Synthetic biology, 'modified metabolism' and biotechnology for production of advanced biofuel molecules

A range of biotechnology techniques, already used in development of therapeutics and bioproducts, are being developed or investigated for production of advanced biofuel molecules or improvement of energy crops. These techniques involve complex biology at the cellular and genetic level, and require detailed explanation. As a broad introduction to the topic, this page includes selected links to advanced biofuels R&D and demonstration projects covering:

  • Synthetic biology - "designing" microorganisms that are able to produce novel fuel molecules or improve process efficiency by combining characteristics in a single microbe or overcome process issues (for example, improving tolerance to a toxic metabolic byproducts). See Joint BioEnergy Institute

    In April 2014, NYU Langone Medical Center’s Institute for Systems Genetics, announced the synthesis of the first functional chromosome in yeast, paving the way for production of novel biochemicals, including advanced biofuels, in future. See Sc2.0

  • Metabolic modification - optimising cell processes to increase production of a specific substance that can be more readily converted to advanced biofuel molecules.
  • The journal Biotechnology for Biofuels is a good source of information and recent research articles in this area.
  • Download the EBTP pdf Value Chain Fact Sheet #6 Sugar to Hydrocarbons (246 Kb)

R&D&D on biotechnological pathways to advanced biofuels

See also Process Innovation page for commercial use of biotech to improve productivity of ethanol production.

Total Amyris Biosolutions

Amyris Inc is developing technology based on synthetic biology to produce added value biofuels (renewable biodiesel and jet fuel) from sugar cane, and has established 'joint undertakings' for production facilities in Brazil.

In December 2013 Total and Amyris, announced a joint venture 'Total Amyris BioSolutions BV' to produce and sell advanced biofuels. In August 2013, Amyris announced increased production and sales of farnesene from its facility in Brazil. The company also announced that it has secured further funding worth $60m. In October 2012 Amyris entered a collaboration agreement with Microbiogen Pty Ltd for improvement of base yeast strains for biofuels production. On 19 June 2012, Azul Brazilian Airlines, completed a demonstration flight using jet fuel produced from sugarcane using Amyris technology.

"On 30 November 2011, Total and Amyris announced an agreement to expand their ongoing research and development collaboration to accelerate the deployment of Biofene® and develop renewable diesel based on this molecule produced from plant sugars. The ambitious R&D programme, launched in 2010 and managed jointly by researchers from both companies, aims to develop the necessary stages to bring the next generation renewable fuels to market at commercial scale. Total has committed to contribute $105 million in funding for an existing $180 million program. In addition, Total and Amyris have agreed to form a 50-50 joint venture company that will have exclusive rights to produce and market renewable diesel and jet fuel worldwide, as well as non-exclusive rights to other renewable products such as drilling fluids, solvents, polymers and specific biolubricants. The venture aims to begin operations in the first quarter of 2012" [Source: Amyris Press Release].

In July 2011 Amyris Brasil S.A., a subsidiary of Amyris, Inc., announced it will begin supplying up to 160 city buses in the Brazilian city of São Paulo with Amyris renewable diesel derived from sugarcane (Diesel de Cana™). Vehicle manufacturers in Brazil have issued warranties for the use of 10% Amyris renewable diesel blends in Brazil. The renewable diesel derived from plant-based sugars does not require engine or infrastructure modifications.

In March 2012, it was announced Amyris fuels would be used for performance testing with VW TDI Clean Diesel Technology.

Global Bioenergies / Audi

In January 2014, the French company Global Bioenergies announced a collaboration with Audi to develop isobutene-derived isooctane (for gasoline engines) via synthetic biology. An initial pilot plant was supported by a 5.2m Euro investment under the Investissemements d'avenir programme. Global Bioenergies is now developing a pre-commercial pilot plant at the Fraunhofer CBP in Leuna near Leipzig, Germany, to produce high-purity isobutene.

Joule / Audi

Joule has partnered with Audi AG to accelerate the commercialization of its biofuels, Sunflow-E (ethanol) and Sunflow-D (diesel). In April 2013, Joule announced advances in the development of Sunflow-G  and Sunflow- J processes for gasoline and jet fuel, respectively. In September 2012 Joule announced the commissioning of its first SunSprings™ demonstration plant in Hobbs, New Mexico, where the company will prove its scalable platform for advanced biofuel production. The Joule process uses optimized microorganisms that act as living catalysts to produce fuel, rather than first producing biomass and later extracting lipids or sugars for subsequent multi-step conversion into fuel. Joule says that its process uses a fraction of the land and capital investment required for algae-derived or agricultural biofuels. "Joule aims to show that its uniquely modular system can achieve replicable results whether installed across one or thousands of acres – opening the door to near-term deployment by eliminating scale-up costs and risks that have hamstrung biofuels for years." [Source: Joule website].

Renewable Energy Group Inc.

In January 2014, LS9 was acquired for ~$61m by Renewable Energy Group Inc. LS9 is engineering a wide range of DesignerMicrobes™ that are used in a proprietary 1-step fermentation process to produce renewable fuels and sustainable chemicals. The technology enables the rapid and widespread adoption of renewable transportation fuels. Patent-pending UltraClean™ fuels are custom engineered to have higher energetic content than ethanol or butanol; to have fuel properties that are essentially indistinguishable from those of gasoline, diesel, and jet fuel; and to be distributed in existing pipeline infrastructure and run in any vehicle. [Source: LS9].

