Production and induction of manganese peroxidase isozymes in a white-rot fungus Pleurotus ostreatus

• Published in: Applied microbiology and biotechnology Vol. 65; no. 3; pp. 287 - 294
• Main Authors: Kamitsuji, H, Honda, Y, Watanabe, T, Kuwahara, M
• Format: Journal Article
• Language: English
• Published: Berlin Springer 01.08.2004; Springer Nature B.V
• Subjects: Amino Acid Sequence; Amino acids; Biological and medical sciences; Biology of microorganisms of confirmed or potential industrial interest; biosynthesis; Biotechnology; Culture Media; Enzymes; Fundamental and applied biological sciences. Psychology; Gene Expression Regulation, Enzymologic; Gene Expression Regulation, Fungal; Isoenzymes - chemistry; Isoenzymes - genetics; Isoenzymes - isolation & purification; Isoenzymes - metabolism; ligninolytic microorganisms; Manganese; Miscellaneous; Mission oriented research; Molecular Sequence Data; Peptones; Peroxidases - chemistry; Peroxidases - genetics; Peroxidases - isolation & purification; Peroxidases - metabolism; Pleurotus - enzymology; Pleurotus - genetics; Pleurotus - growth & development; Pleurotus ostreatus; Reverse Transcriptase Polymerase Chain Reaction; Sequence Analysis, DNA; Transcription, Genetic; white-rot fungi; Yeasts; Fungi; Pleurotus ostreatus; Purification; Peroxidases; Enzyme; Induction; Isozyme; Manganese peroxidase; Production; Basidiomycetes; Oxidoreductases; Thallophyta
• ISSN: 0175-7598; 1432-0614
• DOI: 10.1007/s00253-003-1543-9

The production of MnP by Pleurotus ostreatus in different liquid cultures was investigated. The highest level of activity was observed after 8 days of culture in peptone-glucose-yeast extract medium (PGY), whereas maximal activity was achieved after 30 days in glucose-yeast extract medium (GY). MnP was purified to homogeneity from PGY (designated MnP-PGY) and GY (MnP-GY). The isoelectric points of MnP-PGY and MnP-GY were 3.77 and 4.06, respectively. The molecular mass of both enzymes was 42 kDa. Analysis of the N-terminal amino acid sequence of purified MnPs and nucleotide sequence of cloned mnp indicated that MnP-GY has VTCATGQTTANE at the N-terminus, whereas MnP-PGY has ATCADGRTTANA. A putative exposed tryptophan residue (W170) was found in MnP-GY. Both isozymes oxidized veratryl alcohol, although the K m of MnP-GY was lower than that of MnP-PGY. Thus, the presence of peptone in the medium affected the production of MnP isozymes. Reverse transcription-polymerase chain reaction (RT-PCR) analysis indicated that the synthesis of MnP isozymes is controlled by culture conditions at the transcriptional level.