Metabolic engineering of clostridium cellulovorans to improve butanol production by consolidated bioprocessing.
File(s)acssynbio.9b00331.pdf (4.47 MB)
Accepted version
Author(s)
Wen, Zhiqiang
Ledesma-Amaro, Rodrigo
Lu, Minrui
Jin, Mingjie
Yang, Sheng
Type
Journal Article
Abstract
Clostridium cellulovorans DSM 743B can produce butyrate when grown on lignocellulose, but it can hardly synthesize butanol. In a previous study, C. cellulovorans was successfully engineered to switch the metabolism from butyryl-CoA to butanol by overexpressing an alcohol aldehyde dehydrogenase gene adhE1 from Clostridium acetobutylicum ATCC 824; however, its full potential in butanol production is still unexplored. In the study, a metabolic engineering approach based on a push-pull strategy was developed to further enhance cellulosic butanol production. In order to accomplish this, the carbon flux from acetyl-CoA to butyryl-CoA was pulled by overexpressing a trans-enoyl-coenzyme A reductase gene (ter), which can irreversibly catalyze crotonyl-CoA to butyryl-CoA. Then an acid reassimilation pathway uncoupled with acetone production was introduced to redirect the carbon flow from butyrate and acetate toward butyryl-CoA. Finally, xylose metabolism engineering was implemented by inactivating xylR (Clocel_0594) and araR (Clocel_1253), as well as overexpressing xylT (CA_C1345), which is expected to supply additional carbon and reducing power for CoA and butanol synthesis pathways. The final engineered strain produced 4.96 g/L of n-butanol from alkali extracted corn cobs (AECC), increasing by 235-fold compared to that of the wild type. It serves as a promising butanol producer by consolidated bioprocessing.
Date Issued
2020-01-24
Date Acceptance
2020-01-01
Citation
ACS Synthetic Biology, 2020, 9 (2), pp.304-315
ISSN
2161-5063
Publisher
American Chemical Society
Start Page
304
End Page
315
Journal / Book Title
ACS Synthetic Biology
Volume
9
Issue
2
Copyright Statement
© 2020 American Chemical Society. This document is the Accepted Manuscript version of a Published Work that appeared in final form in ACS Synthetic Biology, after peer review and technical editing by the publisher. To access the final edited and published work see https://doi.org/10.1021/acssynbio.9b00331
Identifier
https://www.ncbi.nlm.nih.gov/pubmed/31940438
Subjects
Clostridium
butanol
carbon flux
consolidated bioprocessing
push−pull strategy
Publication Status
Published
Coverage Spatial
United States
Date Publish Online
2020-01-24