Telke, Amar A. Ghatge, Sunil S. Kang, Seo-Hee Thangapandian, Sundarapandian Lee, Keun-Woo Shin, Hyun-Dong Um, Youngsoon Kim, Seon-Won 2024-01-20T15:00:52Z 2024-01-20T15:00:52Z 2021-09-05 2012-05 0960-8524 https://pubs.kist.re.kr/handle/201004/129280 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. (C) 2012 Elsevier Ltd. All rights reserved. English Elsevier BV Construction and characterization of chimeric cellulases with enhanced catalytic activity towards insoluble cellulosic substrates Article 10.1016/j.biortech.2012.02.066 1 Bioresource Technology, v.112, pp.10 - 17 Bioresource Technology 112 10 17 N scie scopus 000302971200002 2-s2.0-84859211668 Agricultural Engineering Biotechnology & Applied Microbiology Energy & Fuels Agriculture Biotechnology & Applied Microbiology Energy & Fuels Article CARBOHYDRATE-BINDING MODULES MOLECULAR-DYNAMICS HYDROLYSIS DOMAIN ACID SIMULATIONS MECHANISM Alicyclobacillus acidocaldrious Endoglucanase Cellulose binding module Chimeric enzymes Computational modeling