Unique plasmids generated via pUC replicon mutagenesis in an error-prone thermophile derived from Geobacillus kaustophilus HTA426

Applied and Environmental Microbiology
Jyumpei KobayashiHirokazu Suzuki

Abstract

The plasmid pGKE75-catA138T, which comprises pUC18 and the catA138T gene encoding thermostable chloramphenicol acetyltransferase with an A138T amino acid replacement (CATA138T), serves as an Escherichia coli-Geobacillus kaustophilus shuttle plasmid that confers moderate chloramphenicol resistance on G. kaustophilus HTA426. The present study examined the thermoadaptation-directed mutagenesis of pGKE75-catA138T in an error-prone thermophile, generating the mutant plasmid pGKE75(αβ)-catA138T responsible for substantial chloramphenicol resistance at 65°C. pGKE75(αβ)-catA138T contained no mutation in the catA138T gene but had two mutations in the pUC replicon, even though the replicon has no apparent role in G. kaustophilus. Biochemical characterization suggested that the efficient chloramphenicol resistance conferred by pGKE75(αβ)-catA138T is attributable to increases in intracellular CATA138T and acetyl-coenzyme A following a decrease in incomplete forms of pGKE75(αβ)-catA138T. The decrease in incomplete plasmids may be due to optimization of plasmid replication by RNA species transcribed from the mutant pUC replicon, which were actually produced in G. kaustophilus. It is noteworthy that G. kaustophilus was transformed with pGKE75...Continue Reading

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Jun 25, 2013·Applied and Environmental Microbiology·Hirokazu SuzukiToshihisa Ohshima
Nov 10, 2013·Bioscience, Biotechnology, and Biochemistry·Hirokazu SuzukiToshihisa Ohshima
Oct 19, 2014·Applied and Environmental Microbiology·Hirokazu SuzukiKatsumi Doi
Mar 19, 2015·Applied Microbiology and Biotechnology·Jyumpei KobayashiHirokazu Suzuki

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Citations

Jan 20, 2018·The Journal of Antibiotics·Hirokazu SuzukiTakashi Ohshiro
Oct 13, 2018·Applied Microbiology and Biotechnology·Hirokazu Suzuki
Nov 9, 2019·Extremophiles : Life Under Extreme Conditions·Ryota KurashikiHirokazu Suzuki
Oct 17, 2019·Frontiers in Microbiology·Maria S RamirezMarcelo E Tolmasky
Apr 13, 2021·Frontiers in Microbiology·Hirokazu SuzukiTakashi Ohshiro

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