Clostridium autoethanogenum isopropanol production via native plasmid pCA replicon

Robert Nogle; Shilpa Nagaraju; Sagar M. Utturkar; Richard J. Giannone; Vinicio Reynoso; Ching Leang; Robert L. Hettich; Wayne P. Mitchell; Sean D. Simpson; Michael C. Jewett; Michael Köpke; Steven D. Brown

doi:10.3389/fbioe.2022.932363

Clostridium autoethanogenum isopropanol production via native plasmid pCA replicon

Robert Nogle, Shilpa Nagaraju, Sagar M. Utturkar, Richard J. Giannone, Vinicio Reynoso, Ching Leang, Robert L. Hettich, Wayne P. Mitchell, Sean D. Simpson, Michael C. Jewett, Michael Köpke, Steven D. Brown

Bioanalytical Mass Spectrometry

Research output: Contribution to journal › Article › peer-review

5 Scopus citations

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.

Original language	English
Article number	932363
Journal	Frontiers in Bioengineering and Biotechnology
Volume	10
DOIs	https://doi.org/10.3389/fbioe.2022.932363
State	Published - Aug 5 2022

Funding

This material by the Clostridium foundry for biosystem designs (cBioFAB) is based upon the work supported by the U.S. Department of Energy, Office of Biological and Environmental Research in the DOE Office of Science under Award Number DE-SC0018249. Oak Ridge National Laboratory is managed by UT-Battelle LLC for the DOE under contract DE-AC05-00OR22725. The funder had no role in the design of the study and collection, analysis, and interpretation of data and in writing the manuscript.

Funders	Funder number
U.S. Department of Energy	DE-AC05-00OR22725
Office of Science	DE-SC0018249
Biological and Environmental Research
Oak Ridge National Laboratory
UT-Battelle

Keywords

acetogen
biofuel
clostridia
ethanol
genome
syngas
synthetic biology

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.3389/fbioe.2022.932363

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Nogle, R., Nagaraju, S., Utturkar, S. M., Giannone, R. J., Reynoso, V., Leang, C., Hettich, R. L., Mitchell, W. P., Simpson, S. D., Jewett, M. C., Köpke, M., & Brown, S. D. (2022). Clostridium autoethanogenum isopropanol production via native plasmid pCA replicon. Frontiers in Bioengineering and Biotechnology, 10, Article 932363. https://doi.org/10.3389/fbioe.2022.932363

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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.",

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AB - 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.

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Clostridium autoethanogenum isopropanol production via native plasmid pCA replicon

Abstract

Funding

Keywords

UN SDGs

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