Novel Enzyme Family Found in Filamentous Fungi Catalyzing trans-4-Hydroxylation of l-Pipecolic Acid |
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| Authors: | Makoto Hibi Ryosuke Mori Ryoma Miyake Hiroshi Kawabata Shoko Kozono Satomi Takahashi Jun Ogawa |
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| Institution: | USDA Forest Products Laboratory;aIndustrial Microbiology, Graduate School of Agriculture, Kyoto University, Kyoto, Japan;bDivision of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kyoto, Japan;cMitsubishi Chemical Group Science and Technology Research Center, Inc., Yokohama, Kanagawa, Japan;dAPI Corporation, Yokohama, Kanagawa, Japan |
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| 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. |
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