Heterologous expression of diadenylate cyclase in the form of inclusion bodies with enzymatic activity
https://doi.org/10.29235/1561-8323-2022-66-5-509-516
Abstract
Using the DNA recombination technique, a new bacterial strain Escherichia coli DAC-22 was derived, whose cells are able to carry out the heterologous expression of Bacillus thuringiensis diadenylate cyclase – the enzyme catalyzing the reaction of adenosine-5′-triphosphate (ATP) transformation into cyclic 3′,5′-diadenylate (cyclo-di-AMP). To derive the strain, E. coli “Rosetta (DE3) pLysS” cells were originally used as recipients of plasmid pET42a+ with the inserted gene disA encoding diadenylate cyclase of B. thuringiensis. The cells of the recombinant strain are able to produce heterologous diadenylate cyclase localized predominantly (by 90 %) in the fraction of the catalytically active inclusion bodies. The productivity of the new strain with respect to diadenylate cyclase structurally arranged as the inclusion bodies was 720 units/l of cultural fluid. The inclusion bodies formed by the newly engineered strain are intended for use in the technology of producing pharmacologically promising cyclo-di-AMP.
About the Authors
M. A. Vinter
Institute of Microbiology of the National Academy of Sciences of Belarus
Belarus
Belarus
Vinter Margarita A. – Junior Researcher
2, Kuprevich Str., 220141, Minsk
I. S. Kazlouski
Institute of Microbiology of the National Academy of Sciences of Belarus
Belarus
Belarus
Kazlouski Illia S. – Ph. D. (Biology), Researcher
2, Kuprevich Str., 220141, Minsk
A. I. Zinchenko
Institute of Microbiology of the National Academy of Sciences of Belarus
Belarus
Belarus
Zinchenko Anatoliy I. – Corresponding Member, D. Sc. (Biology), Professor, Head of the Laboratory
2, Kuprevich Str., 220141, Minsk
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