title: Purification and characterization of a novel O-methyltransferase from Flammulina velutipes.
abstract: An enzyme catalyzing the methylation of phenolic hydroxyl groups in polyphenols was identified from mycelial cultures of edible mushrooms to synthesize O-methylated polyphenols. Enzyme activity was measured to assess whether methyl groups were introduced into (-)-epigallocatechin-3-O-gallate (EGCG) using SAM as a methyl donor, and (-)-epigallocatechin-3-O-(3-O-methyl)-gallate (EGCG3″Me), (-)-epigallocatechin-3-O-(4-O-methyl)-gallate (EGCG4″Me), and (-)-epigallocatechin-3-O-(3,5-O-dimethyl)-gallate (EGCG3″,5″diMe) peaks were detected using crude enzyme preparations from mycelial cultures of Flammulina velutipes. The enzyme was purified using chromatographic and two-dimensional electrophoresis. The purified enzyme was subsequently analyzed on the basis of the partial amino acid sequence using LC-MS/MS. Partial amino acid sequencing identified the 17 and 12 amino acid sequences, VLEVGTLGGYSTTWLAR and TGGIIIVDNVVR. In database searches, these sequences showed high identity with O-methyltransferases from other mushroom species and completely matched 11 of 17 and 9 of 12 amino acids from five other mushroom O-methyltransferases.
authors.affiliation: a Research & Development-Production Headquarters, Asahi Breweries Limited , Moriya-shi , Japan.
authors.forename: Masanobu
authors.initials: M
authors.lastname: Kirita
authors.forename: Yoshihisa
authors.initials: Y
authors.lastname: Tanaka
authors.forename: Motoyuki
authors.initials: M
authors.lastname: Tagashira
authors.forename: Tomomasa
authors.initials: T
authors.lastname: Kanda
authors.forename: Mari
authors.initials: M
authors.lastname: Maeda-Yamamoto
mesh_terms: D000595:Amino Acid Sequence; D002392:Catechin / Q000031:analogs & derivatives / Q000378:metabolism; D055393:Flammulina / Q000201:enzymology*; D006863:Hydrogen-Ion Concentration; D008780:Methyltransferases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D008969:Molecular Sequence Data; D017421:Sequence Analysis; D013696:Temperature
chemical_list: D002392:Catechin; C045651:epigallocatechin gallate; D008780:Methyltransferases
keywords: (−)-epigallocatechin-3-O-gallate; EGCG; Flammulina velutipes; O-methyltransferase