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Information × Registration Number 0221U000219, 0117U000657 , R & D reports Title Investigation of the mechanisms of tumor growth inhibition through limiting of carbohydrate supply, inhibition of glycolysis and gluconeogenesis and induction of arginine starvation popup.stage_title Head Sybirnyi Andrii A., Registration Date 03-01-2021 Organization Institute of Cell Biology popup.description2  The CYB2 gene encoding flavocytochrome b2 of the yeast Ogataea polymorpha was cloned on the vectors of inductive expression systems with the inclusion of a (His)6–fragment into the recombinant protein. A recombinant overproducer of this enzyme was constructed using Escherichia coli cells as a host microorganism with a productivity of 25-50 mg/L culture. A one-step method for isolation and purification of flavocytochrome b2 from recombinant E. coli cells in the form of inclusion bodies was developed. A renaturated catalytically active enzyme preparation with a specific activity of 1-2 μmol·min-1·mg-1 protein was obtained after solubilization of inclusion bodies and protein refolding in the presence of 0.3 % N-lauroylsarcosine or 0.1% sodium dodecyl sulfate. Preparative amounts of the purified flavocytochrome b2 from the recombinant yeast O. polymorpha (cell-free extracts) and E. coli cells (solubilized inclusion bodies) were obtained. A highly purified preparation of human arginase I was obtained from cells of the recombinant yeast S. cerevisiae overproducer. To detect L-Arg, we have used previously developed by us bi-enzymatic fluorimetric method, based on the reaction of urea (the cleavage product of L-Arg by arginase I) with 2,3-butanedione monooxime. Commercial urease was used to hydrolyze the endogenous urea. The developed method is characterized by a high threshold sensitivity for the determination of L-Arg (45 nM), as well as a wide range of linearity: 57 nM - 200 μM. Enzymatic analysis of glucose, lactic acid and L-arginine using recombinant enzymes (flavocytochrome b2 and arginase I) in the blood of animals (mice) with transplanted tumors under conditions of inhibition of glycolysis and/or gluconeogenesis was performed.According to the results of research in 2020, 9 works were published (7 international articles, 2 chapters in international monographs) and 2 abstracts. The total impact factor of publications is 19.70. Product Description popup.authors Ivash Mariya F Gayda Galyna Z. Demkiv Olha M. Zakalska Oksana M. Zakalskiy Andriy Ye. Prokopiv Tetyana M. Pshoniuk Bogdan V Serkiz Roman Ya. Smutok Oleh V. Stasiuk Nataliia Ye. popup.nrat_date 2021-01-03 Close