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Information × Registration Number 0213U004399, 0113U002866 , R & D reports Title Novel nanosized elements of biosensors based on oxidoreductases. popup.stage_title Head Gonchar Mykhailo Vasylyovych, Кандидат педагогічних наук Registration Date 12-12-2013 Organization Institute of cell biology, National Academy of Sciences of Ukraine popup.description2 Report about scientific project: 31 p., 1 table, 38 references. The object of investigation: oxido reductases of analytical importance: glucose oxidase (GO) of fungi Penicillium, methylamine oxidase (AMO) isolated from recombinant yeast Saccharomyces cerevisiae, horse radish peroxidase, nanoparticles of gold (Au-NPs) and silver(Ag-NPs). The aim of the work: a development of a new nano-sized bio-recognition elements for mediator-less biosensors on glucose and methylamin. These biosensors of “third generation” on glucose and methylamine will be constructed on the base of GO and AMO, co-immobilized with peroxidase on the surface of functionalized NPs. Synthesized bio-recognizing membranes of nano-sized range will be studied in the light of their structure and catalytic properties. The aim of the 1-st stage investigations: to develop the technology of AMO isolation from recombinant yeast S. cerevisiae and to characterize purified enzyme; to synthesize and study the structure of NPs; to study the methods of their bio-functionalization by protein molecules. The methods of the investigation: microbiological, biochemical, physico-chemical, enzymatic, genetic, gene-engineering. As result of the 1-st stage: expressive plasmid vector, that contains AMO gene of Hansenula polymorpha and tag-fragment, (His)6, under control of CUP1 yeast promoter, was constructed; effective method of recombinant AMO isolation from the transformant’s cells of S.cerevisiae and purification by affinity chromatography on Ni-NTA-agarose was developed, as result, highly purified preparations of AMO were obtained; Ag-NPs and Au-NPs were synthesized through reduction of silver nitrate by glucose and reduction of tetrachloroauric acid by sodium borohydride; nanosizes of synthesized NPs and Au-nanorods were proved by scanning and transmission microscopy, atomic force microscopy and small angle X-ray scattering diffraction methods; Ag-NPs and Au-NPs, activated by two methods - with 16-mercaptohexadecanoic acid and cisteamine usage, were obtained. All investigations have been carried out according to the plan. As result of the 1-st stage study, 1 paper was accepted (in press) and abstracts were published. METHYLAMINE OXIDASE, NANOPARTICLES OF NOBLE METALS, METHODS OF NANOPARTICLES’S FUNCTIONALIZATION. Product Description popup.authors Гайда Галина Зуфарівна Закальська Оксана Мирославівна Закальський Андрій Євстахович Стасюк Наталія Євгенiвна popup.nrat_date 2020-04-02 Close