Biosynthesis of brazzein using the bacterial cell-free protein synthesis system
https://doi.org/10.29235/1561-8323-2020-64-1-71-77
Abstract
Feasibility of producing brazzein plant sweetener by the cell-free biosynthesis procedure as an alternative to the classical submerged fermentation method was assessed in the present investigation. Chimeric RNA polymerase of T7 bacteriophage, S30-cell extract of Escherichia coli and multicopy plasmid vector pET42mut with the inserted brazzein gene were included into the protein synthesis. The completed research resulted in the first successful demonstration of the sweet protein biosynthesis in the cell-free system. The volumetric brazzein yield under optimized process conditions was 2 mg/ml of the reaction mixture, exceeding 57 times the maximum values that had been achieved in the previous studies applying whole-cell expression systems.
About the Authors
I. S. Kazlouski
Institute of Microbiology, National Academy of Sciences of Belarus
Belarus
Belarus
Kazlouski Illia S. - Master of Biology, Researcher.
2, Kuprevich Str., 220141, MinskI. V. Belskaya
Republican Research and Practical Center for Epidemiology and Microbiology
Belarus
Belarus
Belskaya Ina V. - Master of Biology, Junior researcher.
23, Filimonov Str., 220114, Minsk
А. I. Zinchenko
Institute of Microbiology, 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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