Authors: Tatiana Semashko Raisa Mikhailova Almira Ramanaviciene Arunas Ramanavicius
Publish Date: 2013/08/31
Volume: 171, Issue: 7, Pages: 1739-1749
Abstract
Glucose oxidase GOx from Penicillium funiculosum 461 was purified using stepbystep ultrafiltration and it was characterized by spectrophotometric and spectrofluorometric methods It was shown that spectra of GOx produced by P funiculosum are typical for flavoproteins Absorption spectrum has distinct peaks at 380 and 457 nm excitation spectrum at 373 and 447 nm and emission spectrum at 530 and 562 nm The pH correlation of enzyme activity and catalytic characteristics in various buffer systems phosphate pH 50–90 citrate pH 30–50 citratephosphate pH 30–90 and universal pH 30–90 were registered It was determined that the GOx is the most efficiently interacting with substrate glucose in phosphate buffer at pH 70 with k cat/K m = 21825 M−1 s−1 Interaction of several different redox mediators 910phenantroline56dione 910phenanthrenequinone Nmethylphenazonium methyl sulfate ferrocene ferrocenecarboxylic acid αmethylferrocenemethanol ferrocenecarboxaldehyde with GOx from P funiculosum was investigated by evaluation of the difference in fluorescence emission intensity of FADoxidized and FADH2reduced forms It was found that 910phenantroline56dione and 910phenanthrenequinone are the best redox mediators for this type of GOxThe authors will thank the State Committee on Science and Technology of the Republic of Belarus Belarusian Republican Foundation for Fundamental Research project number B11LIT012 and Lithuanian Scientific Council grant no TAP30/2011 for financial support
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