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Competitive inhibition of cellobiohydrolase I by manno-oligosaccharides

Paper ID Volume ID Publish Year Pages File Format Full-Text
16970 42627 2015 7 PDF Available
Title
Competitive inhibition of cellobiohydrolase I by manno-oligosaccharides
Abstract

•Both mannan and MOS inhibited the hydrolysis of cellulose by cellulases.•Strong inhibitory effects of mannan and MOS on Cel7A (CBHI) were observed.•MOS were competitive inhibitors of CBHI from T. aurantiacus.•M2 exhibited a stronger inhibitory effect than M3 and M4 on the CBHI.•β-Glucosidase in cellulase preparation played an important role in mannan hydrolysis.

In the hydrolysis of softwood, significant amounts of manno-oligosaccharides (MOS) are released from mannan, the major hemicelluloses in softwood. However, the impact of MOS on the performance of cellulases is not yet clear. In this work, the effect of mannan and MOS in cellulose hydrolysis by cellulases, especially cellobiohydrolase I (CBHI) from Thermoascus aurantiacus (Ta Cel7A), was studied. The glucose yield of Avicel decreased with an increasing amount of added mannan. Commercial cellulases contained mannan hydrolysing enzymes, and β-glucosidase played an important role in mannan hydrolysis. Addition of 10 mg/ml mannan reduced the glucose yield of Avicel (at 20 g/l) from 40.1 to 24.3%. No inhibition of β-glucosidase by mannan was observed. The negative effects of mannan and MOS on the hydrolytic action of cellulases indicated that the inhibitory effect was at least partly attributed to the inhibition of Ta Cel7A (CBHI), but not on β-glucosidase. Kinetic experiments showed that MOS were competitive inhibitors of the CBHI from T. aurantiacus, and mannobiose had a stronger inhibitory effect on CBHI than mannotriose or mannotetraose. For efficient hydrolysis of softwood, it was necessary to add supplementary enzymes to hydrolyze both mannan and MOS to less inhibitory product, mannose.

Keywords
βG, glucosidase; CBH, cellobiohydrolase; CEL, cellulase; DM, dry matter; EG, endoglucanase; ESI-MS, Electrospray ionization-mass spectrometry; Km, Michaelis constant; M2, mannobiose; M3, mannotriose; M4, mannotetraose; MOS, manno-oligosaccharides; pNPC, p
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Competitive inhibition of cellobiohydrolase I by manno-oligosaccharides
Publisher
Database: Elsevier - ScienceDirect
Journal: Enzyme and Microbial Technology - Volume 68, January 2015, Pages 62–68
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering