Meganucleases, the End of GMO plants?

By Daniela Sciaky, Ph.D.

Advancements in DNA sequencing technology have produced a large library of completed plant genome sequences. These sequences are an important step to the introduction of desirable characteristics into plants such as soybean and corn, including those traits important to the production of biofuels. Coupled with these sequences are advancements in the technology to breed plants for these traits.

Cellectis plant sciences, a subsidiary of Cellectis (Alternext: ALCLS) designs meganucleases that can be used to make targeted gene modifications. Recently the company announced the licensing of PureIntro® Agrobacterium transformation technology for monocotyledonous plants such as corn and rice from Japan Tobacco Inc. and Aerosol Beam Injector technology from Midwest Oilseeds Inc. These tools will be used by Cellectis plant sciences to modify plant cells by helping to deliver meganucleases into plant cells. Likewise, Precision BioSciences has also developed meganuclease technology for plant genome modification.

Meganucleases are enzymes that recognize specific strings of DNA sequence containing more than 12 nucleotides. These specific sequences occur in the plant genome rarely, no more than two times. Cellectis plant sciences, as well as Precision BioSciences, design meganucleases that can make targeted gene modifications without deleteriously affecting other genes. The plant’s DNA replication apparatus is used in conjunction with the meganucleases to introduce modification of specific genes.

The advantages of this technique are enormous and have implications for the development of GMO crops. Because endogenous genes are targeted for modification, no foreign sequences are retained or can be removed after the modification has taken place. Expression of the modified gene is not affected by any random placement in the genome as might occur with other modification systems. The time to achieve regulatory approval of the modified plant can potentially be reduced.

Many of the commercial plant sciences companies have become interested in the technology. Precision BioSciences is collaborating with DuPont, Bayer CropSciences and BASF Plant Sciences while BASF, Bayer CropSciences, Pioneer, Limagrain and Monsanto are reported to use Cellectis plant sciences technology. However these two technology companies appear to be embroiled in a series of patent disputes. On October 28^th Precision BioSciences announced that the US Patent and Trademark Office (PTO) had issued final decisions rejecting the claims of four Cellectis patents. Whatever the outcome, expect to see more companies interested in this technology.

Big Oil, Big Food, Small Biofuels

By Robin J. Johnson, Ph.D.

In her excellent analysis of biofuels' prospects in the 112th Congress, biofuels blogger Joanna Schroeder quotes Renewable Fuels Association head, Bob Dinneen as saying, "The ethanol industry, and the domestic biofuel industry as a whole, ought not be asked to unilaterally disarm while extensive government support continues for petroleum companies."

More and more, the line between the biofuels and petroleum industries is becoming blurred. Companies such as Valero*, Petrobras, and Sunoco* own outright, or hold partial stakes in, a number of existing ethanol plants. (Many of these plants were acquired at bargain basement prices following the collapse of the original owners' businesses). In addition, investment in biofuels technology development has become de rigueur for oil companies such as Exxon Mobil*, Chevron*, Shell, ConocoPhillips*, and Marathon Oil*.

Not to be left behind by big oil, big food is part of the biofuels scene. Actually, big food preceded big oil into the biofuels arena. Companies such as Archer Daniels Midland*, Tyson*, and Cargill** have major presences in the biofuels industry, having developed technologies and established production facilities for biofuels. ADM was once the world's largest ethanol producer, a position now held by POET of South Dakota.

With these giants mixing it up, is it any surprise the food versus fuel debate continues to rage and oil and food execs get richer?

Government subsidies for petroleum companies dwarf those for biofuels. The famous volumetric ethanol excise tax credit (VEETC), or blenders' credit, pays oil companies to blend ethanol into gasoline and biodiesel into diesel. (The American Jobs Creation Act of 2004 created the blenders' tax credit for both ethanol and biodiesel. The credit for biodiesel expired December 31, 2009, seriously harming the US biodiesel industry. So much for jobs!)

With big oil claiming three of the top ten spots on the 2010 Fortune 500 list, it's time to revamp biofuels subsidies so they benefit producers who take the lion's share of the risk. Let's reward big oil and big food for innovation and support of biofuels instead of simply paying them for obeying mandates on using it.

