Smutok O. Screening of microbial producers, isolation and bioanalytical application of L-lactate:cytochrome c-oxidoreductase of Hansenula polymorpha

Object: thermostable form of flavocytochrome b2 (FC b2) of H. polymorpha, purification of enzyme and investigation of its characteristics. The aim: a screening of microbial producers of a stable form of FC b2 and developed new enzymatic and biosensor methods of L-lactate assay based on this enzyme. The methods: microbiological, biochemical, amperometrical. A new method of visualization of enzymatic activity of FC b2 in PAAG has been developed. A screening different species and strains of yeasts - potential producers of a stable enzyme has carried out. H. polymorpha 356 has been chosen as the best one. A specific of regulation and some characteristics of FC b2 from H. рolymorpha have been investigated. A new effective scheme of preparative isolation of a high purified FC b2 from the cells of this yeast has been developed. A new photometric method and prototype of amperometric biosensors for L-lactate analysis based on FC b2 from H. polymorpha have been developed. The proposed analytical methods are approbated on the real samples of wine. Area of using: analytical chemistry, biotechnology, clinical diagnostic.