Monday, May 4, 2009
12-29
Using carbohydrate-binding module as molecular probe to map biomass polysaccharides
Yonghua Luo, Qi Xu, Yu-San Liu, Yining Zeng, Michael E. Himmel, and Shi-You Ding. Chemical and Biosciences Center, National Renewable Energy Laboratory, 1617 cole blvd, Golden, CO 80401
Carbohydrate-binding modules (CBMs) are noncatalytic domains found in many carbohydrate-hydrolyzing enzymes, such as the cellulases and hemicellulases. They are thought to function as recognition modules that convey the catalytic modules of these enzymes to the target substrates. We have constructed a library that contains various families of CBMs, each of these CBMs has been tagged with either various fluorescent protein or genetic tag that could be used to conjugate to other fluorophores,such as quantum dots and fluorescence dyes. The fluorescently-labeled CBMs have been applied to map polysacchride distribution of raw and chemically/biologically-pretreated biomass,and further investigeted by semi-quantitative fluorescence microscopy.