Fusion of Carbohydrate Binding Modules to Bifunctional Cellulase to Enhance Binding Affinity and Cellulolytic Activity
| Autor: | Bassam Alkotaini, Beom Soo Kim, Anoth Maharjan |
|---|---|
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
biology Chemistry Biomedical Engineering Bioengineering Cellulase biology.organism_classification Applied Microbiology and Biotechnology Carboxymethyl cellulose 03 medical and health sciences chemistry.chemical_compound Hydrolysis 030104 developmental biology Thermotoga maritima medicine biology.protein Glycoside hydrolase Carbohydrate-binding module Cellulose Biotechnology Thermostability medicine.drug Nuclear chemistry |
| Zdroj: | Biotechnology and Bioprocess Engineering. 23:79-85 |
| ISSN: | 1976-3816 1226-8372 |
| Popis: | 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 (KM), for which KM decreased from 104 to 33.9 ~ 35.1 mg/mL for CMC, and from 115 to 55.5 ~ 80.3 mg/mL for Avicel, respectively. The catalytic efficiency (kcat/KM) also increased from 4.80 to 5.36 ~ 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. |
| Databáze: | OpenAIRE |
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