and many have been skeptical that algae would ever become a significant production route for transportation fuels. Nevertheless, a substantial number of companies have continued plugging away at developing algal biodiesel as a viable fuel, and many new companies have been established for this purpose. The Algal Biomass Organization now has over 170 members (according to their 2009 annual report) and has become a prominent voice for the industry. The industry benefitted from increased media attention through the announcement of several significant business deals and/or research alliances between algae biofuel companies and larger energy companies, not least of which was the research partnership announced in 2009 between ExxonMobil and Synthetic Genomics (described below), as well as the increasing market presence of some of the newer, more innovative algae companies that have attracted venture capital money or other high-profile investments. However, in spite of the increased commercial attention, the economics of producing biodiesel using algae remains a significant concern, as evidenced by the intensive lobbying effort by the industry for a change in U.S. tax policy to allow algae fuel producers to access the $1.01 per gallon production tax credit that is available to manufacturers of other advanced biofuels. As of this writing, legislation to accomplish this has passed both houses of Congress, having passed the Senate in early March 2010, and is awaiting action by a conference committee before it can achieve final approval and be signed into law. 

Along with this increased visibility has come additional skepticism of algae’s potential, most recently a 2010 paper in Environmental Science and Technology from the University of Virginia claiming that use of algae for biofuels entailed greater environmental impacts in energy use, greenhouse gas emissions and water usage than the use of conventional crops. The Algal Biomass Organization has vigorously objected to this study and has issued a press release refuting its conclusions, largely on the basis that the UVA study “was based upon obsolete data and grossly outdated business models, and overlooked tremendous improvements in technology and processes across the [algal biofuel] production cycle”. But there have been other, positive, developments for the algae biofuel industry: in February 2010, the Environmental Protection Agency, in issuing final regulations under the Renewable Fuels Standard, announced its determination that diesel produced from algal oils complies with the 50% greenhouse gas reduction threshold and therefore qualifies as an “advanced biofuel” that could be used to satisfy biodiesel mandates; and in January of this year, the Energy Department announced nearly $80 million in funding for research and development on algae-based fuels, most of which will go to two research consortia In addition, as mentioned in an earlier entry of this blog, there has been a large uptick of interest in the use of biological systems, including algae, for the production of jet fuels, with several airlines conducing highly-publicized test flights using jet biofuels, and with the announcement of several high-profile research collaborations relating to microbial, algal or biomass-based production of jet fuel. 

Without delving too deeply into any of the controversies swirling around the use of algae for biofuels or the  differences of opinion about its prospects, I would note that the companies and technology strategies I’m discussing in this blog provide a good deal of hope that many of the historic shortcomings of algal production systems can be overcome. It is believed that enhancing the efficiency of photosynthesis or algal growth or increasing the output of fatty acids, hydrocarbons and oils by genetic manipulation and/or improved strain selection, along with engineering improvements in the way algae are grown and processed, will begin to make it much more economically feasible to produce diesel or jet fuels using algae. Much attention has also recently been directed at co-localization of algal biofuel plants at or near sources of carbon dioxide release, not only to improve the efficiency and economics of biofuel production but also for environmental benefit as a carbon capture strategy. Many of the companies profiled here, as well as others developing or using innovative non-GMO technology and process engineering improvements are operating pilot facilities that are already producing biodiesel, jet fuels or ethanol using algae, leading to some hope that the corner has been turned commercially. 

There are of course no guarantees that any of the companies profiled here, or any particular technological improvement will succeed where others have failed in the past, but many observers do feel that all this new intellectual brainpower will give us the best chance to date of seeing an economical, profitable algal biofuel industry. So there is reason to be guardedly optimistic, but the next few years should tell the tale, as more companies move to larger scale production and begin to achieve wider market acceptance. 

The following are the companies that are believed to be using or developing algae genetically enhanced in some manner, in biofuel production. Inclusion in this entry of the blog is based on press or Internet reports, and in some cases is not necessarily confirmed on company websites, and not every company listed is using “genetic engineering” technologies as opposed to more traditional methods of modifying or enhancing biological function. Also, it’s worth noting that these companies represent only a subset of the large number of companies now active in the algal biofuels arena – since my blog deals only with uses of advanced biotechnology techniques, I am not mentioning or discussing the dozens of other companies that are using naturally-occurring or other nonmodified algal strains for production of biodiesel or other fuels, even though some of these companies are developing technological innovations in reactor design, process engineering, and other key steps of the fuel production process. 

  • Algenol Biofuels
  • Aurora Biofuels
  • AXI LLC
  • Global Green Solutions
  • Kuehnle AgroSystems
  • Planktonix Corporation
  • Sapphire Energy
  • Solazyme
  • Synthetic Genomics
  • Targeted Growth

The following are brief profiles of companies that are developing engineered or modified algae for the production of biofuels. These profiles have been adapted or excerpted from company websites and/or other publicly available information, and I don’t assume any liability for the accuracy, comprehensiveness or use of the information. 