* indicates member of the Fortune 100 list for 2010

** indicates member of the Forbes top 100 private companies list for 2010

Drop-in Fuels from Butanol

By Robin J. Johnson, Ph.D.

Despite a 55% drop in U.S. venture capital investment in the cleantech sector, new technologies in biofuels continue to appear on the horizon. One of these, drop-in fuels from biobutanol, has gained air time as players such as the U.S. Navy, and French oil giant TOTAL throw their support behind biobutanol development.

On November 3, 2010, Cobalt Technologies of California announced that it signed a Cooperative Research and Development Agreement (CRADA) with the U.S. Navy to develop jet and diesel fuels from biobutanol. Cobalt has developed a process for converting biomass to n-butanol (a butanol isoform) using a Clostridium bacterium that can metabolize both C5 and C6 sugars found in lignocellulosic materials. The technology for converting n-butanol (which has 4 carbons) to jet fuel (a mixture of hydrocarbons containing 6-16 carbons) and diesel fuel (a mixture of hydrocarbons containing 8-21 carbons) was developed by the U.S. Naval Air Warfare Center Weapons Division. Cobalt's biobutanol process employs a chemical process to extract sugars from biomass followed by continuous fermentation. Converting n-butanol to larger hydrocarbons involves a dehydration reaction that converts n-butanol to butene (a.k.a. butylene). Butene reacts with itself to form oligomers. Cobalt opened its pilot facility in Mountain View California in January of this year.

A similar story (though without the U.S. Navy support and with support from TOTAL) exists for a Colorado company, Gevo. Gevo produces isobutanol (another isoform of butanol) from sugar and grain crops using genetically modified yeast for the fermentation step. The company has demonstrated technology to convert isobutanol into aliphatic and aromatic hydrocarbons. Like Cobalt Technologies, Gevo's chemistry depends on an initial dehydration step where isobutanol is converted to isobutylene. Isobutylene can in turn be oligomerized to jet and diesel fuel blend stocks. In August of this year, Gevo filed a registration statement on form S-1 with the Securities and Exchange Commission for a proposed initial public offering of shares of its common stock. In September, the company acquired Agri-Energy's ethanol production plant in Luverne, MN in order to retrofit it for isobutanol production.

While many may decry the widespread use of biofuels in automobiles in the U.S., preferring development of electric cars, it is clear that certain modes of transport (airplanes, trucks, freight trains, freighters) will depend for quite some time upon the energy density contained in chemical bonds. It is for these applications that drop-in biofuels hold the most promise.

Pumping Out the Biofuel

By Daniela Sciaky, Ph.D.

A $50,000 ConocoPhillips Energy Prize was recently awarded to the research team headed by Gregory Stephanopoulos of the Massachusetts Institute of Technology for engineering of a microbe that converts carbohydrates into lipids. The significance of this award is that the patent-pending technology has resulted in the creation of a heterotrophic microbe that, according to Stephanopoulos, has remarkable yields. These yields, which are reported to be close to the theoretical maximum, will help to reduce the cost of producing oil-based biofuels such as biodiesel.

The prize from ConocoPhillips is to be used by the MIT team to explore the use of hydrogen and carbon dioxide as feedstocks for the microbe. One of the schemes proposed by the team sends the carbon dioxide produced by the aerobic (organisms that grow in the presence of oxygen) heterotrophic oil-producing microbe from fermentation to a separate fermentor for use by anaerobic bacteria in the presence of hydrogen, resulting in the production of ethanol. This scheme converts carbohydrate and hydrogen to both oil and ethanol.

In another scheme anaerobic bacteria or bacteria that grow in the absence of oxygen are used to fix carbon dioxide with hydrogen to produce the feedstock for use by the aerobic heterotrophic oil-producing microbe. This scheme converts hydrogen to oil for production of high-density biofuels. The technology proposed in these schemes has been proven in different contexts to varying degrees. Existing facilities can be used to test the scalability of the technology.

