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Purification and characterization of catechol 1, 2-dioxygenase from ＂Rhodococcus＂ sp. NCIM 2891
並列摘要
The main aim of this study was to purify Catechol 1, 2 dioxygenase from phenol utilizing Rhodococcus sp. NCIM 2891 which is basically an organism used wide for desulfurization purposes like desulfurization of diesel. The organism was cultivated in minimal medium supplemented with 500mg/L phenol, as a sole carbon source. The temperature of incubation was 30℃ at pH 7.0±0.2. The growth and phenol degradation studies show that organism was able to degrade the phenol completely within 120h. The stationary phase of growth of the organism, however, started after 72h. This implies that the degradation activities starts from late log phase and carries on till late stationary phase of growth. The enzyme responsible for phenol degradation was catechol 1, 2 dioxygenase. The optimal activity of catechol 1, 2 dioxygenase was observed at 500mg/L phenol concentration. The enzyme was purified 2.34 fold by ion exchange chromatography. The Km was found to be 5μmole with V_max of 62.5U/mg of protein. The optimal pH and temperature of the enzyme was 7.5 and 30℃ respectively. The enzyme activity was completely inhibited by Cu^++, Hg^++ and Fe^+++ metal ions. The SDS-PAGE analysis reveals that the enzyme has a molecular weight of 30k Dalton.
延伸閱讀
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