期刊
BIOTECHNOLOGY AND BIOPROCESS ENGINEERING
卷 23, 期 1, 页码 79-85出版社
KOREAN SOC BIOTECHNOLOGY & BIOENGINEERING
DOI: 10.1007/s12257-018-0011-4
关键词
bifunctional cellulase; carbohydrate binding module; cellulose hydrolysis; fusion enzyme
资金
- National Research Foundation of Korea [NRF-2013R1A2A2A01067117, NRF-2017R1A2B4002371]
- National Research Foundation of Korea [2017R1A2B4002371, 21A20130000013] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)
Bifunctional cellulase (glycoside hydrolase 5, GH5) from Bacillus sp. D04 having both endo- and exoglucanase activities was fused with two types of carbohydrate binding modules (CBMs). CBM3 from Bacillus sp. D04 and CBM9 from Thermotoga maritima Xyn10A were added to GH5 to hydrolyze microcrystalline cellulose (Avicel) as well as water-soluble cellulose (carboxymethyl cellulose, CMC). The optimum temperature of GH5 was 50oC, while it increased to 60oC for the fusion GH5-CBM3 and GH5-CBM9, indicating that addition of CBM increased the thermostability of the enzyme. Addition of CBM3 and CBM9 enhanced the GH5 affinity (K (M)), for which K (M) decreased from 104 to 33.9 similar to 35.1 mg/mL for CMC, and from 115 to 55.5 similar to 80.3 mg/mL for Avicel, respectively. The catalytic efficiency (k (cat)/K (M)) also increased from 4.80 to 5.36 similar to 6.46 (mL/mg)/sec for CMC, and from 1.77 to 2.40 4.45 (mL/mg)/sec for Avicel, respectively, by addition of CBM3 and CBM9.
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