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Cloning and characterization of a novel O-methyltransferase from Flammulina velutipes that catalyzes methylation of pyrocatechol and pyrogallol structures in polyphenols.
- Source :
-
Bioscience, biotechnology, and biochemistry [Biosci Biotechnol Biochem] 2015; Vol. 79 (7), pp. 1111-8. Date of Electronic Publication: 2015 Mar 10. - Publication Year :
- 2015
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Abstract
- A novel O-methyltransferase gene was isolated from Flammulina velutipes. The isolated full-length cDNA was composed of a 690-nucleotide open reading frame encoding 230 amino acids. A database search revealed that the deduced amino acid sequence was similar to those of other O-methyltransferases; the highest identity was only 61.8% with Laccaria bicolor. The recombinant enzyme was expressed by Escherichia coli. BL21 (DE3) was assessed for its ability to methylate (-)-epigallocatechin-3-O-gallate (EGCG). LC-TOF-MS and NMR revealed that the enzyme produced five kinds of O-methylated EGCGs: (-)-epigallocatechin-3-O-(3-O-methyl)gallate, (-)-epigallocatechin-3-O-(4-O-methyl)gallate, (-)-epigallocatechin-3-O-(3,4-O-dimethyl)gallate, (-)-epigallocatechin-3-O-(3,5-O-dimethyl)gallate, and (-)-4'-O-methylepigallocatechin-3-O-(3,5-O-dimethyl)gallate. The substrate specificity of the enzyme for 20 kinds of polyphenols was assessed using the crude recombinant enzyme of O-methyltransferase. This enzyme introduced methyl group(s) into polyphenols with pyrocatechol and pyrogallol structures.
- Subjects :
- Amino Acid Sequence
Catechin analogs & derivatives
Catechin chemistry
Catechin metabolism
Catechol O-Methyltransferase genetics
Catechol O-Methyltransferase metabolism
Catechols chemistry
Catechols metabolism
Cloning, Molecular
Escherichia coli genetics
Flammulina genetics
Fungal Proteins genetics
Fungal Proteins metabolism
Gallic Acid analogs & derivatives
Gallic Acid metabolism
Methylation
Methyltransferases genetics
Molecular Sequence Data
Molecular Structure
Polyphenols chemistry
Polyphenols metabolism
Pyrogallol chemistry
Recombinant Proteins genetics
Recombinant Proteins metabolism
Substrate Specificity
Flammulina enzymology
Methyltransferases metabolism
Pyrogallol metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 1347-6947
- Volume :
- 79
- Issue :
- 7
- Database :
- MEDLINE
- Journal :
- Bioscience, biotechnology, and biochemistry
- Publication Type :
- Academic Journal
- Accession number :
- 25754602
- Full Text :
- https://doi.org/10.1080/09168451.2015.1015955