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Information × Registration Number 0217U004841, 0116U002208 , R & D reports Title Development of new biosensor and enzymatic methods for determination of the content of creatinine as a biomarker of kidney function and control process of hemodialysis. popup.stage_title Head Gonchar Mykhailo Vasyl'ovych, Кандидат педагогічних наук Registration Date 09-03-2017 Organization Institute of cell biology, National Academy of Sciences of Ukraine popup.description2 The highly sensitive express method for assay of creatinine deiminase (CDI ) activity with fluorometric detection of final product was developed and collection of mould fungi (10 species) were analysed for the presence of CDI. It was shown that the highest activity of CDI in the medium containing creatinin exhibit the fungi strains Botrytis allii and Chaetomium globosum. CodA gene of Corynebacterium glutamicum was cloned on the vectors of the inducible expression systems with introduction into a structure of recombinant plasmid (His)6-tag. Recombinant strain E. coli overproducing CDI with the productivity 20-50 mg/L of culture was constructed. An one stage method of purification of recombinant enzyme using metal affinity chromatography was developed and the highly purified enzyme preparations with specific activity 22 mkmol·min- 1·mg-1 of protein were obtained. The physical and chemical properties of recombinant CDI were studied. It was shown that an enzyme is termostable, has a high temperature optimum (75 °С) and is stabile in wide range of рН (6,0-10,5). Product Description popup.authors Іваш М.Ф. Гайда Г.З. Демків О.М. Закальська О.М. Закальський А.Є. Куцяба В.І. Лаврик М.І. Прокопів Т.М. Синенька М.М. Смуток О.В. Стасюк Н.Є. popup.nrat_date 2020-04-02 Close