title: Novel Enzyme Family Found in Filamentous Fungi Catalyzing trans-4-Hydroxylation of L-Pipecolic Acid.
abstract: Hydroxypipecolic acids are bioactive compounds widely distributed in nature and are valuable building blocks for the organic synthesis of pharmaceuticals. We have found a novel hydroxylating enzyme with activity toward L-pipecolic acid (L-Pip) in a filamentous fungus, Fusarium oxysporum c8D. The enzyme L-Pip trans-4-hydroxylase (Pip4H) of F. oxysporum (FoPip4H) belongs to the Fe(II)/α-ketoglutarate-dependent dioxygenase superfamily, catalyzes the regio- and stereoselective hydroxylation of L-Pip, and produces optically pure trans-4-hydroxy-L-pipecolic acid (trans-4-L-HyPip). Amino acid sequence analysis revealed several fungal enzymes homologous with FoPip4H, and five of these also had L-Pip trans-4-hydroxylation activity. In particular, the homologous Pip4H enzyme derived from Aspergillus nidulans FGSC A4 (AnPip4H) had a broader substrate specificity spectrum than other homologues and reacted with the L and D forms of various cyclic and aliphatic amino acids. Using FoPip4H as a biocatalyst, a system for the preparative-scale production of chiral trans-4-L-HyPip was successfully developed. Thus, we report a fungal family of L-Pip hydroxylases and the enzymatic preparation of trans-4-L-HyPip, a bioactive compound and a constituent of secondary metabolites with useful physiological activities.
authors.affiliation: Industrial Microbiology, Graduate School of Agriculture, Kyoto University, Kyoto, Japan.
authors.forename: Makoto
authors.initials: M
authors.lastname: Hibi
authors.affiliation: Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kyoto, Japan.
authors.forename: Ryosuke
authors.initials: R
authors.lastname: Mori
authors.affiliation: Mitsubishi Chemical Group Science and Technology Research Center, Inc., Yokohama, Kanagawa, Japan.|API Corporation, Yokohama, Kanagawa, Japan.
authors.forename: Ryoma
authors.initials: R
authors.lastname: Miyake
authors.affiliation: Mitsubishi Chemical Group Science and Technology Research Center, Inc., Yokohama, Kanagawa, Japan.|API Corporation, Yokohama, Kanagawa, Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Kawabata
authors.affiliation: Industrial Microbiology, Graduate School of Agriculture, Kyoto University, Kyoto, Japan.
authors.forename: Shoko
authors.initials: S
authors.lastname: Kozono
authors.affiliation: Industrial Microbiology, Graduate School of Agriculture, Kyoto University, Kyoto, Japan.
authors.forename: Satomi
authors.initials: S
authors.lastname: Takahashi
authors.affiliation: Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kyoto, Japan ogawa@kais.kyoto-u.ac.jp.
authors.forename: Jun
authors.initials: J
authors.lastname: Ogawa
mesh_terms: D055162:Biocatalysis; D049308:Dioxygenases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D005670:Fusarium / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D006900:Hydroxylation; D005810:Multigene Family; D010875:Pipecolic Acids / Q000737:chemistry / Q000378:metabolism*; D013379:Substrate Specificity
chemical_list: D005656:Fungal Proteins; D010875:Pipecolic Acids; D049308:Dioxygenases; C031345:pipecolic acid