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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Pengpai, Zhang Sheng, Hu Lehe, Mei Yinlin, Lei Zhihua, Jin Guixiang, Hu |
| Description | Author Affiliation: Pengpai Z ( Institute of Bioengineering, Zhejiang University, Hangzhou, 310027, Zhejiang, People's Republic of China.) |
| Abstract | Cytochrome P450 BM-3 (A74G/F87V/L188Q) could catalyze indole to produce indigo. To further improve this capability, random mutagenesis was performed on the heme domain of P450 BM-3 (A74G/F87V/L188Q) with error-prone PCR. A single mutant V445A was selected out from the error-prone library and exhibited the highest specific activity toward indole among the mutants obtained. The kinetic parameters of V445A were also highly improved. Compared with the parent enzyme, the turnover rate (k cat) of V445A was increased by 7.5 times, while its K m value decreased by 9.2 %. Consequently, the catalytic efficiency (k cat/K m) of V445A was raised to 8.2 times than that of the parent enzyme. Moreover, alanine was confirmed as the best amino acid substitution by saturated mutagenesis in Val445 position. Three-dimensional structure analysis was also used to rationalize the effect on the enzyme properties of the mutation. This study showed that random mutagenesis was efficient to identify mutants with potential values in industry and increased our insight into P450 BM-3. |
| File Format | HTM / HTML |
| ISSN | 02732289 |
| Issue Number | 1 |
| Volume Number | 171 |
| e-ISSN | 15590291 |
| Journal | Applied Biochemistry and Biotechnology |
| Language | English |
| Publisher | Springer |
| Publisher Date | 2013-09-01 |
| Publisher Place | United States |
| Access Restriction | One Nation One Subscription (ONOS) |
| Subject Keyword | Discipline Biochemistry Discipline Biotechnology Bacterial Proteins Genetics Metabolism Biocatalysis Cytochrome P-450 Enzyme System Directed Molecular Evolution Indoles Nadph-ferrihemoprotein Reductase Chemistry Electron Transport Gene Library Hydroxylation Models, Molecular Mutagenesis, Site-directed Mutation Protein Conformation Stereoisomerism Substrate Specificity Journal Article Research Support, Non-u.s. Gov't |
| Content Type | Text |
| Resource Type | Article |
| Subject | Medicine Molecular Biology Environmental Engineering Biochemistry Bioengineering Applied Microbiology and Biotechnology Biotechnology |
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