Algenol Biofuels Inc. is pursuing a strategy different from most other algal biofuel companies: it is developing technology to use metabolically enhanced algae to produce ethanol, in a process it calls its DIRECT TO ETHANOL™ technology. Algenol has engineered cyanobacteria to express the enzymes pyruvate decarboxylase and alcohol dehydrogenase, allowing the algae to convert the common Krebs cycle molecule pyruvate first to acetaldehyde and then to ethanol, linking ethanol production to the photosynthesis that drives algal growth and metabolism. The enhanced algae are grown in proprietary Capture TechnologyTM bioreactors. These bioreactors are contained and sealed units that hold algae in the bioreactor, prevent contamination, maximize ethanol recovery and allow for fresh water recovery. Algenol claims to have the most advanced 3rd generation biofuels technology and the company says its technology is capable of producing industrial-scale, low-cost ethanol using algae, sunlight, CO2, and seawater, without the use of food, farmland, or fresh water. 

The initial proof of concept was generated by Dr. John Coleman at the University of Toronto between 1989 and 1999. Since then, the process has been refined to allow algae to tolerate high heat, high salinity, and the alcohol levels present in ethanol production. The company says they have access to over 100,000 useable species of blue-green algae with rapid growth cycles, high photosynthesis efficiency, large sugar storage attributes for use with its Direct to EthanolTM process. The algae are metabolically enhanced to produce ethanol while being resistant to high temperature, high salinity, and high ethanol levels, which were previous barriers to ramping to commercial scale volumes. 

Algenol says its prototype production strains can produce ethanol at a rate of 6,000 gallons/acre/year, and the company was expecting yields to have improved to 10,000 gallons/acre/year by the end of 2009. With further refinement, Algenol says that the algae cells have the potential to increase production rates to 20,000 gallons/acre/year in the future. Algenol has been building a pilot plant in Freeport, Texas, in collaboration with Dow Chemical, and in December 2009, Algenol received a $25 million “stimulus” grant to continue working with Dow and Georgia Tech to build this pilot biorefinery. More recently, the company received a $10 million incentive from Lee County, Florida for Algenol to move into a 43,000 square foot facility near Fort Myers, Florida that will serve as company headquarters beginning in May 2010 and will serve as a pilot production plant, anticipated to be able to produce 300,000 gallons of ethanol per year, which is reported to be three times the capacity of the Freeport plant under construction. Ethanol production from the new plant is expected by August 2010. 

Algenol has entered into a license agreement with Sonora Fields S.A.P.I. de C.V., a wholly owned subsidiary of Biofields S.A.P.I. de C.V. in Mexico and is currently working closely with Sonora Fields and both local and federal Mexican governments to commercialize the DIRECT TO ETHANOL™ process, with commercial sales expected in Sonora, Mexico by the end of 2010. 

Aurora Biofuels generates biodiesel from optimized algae in a patented production process. The company has developed a cost-competitive, scalable method for fuel generation using robust, highly-productive custom algae strains in open pond systems, which the company is rapidly commercializing. Aurora has successfully operated a pilot facility since August of 2007, and is developing a 20-acre demonstration plant that was expected to be completed in 2009, that will further showcase the company’s ability to produce biofuel at scale. Aurora expects to begin commercial production in 2012. The biodiesel produced from the pilot facility has successfully passed ASTM standards, and the consistency of fuel production over the trial period marks a success in perfecting the company’s chosen pathway of low-cost open pond algae production systems. The achievement showcases the company’s pathway to cost-effectively producing consistent-quality biofuels from algae in industrial-scale volumes, and points to the company’s readiness to pursue the commercialization of its biofuel process. 

Aurora Biofuels combines the latest in biotechnology and process engineering to create a pathway for the cost-effective mass production of algae based biofuels. Aurora’s scientific research team has screened a myriad of strains in search of microalgae that outperform others in terms of oil production and yield. Aurora has further bred its select portfolio to maximize fuel-production performance and to be cost-effective at scale. 

The Aurora process does not compete for agricultural resources. Aurora has a minimal environmental footprint: the system, which is extremely efficient in terms of land usage, uses salt water in its ponds and can be built on nonarable land. The company says that its process can produce an equivalent amount of fuel as sugar-based fuels, using approximately 1/25th the land space, and that the company’s approach is 70-100 times more productive than  oil-rich agricultural crops, . Aurora’s algae feed on carbon dioxide and sequester 90 percent of the CO2 fed into their environment. Because of this ability, Aurora Biofuels hopes to become an important player in the emerging cap-and-trade market by converting carbon producers’ waste emissions into algae feedstock. 

In June 2008, Aurora announced that it completed a second round of funding, raising $20 million. The round was led by existing investor Oak Investment Partners. Previous Series A investors Gabriel Venture Partners and Noventi also participated in the round. In August 2009, Aurora announced that it succeeded in optimizing its base algae strains to more than double CO2 consumption and fuel production, using a proprietary process which allows for the superior selection and breeding of non-transgenic algae. The company has proven these results in an outdoor open system, with the result that these algae strains can produce more than twice the amount of oil. 

AXI, LLC has been created by the venture capital firm Allied Minds and the University of Washington to commercialize novel technology that creates improved strains of algae for the production of biofuels. Allied Minds says that new algae strains will bridge the gap between the promise of clean energy generation and the reality of economical biofuel production systems. AXI is initially targeting the development of unique physiological traits that permit efficient growth and processing of algae using proprietary techniques and supported by 25 years of primary phycology research. 

The research team at the Cattolico Laboratory at the University of Washington (AXI’s academic founders) has been investigating the physiology of algae for more than 25 years. The result of this work is the discovery of several unique and advantaged varieties of algae, and, more importantly, the ability to customize and optimize virtually any species of algae to maximize its commercial usefulness. In January 2010, AXI was part of a consortium – the National Alliance for Advanced Biofuel and Bio-products (NAABB) – that was awarded a $44 million grant in federal stimulus money from the U.S. Department of Energy. The consortium will use the funds to develop a systems approach for sustainable commercialization of algal biofuels, and AXI will contribute at the front end of this initiative with its expertise in algal biology. 

