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Enhancement of acetyl xylan esterase activity on cellulose acetate through fusion to a family 3 cellulose binding module

Paper ID Volume ID Publish Year Pages File Format Full-Text
16856 42618 2015 7 PDF Available
Title
Enhancement of acetyl xylan esterase activity on cellulose acetate through fusion to a family 3 cellulose binding module
Abstract

•A cellulose-binding module (CBM) was fused to an acetyl xylan esterase (AnAXE).•CBM fusion increased AnAXE binding to cellulose and cellulose acetate.•CBM fusion increased AnAXE action on cellulose acetate but not cellobioseoctaacetate.•CBM fusion did not affect protein yield or catalytic efficiency on soluble substrates.•CBM fusion did not affect the regioselectivity of AnAXE.

The current study investigates the potential to increase the activity of a family 1 carbohydrate esterase on cellulose acetate through fusion to a family 3 carbohydrate binding module (CBM). Specifically, CtCBM3 from Clostridium thermocellum was fused to the carboxyl terminus of the acetyl xylan esterase (AnAXE) from Aspergillus nidulans, and active forms of both AnAXE and AnAXE–CtCBM3 were produced in Pichia pastoris. CtCBM3 fusion had negligible impact on the thermostability or regioselectivity of AnAXE; activities towards acetylated corncob xylan, 4-methylumbelliferyl acetate, p-nitrophenyl acetate, and cellobiose octaacetate were also unchanged. By contrast, the activity of AnAXE–CtCBM3 on cellulose acetate increased by two to four times over 24 h, with greater differences observed at earlier time points. Binding studies using microcrystalline cellulose (Avicel) and a commercial source of cellulose acetate confirmed functional production of the CtCBM3 domain; affinity gel electrophoresis using acetylated xylan also verified the selectivity of CtCBM3 binding to cellulose. Notably, gains in enzyme activity on cellulose acetate appeared to exceed gains in substrate binding, suggesting that fusion to CtCBM3 increases functional associations between the enzyme and insoluble, high molecular weight cellulosic substrates.

Keywords
Acetyl xylan esterase; Cellulose acetate; Hemicellulose; Carbohydrate binding module; Protein engineering
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Enhancement of acetyl xylan esterase activity on cellulose acetate through fusion to a family 3 cellulose binding module
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Publisher
Database: Elsevier - ScienceDirect
Journal: Enzyme and Microbial Technology - Volumes 79–80, November 2015, Pages 27–33
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us