skip to main content
DOE PAGES title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Development of a plasmid-based expression system in Clostridium thermocellum and its use to screen heterologous expression of bifunctional alcohol dehydrogenases (adhEs)

Abstract

Clostridium thermocellum is a promising candidate for ethanol production from cellulosic biomass, but requires metabolic engineering to improve ethanol yield. A key gene in the ethanol production pathway is the bifunctional aldehyde and alcohol dehydrogenase, adhE. To explore the effects of overexpressing wild-type, mutant, and exogenous adhEs, we developed a new expression plasmid, pDGO144, that exhibited improved transformation efficiency and better gene expression than its predecessor, pDGO-66. This new expression plasmid will allow for many other metabolic engineering and basic research efforts in C. thermocellum. As proof of concept, we used this plasmid to express 12 different adhE genes (both wild type and mutant) from several organisms. Ethanol production varied between clones immediately after transformation, but tended to converge to a single value after several rounds of serial transfer. The previously described mutant C. thermocellum D494G adhE gave the best ethanol production, which is consistent with previously published results.

Authors:
; ; ; ; ; ;
Publication Date:
Research Org.:
Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23)
OSTI Identifier:
1250131
Alternate Identifier(s):
OSTI ID: 1286889
Grant/Contract Number:  
AC05–00OR22725; AC02–05CH11231; AC05-00OR22725
Resource Type:
Published Article
Journal Name:
Metabolic Engineering Communications
Additional Journal Information:
Journal Name: Metabolic Engineering Communications Journal Volume: 3 Journal Issue: C; Journal ID: ISSN 2214-0301
Publisher:
Elsevier
Country of Publication:
Netherlands
Language:
English
Subject:
60 APPLIED LIFE SCIENCES

Citation Formats

Hon, Shuen, Lanahan, Anthony A., Tian, Liang, Giannone, Richard J., Hettich, Robert L., Olson, Daniel G., and Lynd, Lee R. Development of a plasmid-based expression system in Clostridium thermocellum and its use to screen heterologous expression of bifunctional alcohol dehydrogenases (adhEs). Netherlands: N. p., 2016. Web. doi:10.1016/j.meteno.2016.04.001.
Hon, Shuen, Lanahan, Anthony A., Tian, Liang, Giannone, Richard J., Hettich, Robert L., Olson, Daniel G., & Lynd, Lee R. Development of a plasmid-based expression system in Clostridium thermocellum and its use to screen heterologous expression of bifunctional alcohol dehydrogenases (adhEs). Netherlands. doi:10.1016/j.meteno.2016.04.001.
Hon, Shuen, Lanahan, Anthony A., Tian, Liang, Giannone, Richard J., Hettich, Robert L., Olson, Daniel G., and Lynd, Lee R. Thu . "Development of a plasmid-based expression system in Clostridium thermocellum and its use to screen heterologous expression of bifunctional alcohol dehydrogenases (adhEs)". Netherlands. doi:10.1016/j.meteno.2016.04.001.
@article{osti_1250131,
title = {Development of a plasmid-based expression system in Clostridium thermocellum and its use to screen heterologous expression of bifunctional alcohol dehydrogenases (adhEs)},
author = {Hon, Shuen and Lanahan, Anthony A. and Tian, Liang and Giannone, Richard J. and Hettich, Robert L. and Olson, Daniel G. and Lynd, Lee R.},
abstractNote = {Clostridium thermocellum is a promising candidate for ethanol production from cellulosic biomass, but requires metabolic engineering to improve ethanol yield. A key gene in the ethanol production pathway is the bifunctional aldehyde and alcohol dehydrogenase, adhE. To explore the effects of overexpressing wild-type, mutant, and exogenous adhEs, we developed a new expression plasmid, pDGO144, that exhibited improved transformation efficiency and better gene expression than its predecessor, pDGO-66. This new expression plasmid will allow for many other metabolic engineering and basic research efforts in C. thermocellum. As proof of concept, we used this plasmid to express 12 different adhE genes (both wild type and mutant) from several organisms. Ethanol production varied between clones immediately after transformation, but tended to converge to a single value after several rounds of serial transfer. The previously described mutant C. thermocellum D494G adhE gave the best ethanol production, which is consistent with previously published results.},
doi = {10.1016/j.meteno.2016.04.001},
journal = {Metabolic Engineering Communications},
number = C,
volume = 3,
place = {Netherlands},
year = {2016},
month = {12}
}

Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record
DOI: 10.1016/j.meteno.2016.04.001

Save / Share:

Works referencing / citing this record:

Engineering Clostridium for improved solvent production: recent progress and perspective
journal, May 2019

  • Cheng, Chi; Bao, Teng; Yang, Shang-Tian
  • Applied Microbiology and Biotechnology, Vol. 103, Issue 14
  • DOI: 10.1007/s00253-019-09916-7

Engineering Clostridium for improved solvent production: recent progress and perspective
journal, May 2019

  • Cheng, Chi; Bao, Teng; Yang, Shang-Tian
  • Applied Microbiology and Biotechnology, Vol. 103, Issue 14
  • DOI: 10.1007/s00253-019-09916-7