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Shanmugam, K. T.

Publications and source records attributed to Shanmugam, K. T..

Principles and practice of designing microbial biocatalysts for fuel and chemical production

Abstract The finite nature of fossil fuels and the environmental impact of its use have raised interest in alternate renewable energy sources. Specifically, nonfood carbohydrates, such as lignocellulosic biomass, can be used to produce next generation biofuels, including cellulosic ethanol and other nonethanol fuels like butanol. However, currently there is no native microorganism that can ferment all lignocellulosic sugars to fuel molecules. Thus, research is focused on engineering improved microbial biocatalysts for production of liquid fuels at high productivity, titer, and yield. A clear understanding and application of the basic principles of microbial physiology and biochemistry are crucial to achieve this goal. In this review, we present and discuss the construction of microbial biocatalysts that integrate these principles with ethanol-producing Escherichia coli as an example of metabolic engineering. These principles also apply to fermentation of lignocellulosic sugars to other chemicals that are currently produced from petroleum.

09 BIOMASS FUELS↗

A Combined Thermochemical and Microbial Process for Recycling Polylactic Acid Polymer to Optically Pure L-Lactic Acid for Reuse

Polylactic acid polymer (PLA) produced from renewable resources can be recycled at the end of life to constituent monomer, optically pure lactic acid (LA), by a combination of chemical and biological processes. Efficient application of this closed loop of LA-PLA plastics-LA can minimize accumulation of plastics waste that pollute land and oceans. Temperature-dependent hydrolysis of PLA in water to LA follows apparent first order decay kinetics after a short lag. A modified Gompertz equation can explain the overall hydrolysis process. Alkali increased the rate of hydrolysis of PLA and reduced the length of lag period compared to water alone. The stoichiometry of base added to LA released was 1.0. The highest lactic acid yield was 0.95 g g -1 of PLA. D-LA in the syrup obtained after hydrolysis of PLA-plastics was removed using an engineered Escherichia coli to produce a L-LA syrup with an optical purity ≥ 99%. Here, these results show that thermochemical hydrolysis of PLA-based plastics to LA with optimum amount of base followed by bio-based purification to L-LA is an effective method of recycling PLA-plastics for reuse.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

US-India Consortium for Development of Sustainable Advanced Lignocellulosic Biofuel Systems

This multi-institutional project had as it main objectives to develop a fully integrated platform for the production of advanced biofuels, specifically butanol, from sustainably produced switchgrass and biomass sorghum and to develop economic models that could guide the establishment of a crop to fuel value chain. The project was organized into three work packages: (1) WP1 Feedstock Development and Supply. Activities in this WP resulted in the development of improved biomass sorghum cultivars and biomass hybrids that generate high yields of biomass with limited inputs of water, fertilizer and agri-chemicals and that display less recalcitrance to enzymatic saccharification at the biorefinery; the identification of physiological responses and genes associated with sorghum’s tolerance to water logging as the basis for the development of biomass sorghums that can be cultivated on land prone to flooding; the identification of water-logging tolerant switchgrass genotypes among existing germplasm; the identification of management practices that maximize switchgrass biomass yield (fertilizer applications, harvest regimens) on commercial farms on different types of soil in Missouri. (2) WP2 Biorefinery Technologies. This WP included the development of a recombinant E. coli strain able to convert fermentable sugars derived from biomass sorghum and switchgrass to butyrate as a precursor for the advanced biofuel butanol. Butanol is toxic to most microbes, so that yields tend to be low. In contrast, butyrate can be produced in high titer and high yield by several naturally occurring bacteria, but these species are strictly anaerobic, which complicates commercial production. Our strategy involved developing a recombinant strain of E. coli able to produce high yields and high titers of butyrate. The butyrate can then be reduced to butanol via chemical conversion. In addition, a techno-economic analysis of converting sorghum and switchgrass biomass to ethanol was conducted based on data obtained at a pilot biorefinery. These data combined with small-scale butanol production data were used as the basis for a techno-economic analysis of biomass-to-butanol conversion. Furthermore, the biorefinery stillage was evaluated as a source of fertilizer or soil amendment. In contrast to the use of biochar as a soil amendment, stillage resulted in enhanced microbial activity in the soil and release of CO2. Stillage was determined to be a suitable feedstock for biogas production in an anaerobic digester. Biorefinery lignin can be blended with polymers to create novel UV-resistant materials promising for application in outer space, and as a source of organic acids for use in plastics. These applications offer opportunities to create value from the biorefinery residues. (3) WP3. This WP focused on development of economic models that can help define the most successful strategy for implementing biofuel production from bioenergy crops. Southeastern Missouri was determined to be the best location for a bio-butanol facility that processes biomass crops, due to the abundance of feedstock within a small radius of the facility. The most effective transportation method is by rail car. Feedstock availability was considered a more important driver for commercial success than the availability of a butanol market. The average cost of producing switchgrass biomass was calculated to be $65/ton assuming a biomass yield of 8 tons per hectare (the upper end of the yield in the commercial trials in WP2). Lower yields drive up costs, but improved genotypes will conversely reduce costs. Based on farmer surveys, conducting university extension activities was determined to positively influence farmers’ willingness to consider switchgrass. Other factors included experience with pasture land and forest land, and the availability of policies that provide a safety net to the farmers in case of instability in the biofuels market. This instability can be mitigated in part by ensuring adequate biomass supplies through diversification. The project supported the training of 20 graduate students and resulted in 57 peer-reviewed journal articles (with eight additional in review or in progress), eight book chapters and five patents.

09 BIOMASS FUELS↗

Metabolic engineering of Escherichia coli for the production of butyric acid at high titer and productivity

Background: Several anaerobic bacteria produce butyric acid, a commodity chemical with use in chemical, pharmaceutical, food and feed industries, using complex media with acetate as a co-product. Butyrate titer of various recombinant Escherichia coli did not exceed 10 g l -1 in batch fermentations in any of the media tested. Results: A recombinant E. coli (strain LW393) that produced butyrate as the major fermentation product was constructed with genes from E. coli, Clostridium acetobutylicum and Treponema denticola. Strain LW393 produced 323±6 mM (28.4±0.4 g l -1 ) butyric acid in batch fermentations in mineral salt medium with glucose as C source at a yield of 0.37±0.01 g (g glucose consumed) -1 . Butyrate accounted for 90% of the total products produced by the culture. Supplementing this medium with yeast extract further increased butyric acid titer to 375±4 mM. Average volumetric productivity of butyrate with xylose as C source was 0.89±0.07 g l -1 h -1 . Conclusions: The butyrate titer reported in this study is about 2.5–3-times higher than the values reported for other recombinant E. coli and this is achieved in mineral salt medium with an expectation of lower purifcation and production cost of butyrate.

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