Are there any companies proposing or commercializing a similar approach? As discussed in a previous post “Algae, the Dark Side of Solazyme’s Success”, Solazyme has successfully used the heterotrophic concept to produce algal-based renewable naval distillate fuel for the US Navy. The company continues to explore the use of cheap sources of fixed carbon such as conversion of biomass to sugars for fermentation by heterotrophic microorganisms. Qteros has developed the Q Microbe also known as Clostridium phytofermentans, capable of digesting a variety of cellulosic feedstocks as a source of fermentable sugars. The organism can then ferment these sugars to produce ethanol. Qteros reports that it has significantly improved the yield and titre of ethanol produced by optimizing the microbe’s metabolism.

The key to success is that the heterotrophic microbe developed by the MIT team has yields close to theoretical maximum.  Historically, production of oil from oleaginous microorganisms, organisms whose lipid content is more than 20% of their biomass, has been prohibitively expensive, ranging from $1000 to $2400 per ton. High costs result from the need to supply microorganisms with a carbon source and the slow cell growth rate during fermentation resulting in high lipid accumulation with very few cells. Only when these costs become competitive with production of biofuels from oilseeds or sugar/starch-based feedstocks will production of biofuels from heterotrophic microorganisms gain favor.

Ethanol, Why Brazil and Not the United States ….Yet?

By Daniela Sciaky, Ph.D.

Ethanol as a transportation fuel can be a dirty word in the United States when it comes to fuel efficiency. The energy content of ethanol is less than gasoline resulting in lower gas mileage. However ethanol has a higher octane rating than gasoline and if engines are tuned to use ethanol then much better performance is delivered by the use of ethanol.

What technology is available to take into account ethanol’s advantage? Sturman Industries hosted a technology summit in late October that brought the renewable fuel industry together with engine designers. Sturman proposed to replace the camshaft-controlled valves traditionally found in internal combustion engines with computer-controlled valves whose performance can be adjusted on the fly. This technology results in maximizing the performance of any fuel used by the engine.

Scania, a European engine and vehicle manufacturer, has pioneered many efforts to develop engines for ethanol and other renewable fuel powered vehicles. In October the company was awarded a 3.3 million EUR grant by the Strategic Vehicle Research and Innovation Initiative in Sweden to develop an engine for heavy commercial vehicles capable of running on biofuels. The company will focus on developing technology for both alcohol and methane gas-based fuels.

However, in case people haven’t noticed Kia Motors just unveiled the Kia Soul Flex, a car that can run on 100% ethanol, 100% gasoline or an ethanol-gasoline mixture. According to Kia the Soul Flex exhibits a 44% improvement in fuel efficiency. Power and torque are also improved compared to the existing gasoline model. The Soul Flex has a newly adapted fuel injection and catalyst system as well as a fuel tank assistance system that aids engine ignition at low temperatures. Unfortunately for the US, the car is only being introduced to Brazil.

Acceptance of ethanol in the US is still a long way off. Brazil, because of its adoption of alternative fuels has become a magnet for the adoption of new technologies that are still in the proposal/testing stages in the US. Ethanol will fail to reach its full potential in the US unless these innovative technologies are also adopted in the US.

Petrobras Cellulosic Ethanol Project - Who's on First?

By Robin J. Johnson, Ph.D.

In the ongoing quest by Petrobras to enter the cellulosic ethanol field, at least two companies appear to be involved. Novozymes of Denmark and KL Energy of South Dakota have both signed agreements to develop cellulosic ethanol technology for sugarcane bagasse.

The relationship between the three companies isn't clear. Petrobras' ethanol production activities are handled through its subsidiary, Petrobras Biocombustivel, which plans to hold a 45.7% equity interest in Acucar Guarani, the fourth largest sugarcane processor in Brazil, and a 49% equity stake in Nova Fronteira Bioenergia S.A., a joint venture with Grupo Sao Martinho.

Novozymes is a major supplier of enzymes for (among other things) biofuel production. The company is a leader in cellulosic ethanol enzyme technology.

KL Energy claims to have a proprietary process for producing cellulosic ethanol. Their technology includes thermal and mechanical pretreatment and enzymatic hydrolysis of cellulosic materials and fermentation of both C5 and C6 sugars.

