Production and purification of remazol brilliant blue R decolorizing peroxidase from the culture filtrate of Pleurotus ostreatus

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Title
Production and purification of remazol brilliant blue R decolorizing peroxidase from the culture filtrate of Pleurotus ostreatus
Author(s)
Kwang-Soo Shin; Ill-Kyoon Oh; Chang Jin Kim
Bibliographic Citation
Applied and Environmental Microbiology, vol. 63, no. 5, pp. 1744-1748
Publication Year
1997
Abstract
An extracellular H2O2-requiring Remazol brilliant blue R (RBBR) decolorizing enzymatic activity was found in the culture medium of Pleurotus ostreatus. The enzymatic activity was maximally obtained in idiophase, and the optimum C/N ratio was 24. High C/N ratios repressed the enzymatic activity, and addition of veratryl alcohol had no effect on the production of enzyme. The enzyme was purified by ammonium sulfate fractionation, Sephacryl S-200 HR chromatography, DEAE Sepharose CL-6B chromatography, and Mono Q chromatography. The purification of RBBR decolorizing peroxidase, as judged by the final specific activity of 6.00 U/mg, was 54.5-fold, with a yield of 9.9%. The molecular mass of the native enzyme determined by gel permeation chromatography was found to be about 73 kDa. Sodium dodecyl sulfate- polyacrylamide gel electrophoresis revealed that the enzyme was a monomer with a molecular mass of 71 kDa. The enzyme was optimally active at pH 3.0 to 3.5 and at 25°C. Under standard assay conditions, the apparent K(m) values of the enzyme toward RBBR and H2O2 were 10.99 and 32.97 μM, respectively. The enzyme had affinity toward various phenolic compounds and artificial dyes, and it was inhibited by Na2S2O5, potassium cyanide, NaN3, and cysteine. The absorption spectrum of the enzyme exhibited maxima at 407, 510, and 640 nm. The addition of H2O2 to the enzyme resulted in an absorbance decrease at 407 and 510 nm.
ISSN
0099-2240
Publisher
Amer Soc Microb
Type
Article
Appears in Collections:
1. Journal Articles > Journal Articles
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