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Thermotolerant Methanotrophic Bacteria from Sediments of the River Chernaya, Crimea, and Assessment of Their Growth Characteristics

Abstract

The technology of single-cell protein production from natural gas is based on using thermotolerant methanotrophic bacteria with high growth rates on methane. So far, the spectrum of strains used for industrial purposes was restricted to members of the genus Methylococcus. This poses limitations to further development of this technology and fuels the search for new cultures of fast-growing methanotrophs. The later task was addressed in the present work by analyzing the sediment samples of the Chernaya River, Crimea. Molecular analysis of the microbial community composition in the sediment revealed Gammaproteobacteria as the predominant group (33‒42% of all retrieved 16S rRNA gene fragments), as well as Chloroflexi, Actinobacteria, Alphaproteobacteria, Bacteroidota, and Acidobacteria as other numerically significant community members. The methanotrophic enrichment culture obtained from the sediment contained bacteria of the genus Methylomonas as the major component, with the relative abundance of up to 60% of all 16S rRNA gene fragments. The use of various strategies for methanotroph isolation resulted in obtaining three isolates of target bacteria of the genera Methylomonas, Methylomagnum and Methylocystis. The optimal growth temperatures of these isolates were 25, 35, and 40°С, respectively. The highest specific growth rate in batch culture, 0.21 h−1, was determined for Methylocystis sp. Kr9, which displayed 99.22 and 99.13% 16S rRNA gene sequence similarity to the type strains of two Methylocystis species, Methylocystis echinoides IMET 10491T and Methylocystis parvus OBBPT, respectively.

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Funding

The work was supported by the Russian Federation Ministry of Science and Higher Education.

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Contributions

SND and NVP proposed the outline of the work. TVM and AIM collected the samples and determined their physicochemical parameters. IYuO and OVD carried out molecular analysis of microbial diversity and isolation and identification of methanotrophic strains. RZS determined the growth characteristics of the isolates. IYuO, NVP, and SND wrote the text of the article. All authors participated in discussion of the results.

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Correspondence to S. N. Dedysh.

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The authors declare that they have no conflict of interest. This article does not contain any studies involving animals or human participants performed by any of the authors.

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Translated by P. Sigalevich

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Oshkin, I.Y., Danilova, O.V., Suleimanov, R.Z. et al. Thermotolerant Methanotrophic Bacteria from Sediments of the River Chernaya, Crimea, and Assessment of Their Growth Characteristics. Microbiology 90, 588–597 (2021). https://doi.org/10.1134/S0026261721050131

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Keywords:

  • thermotolerant methanotrophic bacteria
  • river sediments
  • Methylomonas
  • Methylomagnum
  • Methylocystis
  • growth on methane