Is Novozymes supplying enzymes for KL Energy's process to be integrated in a Petrobras sugar mill? Here's what we know.

In August of 2010, Petrobras, and KL Energy Corporation signed a joint development agreement for technology to turn sugarcane bagasse into ethanol. The 18-month project includes an investment by Petrobras of $11 million to adapt KL Energy’s demonstration facility in Upton, Wyoming to use bagasse. The agreement provides Petrobras with the option to license KL Energy's technology for use within Petrobras Group assets. Integration of KL Energy technology into one of Petrobras’ Brazilian sugarcane mills is scheduled to go online sometime in 2013.

On October 15, 2010 Novozymes and Petrobras announced an agreement to develop a process to make ethanol from sugarcane bagasse.

On October 21, 2010, KL Energy announced it had re-acquired all third party technology licensing agreements in order to facilitate commercialization of its technology with its strategic partners. On September 1, 2010 KL Energy signed a Master Coillaboration Agreement with add blue Consultoria Ltda. of Brazil. The two companies plan to set up a new company to handle business in Brazil.

KL Energy lists Petrobras as one of its partners on its website, but websites aren't always kept current. The Novozymes press release doesn't mention any other partners, though any cellulosic ethanol plant would have to use some form of pretreatment and fermentation technology. It will be interesting to see how all these arrangements shake out and are revealed over time.

Brazil's Braskem Invests in "Green" Plastics to Supply J&J, P&G, Others

THE VIEW FROM BRAZIL

By Henrique Oliveira

Braskem, the Brazilian petrochemical company with 29 operations in the US and Brazil, started production on its lines of “green plastics” barely a month ago and has already announced the construction of a new plant, scheduled to commence operations in 2013. Braskem plans to invest USD 100 million to produce 30,000 tonnes of plastic per year.

The project for the new plant will be officially presented today at the International Trade Fair for Plastics and Rubber, known as the “K Trade Fair", held in Germany. According to Brazilian media, Braskem has plans to become the world leader in sustainable chemistry by 2020.

The company kicked off R&D efforts in the field in 2005 and, two years later, announced the construction of a plant in the city of Triunfo, in Brazil’s southernmost state, Rio Grande do Sul. The plant, which cost USD 500 million, was inaugurated in September and has the capacity to produce 200,000 tonnes of polyethylene.

Even before the plant opened, Braskem already had twenty contracts with companies interested in its “green” plastics. The first products made from the resin are expected to reach consumers by January 2011. Johnson & Johnson, for instance, will use it in its Sundown sunscreen packaging, while P&G will produce bottles for its Pantene product line.

First-Generation Ethanol Technology Continues to Move Forward

By Robin J. Johnson, Ph.D.

In 1996 the ethanol yield from corn was 2.5 gallons per bushel. Today, a dry mill corn ethanol plant can produce up to 2.8 gallons per bushel. In Brazil, the ethanol yield from sugarcane has risen from 375 gallons per acre per year in 1975 to 870 gallons per acre per year in 2006. Technological innovation leading to these yield improvements is a hallmark of the ethanol industry.

On October 26, 2010, a California company called iDiverse announced successful modification of ethanol-producing yeasts, allowing them to tolerate stresses that occur in fuel ethanol production. A visit to the iDiverse website (which appears to have been last updated in mid-2007) reveals that the company is working to commercialize genetic engineering technology developed at and licensed from the University of Nebraska - Lincoln. The technology can be used to strengthen stress tolerance not only in yeast, but in a variety of other types of cells as well. Their various genetic constructs protect plants against diseases and environmental stresses and protect yeasts against oxidative stress. The company states that technology is presently at the proof-of-concept stage and that the second-generation is in development.

When iDiverse's technology becomes ready-for-prime-time, it could increase ethanol yield from each bushel of corn processed. In the mean time, other technological improvements may have an even larger impact on the efficiency of ethanol production. Technological innovations such as improved enzymes, corn varieties optimized for ethanol production, feedstock fractionation, and low heat fermentation are all being examined in order to increase ethanol yield. Another California company, EdeniQ, uses three-pronged approach to increasing corn ethanol yields that includes an improved yeast, a better milling device, and improved enzymes.

