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Title: Engineering electron metabolism to increase ethanol production in Clostridium thermocellum

Journal Article · · Metabolic Engineering
 [1]; ORCiD logo [1];  [1];  [1];  [1];  [1];  [2];  [1]
  1. Dartmouth College, Hanover, NH (United States); Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
  2. Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)

The NfnAB (NADH-dependent reduced ferredoxin: NADP+ oxidoreductase) and Rnf (ion-translocating reduced ferredoxin: NAD+ oxidoreductase) complexes are thought to catalyze electron transfer between reduced ferredoxin and NAD(P)+. Efficient electron flux is critical for engineering fuel production pathways, but little is known about the relative importance of these enzymes in vivo. In this study we investigate the importance of the NfnAB and Rnf complexes in Clostridium thermocellum for growth on cellobiose and Avicel using gene deletion, enzyme assays, and fermentation product analysis. The NfnAB complex does not seem to play a major role in metabolism, since deletion of nfnAB genes had little effect on the distribution of fermentation products. By contrast, the Rnf complex appears to play an important role in ethanol formation. Deletion of rnf genes resulted in a decrease in ethanol formation. Overexpression of rnf genes resulted in an increase in ethanol production of about 30%, but only in strains where the hydG hydrogenase maturation gene was also deleted.

Research Organization:
Dartmouth College, Hanover, NH (United States); Oak Ridge National Laboratory (ORNL), Oak Ridge, TN (United States)
Sponsoring Organization:
USDOE Office of Science (SC), Biological and Environmental Research (BER)
Grant/Contract Number:
AC05-00OR22725; 4000115284
OSTI ID:
1330675
Alternate ID(s):
OSTI ID: 1394381; OSTI ID: 1414311
Journal Information:
Metabolic Engineering, Journal Name: Metabolic Engineering; ISSN 1096-7176
Publisher:
ElsevierCopyright Statement
Country of Publication:
United States
Language:
English
Citation Metrics:
Cited by: 44 works
Citation information provided by
Web of Science

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Expressing the Thermoanaerobacterium saccharolyticum pforA in engineered Clostridium thermocellum improves ethanol production journal September 2018
Enhanced ethanol formation by Clostridium thermocellum via pyruvate decarboxylase journal October 2017
Development of a core Clostridium thermocellum kinetic metabolic model consistent with multiple genetic perturbations journal May 2017
Metabolic engineering of Clostridium thermocellum for n-butanol production from cellulose journal July 2019
Clostridium thermocellum LL1210 pH homeostasis mechanisms informed by transcriptomics and metabolomics journal April 2018
Engineering Clostridium for improved solvent production: recent progress and perspective journal May 2019
Challenges and Advances for Genetic Engineering of Non-model Bacteria and Uses in Consolidated Bioprocessing journal October 2017
Clostridium sp. as Bio-Catalyst for Fuels and Chemicals Production in a Biorefinery Context journal November 2019
Metabolome analysis reveals a role for glyceraldehyde 3-phosphate dehydrogenase in the inhibition of C. thermocellum by ethanol journal November 2017
Anaerobic membrane gas extraction facilitates thermophilic hydrogen production from Clostridium thermocellum journal January 2018
Engineering microbes for direct fermentation of cellulose to bioethanol journal February 2018
Unique genetic cassettes in a Thermoanaerobacterium contribute to simultaneous conversion of cellulose and monosugars into butanol journal March 2018
Expression of a heat-stable NADPH-dependent alcohol dehydrogenase from Thermoanaerobacter pseudethanolicus 39E in Clostridium thermocellum 1313 results in increased hydroxymethylfurfural resistance journal March 2017
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