Expression of heterologous PEP-carboxylase in Methylococcus capsulatus MIR: the influence on growth characteristics and amino acid composition of methanotrophic biomass

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Abstract

Methylococcus capsulatus MIR is a strain of methanotrophic bacteria that is potentially suitable for producing feed protein and other value-added products from methane. Genomic analysis did not reveal in Mc. capsulatus MIR known pathways of C3-carboxylation, necessary for the replenishment of intermediates of the tricarboxylic acid cycle and the full functioning of metabolism. The pepc gene encoding PEP carboxylase in Methylomonas rapida 12 was introduced into Mc. capsulatus MIR cells on a plasmid under the control of a medium-strength promoter. Expression of heterologous PEP carboxylase led to an increase in the content of glutamate, glycine and lysine in the cells of the recombinant strain, but did not increase to the growth rate of the culture. Consequently, the introduction of heterologous PEP carboxylase, which carries out the C3-carboxylation reaction, helps to increase the nutritional value of the methanotroph biomass.

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About the authors

S. Y. But

Research Center of Biotechnology, Russian Academy of Sciences; Pushchino Scientific Center for Biological Research, Russian Academy of Sciences

Author for correspondence.
Email: sergeybut20063@gmail.com

Winogradsky Institute of Microbiology; Institute of Biochemistry and Physiology of Microorganisms

Russian Federation, Moscow, 119071; Moscow region, Pushchino, 142290

O. N. Rozova

Research Center of Biotechnology, Russian Academy of Sciences; Pushchino Scientific Center for Biological Research, Russian Academy of Sciences

Email: sergeybut20063@gmail.com

Winogradsky Institute of Microbiology; Institute of Biochemistry and Physiology of Microorganisms

Russian Federation, Moscow, 119071; Moscow region, Pushchino, 142290

S. V. Chistyakova

Pushchino Scientific Center for Biological Research, Russian Academy of Sciences

Email: sergeybut20063@gmail.com

Institute of Biochemistry and Physiology of Microorganisms

Russian Federation, Moscow region, Pushchino, 142290

D. S. Potapova

Tula State University

Email: sergeybut20063@gmail.com
Russian Federation, Tula, 300012

V. N. Khmelenina

Pushchino Scientific Center for Biological Research, Russian Academy of Sciences

Email: sergeybut20063@gmail.com

Institute of Biochemistry and Physiology of Microorganisms

Russian Federation, Moscow region, Pushchino, 142290

I. I. Mustakhimov

Research Center of Biotechnology, Russian Academy of Sciences; Pushchino Scientific Center for Biological Research, Russian Academy of Sciences

Email: sergeybut20063@gmail.com

Winogradsky Institute of Microbiology; Institute of Biochemistry and Physiology of Microorganisms

Russian Federation, Moscow, 119071; Moscow region, Pushchino, 142290

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Supplementary files

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2. Fig. 1. Growth curves of strains MIR/pAWP-Pphac (1) and MIR/pAWP-pepc (2).

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3. Fig. 2. Accumulation of amino acids by cells of the MIR/pAWP-Pphac (1) and MIR/pAWP-pepc (2) strains. Bars represent the standard deviation for three independent experiments. Student's t-test was used to determine the statistical significance of differences; *p < 0.05.

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