Engineering the N-terminal end of CelA results in improved performance and growth of Caldicellulosiruptor bescii on crystalline cellulose

Kim, Sun -Ki; Chung, Daehwan; Himmel, Michael E.; Bomble, Yannick J.; Westpheling, Janet

doi:10.1002/bit.26242

Title: Engineering the N-terminal end of CelA results in improved performance and growth of Caldicellulosiruptor bescii on crystalline cellulose

Journal Article · Mon Dec 26 00:00:00 EST 2016 · Biotechnology and Bioengineering

DOI:https://doi.org/10.1002/bit.26242· OSTI ID:1349022

Kim, Sun -Ki ^[1]; Chung, Daehwan ^[2]; Himmel, Michael E. ^[2]; Bomble, Yannick J. ^[2]; Westpheling, Janet ^[1]

Univ. of Georgia, Athens, GA (United States); Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); National Renewable Energy Lab. (NREL), Golden, CO (United States)

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. © 2016 Wiley Periodicals, Inc.

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Research Organization:: National Renewable Energy Laboratory (NREL), Golden, CO (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Biological and Environmental Research (BER)

Grant/Contract Number:: AC36-08GO28308

OSTI ID:: 1349022

Alternate ID(s):: OSTI ID: 1401885

Report Number(s):: NREL/JA-2700-67747

Journal Information:: Biotechnology and Bioengineering, Vol. 114, Issue 5; ISSN 0006-3592

Publisher:: WileyCopyright Statement

Country of Publication:: United States

Language:: English

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Cited by: 21 works

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