What does this mean for the fuel ethanol industry? Increased yield from a bushel of corn or hectare of sugarcane naturally increases the profitability of an ethanol plant. It also means that less of the world's arable land will be needed to grow fuel feedstocks, and that is a very good thing.

Algal Products from Carpet Wastewater May Reduce Costs, Increase Revenues

By Daniela Sciaky, Ph.D.

Algae have been touted as feedstocks for biofuel production based on their potential to produce massive quantities of oil. However algae have a high demand for water, as high as 11 to 13 million liters per hectare when cultivated in open ponds. Another promise of algae is their use for the bioremediation of wastewater or water that has been contaminated with animal waste, industrial waste or other sources. Cultivation of algae on just 50% of non-agriculturally consumed water could potentially produce about 250 million tons of algal biomass that could be used to produce 37 million tons of oil. The wastewater, after algal cultivation, is also rendered clean enough for other uses including use by the agricultural industry.

A recently published patent application, US 20100267122 as well as several other publications from scientists associated with The University of Georgia Biorefining and Carbon Cycling Program, demonstrate the cultivation of mixed algal populations on wastewater containing carpet mill effluents and municipal sewage. Consortia of native algal species isolated from carpet wastewater are capable of removing over 96% of the nutrients found in treated wastewater. Lipid content of the algal consortium amounts to 6.82% and the biomass produced approaches 9.2–17.8 tons per hectare per year. About 63.9% of the oil from these algal populations can be used to produce biodiesel, while the algal biomass can be used to produce biomethane. More studies are still needed to bring down the costs of biofuel production using this system.

The implications of these studies to the US carpet industry are enormous since this industry must meet stringent wastewater quality discharge regulations. The carpet industry in Dalton, Georgia generates 100–115 million liters of wastewater per day and is highly motivated to reduce the costs of waste management. The discharge, when viewed as a resource, may help to not only reduce costs but to produce another source of income for the industry.

Is the US Defense Department Leading the Biofuel Bandwagon?

By Robin J. Johnson, Ph.D.

The US Department of Defense's focus on renewable fuels represents a huge opportunity for companies in the biofuels sector. (In 2008, it was reported that the US military bought 130 million barrels of petroleum oil per year.) In Norfolk, Virginia, the US Navy has begun testing an experimental Riverine Command Boat, which will run on a fuel blend containing 50% algae-based renewable diesel fuel and 50% conventional Navy distillate fuel, NATO F-76. The renewable diesel is provided by algae producer, Solazyme, in partnership with Honeywell subsidiary, UOP.

Contrary to some reports, the algae biofuel used in the testing is not biodiesel, but a drop-in fuel made by hydroprocessing of plant oils. The hydroprocessing technology developed by UOP is similar to processes used in conventional oil refineries. By adjusting catalysts and reaction conditions, the process can produce mid-distillate fuels as well as jet fuels. In 2009, Air New Zealand, Continental Airlines, Japan Airlines and KLM Royal Dutch Airlines all successfully tested UOP's aviation biofuel, Bio-SPK (bio-synthetic paraffinic kerosene). In April of this year, the Navy tested renewable jet fuel in an F/A-18 Super Hornet aircraft.

Hydroprocessed biofuel has several advantages over conventional biodiesel fuel. Biodiesel is made up of esters of fatty acids and a small alcohol, most often methanol. It is a suitable replacement for diesel fuel, but does have a tendency to gel at cold temperatures and can also undergo oxidation. The extent of the problems caused by these drawbacks is determined to a large extent by the type of oil used in the processing. Oils with long, saturated fatty acids are more susceptible to gelling, but fairly resistant to oxidation. The opposite is true of oils containing shorter, unsaturated fatty acids. The molecules that make up hydroprocessed biofuels are similar to those found in petroleum-based fuels, making the biofuels more fully compatible with existing infrastructure. A main drawback with hydroprocessed fuels is that it may not be easy or cost-effective to generate the hydrogen required from renewable sources.

