Clostridium autoethanogenum isopropanol production via native plasmid pCA replicon
Abstract
Clostridium autoethanogenum is a model gas-fermenting acetogen for commercial ethanol production. It is also a platform organism being developed for the carbon-negative production of acetone and isopropanol by gas fermentation. We have assembled a 5.5 kb pCA plasmid for type strain DSM10061 (JA1-1) using three genome sequence datasets. pCA is predicted to encode seven open-reading frames and estimated to be a low-copy number plasmid present at approximately 12 copies per chromosome. RNA-seq analyses indicate that pCA genes are transcribed at low levels and two proteins, CAETHG_05090 (putative replication protein) and CAETHG_05115 (hypothetical, a possible Mob protein), were detected at low levels during batch gas fermentations. Thiolase (thlA), CoA-transferase (ctfAB), and acetoacetate decarboxylase (adc) genes were introduced into a vector for isopropanol production in C. autoethanogenum using the native plasmid origin of replication. The availability of the pCA sequence will facilitate studies into its physiological role and could form the basis for genetic tool optimization.
- Authors:
-
- LanzaTech Inc., Skokie, IL (United States)
- Purdue Univ., West Lafayette, IN (United States)
- Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
- Northwestern Univ., Evanston, IL (United States)
- Publication Date:
- Research Org.:
- Oak Ridge National Laboratory (ORNL), Oak Ridge, TN (United States); Northwestern Univ., Evanston, IL (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC), Biological and Environmental Research (BER)
- OSTI Identifier:
- 1883782
- Alternate Identifier(s):
- OSTI ID: 1887274; OSTI ID: 1998587
- Grant/Contract Number:
- AC05-00OR22725; SC0018249
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Frontiers in Bioengineering and Biotechnology
- Additional Journal Information:
- Journal Volume: 10; Journal Issue: na; Journal ID: ISSN 2296-4185
- Publisher:
- Frontiers Media S.A.
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 09 BIOMASS FUELS; acetogen; syngas; genome; clostridia; biofuel; ethanol; synthetic biology
Citation Formats
Nogle, Robert, Nagaraju, Shilpa, Utturkar, Sagar M., Giannone, Richard J., Reynoso, Vinicio, Leang, Ching, Hettich, Robert L., Mitchell, Wayne P., Simpson, Sean D., Jewett, Michael C., Köpke, Michael, and Brown, Steven D. Clostridium autoethanogenum isopropanol production via native plasmid pCA replicon. United States: N. p., 2022.
Web. doi:10.3389/fbioe.2022.932363.
Nogle, Robert, Nagaraju, Shilpa, Utturkar, Sagar M., Giannone, Richard J., Reynoso, Vinicio, Leang, Ching, Hettich, Robert L., Mitchell, Wayne P., Simpson, Sean D., Jewett, Michael C., Köpke, Michael, & Brown, Steven D. Clostridium autoethanogenum isopropanol production via native plasmid pCA replicon. United States. https://doi.org/10.3389/fbioe.2022.932363
Nogle, Robert, Nagaraju, Shilpa, Utturkar, Sagar M., Giannone, Richard J., Reynoso, Vinicio, Leang, Ching, Hettich, Robert L., Mitchell, Wayne P., Simpson, Sean D., Jewett, Michael C., Köpke, Michael, and Brown, Steven D. Fri .
"Clostridium autoethanogenum isopropanol production via native plasmid pCA replicon". United States. https://doi.org/10.3389/fbioe.2022.932363. https://www.osti.gov/servlets/purl/1883782.
@article{osti_1883782,
title = {Clostridium autoethanogenum isopropanol production via native plasmid pCA replicon},
author = {Nogle, Robert and Nagaraju, Shilpa and Utturkar, Sagar M. and Giannone, Richard J. and Reynoso, Vinicio and Leang, Ching and Hettich, Robert L. and Mitchell, Wayne P. and Simpson, Sean D. and Jewett, Michael C. and Köpke, Michael and Brown, Steven D.},
abstractNote = {Clostridium autoethanogenum is a model gas-fermenting acetogen for commercial ethanol production. It is also a platform organism being developed for the carbon-negative production of acetone and isopropanol by gas fermentation. We have assembled a 5.5 kb pCA plasmid for type strain DSM10061 (JA1-1) using three genome sequence datasets. pCA is predicted to encode seven open-reading frames and estimated to be a low-copy number plasmid present at approximately 12 copies per chromosome. RNA-seq analyses indicate that pCA genes are transcribed at low levels and two proteins, CAETHG_05090 (putative replication protein) and CAETHG_05115 (hypothetical, a possible Mob protein), were detected at low levels during batch gas fermentations. Thiolase (thlA), CoA-transferase (ctfAB), and acetoacetate decarboxylase (adc) genes were introduced into a vector for isopropanol production in C. autoethanogenum using the native plasmid origin of replication. The availability of the pCA sequence will facilitate studies into its physiological role and could form the basis for genetic tool optimization.},
doi = {10.3389/fbioe.2022.932363},
journal = {Frontiers in Bioengineering and Biotechnology},
number = na,
volume = 10,
place = {United States},
year = {Fri Aug 05 00:00:00 EDT 2022},
month = {Fri Aug 05 00:00:00 EDT 2022}
}
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