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Title: Enhanced ethanol formation by Clostridium thermocellum via pyruvate decarboxylase

Here, pyruvate decarboxylase (PDC) is a well-known pathway for ethanol production, but has not been demonstrated for high titer ethanol production at temperatures above 50 °C. Here we examined the thermostability of eight PDCs. The purified bacterial enzymes retained 20% of activity after incubation for 30 min at 55 °C. Expression of these PDC genes, except the one from Zymomonas mobilis, improved ethanol production by Clostridium thermocellum. Ethanol production was further improved by expression of the heterologous alcohol dehydrogenase gene adhA from Thermoanaerobacterium saccharolyticum. Finally, the best PDC enzyme was from Acetobactor pasteurianus. A strain of C. thermocellum expressing the pdc gene from A. pasteurianus and the adhA gene from T. saccharolyticum was able to produce 21.3 g/L ethanol from 60 g/L cellulose, which is 70% of the theoretical maximum yield.
Authors:
ORCiD logo [1] ;  [2] ;  [1] ;  [1] ;  [1] ; ORCiD logo [3] ; ORCiD logo [3] ;  [1] ;  [1]
  1. Dartmouth College, Hanover, NH (United States); Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
  2. Dartmouth College, Hanover, NH (United States)
  3. Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
Publication Date:
Grant/Contract Number:
AC05-00OR22725
Type:
Accepted Manuscript
Journal Name:
Microbial Cell Factories
Additional Journal Information:
Journal Volume: 16; Journal Issue: 1; Journal ID: ISSN 1475-2859
Publisher:
BioMed Central
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)
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; Consolidated bioprocessing; Clostridium thermocellum; Cellulosic ethanol; Pyruvate decarboxylase; Alcohol dehydrogenase
OSTI Identifier:
1474857

Tian, Liang, Perot, Skyler J., Hon, Shuen, Zhou, Jilai, Liang, Xiaoyu, Bouvier, Jason T., Guss, Adam M., Olson, Daniel G., and Lynd, Lee R.. Enhanced ethanol formation by Clostridium thermocellum via pyruvate decarboxylase. United States: N. p., Web. doi:10.1186/s12934-017-0783-9.
Tian, Liang, Perot, Skyler J., Hon, Shuen, Zhou, Jilai, Liang, Xiaoyu, Bouvier, Jason T., Guss, Adam M., Olson, Daniel G., & Lynd, Lee R.. Enhanced ethanol formation by Clostridium thermocellum via pyruvate decarboxylase. United States. doi:10.1186/s12934-017-0783-9.
Tian, Liang, Perot, Skyler J., Hon, Shuen, Zhou, Jilai, Liang, Xiaoyu, Bouvier, Jason T., Guss, Adam M., Olson, Daniel G., and Lynd, Lee R.. 2017. "Enhanced ethanol formation by Clostridium thermocellum via pyruvate decarboxylase". United States. doi:10.1186/s12934-017-0783-9. https://www.osti.gov/servlets/purl/1474857.
@article{osti_1474857,
title = {Enhanced ethanol formation by Clostridium thermocellum via pyruvate decarboxylase},
author = {Tian, Liang and Perot, Skyler J. and Hon, Shuen and Zhou, Jilai and Liang, Xiaoyu and Bouvier, Jason T. and Guss, Adam M. and Olson, Daniel G. and Lynd, Lee R.},
abstractNote = {Here, pyruvate decarboxylase (PDC) is a well-known pathway for ethanol production, but has not been demonstrated for high titer ethanol production at temperatures above 50 °C. Here we examined the thermostability of eight PDCs. The purified bacterial enzymes retained 20% of activity after incubation for 30 min at 55 °C. Expression of these PDC genes, except the one from Zymomonas mobilis, improved ethanol production by Clostridium thermocellum. Ethanol production was further improved by expression of the heterologous alcohol dehydrogenase gene adhA from Thermoanaerobacterium saccharolyticum. Finally, the best PDC enzyme was from Acetobactor pasteurianus. A strain of C. thermocellum expressing the pdc gene from A. pasteurianus and the adhA gene from T. saccharolyticum was able to produce 21.3 g/L ethanol from 60 g/L cellulose, which is 70% of the theoretical maximum yield.},
doi = {10.1186/s12934-017-0783-9},
journal = {Microbial Cell Factories},
number = 1,
volume = 16,
place = {United States},
year = {2017},
month = {10}
}

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