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Purification and partial characterization of manganese peroxidase from immobilized Phanerochaete chrysosporium

Paper ID Volume ID Publish Year Pages File Format Full-Text
36742 45146 2004 8 PDF Available
Title
Purification and partial characterization of manganese peroxidase from immobilized Phanerochaete chrysosporium
Abstract

Solid-state culture of the white-rot fungus Phanerochaete chrysosporium BKMF-1767 (ATCC 24725) has been carried out, using an inert support, polystyrene foam. Suitable medium and culture conditions have been chosen to favor the secretion of manganese peroxidase (MnP). The enzyme was isolated and purified from immobilized P. chrysosporium and partially characterized. Partial protein precipitation in crude enzyme was affected using ammonium sulphate, polyethylene glycol, methanol, and ethanol methods. Fractionation of MnP was performed by DEAE-Sepharose ion exchange chromatography followed by Ultragel AcA 54 gel filtration chromatography. This purification attained 23.08% activity yield with a purification factor of 5.8. According to data on gel filtration chromatography and sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE), the molecular weight of the enzyme was 45000±1000 Da. The optimum pH and temperature of purified MnP were 4.5 and 30 °C, respectively. This enzyme was stable in the pH range 4.5–6.0, at 25 °C and also up to 35 °C at pH 4.5 for 1 h incubation period. MnP activity was inhibited by 2 mM NaN3, ascorbic acid, β-mercaptoethanol and dithreitol. The Km values of MnP for hydrogen peroxide and 2.6-dimetoxyphenol were 71.4 and 28.57 μM at pH 4.5, respectively. The effects of possible inhibitors and activators of enzyme activity were investigated.

Keywords
Solid state fermentation; Immobilized Phanerochaete chrysosporium; MnP; Purification; Characterization
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Purification and partial characterization of manganese peroxidase from immobilized Phanerochaete chrysosporium
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Publisher
Database: Elsevier - ScienceDirect
Journal: Process Biochemistry - Volume 39, Issue 12, 29 October 2004, Pages 2061–2068
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us