Clostridium autoethanogenum isopropanol production via native plasmid pCA replicon

Autor: 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
Jazyk: angličtina
Rok vydání: 2022
Předmět:
Zdroj: Frontiers in Bioengineering and Biotechnology, Vol 10 (2022)
Druh dokumentu: article
ISSN: 2296-4185
DOI: 10.3389/fbioe.2022.932363
Popis: 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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