Electrotransformation of thermophilic bacterium Caldimonas manganoxidans

J Microbiol Methods. 2022 Jan:192:106375. doi: 10.1016/j.mimet.2021.106375. Epub 2021 Nov 15.

Abstract

Caldimonas manganoxidans is a Gram-negative, thermophilic, bioplastic-producing bacterium that is a promising strain to overcome the drawbacks of existing bioplastic manufacturing methods. However, genetic manipulation of this species has not previously been studied. Here, we developed an optimized electrotransformation protocol for C. manganoxidans by screening conditions, including the bacterial growth phase, electroporation buffer, pulse strength, and recovery time. The optimized transformation protocol obtained (3.1 ± 0.78) × 108 colony-forming units/μg DNA of plasmid pBBR1MCS-2. High transformation efficiency was observed when using plasmid DNA isolated from C. manganoxidans. The DNA methylases of Escherichia coli did not affect the transformation efficiency of C. manganoxidans. The electrotransformation technique proposed here will be beneficial for the genetic manipulation of thermophilic Caldimonas species.

Keywords: Caldimonas manganoxidans; Electrotransformation; Genetic manipulation; Thermophilic bacterium; Transformation efficiency.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Comamonadaceae / genetics*
  • DNA, Bacterial / genetics
  • Electroporation / methods*
  • Plasmids / genetics
  • Transformation, Bacterial / genetics*

Substances

  • DNA, Bacterial

Supplementary concepts

  • Caldimonas manganoxidans