title: Novel fungal phenylpyruvate reductase belongs to d-isomer-specific 2-hydroxyacid dehydrogenase family.
abstract: We discovered the phenyllactate (PLA)-producing fungal strain Wickerhamia fluorescens TK1 and purified phenylpyruvate reductase (PPR) from fungal cell-free extracts. The PPR used both NADPH and NADH as cofactors with more preference for the former. The enzyme reaction as well as the fungal culture produced optically active d-PLA. The gene for the PPR (pprA) was cloned and expressed in Escherichia coli cells. Purified preparations of both native and recombinant PPR used hydroxyphenylpyruvate, glyoxylate and hydroxypyruvate as substrates but not pyruvate, oxaloacetate or benzoylformate. The predicted PPR protein had sequence similarity to proteins in the d-isomer-specific 2-hydroxyacid dehydrogenase family. Phylogenetic analyses indicated that the predicted PPR protein together with fungal predicted proteins constitutes a novel group of glyoxylate/hydroxypyruvate reductases. The fungus efficiently converted phenylalanine and phenylpyruvate to d-PLA. These compounds up-regulated the transcription of pprA, suggesting that it plays a role in fungal phenylalanine metabolism.
authors.affiliation: Graduate School of Life and Environmental Sciences, University of Tsukuba, Tsukuba, Ibaraki, Japan.
authors.forename: Taiki
authors.initials: T
authors.lastname: Fujii
authors.forename: Motoyuki
authors.initials: M
authors.lastname: Shimizu
authors.forename: Yuki
authors.initials: Y
authors.lastname: Doi
authors.forename: Tomoya
authors.initials: T
authors.lastname: Fujita
authors.forename: Takashi
authors.initials: T
authors.lastname: Ito
authors.forename: Daisuke
authors.initials: D
authors.lastname: Miura
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Wariishi
authors.forename: Naoki
authors.initials: N
authors.lastname: Takaya
mesh_terms: D000429:Alcohol Oxidoreductases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D015971:Gene Expression Regulation, Enzymologic; D015966:Gene Expression Regulation, Fungal; D008969:Molecular Sequence Data; D005810:Multigene Family; D010667:Phenylpyruvic Acids / Q000378:metabolism; D010802:Phylogeny; D050743:Pyruvate Synthase / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D004718:Saccharomycetales / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism
chemical_list: D010667:Phenylpyruvic Acids; D000429:Alcohol Oxidoreductases; C021379:D-2-hydroxyacid dehydrogenase; D050743:Pyruvate Synthase; C031606:phenylpyruvic acid