The culture temperature affects the properties of the Yersinia pseudotuberculosis porin inclusion bodies and the structure of the recombinant porin

Authors

  • Valentina Alexandrovana KHOMENKO G.B. Elyakov Pacific Institute of Bioorganic Chemistry, FEB RAS, Vladivostok, Russia
  • Evgeny Viktorovich SIDORIN G.B. Elyakov Pacific Institute of Bioorganic Chemistry, FEB RAS, Vladivostok, Russia
  • Svetlana Ivanovna BAKHOLDINA G.B. Elyakov Pacific Institute of Bioorganic Chemistry, FEB RAS, Vladivostok, Russia
  • Nadezhda Uryevna CHERNYSHEVA G.B. Elyakov Pacific Institute of Bioorganic Chemistry, FEB RAS, Vladivostok, Russia
  • Natalya Yurievna KIM G.B. Elyakov Pacific Institute of Bioorganic Chemistry, FEB RAS, Vladivostok, Russia
  • Marina Petrovna ISAEVA G.B. Elyakov Pacific Institute of Bioorganic Chemistry, FEB RAS, Vladivostok, Russia
  • Tamara Fedorovna SOLOV’EVA G.B. Elyakov Pacific Institute of Bioorganic Chemistry, FEB RAS, Vladivostok, Russia

Keywords:

inclusion bodies, recombinant porin, Yersinia pseudotuberculosis, porin folding intermediates, circular dichroism (CD) spectroscopy, dynamic light scattering

Abstract

The structure of inclusion bodies (IBs), in addition to the nature of the protein that forms them, depends on a number of parameters of the expression process, including the growth temperature of the cells of the producer strain. In this work we studied the effect of cell cultivation temperature on the properties and structure of Yersinia pseudotuberculosis porin IBs. The inclusion bodies synthesized in E. coli at different temperatures (37, 30 and 18 ° C) were isolated, purified and characterized. Using dynamic light scattering and optical spectroscopy, the solubility of IBs in SDS and urea and the state of the recombinant porin in the denaturants solutions (particle size, degree of polydispersity, secondary structure) were investigated. It was found that lowering the cell growth temperature resulted in a decrease in the stability of IBs in aqueous solutions of urea and SDS and an increase in the proportion of native-like conformations in IBs but also led to an increase in sensitivity of the recombinant porin to chemical denaturation. Mild methods of solubilization of IBs formed at reduced temperature are necessary for the preservation of the native-like porin structure.

Author Biographies

Valentina Alexandrovana KHOMENKO, G.B. Elyakov Pacific Institute of Bioorganic Chemistry, FEB RAS, Vladivostok, Russia

PhD, Senior Researcher

Evgeny Viktorovich SIDORIN, G.B. Elyakov Pacific Institute of Bioorganic Chemistry, FEB RAS, Vladivostok, Russia

PhD, Researcher

Svetlana Ivanovna BAKHOLDINA, G.B. Elyakov Pacific Institute of Bioorganic Chemistry, FEB RAS, Vladivostok, Russia

PhD, Senior Researcher

Nadezhda Uryevna CHERNYSHEVA, G.B. Elyakov Pacific Institute of Bioorganic Chemistry, FEB RAS, Vladivostok, Russia

Graduate student

Natalya Yurievna KIM, G.B. Elyakov Pacific Institute of Bioorganic Chemistry, FEB RAS, Vladivostok, Russia

Researcher

Marina Petrovna ISAEVA, G.B. Elyakov Pacific Institute of Bioorganic Chemistry, FEB RAS, Vladivostok, Russia

PhD, The Head of The Laboratory

Tamara Fedorovna SOLOV’EVA, G.B. Elyakov Pacific Institute of Bioorganic Chemistry, FEB RAS, Vladivostok, Russia

DSc, Principal Researche

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Published

2019-07-17

How to Cite

KHOMENKO, V. A., SIDORIN, E. V., BAKHOLDINA, S. I., CHERNYSHEVA, N. U., KIM, N. Y., ISAEVA, M. P., & SOLOV’EVA, T. F. (2019). The culture temperature affects the properties of the Yersinia pseudotuberculosis porin inclusion bodies and the structure of the recombinant porin. Vestnik of the Far East Branch of the Russian Academy of Sciences, (6s), 109–111. Retrieved from http://vestnikdvo.ru/index.php/vestnikdvo/article/view/351

Issue

No. 6s (2018): 6 Supplement

Section

Статьи