• Applied Biological Chemistry
• /
• v.47 no.1
• /
• pp.22-26
• /
• 2004

The ligninolytic basidiomycete, Pleurotus ostreatus K-2946, was produced a manganese peroxidase (MnP) activity when grown in liquid culture with glucose-yeast-peptone (G-Y-P) medium. However, lignin peroxidase (LiP) was not detected in this culture medium. The purification progress of MnP was purified that included chromatography on Sepharose CL-6B, Superdex 75 prep grade and Mono-Q. MnP purified by column chromatography, was 36400 dalton and a pI of 3.95. The optimal pH and temperature of the purified MnP activity were 5.0 and $55^{\circ}C$. The characteristics of MnP produced was quite similar to those of MnP 3 isoenzyme produced by other strains of P. ostreatus.

• Journal of Mushroom
• /
• v.17 no.4
• /
• pp.185-190
• /
• 2019

Pleurotus ostreatus No.42, known as the ligninolytic basidiomycetes, showed production of MnP and Lac, but did not show any LiP acitivity in static culture, grown in GPYW liquid medium. Maximum production of MnP (80U/flask) was observed on day 11 of culturing in this medium. Chromatographic purification of MnP included the use of Sepharose CL-6B and Mono-Q. The major MnP isozyme purified by column chromatography was observed to be a 36.4 KDa (single band on SDS PAGE). The 19-amino acid sequence from the N-terminal was determined by protein sequencing to be ATCADGRTTANAACCVLFP. The N-terminal sequence of the major MnP isozyme of P. ostreatus No.42 was found to be the same as a previously reported sequence of an MnP3 isozyme from this fungus.

• Journal of Mushroom
• /
• v.19 no.4
• /
• pp.316-321
• /
• 2021

Ligninolytic enzymes were produced by Pleurotus ostreatus No.42, cultivated in a new kind of bioreactor that has a rotating draft tube with a helical ribbon. Maximum laccase (Lac) production (about 8,200 U/bioreactor) was reached after 3 days of incubation, then production decreased. Production of manganese peroxidase (MnP) in this fermenter reached a maximum level of about 8,400 U/bioreactor after 6 days of incubation. Lignin peroxidase (LiP) was not detected under these growth conditions. These results indicate that the rotary draft tube bioreactor (RTB) is compatible with large scale production of ligninolytic enzymes. MnP produced under these fermentation conditions was purified via a multistep process that included chromatography on Sepharose CL-6B, prep grade Superdex 75, and Mono-Q. This major isoenzyme was confirmed to have an apparent molecular weight of 36,400 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and its isoelectric point (IEF) was determined to be 3.95. N-terminal sequencing of the major isoenzyme from this fermentation was identical to that reported for an MnP3 isoenzyme isolated under different cultivation conditions, including stationary and shaking culture.