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United States Department of Agriculture

Agricultural Research Service

Research Project: IMPROVING BIOCHEMICAL PROCESSES FOR THE PRODUCTION OF SUSTAINABLE FUELS AND CHEMICALS

Location: Renewable Product Technology Research

2011 Annual Report


1a. Objectives (from AD-416)
The broad goal of this project is to improve the biochemical processes for converting agricultural materials to fuels and chemicals, which will consequently enable cost reductions and increase profitability of biorefining. Research will focus on significant technical challenges that must be overcome to achieve cost-competitive conversion of agricultural feedstocks to biofuels. Objective 1: Develop commercially-viable technologies based on novel deconstruction enzymes for hydrolyzing ligno-cellulose. Objective 2: Develop, via genetic engineering, strains of pentose-utilizing gram-positive bacteria that enable commercially-viable processes for producing fuel-grade ethanol or butanol from ligno-cellulosic hydrolyzates. Objective 3: Develop commercially-preferred methods for preventing, detecting, controlling, and/or correcting economically-harmful microbial contamination in ethanol production facilities. Objective 4: Develop, via high-throughput methods, enzyme and/or microbial-based technologies that enable new commercially-viable coproducts from ethanol fermentations.


1b. Approach (from AD-416)
The growth and sustainability of bioenergy production in the United States (U.S.) are hindered by a number of technical and commercial barriers. Biochemical conversion of lignocellulosic biomass to fuels and chemicals is technically feasible, but inefficiencies in the process make it economically impractical. Accomplishing the objectives will help overcome significant technical challenges to producing sustainable fuels and chemicals from agricultural feedstocks.


3. Progress Report
This report documents accomplishments for the research project 3620-41000-135-00D, entitled Improving Biochemical Processes for the Production of Sustainable Fuels and Chemicals. Research focuses on four objectives: 1. Develop novel deconstruction enzymes for hydrolyzing lignocellulose; 2. Develop Gram-positive bacteria for producing biofuels from lignocellulosic hydrolysates; 3. Develop methods for controlling bacterial contamination of fuel ethanol fermentations; 4. Develop technologies to enable new coproducts from biofuel production. In FY 2011, Agricultural Research Service (ARS) scientists in Peoria, IL, made significant progress toward these objectives, as demonstrated by the following accomplishments. Native enzymes that are capable of releasing sugar from lignocellulosic biomass were purified and characterized from fungal species of Acremonium. Several genes encoding hydrolytic enzymes were identified in the genome of the fungus Fusarium, and recombinant forms of these enzymes were expressed in Escherichia coli. Strains of lactic acid bacteria that could tolerate growth in the presence of butanol were isolated. These butanol-tolerant strains are good candidates for genetic engineering to express the butanol production pathway. Bacterial contaminants from commercial dry-grind and wet-mill ethanol facilities have been isolated and identified. Strains that contaminate sugar processing facilities were characterized with respect to polysaccharide production and biofilm formation. A study to determine the fate of antibiotics used to control contamination during ethanol production was conducted at the pilot plant facilities available at the National Corn-to-Ethanol Research Center, Edwardsville, IL. Diverse strains of fungi were screened for production of the enzyme laccase, which has application in conversion of lignin into valuable bioproducts.


4. Accomplishments


Review Publications
Bischoff, K.M., Callaway, T.R., Edrington, T.S., Genovese, K.J., Crippen, T.L., Nisbet, D.J. 2010. Antimicrobial use: Alternatives. In: Ullrey, D.E., Baer, C.K., Pond, W.G., editors. Encyclopedia of Animal Science, Second Edition. Boca Raton, Fl: CRC Press. 1(1):43-45.

Liu, S., Li, Y., Azaizeh, H., Cui, F., Tafesh, A., Bischoff, K.M. 2010. Production of value-added products by lactic acid bacteria. In: Hou, C.T., Shaw, J.-F., editors. Biocatalysis and Molecular Engineering. New York: John Wiley and Sons. p. 421-435.

Rich, J.O., Bischoff, K.M., Leathers, T.D., Cote, G.L., Liu, S. 2010. Lactic acid bacteria – Friend or Foe? Lactic acid bacteria in the production of Polysaccharides and fuel ethanol. KKU Research Journal. 15(5):424-435.

Liu, S., Bischoff, K.M., Qureshi, N., Hughes, S.R., Rich, J.O. 2010. Functional expression of the thiolase gene THl from Clostridium beijerinckii P260 in Lactococcus lactis and Lactobacillus buchneri. New Biotechnology. 27(4):283-288.

Rastogi, G., Bhalla, A., Adhikari, A., Bischoff, K.M., Hughes, S.R., Christopher, L.P., Sani, R.K. 2010. Characterization of thermostable cellulases produced by Bacillus and Geobacillus strains. Bioresource Technology. 101(22):8798-8806.

Rich, J.O., Leathers, T.D., Nunnally, M.S., Bischoff, K.M. 2011. Rapid evaluation of the antibiotic susceptibility of fuel ethanol contaminant biofilms. Bioresource Technology. 102(2):1124-1130.

Hughes, S.R., Moser, B.R., Harmsen, A.J., Bischoff, K.M., Jones, M.A., Pinkelman, R., Bang, S.S., Tasaki, K., Doll, K.M., Qureshi, N., Liu, S., Saha, B.C., Jackson Jr, J.S., Cotta, M.A., Rich, J.O., Caimi, P. 2010. Production of Candida antaractica Lipase B gene open reading frame using automated PCR gene assembly protocol on robotic workcell and expression in ethanologenic yeast for use as resin-bound biocatalyst in biodiesel production. Journal of the Association for Laboratory Automation. 16(1):17-37. DOI: 10.1016/j.jala.2010.04.002.

Songstad, D., Lakshmanan, P., Chen, J., Gibbons, W., Hughes, S.R., Nelson, R. 2011. Historical perspective of biofuels: Learning from the past to rediscover the future. In: Tomes, D., Lakshmanan, P., Songstad, D., editors. Biofuels. New York Dordrecht Heidelberg London: Springer. p. 1-8.

Gibbons, W., Hughes, S.R. 2011. Integrated biorefineries with engineered microbes and high-value co-products for profitable biofuels production. In: Tomes, D., Lakshmanan, P., Songstad, D., editors. Biofuels. New York Dordrecht Heidelberg London: Springer. p. 265-284.

Rich, J.O., Manitchotpisit, P., Peterson, S.W., Leathers, T.D. 2011. Laccase production by diverse phylogenetic clades of Aureobasidium pullulans. Rangsit Journal of Arts and Sciences. 1(1):41-47.

Walton, S.L., Bischoff, K.M., Van Heiningen, A.R., Van Walsum, G. 2010. Production of lactic acid from hemicellulose extracts by Bacillus coagulans MXL-9. Journal of Industrial Microbiology and Biotechnology. 37:823-830.

Liu, S., Leathers, T.D., Copeland, A., Chertkov, O., Goodwin, L., Mills, D.A. 2011. Complete genome sequence of Lactobacillus buchneri NRRL B-30929, a novel strain from a commercial ethanol plant. Journal of Bacteriology. 193(15):4019-4020. DOI: 10.1128/JB.05180-11.

Last Modified: 10/17/2017
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