Global Green Solutions Inc. (GGRN), with operations in North America, Europe, and South Africa, develops and implements ecotechnology solutions for renewable energy generation. Founded in 2006, GGRN has developed two innovative biomass technologies; Greensteam, a high-efficiency combustion system that generates industrial steam and electrical power from waste biomass; and Global Green Algae, a self-contained algae growing system which produces biofuel feedstock.

In the Global Green Algae program, the company looks to increase future biomass fuel production and resources by rapidly growing algae for production of biofuels and co-products. The residual biomass after extraction of the algal oil contains chemical elements which are suitable for animal feed, agriculture, specialty chemicals, cosmetics, nutraceutical and pharmaceutical applications. The Company is carrying out a two-stage program: Stage 1 is a two-year R&D program including construction of a facility for algae bioscience research, and the development and testing of algae photobioreactor growing processes. The Stage 2 R&D program will be focused on the algae growing process economic and operational sustainability issues. This will require an integrated program of algae transgenic technology and physiology and advanced process and materials engineering. GGRN is seeking strategic partners in North America and Europe for the Stage 2 R&D program. 

Kuehnle AgroSystems is an algae seedstock and strain development company that develops algae by traditional and GMO strategies for the renewable fuel and chemical markets, as well as strains for aquaculture. The company, based in Honolulu, HI, produces customized EliteAlgaeTM and MightyMealTM strains of algae and is also providing algae strains for several federally-funded algae biofuels programs. Kuehnle is developing modified algae for closed bioreactor systems, creating platform technology to enable genetic manipulation of multiple species of algae, including custom strain development for end user clients. The company also maintains a collection of native Hawaiian species that are being used for open pond biofuel projects.

Planktonix Corporation is utilizing a coalition-based approach for research, development, deployment and commercialization of microcrop-based (green algae and cyanobacteria) biofuels production. The company maintains three main bioenergy technology development initiatives: (1) algal and cyanobacterial biomass-to-lipid biodiesel/biocrude production (with a 100,000 GPY pilot production facility planned with coalition partners, targeted to break ground in the summer of 2010); (2) streamlined bacterial fermentation of waste biomass (corn stover and switchgrass) to biobutanol; and (3) a biomass-free direct production technology to produce biobutanol and co-products without any requiring any refining or fermentation steps, utilizing bioengineered strains of oxyphotobacteria through a modified photosynthetic biochemical pathway process. 

Planktonix has formed a strong coalition of academic, industry, non-profit and National Laboratory partners to develop solutions to these very compelling bioenergy challenges. The coalition, called the National PhytoFuels Energy Innovation Hub (NPEIH), includes institutions such as Brookhaven National Laboratory, Old Dominion University, South Dakota State University, Ohio State University, SunsOil LLC, and Emery Energy Corporation, among others. 

Planktonix and its partners have developed algal biofuels processes that dramatically reduce greenhouse gas emissions, reduce the lead time from start-up to commercial-scale production, optimize “lowest cost” advanced biofuels production, reduce costs using a systems approach. Planktonix has identified seven target species of phytoplankton with unique characteristics ideal for biofiltration or biofuels production. The Planktonix process relies on waste materials and microcrops to create several types of biofuel. Through microcrop biotechnology, Planktonix and its partners will use utility-scale installations to reduce toxins while using existing infrastructure to develop, deliver, and store biofuels. Through algal biotechnology, Planktonix and its partners will optimize the algal species and growth conditions that can reduce greenhouse gas emissions significantly.   

Sapphire Energy is focused on the entire “pond to pump” value chain and is developing technology spanning the entire algae-to-fuel process. The company is developing industrial algae strains through synthetic biology and breeding techniques and are building the technologies and systems for CO2 utilization, cultivation, harvesting and refining. The algae and processes developed are field tested at a New Mexico research and development center where all the processes — from biology to cultivation to harvest and extraction — can be performed at a pilot scale. These processes result in a product called Green Crude which can be refined into gasoline, diesel or jet fuel. 

Sapphire’s goal is to become the world’s leading producer of renewable petrochemical products, based on a molecular platform that converts sunlight and CO2 into renewable, carbon-neutral alternatives to conventional fossil fuels. Sapphire uses photosynthetic microorganisms to produce a renewable, high-value replacement for fossil fuel petroleum, using only sunlight, CO2 and non-potable water, and which can be produced in large scale on non-arable land. The goal is to develop biological production methods for the production of transportation fuels, including high-octane gasoline, to ASTM certification standards. The company’s final products will have the same chemical composition as gasoline and will be completely compatible with the existing refining, distribution and fleet infrastructure. 

In 2008, Sapphire successfully produced 91-octane gasoline from algae that fully conforms to ASTM certification standards. In 2009, the company participated in a test flight using algae-based jet fuel in a Boeing 737-800 twin-engine aircraft, and that same year, Sapphire provided the fuel for the world’s first cross-country tour of a gasoline vehicle powered with a complete drop-in replacement fuel containing a mixture of hydrocarbons refined directly from algae-based Green Crude. In 2010, the company expects to break ground for an Integrated Algal Bio-Refinery in Southern New Mexico, a project that was awarded more than $100 million in federal “stimulus” grant money from the American Reinvestment and Recovery Act through the U.S. Department of Energy and a loan guarantee from the U.S. Department of Agriculture Bio-refinery Assistance Program. 

