PRODUCTION OPTIMIZATION CHARACTERIZATION OF AZOREDUCTASE FROM PSEUDOMONAS OLEOVORANS PAMD1 STUDIES ON THERMOSTABILIZATION AND APPLICATION IN BIOSENSOR

Abstract

Industrial pollution is one of the problems presently facing throughout world and newlineseveral efforts are being vigorously pursued to control it. Toxic effluents containing azo newlinedyes are discharged from various industries and they adversely affect the water resources, newlinesoil fertility, and ecosystem integrity. Azo dyes are aromatic chromophore moieties newlinelinked by (-N--N-), represent the largest class of dyes used in textile processing and other newlineindustries. The release of these compounds into the environment is undesirable because newlineof their toxic nature. Removal of azo dyes from wastewater is a challenging task for the newlineresearchers. Therefore, this research was aimed to optimize the azo dye reduction by newlineisolating new bacterial species capable of secreting azo reductive enzyme, investigation newlineinto the production, characterization and further by consequently applying and obtained newlineinsights. newlineThe isolated bacteria Pseudomonas oleovorans PAMDJ produced azoreductase newlineas the prime azo reductive enzyme during the dye decolorization process. Azoreductase newlinecatalytic activity was optimized by statistical based experiment, Plackett-Burman design newlinefollowed by Response surface methodology. Dye and NADH were found as potential newlineinducer on azoreductase production. The optimal percentage degradation of azo dyes of newlinevariables up to 85.18 % was achieved by dye (200 mgl-1), peptone (6.44 g11), NADH newline(1.14 mM) and glucose (2.07 g1-1), respectively. newlineProduction optimization, characterization of azoreductase from. newlineThe produced intracellular azoreductase under optimized conditions was purified newlineup to 9 fold with a recovery of 16 %. The purified azoreductase was monomeric with an newlineapparent molecular mass of 29 KDa having an optimum pH and temperature of 7.0 and newline35 °C respectively. The degradation product of Orange II azo dye sulphanilic acid was newlineanalyzed using HPLC with reference to corresponding standards and the FTIR analysis newlinealso proves the degradation of the functional group of azo dye in decolorization process. newlineToxicity analysis were carried