Design of Lactococcus lactis strains producing S protein or receptor-binding domain of the SARSCoV-2 virus

  • Olesia V. Evdokimova Institute of Microbiology, National Academy of Sciences of Belarus, 2 Akademika Kuprevicha Street, Minsk 220084, Belarus
  • Artur E. Akhremchuk Institute of Microbiology, National Academy of Sciences of Belarus, 2 Akademika Kuprevicha Street, Minsk 220084, Belarus
  • Katsiaryna U. Akhremchuk Institute of Microbiology, National Academy of Sciences of Belarus, 2 Akademika Kuprevicha Street, Minsk 220084, Belarus
  • Dmitri O. Dormeshkin Institute of Bioоrganic Chemistry, National Academy of Sciences of Belarus, 5 Akademika Kuprevicha Street, 2 building, Minsk 220084, Belarus
  • Leonid N. Valentovich Institute of Microbiology, National Academy of Sciences of Belarus, 2 Akademika Kuprevicha Street, Minsk 220084, Belarus

Abstract

We obtained recombinant bacterial strains of Lactococcus lactis containing expression plasmids with fragments of the SARS-CoV-2 virus genome. The pNZ::spike vector construction contains the complete coding sequence of the S protein gene of the SARS-CoV-2 virus, the vector constructions pNZ::mini-spike and pNZ::HA-spike contain s gene fragments with different codon composition and size, which translated into the receptor-binding domain. Induction of s gene fragment expression in cells of the obtained strains by nisin (1 ng/ml) is followed by the synthesis of proteins specifically binding to commercial antibodies against the receptor-binding domain of the SARS-CoV-2 virus. Recombinant proteins produced by the bacteria L. lactis pNZ::spike are represented by several fractions, a molecular mass of the major fraction is about 150 kDa, which coincides with the theoretically calculated molecular mass of the full-length S protein. The bacteria L. lactis pNZ::HA-spike synthesised the recombinant protein with a molecular mass of approximately 23 kDa. In L. lactis pNZ::mini-spike cells the target protein is represented by a major fraction with a molecular mass of about 35 kDa. The produced recombinant proteins have cellular localisation.

Author Biographies

Olesia V. Evdokimova, Institute of Microbiology, National Academy of Sciences of Belarus, 2 Akademika Kuprevicha Street, Minsk 220084, Belarus

researcher at the laboratory «Centre of analytical and genetic engineering research»

Artur E. Akhremchuk, Institute of Microbiology, National Academy of Sciences of Belarus, 2 Akademika Kuprevicha Street, Minsk 220084, Belarus

researcher at the laboratory «Centre of analytical and genetic engineering research»

Katsiaryna U. Akhremchuk, Institute of Microbiology, National Academy of Sciences of Belarus, 2 Akademika Kuprevicha Street, Minsk 220084, Belarus

PhD (biology); senior researcher at the laboratory «Centre of analytical and genetic engineering research»

Dmitri O. Dormeshkin, Institute of Bioоrganic Chemistry, National Academy of Sciences of Belarus, 5 Akademika Kuprevicha Street, 2 building, Minsk 220084, Belarus

PhD (chemistry); leading researcher at the laboratory of molecular diagnostics and biotechnology

Leonid N. Valentovich, Institute of Microbiology, National Academy of Sciences of Belarus, 2 Akademika Kuprevicha Street, Minsk 220084, Belarus

PhD (biology), docent; head of the labo­ ratory «Centre of analytical and genetic engineering research»

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Published
2024-10-25
Keywords: lactic acid bacteria, Coronavirus, spike protein, receptor-binding domain, heterologous gene expression
Supporting Agencies The authors would like to thank the staff of the Republican Research and Practical Centre for Epidemiology and Microbiology doctor of science (medicine), full professor T. V. Amvrosieva and I. V. Belskaya, as well as the leading researcher at the Institute of Microbiology of the National Academy of Sciences of Belarus, PhD (biology), docent I. S. Kazlouski for provided plasmids.
How to Cite
Evdokimova, O. V., Akhremchuk, A. E., Akhremchuk, K. U., Dormeshkin, D. O., & Valentovich, L. N. (2024). Design of Lactococcus lactis strains producing S protein or receptor-binding domain of the SARSCoV-2 virus. Experimental Biology and Biotechnology, 3, 37-49. Retrieved from https://journals.bsu.by/index.php/biology/article/view/6502
Issue
No 3 (2024): Experimental Biology and Biotechnology
Section
Biotechnology and Microbiology

Copyright (c) 2024 Experimental Biology and Biotechnology

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