Solazyme, Inc., a renewable oil and bioproducts company, uses algal biotechnology to renewably produce clean fuels, chemicals, foods and health science products. Solazyme’s advanced and proprietary technology uses algae to produce oils and biomaterials in standard fermentation facilities quickly, cleanly, cost effectively, and at large scale. These natural oils and biomaterials are tailored, not only for fuel production, but also as replacements for fossil-derived petroleum and a variety of natural plant oils and compounds. 

Solazyme produces an oil-based fuel, Soladiesel®, at industrial manufacturing scale with production capabilities currently in the tens of thousands of gallons, and plans to ramp up production for demonstration and pre-commercialization purposes. The company’s indirect photosynthesis process uses microalgae to convert biomass directly into oil and other biomaterials, a process that can be performed in standard commercial fermentation facilities cleanly, quickly, and at low cost and large scale. The company has manufactured thousands of gallons of oil and hundreds of tons of biomaterials that are tailored not only for biofuel production, but also as replacements for fossil petroleum and plant oils and compounds in a diverse range of products from oleochemicals to cosmetics and foods. 

Among corporate highlights, the company has obtained a $57 million third round of funding from venture groups including Braemar Energy Ventures, Lightspeed Venture Partners, VantagePoint, Roda Group, and Harris & Harris. The company was also selected by the U.S. Department of Defense to research, develop, and demonstrate commercial scale production of algae-derived F-76 Naval Distillate fuel for testing and fuel certification to demonstrate it meets all military specifications and functional requirements. The contract includes both R&D and fuel delivery components and calls for delivery of over 20,000 gallons of Soladiesel F-76 fuel to the Navy for compatibility testing over the next year. In December 2009 Solazyme announced that it had received a $21.8 million federal grant to build its first integrated biorefinery in rural Riverside, Pennsylvania, marking a major step toward commercial scale production of algal based fuel. The project will enhance the national infrastructure investment in biofuels as an alternative to fossil fuels and will create or preserve many green jobs. The project funding was announced today by the U.S. Department of Energy (DOE).

Synthetic Genomics Inc. was founded to commercialize genomic-driven technologies, and is based on the pioneering research of its founders J. Craig Venter, Ph.D., Nobel Laureate Hamilton O. Smith, M.D., and the leading scientific teams they have assembled. The company’s scientific capabilities encompass areas such as environmental genomics, microbiology, biochemistry, bioinformatics, plant genomics, genome engineering, synthetic biology, and climate change. In addition to its own in-house research, the company sponsors more basic research at the J. Craig Venter Institute, a not-for-profit organization with more than 400 scientists and staff working on a variety of genomic research and policy fronts.

SGI is working in three broad projects areas of renewable fuels and chemicals (alliance with ExxonMobil Research and Engineering Company to develop algal biofuels), microbial-enhanced hydrocarbon recovery (collaboration with BP), and sustainable agricultural products (collaboration with Asiatic Centre for Genome Technology [ACGT]). In the fuels area, SGI and ExxonMobil, the last of the oil majors to commit to a major investment in biofuels, announced that its Research and Engineering unit will invest $300 million into in-house algae research, and up to an additional $300 million in La Jolla-based Synthetic Genomics, the genetics firm founded by J. Craig Venter that has been working on algae-to-energy research since 2005. SGI and ExxonMobil have established a multi-year research and development strategic alliance focused on exploring the most efficient and cost effective ways to produce next generation biofuels using photosynthetic algae. In this partnership, SGI will continue its work to discover and develop superior strains of algae using leading edge genomic technologies. ExxonMobil’s engineering and scientific expertise will be utilized throughout the program, from the development of systems to increase the scale of algae production through to the manufacturing of finished fuels. 

SGI is also applying its cutting-edge synthetic biology tools to develop new microbial solutions to convert lignocellulosic biomass into advanced fuels and chemicals in a simple and cost-effective, one-step conversion process, known as Consolidated Bio-processing. The alliance with ACGT is aimed at improving oil crops such as Jatropha using genomics and the company’s other advanced technologies. 

Targeted Growth, Inc. (TGI) is a crop biotechnology company focused on developing products with enhanced yield and improved quality for the agriculture and energy industries. Founded in 1998, TGI has developed a technology portfolio based on the principle that regulating cell cycle processes can directly and significantly enhance plant yields. Much of TGI’s activities are devoted to modifying plant species as superior biofuel feedstocks, but the company also maintains an active research program in algae. The company is reportedly focusing on cultivation and genetic engineering of cyanobacteria (blue-green algae) algae strains for use in production of renewable fuels. I will include a more complete profile of Targeted Growth in the next series of blog entries, focusing on the companies modifying plants for biofuel use. 

D. Glass Associates, Inc. is a consulting company specializing in several fields of biotechnology. David Glass, Ph.D. is a veteran of nearly thirty years in the biotech industry, with expertise in patents, technology licensing, industrial biotechnology regulatory affairs, and market and technology assessments. This blog provides back-up and expanded content to complement a presentation Dr. Glass made at the EUEC 2010 conference on February 2, 2010 entitled “Prospects for the Use of Genetic Engineering in Biofuel Production.” The slides from that presentation are available at www.slideshare.net/djglass99.

