Abstract
The genome of aerobic methanotroph Methylococcus capsulatus Bath possesses genes of three biochemical pathways of C1-carbon assimilation: the ribulose monophosphate cycle, the Calvin–Benson–Bassham cycle, and the partial serine cycle. Numerous studies have demonstrated that during methanotrophic growth cells of Methylococcus capsulatus Bath express key enzymes of these routes. In this study, the role of the serine cycle key enzymes, serine-glyoxylate aminotransferase (Sga) and malyl-CoA lyase (Mcl) in metabolism of Methylococcus capsulatus Bath was investigated by gene inactivation. The Δsga mutant obtained by double homologous recombination showed a prolonged lag phase, and after the lag period, the growth rate became similar to that of the wild type strain. The elevated intracellular levels of glutamate, serine, glycine, alanine, methionine, leucine, and succinate suggested significant metabolic changes in the mutant cells. Deletion of the mcl gene resulted in very poor growth and glycine only partially improved growth of the mutant strain. Cells of Δmcl mutant possess lower content of histidine, but enhanced level of alanine, leucine, and lysine than those of the wild type strain. Our data imply the importance of the serine cycle enzymes in metabolism of the methanotroph as well as relationships of the three C1 assimilation pathways in the gammaproteobacterial methanotrophs.
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Funding
The work was supported by the Russian Foundation for Basic Research # 20–04-00493 and by a grant for the Development of genomic editing technologies for innovation in industrial biotechnology (Grant No. 075-15-2021-1071) from the Ministry of Science and Higher Education of the Russian Federation.
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SVE and SYB carried out the experiments; SYB and VNK analyzed the data; SYB, VNK and IIM prepared the manuscript. All authors read, reviewed and approved the final manuscript.
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Egorova, S.V., Khmelenina, V.N., Mustakhimov, I.I. et al. The Role of Serine-Glyoxylate Aminotransferase and Malyl-CoA Lyase in the Metabolism of Methylococcus capsulatus Bath. Curr Microbiol 80, 311 (2023). https://doi.org/10.1007/s00284-023-03426-6
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DOI: https://doi.org/10.1007/s00284-023-03426-6