M1
Fundamental understanding of the recalcitrant cell wall and soluble enzymes present in hydrolysate at different stages of enzyme hydrolysis
Monday, April 28, 2014
Exhibit/Poster Hall, lower level (Hilton Clearwater Beach)
Christa Gunawan1, Mingjie Jin2, Leonardo Sousa3, Sivakumar Pattathil4, Seema Singh5, Rebecca Garlock Ong1, Bruce Dale3 and Venkatesh Balan6, (1)Biomass Conversion Research Laboratory, Deparment of Chemical Engineering and Materials Science, DOE Great Lakes Bioenergy Research Center, Michigan State University, Lansing, MI, (2)Chemical Engineering and Materials Science,Great Lakes Bioenergy Research Center (GLBRC), Michigan State University, Lansing, MI, (3)Department of Chemical Engineering and Materials Science, Michigan State University, DOE Great Lakes Bioenergy Research Center, Lansing, MI, (4)Complex Carbohydrate Research Center, University of Georgia, Athens, GA, (5)Deconstruction Division, Joint BioEnergy Institute, Emeryville, CA, (6)Department of Chemical Engineering and Materials Science,, Michigan State University and University of Pune, Lansing, MI
Ammonia based pretreatments, including AFEX and Extractive Ammonia (EA), do not extensively hydrolyze plant cell walls, and thus a complex set of enzymes are required to effectively break down and generate simple sugars. Fundamental understanding of the nature of these cell walls at different stages of enzyme hydrolysis can lead to designing a more effective enzyme cocktail, thereby leading to improved sugar release.
Corn stover will be treated using AFEX and EA and hydrolyzed at 20% solid loading using commercial enzymes. The unhydrolyzed solids (UHS) will be collected at various time points and analyzed for their polysaccharides and lignin content. In addition, glycome profiling will be used to determine the composition and extractability of the glycans present in UHS. The cell wall architecture and composition will be analyzed by Raman Imaging. These analyses will help us identify the recalcitrant portion of plant cell walls that are difficult to hydrolyze and therefore could give us clues about which enzyme activities are required to deconstruct the cell walls. In addition to these experiments, 2D-Quant analysis will be used to determine the amount of enzymes that are present in the hydrolysate. This experiment will determine the binding affinity of enzymes at different stages of hydrolysis and help us identify methods that could effectively recover and reuse them in the subsequent cycles.