Companies Developing Modified Microorganisms for Manufacture of Biofuel Enzymes

Of all the industrial sectors I’ll be profiling in this blog, the industrial enzyme sector is by far the most mature, and the one that is to the greatest extent already a critical part of today’s transportation fuel industry. It also benefits from being part of a larger, longstanding sector of the bio-economy that for decades has been producing pure or partially-purified preparations of enzymes for use in various industries primarily including food and beverage processing. 

Enzymes are the protein catalysts that drive virtually all the biochemical reactions that take place in living cells: enzymes and the genes that encode them are the key to almost all the genetic modifications that are discussed in this blog. Like most proteins, each enzyme in every living cell is encoded by a single gene (although technically, many enzymes are built of multiple subunits, and each subunit is usually encoded by its own gene). Much of the genetic modification discussed elsewhere in this blog is directed towards transferring genes into desired “host” organisms to give them the ability to express enzymes not naturally found in that host, giving the host new biochemical powers (particularly when multiple enzyme-encoding genes are spliced into the host). However, the companies producing enzymes for industrial use follow a different strategy. These companies seek to identify or create microbial strains that can synthesize very high levels of a given enzyme (so-called “overexpression”), which are often secreted from the host organism, but which in any case are purified after the microbes are fermented at large scale. Pure or partially pure preparations of such enzymes are then sold as commercial products for various industrial uses. Historically, enzyme manufacturers have relied on naturally occurring or classically mutated strains, but more recently these companies have turned to genetic engineering to enable the creation of host organisms capable of high level expression of desired enzymes. 

Enzymes have been produced and sold for various industrial purposes for decades, and today industrial enzymes represent a major sector of the life sciences industry. Market estimates place annual global revenues as approaching $3 billion in the early years of the current decade (a 2007 BCC Research report estimated the 2007 market at $2.3 billion, and projected an increase to over $2.7 billion by 2012; and a report from Global Industry Analysts, Inc. estimated that the world market will exceed $2.9 billion by 2012; there are also estimates that are even higher). The sector of this market that includes the enzymes used in biofuel production is harder to estimate, but is widely considered to be one of the more rapidly growing segments of the industry. 

Industrial enzymes are currently an integral part of most existing cellulosic ethanol production processes. Unlike sugar-based ethanol feedstocks like sugar cane and corn, cellulosic biomass has a more complex molecular structure, composed not only of cellulose, which is a simple homopolymer of glucose molecules, but also of hemicellulose  (heteropolymers of 6- and 5-carbon sugars) and lignins. Few if any naturally-occurring ethanol-fermenting organisms possess the ability to break down these more complex molecules, and so it is common in most existing ethanol plants for the cellulosic feedstock to be pretreated using a combination of enzyme preparations to degrade the long polymers into sugars that are more readily utilized by natural ethanol-fermenting organisms. 

At one time, the cost of enzymes made up a significant percentage of the overall cost of cellulosic ethanol – reportedly as much as 50% of the production cost, at a time when cellulosic ethanol cost $5-10 per gallon (most observers feel that the target production price for cellulosic to be competitive with corn ethanol is between $1-2 per gallon). The advances that have been made by the companies described below and by other researchers have substantially cut the cost of manufacturing enzymes for cellulosic ethanol, to the point where new product announcements by Novozymes and Genencor in February 2010 (described below) brought widespread speculation that enzyme costs were starting to get low enough to realistically expect these target costs to be met early in the decade.  

The market for biofuel enzymes is currently dominated by big players – companies that have been in the enzyme business for decades. However, some of the companies profiled below are smaller, start-up companies or are otherwise new to the industrial enzyme industry. It seems clear that the larger companies have a substantial edge in the market, due to their existing infrastructure, sales force, knowledge and experience with the market, and other advantages. Often, smaller companies in any given market would be more prone to innovation than would the larger companies, and while that may be true here, it should be said that some of the established companies, particularly Danisco Genencor and Novozymes, have top-notch molecular biologists on their research staffs and conduct cutting edge research that smaller companies may have trouble rivaling. Although there may be an ongoing role in the market for niche players, it is likely that the larger companies will continue to dominate for the foreseeable future.

An intriguing question, however, is whether the need for enzyme pretreatment of cellulosic feedstocks will ever be supplanted by the increased use of some of the more versatile microorganisms that are being developed that will express exogenous enzymes enabling the strains to digest complex carbohydrates on their own. It is surely the long-term hope of many of the microorganism developers that this will take place, resulting in process cost savings by altogether avoiding the need for added enzyme preparations.  However, my gut feeling is that this won’t happen for quite some time, if at all, for several reasons. First, it is too early to know how soon such improved microbes may become widely available, and widely accepted, for use in cellulosic biofuel production. Second, we can anticipate that the demand for cellulosic ethanol will grow dramatically in the coming years, due to the Renewable Fuel Standards in the U.S. and similar aggressive government policies elsewhere in the world, meaning that production capacity may expand rapidly enough so that there is room in the market for both the more versatile bugs as well as traditional microorganisms which will continue to need to be supplemented by enzyme pretreatment of cellulosic biomass.

The following are the companies that are manufacturing enzymes for use in biofuel production. 

  • AB Enzymes
  • Danisco/Genencor
  • DSM
  • Dyadic International
  • Infinite Enzymes*
  • Iogen
  • Medicago*
  • Novozymes
  • Proteus
  • Syngenta*
  • Zymetis

Most of these companies are using microorganisms, enhanced using both traditional methods and through biotechnology, to produce enzymes. However, the companies identified with an asterisk are using modified plant species as the platform to produce enzymes for use in biofuel production, and these companies will be profiled in a later section of the blog. 