100% sugar cane biodiesel (B100) produced by LS9, has been used succesfully in Brazil by Man Latin America, who ran tests in Summer 2013 in trucks with Euro 5 engines. The results, publicised in October 2013, showed a decrease of 15% in NOx emissions, 77% in particulate matter, and 42% in black smoke.

Gevo isobutanol production

See also Gevo (cellulosic yeast strains engineered to produce butanol) on the butanol page.

Use of biotechnology for production of advanced biofuels with algae

Please see the Algae and aqauti cbiomass page for more infromaiton on Synthetic Genomics (engineered algae), Algenol Biofuels, and Solazyme (heterotrophic algae modified to increase oil yield using standard fermentation equipment).

Other recent R&D&D on biotechnology for biofuels

OPX Biotechnology patented its Efficiency Directed Genome Engineering (EDGE) technology platform, which promises to more rapidly, rationally, and robustly engineer microbes and develop cost-effective bioprocesses. In April 2015, the technology was acquired by Cargill. It can be used to accelerate the production of a range of non-food biomaterials, for example fatty acids for conversion into renewable diesel. View the infographic on BioAcrylic production.

In the Netherlands, the 120M Euro Public/Private BE-Basic project is developing advanced genomics technologies and bioprocess engineering for bioproducts, including advanced biofuels.

In the US, the Joint BioEnergy Institute (JBEI) has identified a three-gene cluster in Micrococcus luteus that encodes enymes that catalyze key steps in the conversion of plant sugars into hydrocarbons. When introduced into Escherichia coli, the genes enabled synthesis of long-chain alkene hydrocarbons from glucose.

The JBEI has also conducted research with Lawrence Livermore National Laboratory to introduce tolerance to ionic liquids in bacteria (so they may grow normally in presence or absence of ionic liquids). This facilitates the use ionic of liquid pretreatment of cellulosic biomass, a part of JBEI's biofuel production strategy.

ARPA-E's Electrofuels program in the U.S. (Microorganisms for Liquid Transportation Fuel), involves a number of institues that are researching the use of varous microbes to produce electrofuels from carbon dioxide and water. Electrofuel technology enables electricity from renewable sources to be converted into liquid and gaseous fuels, to facilitate energy storage and use in transport.

The Energy Biosciences Institute (California / Illinois) has identified pathways used by Neurospora crassa to digest cellodextrins and xylodextrins released from plant cell walls by its secreted enzymes. Five N.crassa genes have been introduced to yeast. A hot water pretreatment is required to release the xylose from the plant cells The Institute is involved in a number of other biotech RTD projects relating to biofuels [Source: Energy Biosciences Institute 2015].

Georgia Institute of Technology and the JBEI have engineered a bacterium to synthesize pinene, a hydrocarbon produced by trees that could potentially replace high-energy rocket fuels.

University of Georgia has developed a strain of yeast Saccharomyces cerevisiae strain AJP50 (PCT/US2009/043358) that can produce higher yields of ethanol from pretreated pinewood. Research was funded by C2 Biofuels LLC and DoE [2015].

The Adams Laboratory at University of Georgia is also researching Caldicellulosiruptor bescii, an anaerobic microbe capable of breaking down plant components (cellulose, sugars, and lignin). Promising studies have been carried out using Switchgrass. Several open access papers on Caldicellulosiruptor bescii and related research are available via the Bioechnology for Biofuels journal online.

Researchers at the Colorado School of Mines have engineered Synechococcus sp. PCC 7002 to produce the plant terpenoids limonene (C10H16) and α-bisabolene (C15H24), which can be used as hydrocarbon precursors for biofuels and oher industrially biochemicals. Colorado School of Mines is a partner of the Colorado Center for Biorefining and Bioproducts.

The Energy Biosciences Institute at Berkley is researching microbial production of 1-Undecene and related fuel-like molecules. EBI discovered the first microbial aliphatic medium-chain 1-alkene (MCAE) biosynthetic enzyme, UndA, which was able to convert medium-chain fatty acids (C10-C14) into their corresponding terminal olefins using an oxygen-activating, non-heme iron dependent mechanism. This paves the way pave the way for tailored bioconversion of renewable raw materials to MCAE-based biofuels.

Researchers at UCLA have successfully introduced enzymes to Ralstonia eutropha to enable the microorganism to produce isobutanol and 3-methyl-1-butanol in an electrobioreactor using carbon dioxide as the carbon source and electricity for energy. Formic acid is used as an energy carrier. The research is supported by ARPA-E’s PETRO program.

VG Energy (a majority owned subsidiary of Viral Genetics) is developing commercial applications of Metabolic Disruption Technology MDT in the energy sector. MDT, which was primarily developed to treat diseases, is now being used to increase production of oils from algae and seeds, for use as feedstocks for biodiesel.

Aalto University in Finland is developing microbial methods for production of butanol and has a research project with Neste Oil to convert lignocellulosic wastes to microbial oil (using yeasts and molds). In November 2012 it was announced that a pilot plant would be constructed by Neste Oil to develop a commercial process for production of mircobial oil from wastes using proprietary technology (patents applied for). Waste and residues are first broken down into various sugars, which are used by the microbes to grow and produce oil [Source: Neste Oil and Aalto University websites].

In the UK, Shell and BBSRC have supported research at the University of Exeter to develop strains of E.coli that produce drop-in biodiesel molecules 'on demand'. Currently research is at the lab scale, but could offer potential for future scale-up [Source: University of Exeter Press Release, May 2013].

At the University of Texas in Austin, researchers have developed modified strains of the yeats Yarrowia Lipolytica that produce 90% cell mass as lipids, which can then be converted to biofuel [Source: Nature Communications 5, Article number: 3131 (2014)].