Secretary of the Navy Ray Mabus has ordered the US Navy sea service to use alternative energy sources (e.g., biofuels and nuclear power) for half of its total energy consumption by 2020. The fact that hydroprocessed biofuels have already been successfully tested in military and civilian applications bodes well for this technology. Having the DOD as a customer can’t hurt the economics.

A Tale of Two Fuels from One Feedstock

By Daniela Sciaky, Ph.D.

One problem associated with the production of biofuels from a first-generation feedstock is the un- or under-utilization of the waste products associated with production of that fuel. Scientists at the Universidade Federal de Viçosa in Brazil collaborating with scientists at Iowa State University report in the October 9^th online version of Biomass and Bioenergy that cellulosic waste remaining after the extraction of oil from first generation feedstocks can be used for ethanol production. The ethanol in turn can be used to meet the transesterification demand of the oil for biodiesel production.

Feedstocks tested include soybean, sunflower, Jatropha curcas, palm kernel, castor bean, and cottonseed. After oil extraction, palm biomass yields 108 m³ per km² (approximately 191.6 gallons per acre) of cellulosic ethanol, the highest potential ethanol yield of any of the crops studied based on Brazilian crop yields. Second is Jatropha curcas, with an ethanol yield of 40 m³ per km² or approximately 71 gallons per acre. In comparison the US Department of Agriculture estimates that 180 gallons of ethanol can be produced from one acre of harvested corn stover or 5,500 pounds of dry corn stover. Potentially a total of 3.5 hm³, or 922 million gallons of ethanol, could be produced from soybean oil extraction co-products from Brazil.

The results of this research provide an example of where waste biomass becomes extremely useful. Waste palm biomass remaining after oil extraction can yield more ethanol per acre than harvesting corn stover. In actuality a lot of the corn stover must be retained in the field in order to maintain the integrity of the soil. Therefore production of ethanol from the waste products of other biofuel production holds great promise.

Progress on Cellulosic Ethanol Plugs On

By Robin J. Johnson, Ph.D.

On Monday, October 14, investors learned that INEOS New Planet BioEnergy has received key permits for its commercial scale cellulosic ethanol plant in Indian River County, Florida. Expected to be completed some time in 2012, the plant will convert wastes and energy crops to 8 million gallons of ethanol per year and 6 megawatts of electricity. INEOS New Planet BioEnergy is a joint venture between INEOS Bio and New Planet Energy and is using INEOS Bio's unique combination of thermochemical and biochemical processes to make ethanol.

Many of the currently operating cellulosic ethanol plants use a combination of enzymatic or chemical hydrolysis followed by fermentation using a microorganism. Companies in North America pursuing this technology include Fiberight, AE Biofuels, KL Energy, DuPont Danisco Cellulosic Ethanol, and Iogen. Other companies such as POET and BlueFire Ethanol have similar plants in development. Recently, Brazilian oil giant, Petrobras, teamed up with enzyme powerhouse, Novozymes, to develop a cellulosic ethanol project for processing sugarcane bagasse. (Petrobras also has a joint development agreement for cellulosic ethanol with KL Energy.)

An alternate strategy used by some companies including INEOS Bio, substitutes a thermochemical process, gasification, for the initial hydrolysis step. The product of gasification is synthesis gas (sometimes erroneously called synthetic gas), or syngas. INEOS Bio Ethanol technology employs a patented microorganism, Clostridium ljungdahlii, which is capable of fermenting carbon monoxide and hydrogen in syngas to ethanol. Other companies using gasification as a first step in cellulosic ethanol production include Range Fuels, which broke ground on its commercial plant in 2007, and Enerkem, which has a demonstration-scale plant in operation, a commercial plant under construction, and a commercial plant in the planning stages. Both of these companies use chemical catalytic processes to convert syngas to fuel.

Gasification has several key advantages over typical hydrolysis processes. It is claimed that almost any carbon-containing material can be gasified, though in actuality different materials yield syngas with varying compositions. The reaction itself gives off heat which can be used for drying the starting materials or for generating electricity. Finally, cleaned syngas has a number of other possible uses besides conversion to ethanol. It can be burned to produce heat or catalytically converted to other fuels and chemicals.