The following are brief profiles of companies that are developing engineered or modified microorganisms for the production of industrial enzymes for biofuel production and other uses. These profiles have been adapted or excerpted from company websites and/or other publicly available information, and I don’t assume any liability for the accuracy, comprehensiveness or use of the information. 

AB Enzymes GmbH is one of the world’s oldest and best known companies producing industrial enzymes. AB Enzymes is a member of the ABF Ingredients group of companies, which focuses on high value ingredients for both food and non-food applications. AB Enzymes manufactures and sells enzymes for industrial applications worldwide, including food enzymes for bakery and beverages, enzymes for animal feed, textile technology and the pulp and paper industry. 

AB Enzymes’ focus in the biofuel market is in supplying enzymes to support the use of lignocellulosic biomass to produce transportation fuels. The company believes that abundant forms of low value agricultural residues such as ricestraw, cornstover, wood chips, wheatstraw and baggase will increasingly need to be used for this purpose. The company sees the key challenges to include the availability of low-cost substrate and the reduction of the processing costs of cellulosic biomass, and that  a significant part of this effort will come from the reduction in costs of enzymatic saccharification.  Currently, enzyme dosages and thereby enzyme costs are significantly higher for cellulosic ethanol compared to ethanol production from starch, and such costs need to be reduced dramatically over the next few years to enable success in the use of lignocellulosic biomass for ethanol production.

The wide variety of feedstocks and processes require a tailor-made enzyme approach. AB Enzymes offers this “tailor-made” service which will enable the industry to speed up development processes at a reduced cost. AB Enzymes has been successful in the discovery and production of enzymes for the breakdown of lignocellulosic substrates, using Trichoderma strains as the production host of choice for this application. 

Danisco US Inc., Genencor Division is a leading industrial biotechnology company that develops and markets innovative enzymes and bio-based products. Originally formed in the 1980s as a joint venture between Genentech and Corning Glassworks, Genencor is now a division of the Danish company Danisco. Genencor discovers, develops, manufactures, and delivers eco-friendly, efficient enzyme product solutions for the agri processing, cleaning and textiles, food and feed, consumer, and industrial markets, and is also developing innovative technologies for the biofuels, biodefense, and biosafety industries. The company has recognized expertise and longstanding experience in protein and pathway engineering. The company’s two-pronged strategy includes sales of products through its commercial enzyme business as well as its joint venture with DuPont – DuPont Danisco Cellulosic Ethanol LLC (discussed in an earlier entry of this blog). 

Genencor was one of the first companies to develop expertise in cellulosic ethanol enzyme technology, and the company says it was the first company to come to the market with a large-scale enzyme for biomass-to-ethanol conversion, with the introduction of Accellerase in 2007. In February 2009, Genencor launched Accellerase 1500 enzyme complex, for improved economics and higher yields. Genencor added  three products to its Accellerase enzyme line for smaller scale process developers: Accellerase XY, XC and BG.  

In February 2010, Genencor announced the availability of a new enzyme in its Accllerase line. This product,  Accellerase® DUET, is the latest generation in the company’s line of enzymes used to convert biomass into sugars, a critical step in the production of cellulosic ethanol and other advanced biofuels and biochemicals from non-food feedstocks. The company claims that with improved overall hemicellulase activity, Accellerase® DUET builds on the advances in beta-glucosidase and cellulase activity previously made by Accellerase® 1500. These additional improvements allow Accellerase® DUET to achieve higher sugar and biofuel yields, often at 3-fold lower dosing, and to be feedstock- and pretreatment- flexible. Accellerase® DUET employs a whole broth formulation, which provides nutrients for fermentative organisms and lowers the chemical load introduced into our customers’ processes. Higher performance at lower dose will lead to significant improvements in enzyme cost in use for producers, which is critical to enable the cellulosic biofuels industry. 

Royal DSM N.V. is a publicly listed company, founded in 1902, that creates innovative products and services in life sciences and materials sciences. DSM’s products and services are used globally in a wide range of markets and applications, in end markets that include human and animal nutrition and health, personal care, pharmaceuticals, automotive, coatings and paint, electrical and electronics, life protection and housing. DSM has annual net sales of EUR 9.3 billion and employs some 23,500 people worldwide. The company is headquartered in the Netherlands, with locations on five continents. 

DSM is a leading producer of numerous enzymes, in particular for the food market. With its extensive expertise and know-how in the field of enzyme technology, DSM is increasingly able to replace its chemical process steps by enzymatic processes. These are more efficient, sustainable and cleaner. DSM is a major practitioner of white (industrial) biotechnology, Among its capabilities are the use of advanced micro-organisms, including microbial biocatalysts and an ethanologen capable of co-fermenting C5 and C6 sugars; and enzymatic hydrolysis, e.g. using cellulases and hemicellulases. DSM says that it does not produce any genetically modified products. Although genetically modified micro-organisms are sometimes used in production, the end product is always identical to the natural product. 

Dyadic International, Inc. is a global biotechnology company with the groundbreaking technology that it says “brings nature to the marketplace”. Dyadic is focused on the discovery, development, and manufacturing of novel products derived from the DNA of complex living organisms – including humans – found in the earth’s biodiversity. Using its integrated technology platform, Dyadic develops biological products such as proteins, enzymes, polypeptides and small molecules for applications in large segments of the agricultural, industrial, bioenergy, chemical and biopharmaceutical industries. The company utilizes a proprietary host organism that allows the rapid discovery and expression of eukaryotic genes which can then be used to manufacture unique biological products for commercial applications. 

