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Information × Registration Number 0213U003129, 0109U000118 , R & D reports Title Study of biorecognising properties of micro- and nanosized object based on enzymes and genetically modified cells with the to develop new bioanalytical methods. popup.stage_title Head Gonchar Mykhailo, Кандидат педагогічних наук Registration Date 24-01-2013 Organization Institute of cell biology, National Academy of Sciences of Ukraine popup.description2 Object of study: biorecognition, bioanalytically important enzymes, genetically modified yeast cells, biosensor analysis, enzymatic analysis, L-arginine, L-lactate, methylamine. Aim of the study: To develop a new analytical approach for analysis of L-arginine using recombinant human arginase I (combined with urease), L-lactate - L-lactate: cytochrome c-oxidoreductase and toxic methylamine by the using of recombinant methylamine oxidase. Methods of study: microbiological, genetic engineering, biochemical, physico-chemical and enzymatic. As a result of the work: - plasmid vectors carrying the gene for human liver arginase ARG1 and tag-fragments - GST or histidine cluster (His)6, under the control of yeast inducible CUP1 promoter were constructed; - cheap and effective methods for purification of recombinant arginase by affinity chromatography on macroporous glass with covalently bound ligand were developed; - highly effective methods of immobilization of bioanalytically important enzymes on micro-and nanoparticles of different nature (monodisperse spherical particles based on biocompatible polymers with carboxyl groups, ferromagnetic and gold nanoparticles, monolayer of mercaptohexadecanoic acid on the surface of gold electrodes) using a combination of carbodiimid- and pentafluorophenol methods of COOH groups activation were developed; - the study on optimization of the synthesis of gold, silver and ferromagnetic nanoparticles, their functionalization and covalent conjugation with the appropriate enzyme was done using as a model alcohol oxidase and recombinant enzymes - yeast L-lactate: cytochrome c-oxidoreductase (flavocytochrome b2) and (His)6-tagged yeast metylamine oxidase and human arginase I; - the basis of potentiometric, amperometric and enzymatic methods for the determination of L-arginine in real samples using recombinant arginase was established; - the possibility for increasing catalytic activity by additional input of respective enzyme conjugated with gold nanoparticles into recombinant cells was proved. Product Description popup.authors Гайда А.В. Гайда Г.З. Демків О.М. Закальська О.М. Закальський А.Є. Карковська М.І. Клепач Г.М. Куцяба В.І. Нечай Г.І. Прокопів Т.М. Романова М.Д. Смуток О.В. Стасюк Н.Є. popup.nrat_date 2020-04-02 Close