Engineering the N-Terminal End of CelA Results in Improved Performance and Growth of Caldicellulosiruptor bescii on Crystalline Cellulose

Daehwan Chung, Michael Himmel, Yannick Bomble, Sun-Ki Kim, Janet Westpheling

Research output: Contribution to journalArticlepeer-review

24 Scopus Citations

Abstract

CelA is the most abundant enzyme secreted by Caldicellulosiruptor bescii and has been shown to outperform mixtures of commercially available exo- and endoglucanases in vitro. CelA contains both a glycoside hydrolase family 9 endoglucanase and a glycoside hydrolase family 48 exoglucanase known to be synergistic in their activity, connected by three cellulose-binding domains via linker peptides. Here, repeated aspartate residues were introduced into the N-terminal ends of CelA GH9 and GH48 domains to improve secretion efficiency and/or catalytic efficiency of CelA. Among several constructs, the highest activity on carboxymethylcellulose (CMC), 0.81 ± 0.03 mg/mL was observed for the C. bescii strain containing CelA with 5-aspartate tag at the N-terminal end of GH9 domain—an 82% increase over wild type CelA. In addition, expression of CelA with N-terminal repeated aspartate residues in C. bescii results in a dramatic increase in its ability to grow on Avicel. Biotechnol. Bioeng. 2017;114: 945–950.

Original languageAmerican English
Pages (from-to)945-950
Number of pages6
JournalBiotechnology and Bioengineering
Volume114
Issue number5
DOIs
StatePublished - 2017

Bibliographical note

Publisher Copyright:
© 2016 Wiley Periodicals, Inc.

NREL Publication Number

  • NREL/JA-2700-67747

Keywords

  • biomass deconstruction
  • Caldicellulosiruptior
  • CelA
  • repeated aspartate residues

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