Enhancement of the thermostability and activity of mesophilic Clostridium cellulovorans EngD by in vitro DNA recombination with Clostridium thermocellum CelE
- Authors
- Lee, Chae Yoeng; Yu, Kyung Ok; Kim, Seung Wook; Han, Sung Ok
- Issue Date
- 4월-2010
- Publisher
- SOC BIOSCIENCE BIOENGINEERING JAPAN
- Keywords
- Clostridium cellulovorans; Clostridium thermocellum; Cellulase; Thermostability; in vitro DNA recombination
- Citation
- JOURNAL OF BIOSCIENCE AND BIOENGINEERING, v.109, no.4, pp.331 - 336
- Indexed
- SCIE
SCOPUS
- Journal Title
- JOURNAL OF BIOSCIENCE AND BIOENGINEERING
- Volume
- 109
- Number
- 4
- Start Page
- 331
- End Page
- 336
- URI
- https://scholar.korea.ac.kr/handle/2021.sw.korea/116660
- DOI
- 10.1016/j.jbiosc.2009.10.014
- ISSN
- 1389-1723
- Abstract
- The thermal stability and catalytic activity of endoglucanase (EngD) from mesophilic Clostridium cellulovorans were improved by evolutionary molecular engineering. Thermostable mutants were isolated after staggered extension process (StEP) with celE from thermophilic Clostridium thermocellum performed to conduct family shuffling and overlay screening of the resultant mutant library. The relative activity of the best-evolved clone has been improved of about 2 times higher at 50 degrees C and showed a higher k(cat)/K-m value than its engD parental clone. We determined that these variants had two amino acid substitutions (L157N, Q158E) and confirmed their effects by substituting these amino acids in the parental gene by site-directed mutagenesis. These substitutions resulted in an increase in hydrophilic or charged residues. Our results demonstrate that in vitro recombination is an effective approach to improve the thermostability and enzymatic activity of a mesophilic enzyme. (C) 2009, The Society for Biotechnology, Japan. All rights reserved.
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Collections - College of Engineering > Department of Chemical and Biological Engineering > 1. Journal Articles
- Graduate School > Department of Biotechnology > 1. Journal Articles
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