Cellulose crystallinity--a key predictor of the enzymatic hydrolysis rate.
The enzymatic hydrolysis of cellulose encounters various limitations that are both substrate- and enzyme-related. Although the crystallinity of pure cellulosic Avicel plays a major role in determining the rate of hydrolysis by cellulases from Trichoderma reesei, we show that it stays constant during enzymatic conversion. The mode of action of cellulases was investigated by studying their kinetics on cellulose samples. A convenient method for reaching intermediate degrees of crystallinity with Avicel was therefore developed and the initial rate of the cellulase-catalyzed hydrolysis of cellulose was demonstrated to be linearly proportional to the crystallinity index of Avicel. Despite correlation with the adsorption capacity of cellulases onto cellulose, at a given enzyme loading, the initial enzymatic rate continued to increase with a decreasing crystallinity index, even though the bound enzyme concentration stayed constant. This finding supports the determinant role of crystallinity rather than adsorption on the enzymatic rate. Thus, the cellulase activity and initial rate data obtained from various samples may provide valuable information about the details of the mechanistic action of cellulase and the hydrolysable/reactive fractions of cellulose chains. X-ray diffraction provides insight into the mode of action of Cel7A from T. reesei. In the conversion of cellulose, the (021) face of the cellulose crystal was shown to be preferentially attacked by Cel7A from T. reesei.
School of Chemical and Biomolecular Engineering, Georgia Institute of Technology, Atlanta, GA, USA.
Mélanie
M
Hall
Prabuddha
P
Bansal
Jay H
JH
Lee
Matthew J
MJ
Realff
Andreas S
AS
Bommarius
D000327:Adsorption; D002480:Cellulase / Q000378:metabolism; D002482:Cellulose / Q000737:chemistry* / Q000378:metabolism*; D018360:Crystallography, X-Ray; D006868:Hydrolysis / Q000187:drug effects; D009682:Magnetic Resonance Spectroscopy; D010756:Phosphoric Acids / Q000494:pharmacology; D014242:Trichoderma / Q000201:enzymology*
D010756:Phosphoric Acids; D002482:Cellulose; C030242:phosphoric acid; D002480:Cellulase