Dyadic’s C1 integrated technology platform is expected to enable researchers to identify, select and analyze novel enzymes best suited to convert biomass materials into biofuels. The company believes that its technology may play a significant role in the development of biofuels such as ethanol at costs competitive to oil prices, thereby reducing subsidies and ultimately expanding consumers’ use of these renewable energy sources. Dyadic is using this technology platform along with other proprietary technology to develop biofuels from agricultural byproducts such as corn stover and wheat straw, and the company is using this technology for itself and others. Among its enzyme products for biofuel processing are AlternaFuel® 100P,  a powdered fungal hemicellulase enzyme complex, and AlternaFuel® 200P, a powdered fungal cellulase enzyme complex, both produced from Trichoderma longibrachiatum, that can be used for the degradation of various lignocellulosic biomass substrates for conversion of biomass to glucose. 

Dyadic has a non-exclusive license agreement with Abengoa Bioenergy for certain Dyadic patent rights and know-how relating to its C1 Technology Platform. The license agreement provides for facility fees and royalties to be paid to Dyadic upon commercialization.  In November 2008: Dyadic and Codexis announced a license agreement covering use of Dyadic’s C1 expression system for large-scale production of enzymes in certain fields including biofuels and chemical and pharmaceutical intermediate production. The agreement includes an upfront payment by Codexis of $10 million provided that certain performance criteria are satisfied. Additional financial terms were not disclosed.

Iogen Corporation is a biotechnology firm specializing in cellulosic ethanol.  Iogen also develops, manufactures and markets enzymes used to modify and improve the processing of natural fibers within the textile, animal feed, and pulp and paper industries. Iogen owns and operates a large-scale state-of-the-art enzyme manufacturing facility in Ottawa, Canada. Iogen’s line of enzymes for cellulosic ethanol production is currently being used in the company’s demonstration plant, which has been operational at its Ottawa headquarters since 2004. At full capacity the plant is designed to process about 20-30 tonnes per day of feedstock, and to produce approximately 5,000 – 6,000 liters of cellulosic ethanol per day. But the company’s enzymes are also expected be available for sale in conjunction with technology licenses for the cellulosic ethanol facilities in the future. 

Iogen’s major partner in its ethanol business is Royal Dutch Shell, which first invested in Iogen in 2002. Shell subsequently increased its ownership stake in Iogen’s technology to 50 percent in 2007. Under an expanded agreement with Shell, Codexis will optimize the efficiency of Iogen Energy’s cellulosic ethanol catalysts, as well as developing new to convert biomass directly into green gasoline or green diesel. In October 2008, Iogen announced that it had commenced shipments of a 47,000 gallon cellulosic ethanol order from Shell. In January 2010, the company announced that its cellulosic ethanol production in 2009 topped 581,000 liters, more than doubling the firm’s 2008 fuel production, and surpassing the one million liter mark in cumulative production since 2004. Iogen is currently planning to construct a larger capacity plant a former pulp mill in Prince Albert, Saskatchewan, with financial assistance from the provincial and federal governments.  Iogen representatives have said that they hoped to launch the 23 Mgy plant north of Saskatoon by 2011, using wheat straw and other cellulose as feedstock. Iogen is backed in the venture by Shell, Goldman Sachs and Petro Canada. 

Iogen appears to be the only company that has submitted notifications to Environment Canada under the Canadian Environmental Protection Act for the manufacture of biofuel enzymes using engineered microorganisms (I’ll have more to say about these regulations and their impact on biofuel production in later entries in the blog). These notifications include the following: 

  • NSN (New Substance Notification) #6823: Commercial production of a novel thermophilic xylanase enzyme by the genetically engineered strain Trichoderma longibrachiatum RM4-100.
  • NSN# 11017: Trichoderma reesei 1391A, expressing of a novel xylanase II enzyme with enhanced thermal stability and a selectable marker.
  • NSN # 11909: Commercial production of a β-glucosidase enzyme by genetically engineered Trichoderma reesei P59G.
  • NSN # 12961: Commercial production of a novel thermophilic and alkalophilic xylanase II (xln2) enzyme by genetically engineered Trichoderma reesei P210A.
  • NSN # 13912: Commercial production of a thermophilic/ alkalophilic xylanase II enzyme by genetically engineered strain Trichoderma reesei P345A.

Novozymes A/S says it is the world leader in what it calls “bioinnovation”, creating biological solutions to improve its customers’ businesses. With over 700 products used in 130 countries, Novozymes’ bioinnovations improve industrial performance and safeguard the world’s resources by offering superior and sustainable solutions for tomorrow’s ever-changing marketplace. Novozymes claims to offer the leading technology platform for biofuel production.

Among relevant developments since 2008, Novozymes has launched several new enzyme preparations, including the Cellic product family, the company’s first commercial enzymes for cellulosic bioethanol; Spirizyme Ultra and Spirizyme HS for saccharification, which are expected to  provide yield increases for starch bioethanol production; and Liquozyme SC 4X for liquefaction, for use in  starch bioethanol production. In early 2009, Novozymes broke ground on a new $200 million production facility in Blair, Nebraska, to meet demand for enzymes for the production of first and second generation bioethanol. 

