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Information × Registration Number 0209U008308, 0107U008204 , R & D reports Title Gene and Protein Engineering of Oxidoreductases for Construction of Bionanosized Objects of Analytical Importance. Genetic construction of superproducers, purification, stabilization of the modified enzymes and formation of bioselective membranes on the base of nanocarriers. popup.stage_title Head Gonchar Mykhajlo Vasyl'ovych, Кандидат педагогічних наук Registration Date 26-02-2009 Organization Institute of cell biology, National Academy of Sciences of Ukraine popup.description2 The goal of this project is gene- and protein engineering of oxydoreductases (using model enzymes alcohol oxidases (AOX) and L-lactate oxidoreductase (flavocytochrome b2, FC b2) for construction of bionanoparticles of analytical importance. A novel selection procedure for isolation of the mutant forms of AOX to use it as a suitable bioelement for biosensor technologies has been described. The mutated AOXs (from the strain CA2 and CA4) were characterised by a decreased affinity towards ethanol (KM value is about 0.44; 1.91 і 0.83 mM, respectively). Sequence analysis of AOX mutant alleles discovered numerous point mutations that caused amino acid substitutions and reduction of enzyme affinity to substrate. The gene engineering construction of FC b2 overproduced strain by overexpression of H. polymorpha CYB2 gene has been performed. A method for preliminary screening of the transformants with an increased FC b2 activity in permeabilized cells has been developed. Product Description popup.authors popup.nrat_date 2020-04-02 Close