The Energy Independence and Security Act of 2007 established a 2011 volume target for cellulosic biofuels in the US of 250 million gallons. As of July 2010, the EPA estimated the market availability for cellulosic biofuels in 2011 to be 5-17.1 million gallons. News of progress on cellulosic biofuels plants is certainly welcome in the face of the glacial progress of the industry to date.

Petrobras, Novozymes Ink Deal to Develop Cellulosic Ethanol from Sugarcane Bagasse

THE VIEW FROM BRAZIL

By Henrique Oliveira

Petrobras, Brazil’s state-controlled oil company, and Denmark’s Novozymes have signed a deal to develop a new route for the production of second-generation biofuels from sugarcane bagasse.

The deal comprises the development of enzymes and processes for the production of lignocellulosic ethanol.

The large amount of bagasse available in Brazil makes the sugarcane residue an attractive feedstock. In 2009, 612 million tonnes of sugarcane were milled in the country, according to Conab, a Brazilian government agency. The amount represents a 7% increase year-on-year.

Novozymes has been carrying out research with enzymes capable of converting bagasse into ethanol, while Petrobras has been researching biochemical processes since 2006.

Poul Andersen, head of Novozymes’ bioenergy department, says that the agreement is expected to facilitate the company’s R&D efforts in Brazil, as Petrobras has significant political and financial wherewithal. The company’s political clout is underscored, for instance, by the fact that Dilma Rousseff, chairwoman of Petrobras’ Board until March 2010, is expected by analysts to win Brazil’s presidential elections in a runoff to be held on October 31.

Jatropha, a Feedstock Needing Improvement

By Daniela Sciaky, Ph.D.

SG Biofuels in August 2010 announced the completion of the Jatropha curcas L. sequence and this month announced development of proprietary Jatropha hybrid seed technology. Investors may question the significance of these two announcements in light of the supposed advantages of Jatropha. The surge to use this “wonder” plant for biofuel production has led to a sudden rash of decisions without considering how well an unimproved plant such as Jatropha will respond to plantation growth in suboptimal conditions. SG Biofuels and D1 Oils are just two of the companies seeking to improve Jatropha for plantation growth and for other characteristics such as oil yield using conventional technology and biotechnology.

Jatropha curcas L., a non-edible shrub, is considered to be a drought tolerant plant capable of growing on arid wastelands with little input. Under these conditions Jatropha is still able to yield up to 300 gallons of oil per acre.  Based on these characteristics large numbers of Jatropha plantations have been established in order to produce oil for biofuel production and animal feed.

However Jatropha cultivation on a plantation scale has not been totally successful. One reason for failure of these plantations may be that Jatropha is not naturally found in regions with arid and semi-arid climate conditions nor is it found in regions of less than 944 mm annual precipitation. Recent experiments have also shown that productivity of Jatropha decreases with drought. However Jatropha, unlike many other crops, is able to recover from drought stress.

SG Biofuels has established a Genetic Resource Center to help the company generate hybrids in a manner similar to what has been successful for corn. SG Biofuels, with Life Technologies, has also completed the sequence of Jatropha. The sequence will be used to identify traits to improve fruit yield, pest resistance, flowering capabilities, soil adaption, uniformity and harvesting.

D1 Oils is identifying optimal sites for Jatropha cultivation by examining day length, water availability, latitude, altitude and land use by other crops. Other factors, including plant density, water and fertilizer use, land preparation, intercropping, pest and diseases, pruning regimens and harvesting time and techniques, are under examination. D1 Oils has also established a breeding program in order to develop varieties suitable for plantation cultivation around the world.

On Global Bioenergies, Biochemicals and Bioplastics

By Robin J. Johnson, Ph.D.

Financing agents were undoubtedly pleased when Global Bioenergies of Evry, France, recently announced successful production of isobutene from glucose. The company is using synthetic biology to engineer microorganisms to produce chemicals from biobased sources. Their isobutene technology involves introducing genes that produce enzymes for isobutene biosynthesis. Isobutene has been used in the manufacture of the now-banned gasoline additive, MTBE, and is currently used in the production of isooctane (another fuel additive) and butyl rubber, among other products. The company has exclusive license to technology patented by company cofounder Philippe Marlière.

