公开(公告)号：US20030100744A1

公开(公告)日：2003-05-29

申请号：US10201213

申请日：2002-07-22

Applicant: California Institute of Technology

Inventor： Edgardo T. Farinas ,  Frances H. Arnold ,  Ulrich Schwaneberg ,  Anton Glieder

IPC: C07H021/04 ,  C12N009/02 ,  C12P021/02 ,  C12N005/06

CPC classification number: C12N9/0071 ,  C12P7/02 ,  C12P7/04 ,  C12P7/16 ,  C12P17/02 ,  Y02E50/10 ,  Y02P20/52

Abstract: Nucleic acids encoding cytochrome P450 variants are provided. The cytochrome P450 variants of have a higher alkane-oxidation capability, alkene-oxidation capability, and/or a higher organic-solvent resistance than the corresponding wild-type or parent cytochrome P450 enzyme. A preferred wild-type cytochrome P450 is cytochrome P450 BM-3. Preferred cytochrome P450 variants include those having an improved capability to hydroxylate alkanes and epoxidate alkenes comprising less than 8 carbons, and have amino acid substitutions corresponding to V78A, H236Q, and E252G of cytochrome P450 BM-3. Preferred cytochrome P450 variants also include those having an improved hydroxylation activity in solutions comprising co-solvents such as DMSO and THF, and have amino acid substitutions corresponding to T235A, R471A, E494K, and S1024E of cytochrome P450 BM-3.

Abstract translation: 提供了编码细胞色素P450变体的核酸。 细胞色素P450变体具有比相应的野生型或亲本细胞色素P450酶更高的烷烃氧化能力，烯烃氧化能力和/或较高的有机溶剂耐受性。 优选的野生型细胞色素P450是细胞色素P450 BM-3。 优选的细胞色素P450变体包括具有改善含有少于8个碳原子的烷烃和环氧化烯烃羟基化能力以及对应于细胞色素P450 BM-3的V78A，H236Q和E252G的氨基酸取代的那些变体。 优选的细胞色素P450变体还包括在包含共溶剂如DMSO和THF的溶液中具有改善的羟基化活性的那些，并且具有对应于细胞色素P450 BM-3的T235A，R471A，E494K和S1024E的氨基酸取代。