Monday, April 19, 2010
3-14

Biochemical characterization of thermophilic lignocellulose-degrading enzymes and their potential for biomass bioprocessing

Vasudeo Zambare1, Aditya Bhalla1, Kasiviswanath Muthukumarappan2, Rajesh Sani1, and Lew Christopher1. (1) Center for Bioprocessing Research and Development, South Dakota School of Mines and Technology, 501 East Saint Joseph Street, Rapid City, SD 57702-3995, (2) Agricultural and Biosystems Engineering, South Dakota State University, Brookings, SD 57007

Compost microbial communities are known for their capabilities to adapt to a wide range of environmental conditions when decomposing lignocellulosic materials. Their enzymes have the potential to resist and tolerate harsh pH and temperature conditions, a trait which could be employed to develop a robust process for biomass bioprocessing to fuels and chemicals. In the present study, a thermophilic microbial lignocellulose-degrading microbial consortium (TMC), enriched and isolated from yard waste compost, was used. The TMC crude enzymes (cellulase and xylanase) were produced on lignocellulosic substrates such as thermo-mechanically pretreated prairie cord grass and corn stover. The cellulase and xylanase enzymes showed optimum activity at pH 4 and were stable in a broad range of pH conditions. The optimum temperature of the TMC cellulase was 600C. The TMC xylanase was most active at 700C and had a better thermostability than the TMC cellulase. At 60°C, the half lives of the cellulase and xylanase were 15 and 18 hrs, respectively. The substrate specificity declined in the following descending order: avicel>birchwood xylan>microcrystalline cellulose>filter paper>saw dust>carboxymethyl cellulose. The Km and Vmax values were determined on avicel (cellulase) and birchwood xylan (xylanase). The SDS-PAGE revealed that the TMC produced multiple lignocellulose degrading enzymes (three isozymes of cellulase and xylanase each) with molecular weights in range of 27-75 kDa. The TMC enzymes were used in hydrolysis of PCG and CS and hydrolyzates were fermented to ethanol. The biochemical properties of the TMC crude enzymes were discussed in relation to their potential for biomass bioprocessing to bioethanol.