Unknown

Dataset Information

0

Biocatalytic conversion of avermectin to 4''-oxo-avermectin: improvement of cytochrome p450 monooxygenase specificity by directed evolution.


ABSTRACT: Discovery of the CYP107Z subfamily of cytochrome P450 oxidases (CYPs) led to an alternative biocatalytic synthesis of 4''-oxo-avermectin, a key intermediate for the commercial production of the semisynthetic insecticide emamectin. However, under industrial process conditions, these wild-type CYPs showed lower yields due to side product formation. Molecular evolution employing GeneReassembly was used to improve the regiospecificity of these enzymes by a combination of random mutagenesis, protein structure-guided site-directed mutagenesis, and recombination of multiple natural and synthetic CYP107Z gene fragments. To assess the specificity of CYP mutants, a miniaturized, whole-cell biocatalytic reaction system that allowed high-throughput screening of large numbers of variants was developed. In an iterative process consisting of four successive rounds of GeneReassembly evolution, enzyme variants with significantly improved specificity for the production of 4''-oxo-avermectin were identified; these variants could be employed for a more economical industrial biocatalytic process to manufacture emamectin.

SUBMITTER: Trefzer A 

PROVIDER: S-EPMC1932781 | biostudies-literature | 2007 Jul

REPOSITORIES: biostudies-literature

altmetric image

Publications

Biocatalytic conversion of avermectin to 4''-oxo-avermectin: improvement of cytochrome p450 monooxygenase specificity by directed evolution.

Trefzer Axel A   Jungmann Volker V   Molnár István I   Botejue Ajit A   Buckel Dagmar D   Frey Gerhard G   Hill D Steven DS   Jörg Mario M   Ligon James M JM   Mason Dylan D   Moore David D   Pachlatko J Paul JP   Richardson Toby H TH   Spangenberg Petra P   Wall Mark A MA   Zirkle Ross R   Stege Justin T JT  

Applied and environmental microbiology 20070504 13


Discovery of the CYP107Z subfamily of cytochrome P450 oxidases (CYPs) led to an alternative biocatalytic synthesis of 4''-oxo-avermectin, a key intermediate for the commercial production of the semisynthetic insecticide emamectin. However, under industrial process conditions, these wild-type CYPs showed lower yields due to side product formation. Molecular evolution employing GeneReassembly was used to improve the regiospecificity of these enzymes by a combination of random mutagenesis, protein  ...[more]

Similar Datasets

| S-EPMC1287623 | biostudies-literature
| S-EPMC1287622 | biostudies-literature
| S-EPMC7056670 | biostudies-literature
2018-07-10 | GSE116804 | GEO
| S-EPMC1253577 | biostudies-literature
| S-EPMC2857009 | biostudies-literature
2024-08-31 | GSE215117 | GEO
| PRJEB38781 | ENA
| S-EPMC7658919 | biostudies-literature
| S-EPMC61086 | biostudies-literature