Alkenes represent an important intermediate in chemical synthesis. The market for polymers of alkenes, such as polyethylene and polypropylene, is enormous. These polymers represent only a portion of chemical applications of alkenes. Today, alkenes are largely produced during petroleum refining processes such as catalytic cracking. (In the petrochemical industry, alkenes are referred to as olefins.) Global Energies' technology allows alkenes to be synthesized from renewable sources such as starch, sugar, or cellulosic materials.

According to a recent Marlière patent, alkenes such as isobutene can be produced by enzymatic decarboxylation of 3-hydroxyalkanoic acids. Certain bacteria produce 3-hydroxyalkanoic acids as building blocks for a type of bioplastic called polyhydroxyalkanoic acids or PHA. The bacteria use PHA for storing energy in much the same way animals use fat. (In fact, the biosynthetic pathway for PHA building blocks is related to metabolic pathways for fatty acids). Bacteria that make PHA can use a wide variety of carbon sources for its synthesis. Marlière’s PCT patent application WO 2010/001078 describes a process where the natural metabolic process for making PHA is subverted for the purpose of making alkenes.

Global Bioenergies is not the only company looking at PHA bioplastics as a source of industrial chemicals. The Massachusetts-based company, Metabolix, is also evaluating PHA as a source of industrial chemicals. In 2007, they received $2 million from the National Institute of Standards and Technology (NIST) to develop a commercially viable process for producing biobased chemicals from renewable agricultural sources. Patent information on Metabolix's technology indicates that the starting material for chemical production is the actual PHA bioplastic itself. That is to say, PHA polymer is treated in various ways to yield several types of industrial chemicals.

Algae, the Dark Side of Solazyme's Success

By Daniela Sciaky, Ph.D.

Investors contemplating projects in algal biofuels must have noted that, on September 15, 2010 Solazyme announced the delivery of over 20,000 gallons of algal-based renewable naval distillate fuel to the US Navy and the signing of a contract for delivery of an additional 150,000 gallons. The delivery of 20,000 gallons represents a new milestone in production of this type of biofuel. What has made Solazyme so successful and is their process helping to reduce greenhouse gas emissions? 

Solazyme's process requires heterotrophic growth of algae. Algae are provided with a source of carbon such as sucrose, whereas many companies such as Seambiotic, Solix Biofuels and Sapphire Energy rely on prototrophic growth or use of the photosynthetic process to provide the carbon that the algae rely on for growth. 

To understand Solazyme's success we must examine heterotrophic growth and what Solazyme has done to make the process successful. To grow heterotrophically or in the absence of light, algae need to be able to use a source of fixed carbon, usually in the form of sugar.  Solazyme has isolated and produced algal strains that are able to use multiple forms of fixed carbon including sugars sourced from cellulose such as corn stover or switchgrass and from the byproduct of biodiesel production, glycerol. Algae grown in the dark are able to grow to extremely high density when compared to algae grown using photosynthesis. The company has also engineered these strains to produce more oil than normal; up to 75% of dry weight. 

Most companies attempting to harness algae for biofuel production are depending on the ability of the algae to fix carbon from carbon dioxide (CO₂) using photosynthesis. The Solazyme process is dependent on providing an external source of fixed carbon. Does the Solazyme process help reduce greenhouse gas emissions? A study undertaken by Life Cycle Associates, LLC is reported to have concluded that the lifecycle greenhouse gas emissions for Solazyme's Soladiesel™ are 85 to 92% lower than emissions produced by petroleum-based ultra-low sulfur diesel. Also, the biofuels produced by Solazyme result in a significantly lower carbon footprint than the currently (April 2009) available first-generation biofuels.

Affirmation of Solazyme’s process came from the US Department of Energy that in January awarded Solazyme over $21 million for a pilot-scale biorefinery to demonstrate the heterotrophic algal oil manufacturing process and validate the process’ commercial scale economics.