Extraction of manganese peroxidase produced by Lentinula edodes.
ABSTRACT Lentinula edodes, commonly called shiitake, is considered a choice edible mushroom with exotic taste and medicinal quality. L. edodes grows very well and produces a range of enzymes when cultivated on eucalyptus residues. Development of appropriate experimental procedures for recovery and determination of enzymes became a widely important cash crop. In this work, enzymes produced by L. edodes were extracted using different pH buffer and determined regarding peroxidases and proteases. Lignin peroxidase (LiP) was not detected in the extracts based on veratryl alcohol or azure B oxidation. Proteases were very low while Mn-peroxidases (MnP) predominated. The optimal pH for MnP recovery was 5.0, under agitation at 25 degrees C. The oxidation of phenol red decreased after dark-colored small compounds or ions were eliminated by dialysis. The extract of L. edodes contained components of high molecular weight, such as proteases or high polyphenol, that could be involved in the LiP inactivation. L. edodes sample previously submitted to dialysis was also joined to LiP of Phanerochaete chrysosporium and a total inhibition of LiP was observed.
Article: Coal Depolymerising Activity and Haloperoxidase Activity of Mn Peroxidase from Fomes durissimus MTCC-1173.[show abstract] [hide abstract]
ABSTRACT: Mn peroxidase has been purified to homogeneity from the culture filtrate of a new fungal strain Fomes durissimus MTCC-1173 using concentration by ultrafiltration and anion exchange chromatography on diethylaminoethyl (DEAE) cellulose. The molecular mass of the purified enzyme has been found to be 42.0 kDa using SDS-PAGE analysis. The K(m) values using MnSO(4) and H(2)O(2) as the variable substrates in 50 mM lactic acid-sodium lactate buffer pH 4.5 at 30(°)C were 59 μM and 32 μM, respectively. The catalytic rate constants using MnSO(4) and H(2)O(2) were 22.4 s(-1) and 14.0 s(-1), respectively, giving the values of k(cat)/K(m) 0.38 μM(-1)s(-1) and 0.44 μM(-1)s(-1), respectively. The pH and temperature optima of the Mn peroxidase were 4 and 26(°)C, respectively. The purified MnP depolymerises humic acid in presence of H(2)O(2). The purified Mn peroxidase exhibits haloperoxidase activity at low pH.Bioinorganic Chemistry and Applications 01/2011; 2011:260802. · 0.72 Impact Factor