Construction and characterization of chimeric cellulases with enhanced catalytic activity towards insoluble cellulosic substrates

Title
Construction and characterization of chimeric cellulases with enhanced catalytic activity towards insoluble cellulosic substrates
Authors
Telke, Amar A.Ghatge, Sunil S강서희Thangapandian, Sundarapandian이근우신현동엄영순김선원
Keywords
cellulase; cellulose; enzyme
Issue Date
2012-05
Publisher
Bioresource technology
Citation
VOL 112, 10-17
Abstract
The chimeric proteins viz. CBM3–Cel9A, CBM4–Cel9A and CBM30–Cel9A, are constructed by fusion of family 3, 4, and 30 cellulose binding modules (CBMs) to N-terminus of family 9 endoglucanase (Cel9A) from Alicyclobacillus acidocaldrious. The chimeric enzymes were successfully expressed in Escherichia coli and purified to homogeneity. The chimeric enzymes showed significant increase in Avicel (8–12 folds) and filter paper (7–10 folds) degradation activities compared to Cel9A endoglucanase. Computational protein modeling and simulation on the chimeric enzymes were applied to analyze the fused CBMs effect on the increased insoluble cellulosic substrates degradation activity. Thin layer chromatography analysis of the enzymatic hydrolysis products and distribution of reducing sugars between soluble and insoluble fractions indicated processive cleavage of insoluble cellulosic substrates by the chimeras. The fused CBMs played a critical accessory role for the Cel9A catalytic domain and changed its character to facilitate the processive cleavage of insoluble cellulosic substrates.
URI
http://pubs.kist.re.kr/handle/201004/42417
ISSN
09608524
Appears in Collections:
KIST Publication > Article
Files in This Item:
There are no files associated with this item.
Export
RIS (EndNote)
XLS (Excel)
XML


qrcode

Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.

BROWSE