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Title: Deletion of a single glycosyltransferase in Caldicellulosiruptor bescii eliminates protein glycosylation and growth on crystalline cellulose

Abstract

Protein glycosylation pathways have been identified in a variety of bacteria and are best understood in pathogens and commensals in which the glycosylation targets are cell surface proteins, such as S layers, pili, and flagella. In contrast, very little is known about the glycosylation of bacterial enzymes, especially those secreted by cellulolytic bacteria. Caldicellulosiruptor bescii secretes several unique synergistic multifunctional biomass-degrading enzymes, notably cellulase A which is largely responsible for this organism's ability to grow on lignocellulosic biomass without the conventional pretreatment. It was recently discovered that extracellular CelA is heavily glycosylated. In this work, we identified an O-glycosyltransferase in the C. bescii chromosome and targeted it for deletion. The resulting mutant was unable to grow on crystalline cellulose and showed no detectable protein glycosylation. Multifunctional biomass-degrading enzymes in this strain were rapidly degraded. Furthermore with the genetic tools available in C. bescii, this system represents a unique opportunity to study the role of bacterial enzyme glycosylation as well an investigation of the pathway for protein glycosylation in a non-pathogen.

Authors:
; ; ; ; ; ; ;
Publication Date:
Research Org.:
National Renewable Energy Lab. (NREL), Golden, CO (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23)
OSTI Identifier:
1618735
Alternate Identifier(s):
OSTI ID: 1476874
Report Number(s):
NREL/JA-2700-72558
Journal ID: ISSN 1754-6834; 259; PII: 1266
Grant/Contract Number:  
CBI; AC36-08GO28308
Resource Type:
Published Article
Journal Name:
Biotechnology for Biofuels
Additional Journal Information:
Journal Name: Biotechnology for Biofuels Journal Volume: 11 Journal Issue: 1; Journal ID: ISSN 1754-6834
Publisher:
Springer Science + Business Media
Country of Publication:
Netherlands
Language:
English
Subject:
09 BIOMASS FUELS; bacterial enzymes; glycosylation; lignocellulosic biomass

Citation Formats

Russell, Jordan, Kim, Sun-Ki, Duma, Justin, Nothaft, Harald, Himmel, Michael E., Bomble, Yannick J., Szymanski, Christine M., and Westpheling, Janet. Deletion of a single glycosyltransferase in Caldicellulosiruptor bescii eliminates protein glycosylation and growth on crystalline cellulose. Netherlands: N. p., 2018. Web. doi:10.1186/s13068-018-1266-x.
Russell, Jordan, Kim, Sun-Ki, Duma, Justin, Nothaft, Harald, Himmel, Michael E., Bomble, Yannick J., Szymanski, Christine M., & Westpheling, Janet. Deletion of a single glycosyltransferase in Caldicellulosiruptor bescii eliminates protein glycosylation and growth on crystalline cellulose. Netherlands. doi:10.1186/s13068-018-1266-x.
Russell, Jordan, Kim, Sun-Ki, Duma, Justin, Nothaft, Harald, Himmel, Michael E., Bomble, Yannick J., Szymanski, Christine M., and Westpheling, Janet. Mon . "Deletion of a single glycosyltransferase in Caldicellulosiruptor bescii eliminates protein glycosylation and growth on crystalline cellulose". Netherlands. doi:10.1186/s13068-018-1266-x.
@article{osti_1618735,
title = {Deletion of a single glycosyltransferase in Caldicellulosiruptor bescii eliminates protein glycosylation and growth on crystalline cellulose},
author = {Russell, Jordan and Kim, Sun-Ki and Duma, Justin and Nothaft, Harald and Himmel, Michael E. and Bomble, Yannick J. and Szymanski, Christine M. and Westpheling, Janet},
abstractNote = {Protein glycosylation pathways have been identified in a variety of bacteria and are best understood in pathogens and commensals in which the glycosylation targets are cell surface proteins, such as S layers, pili, and flagella. In contrast, very little is known about the glycosylation of bacterial enzymes, especially those secreted by cellulolytic bacteria. Caldicellulosiruptor bescii secretes several unique synergistic multifunctional biomass-degrading enzymes, notably cellulase A which is largely responsible for this organism's ability to grow on lignocellulosic biomass without the conventional pretreatment. It was recently discovered that extracellular CelA is heavily glycosylated. In this work, we identified an O-glycosyltransferase in the C. bescii chromosome and targeted it for deletion. The resulting mutant was unable to grow on crystalline cellulose and showed no detectable protein glycosylation. Multifunctional biomass-degrading enzymes in this strain were rapidly degraded. Furthermore with the genetic tools available in C. bescii, this system represents a unique opportunity to study the role of bacterial enzyme glycosylation as well an investigation of the pathway for protein glycosylation in a non-pathogen.},
doi = {10.1186/s13068-018-1266-x},
journal = {Biotechnology for Biofuels},
number = 1,
volume = 11,
place = {Netherlands},
year = {2018},
month = {9}
}

Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record
DOI: 10.1186/s13068-018-1266-x

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Cited by: 1 work
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