In February 2010, Novozymes launched its newest products in the Cellic product family, which it says are the first commercially viable enzymes to allow the production of biofuel from agricultural waste, and which might enable cellulosic biofuel to be a cost-competitive alternative to gasoline.  Novozymes says that its new Cellic® CTec2 enzymes enable the biofuel industry to produce cellulosic ethanol at a price below US $2.00 per gallon for the initial commercial-scale plants that are scheduled to be in operation in 2011. This cost is on par with gasoline and conventional ethanol at the current US market prices. The company says that Cellic CTec2 has been proven to work on many different feedstock types, including corn cobs and stalks, wheat straw, sugarcane bagasse, and woodchips.

The company says that extraordinary advances in enzyme development have reduced the enzyme cost for cellulosic ethanol by 80% over the past two years so that enzyme costs are now down to approximately 50 cents per gallon of cellulosic ethanol. Novozymes has allocated unprecedented resources to the project, and the company has also received development grants totaling $29.3 million from the US Department of Energy.

Novozymes has partnered with leading companies in the biofuel industry, such as POET, Greenfield Ethanol, Inbicon, Lignol, ICM, M&G, CTC, COFCO, Sinopec, and PRAJ to help accelerate process technology development and implementation. Coupled with further improvements in enzyme efficiency, Novozymes expects the cost to produce cellulosic biofuel to be further reduced. 

Protéus is a biotechnology company that focuses on the discovery, engineering and manufacturing of proteins of industrial interest, and on the development of innovative protein-based bioprocesses. The efficiency of Proteus’ technology platform has been demonstrated by the successful track record of the company in the life science industry, including healthcare, fine and specialty chemicals, environment and bioenergy. 

Proteus creates bio-based solutions to efficiently convert biomass into energy with dramatically lower energy inputs and net carbon emissions.  The goal is to improve and accelerate the conversion of biomass from a variety of origins, including lignocellulosic feedstock from industrial by-products, agricultural waste, municipal waste, or activated sludge into bioenergy. 

Because of the recalcitrance of biomass to hydrolysis, enhancing and accelerating the conversion of these raw materials into fermentable material is the core challenge for sustainable production of cellulosic fuels. Proteus (i) improves the productivity of existing bio-based processes by improving the performance of the enzymes involved in biomass cracking into fermentable material and (ii) creates and fine-tunes competitive natural or engineered biological cocktails that fit the specific composition of the renewable feedstock available and the characteristics of the energy-producing processes of its clients. 

Among its products are bespoke enzymes for SHF (Separate Enzymatic Hydrolysis and Fermentation), SSF (Simultaneous Saccharification and Fermentation) and for engineering CBP-enabling microorganisms (Consolidated BioProcessing). In January 2009, Proteus announced an agreement with Syngenta to develop enzymes for cellulosic ethanol production.  Syngenta said that the agreement with Proteus will allow it to accelerate its development of optimized enzymes based on Syngenta’s core competency in plant expression. 

Zymetis, Inc. is a biotechnology company dedicated to developing novel enzyme products for the emerging bio-refining industry. Derived from unique organisms, Zymetis products are designed to achieve lower costs, improved yields and higher manufacturing efficiencies in the conversion of cellulosic biomass to usable sugars. Currently, the company is developing the Ethazyme® family of industrial enzymes for use in the production of fuel–grade ethanol from biomass.  This appears to be a change in emphasis for the company, which initially stated its intentions of commercializing the microorganisms themselves as biofuel catalysts. 

Zymetis was formed in 2006 to commercialize technologies discovered by Dr. Steve Hutcheson, professor of cell biology and molecular genetics, and Dr. Ron Weiner, Professor Emeritus, from the University of Maryland. Zymetis genetically modified a rare, cellulose-eating bacterium isolated from the Chesapeake Bay to break down and convert cellulose into sugars for conversion to ethanol. In its first commercial-scale trial in 2008,  the company ran the modified microbe through a series of tests in large fermenters and found that it was able to convert one ton of cellulosic plant fiber into sugar in 72 hours.

According to the company’s website, Zymetis is offering a wide range of specialty enzymes and custom research services. The Ethazyme® family provides flexible and cost–effective enhancements and alternatives for existing enzymatic strategies for biomass saccharification. Ethazyme® products, once optimized, are effective on virtually any biomass and can be tailored to most production processes. Zymetis is currently working with consumer and industrial wastes such as waste paper, agricultural wastes such as corn cob and corn stover, timber sources such as paper mill waste and hybrid poplar and emerging fuel crops such as switchgrass.

In January 2010, the company raised $757,464 out of a total equity offering of $765,000. In February 2010, Zymetis was awarded a grant from the University of Maryland’s Maryland Industrial Partnerships Program of $108,085 to further development of low-cost solvent systems to reduce the crystallinity of native cellulose, reducing the need for enzymes in biomass digestion for the production of ethanol and other biofuels.

D. Glass Associates, Inc. is a consulting company specializing in several fields of biotechnology. David Glass, Ph.D. is a veteran of nearly thirty years in the biotech industry, with expertise in patents, technology licensing, industrial biotechnology regulatory affairs, and market and technology assessments. This blog provides back-up and expanded content to complement a presentation Dr. Glass made at the EUEC 2010 conference on February 2, 2010 entitled “Prospects for the Use of Genetic Engineering in Biofuel Production.” The slides from that presentation are available at www.slideshare.net/djglass99.