Gonchar M. Pathways of energy supply and detoxification in methylotrophic yeasts and their directed modification for development of enzymatic and biosensor analytical systems

Pathways of energy supply of methylotrophic growth and detoxification of methanol intermediates. Directed modification of metabolic reactions in methylotrophic yeasts and application of genetically and chemically modified cells for bioanalytical purposes. Microbiological, genetic, isotopic methods, enzymatic analysis, сhromatography, isoelectrofocusing, NMR-spectroscopy, potentiometric and amperometric methods. A new cyclic pathway of methanol dissimilation and additional systems of detoxification of hydrogen peroxide, formaldehyde, and formic acid have been revealed. The approaches of metabolic engineering have been used for construction of recognizing elements of cell biosensors based on pH-sensitive field effect transistors and oxygen- and hydrogen peroxide-electrodes. A catalase-free overproducer of alcohol oxidase has been constructed. Enzymatic kits for assays of ethanol and glucose ("ALCOTEST", "DIAGLUC", "DIAGLUC-2") have been developed which are produced commercially or in Institute of Cell Bi ology of NAS of Ukraine. Field of application: medicine, biotechnology.