title: New platform for cytochrome p450 reaction combining in situ immobilization on biopolymer.
abstract: We describe an efficienct chemical conversion platform with in situ immobilization of P450-BM3 on poly(3-hydroxybutyrate) granules. Through fusion with phasin, P450-BM3 is easily immobilized on poly(3-hydroxybutyrate) granules in Escherichia coli. In our work, the immobilized P450 exhibited higher stability and catalytic activity compared to free P450 against changes of pH, temperature, and concentrations of urea and ions. Through quick recovery of immobilized enzyme, the P450-P(3HB) complex successfully catalyzed an O-dealkylation reaction several times with maintained activity. Using the robust P450-P(3HB) complex, we performed a P450-catalyzed reaction on a preparative reactor scale (100 mL) and high-level production (12.3 μM) of 7-hydroxycoumarine from 7-ethoxycoumarin could be achieved.
authors.affiliation: Department of Chemical and Biomolecular Engineering, and ‡Department of Materials Science and Engineering, Korea Advanced Institute of Science and Technology , 335 Science Road, Daejeon 305-701, Republic of Korea.
authors.forename: Jae Hyung
authors.initials: JH
authors.lastname: Lee
authors.forename: Dong Heon
authors.initials: DH
authors.lastname: Nam
authors.forename: Sahng Ha
authors.initials: SH
authors.lastname: Lee
authors.forename: Jong Hyun
authors.initials: JH
authors.lastname: Park
authors.forename: Si Jae
authors.initials: SJ
authors.lastname: Park
authors.forename: Seung Hwan
authors.initials: SH
authors.lastname: Lee
authors.forename: Chan Beum
authors.initials: CB
authors.lastname: Park
authors.forename: Ki Jun
authors.initials: KJ
authors.lastname: Jeong
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D003374:Coumarins / Q000378:metabolism; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli; D006863:Hydrogen-Ion Concentration; D006885:Hydroxybutyrates / Q000737:chemistry*; D009251:NADPH-Ferrihemoprotein Reductase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011091:Polyesters / Q000737:chemistry*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013696:Temperature; D014468:Umbelliferones / Q000378:metabolism; D014508:Urea / Q000737:chemistry; D000094122:Polyhydroxybutyrates
chemical_list: D001426:Bacterial Proteins; D003374:Coumarins; D003577:Cytochrome P-450 Enzyme System; D004800:Enzymes, Immobilized; D006885:Hydroxybutyrates; D009251:NADPH-Ferrihemoprotein Reductase; D011091:Polyesters; D011994:Recombinant Proteins; D014468:Umbelliferones; D014508:Urea; C003182:poly-beta-hydroxybutyrate; D000094122:Polyhydroxybutyrates; C017299:7-ethoxycoumarin; C031477:7-hydroxycoumarin; C052423:flavocytochrome P450 BM3 monoxygenases