Search academic texts

Information × Registration Number 0219U001828, 0118U006259 , R & D reports Title "Construction of smart-sensor devices for an analysis of creatinine and ammonium ions on the base of new nanocomposite films. Part 1. Development of an amperometric portable analyzer compatible with modern gadgets for the determination of creatinine and ammonium ions" popup.stage_title Head Gonchar Mykhailo, Кандидат педагогічних наук Registration Date 06-03-2019 Organization Institute of cell biology, National Academy of Sciences of Ukraine popup.description2 The object of research for 2018: Optimization of the technology of isolation, refolding, purification of (His) 6-labeled recombinant creatinine deiminase from overproducing cells and stabilization of the enzyme. Methods of research: microbiological, biochemical, physico-chemical, electrochemical, enzymatic. A recombinant strain of Escherichia coli, capable of overproduction of (His) 6-labeled enzyme forms from Corynebacterium glutamicum, has been developed. The recombinant protein was isolated from the cell-free extract and inclusion bodies, and after reactivation of the protein in the presence of 0.3% N-lauroylsarkosine, the active enzyme was purified in one step using metallocene chromatography on Ni-NTA agarose. Specific activity of the purified enzyme - up to 20 mkmol per minute and mg of protein. The yieldof the active enzyme depends on the isolation scheme, is 20 and 50 mg per 1 liter of microbial culture (from the cell-free extract and solubilized inclusion bodies, respectively). The main physico-chemical and kinetic properties of the (His) 6-labeled enzyme were studied: the molecular weight of the subunit, pH and temperature optimum, kinetic parameters for creatinine, stability, and specificity of the substrate. It is shown that isolated enzyme has an unique temperature stability - with temperature optimum - 75 - 80 0С. The storage stability of the enzyme is also high and can be increased by some additives. As results in 2018 have been published 1 article (with an impact factor of 2.70), 3 theses of reports and submitted 2 articles. Product Description popup.authors А.Є. Закальський М.Ф. Іваш О.В. Смуток О.М. Демків О.М. Закальська Т.М. Прокопів popup.nrat_date 2020-04-02 Close