公开(公告)号：WO03008563A9

公开(公告)日：2003-08-14

申请号：PCT/US0223340

申请日：2002-07-22

Applicant: CALIFORNIA INST OF TECHN ,  FARINAS EDGARDO T ,  GLIEDER ANTON ,  ARNOLD FRANCES H ,  SCHWANEBERG ULRICH

Inventor： FARINAS EDGARDO T ,  GLIEDER ANTON ,  ARNOLD FRANCES H ,  SCHWANEBERG ULRICH

IPC: C12N9/02 ,  C12P7/02 ,  C12P7/04 ,  C12P7/16 ,  C12P17/02 ,  C12N

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。 与相应的野生型或亲本细胞色素P450酶相比，细胞色素P450变体具有更高的烷烃氧化能力，烯烃氧化能力和/或更高的有机溶剂抗性。 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变体包括具有改进的包含少于8个碳的烷烃和环氧化物烯烃的羟化能力并且具有对应于细胞色素P450 BM-3的V78A，H236Q和E252G的氨基酸取代的那些。 优选的细胞色素P450变体还包括在包含共溶剂例如DMSO和THF的溶液中具有改善的羟基化活性并且具有对应于细胞色素P450 BM-3的T235A，R471A，E494K和S1024E的氨基酸取代的那些。

公开(公告)号：WO2003008563A2

公开(公告)日：2003-01-30

申请号：PCT/US2002/023340

申请日：2002-07-22

Applicant: CALIFORNIA INSTITUTE OF TECHNOLOGY ,  FARINAS, Edgardo, T. ,  GLIEDER, Anton ,  ARNOLD, Frances, H. ,  SCHWANEBERG, Ulrich

Inventor： FARINAS, Edgardo, T. ,  GLIEDER, Anton ,  ARNOLD, Frances, H. ,  SCHWANEBERG, Ulrich

IPC: C12N

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。

公开(公告)号：WO2003008563A3

公开(公告)日：2003-01-30

申请号：PCT/US2002/023340

申请日：2002-07-22

Applicant: CALIFORNIA INSTITUTE OF TECHNOLOGY ,  FARINAS, Edgardo, T. ,  GLIEDER, Anton ,  ARNOLD, Frances, H. ,  SCHWANEBERG, Ulrich

Inventor： FARINAS, Edgardo, T. ,  GLIEDER, Anton ,  ARNOLD, Frances, H. ,  SCHWANEBERG, Ulrich

IPC: C12N9/02 ,  C12N1/20 ,  C12N15/00 ,  C12Q1/00 ,  C12Q1/34 ,  C12Q1/68 ,  C12P21/06 ,  C12P21/04 ,  A61K38/44 ,  C07H21/04 ,  C07K1/00 ,  C07K16/00

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.