Determination of product inhibition of CBH1, CBH2, and EG1 using a novel cellulase activity assay.
The hydrolysis of lignocellulosic biomass by degrading enzymes (cellulases) has emerged as a promising process within the bio-ethanol industry. Yet, understanding all the intricacies of how these enzymes work has been a challenging task. Substrate-enzyme interaction in complex feed mixtures, the recalcitrance of the crystalline structure of cellulose and enzyme inactivation by product inhibition, nonproductive binding to lignin, and process stress are only some of the problems standing in the way of creating an effective and efficient process to bio-ethanol production. This study focuses on the product inhibition of cellobiohydrolases and endoglucanases. Here, we present a method of studying product inhibition by measuring the decrease in substrate, utilizing the fluorescent properties of a calcofluor dye.
Du
Faye
F
Genencor, a Danisco Division, 925 Page Mill Road, Palo Alto, CA 94304, USA. faye.du@danisco.com
Wolger
Erin
E
Wallace
Louise
L
Liu
Amy
A
Kaper
Thijs
T
Kelemen
Brad
B
D001557:Benzenesulfonates / Q000737:chemistry / Q000378:metabolism*; D056804:Biofuels; D018533:Biomass; D044602:Cellulases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D057075:Enzyme Assays / Q000379:methods*; D000431:Ethanol / Q000737:chemistry / Q000378:metabolism; D005285:Fermentation; D005456:Fluorescent Dyes / Q000737:chemistry / Q000378:metabolism*; D005656:Fungal Proteins / Q000378:metabolism*; D008031:Lignin / Q000737:chemistry / Q000378:metabolism; D015394:Molecular Structure
D001557:Benzenesulfonates; D056804:Biofuels; D005456:Fluorescent Dyes; D005656:Fungal Proteins; D000431:Ethanol; C007061:C.I. Fluorescent Brightening Agent 28; D008031:Lignin; D044602:Cellulases