Rapid chromatography for evaluating adsorption characteristics of cellulase binding domain mimetics.
The cost of cellulolytic enzymes is one barrier to the economic production of fermentable sugars from lignocellulosic biomass for the production of fuels and chemicals. One functional characteristic of cellulolytic enzymes that improves reaction kinetics over mineral acids is a cellulose binding domain that concentrates the catalytic domain to the substrate surface. We have identified maleic acid as an attractive catalytic domain with pK(a) and dicarboxylic acid structure properties that hydrolyze cellulose while producing minimal degradation of the glucose formed. In this study we report results of a rapid chromatographic method to assess the binding characteristics of potential cellulose binding domains for the construction of a synthetic cellulase over a wide range of temperatures (20 degrees to 120 degrees C). Aromatic, planar chemical structures appear to be key indicators of cellulose adsorption. Indole, the side-chain of the amino acid tryptophan, has been shown to reversibly adsorb to cellulose at temperatures between 30 degrees and 120 degrees C. Trypan blue, a polyaromatic, planar molecule, was shown to be irreversibly adsorbed to cotton cellulose at temperatures of <120 degrees C on the time scale of the experiments. These results confirm the importance of hydrophobic cellulose and the cellulose-binding component of cellulolytic enzymes and cellulolytic enzyme mimetics.
Mosier
Nathan S
NS
Department of Agricultural and Biological Engineering, 500 Central Drive, Purdue University, West Lafayette, IN 47907, USA. mosiern@purdue.edu
Wilker
Jonathan J
JJ
Ladisch
Michael R
MR
D032701:Biomimetics / Q000379:methods*; D020134:Catalytic Domain; D002480:Cellulase / Q000737:chemistry* / Q000378:metabolism; D002482:Cellulose / Q000737:chemistry / Q000378:metabolism*; D002845:Chromatography / Q000379:methods; D007211:Indoles / Q000737:chemistry; D015394:Molecular Structure; D013379:Substrate Specificity; D014343:Trypan Blue / Q000737:chemistry
D007211:Indoles; C030374:indole; D002482:Cellulose; D002480:Cellulase; D014343:Trypan Blue; C109691:microcrystalline cellulose