Study on the expression conditions and primary purification of recombinant P450 BM-3 monooxygenase
The optimum culture conditions of P450 BM-3 expression by recombinant Escherchia coli DH 5α were studied in detail. Using inoculum quantity corresponding to 1% of the culture medium, adding of 0.1mg·L<sup>-1</sup> FeCl<sub>3</sub> of the culture solution and rapid temperature...
Saved in:
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Zhejiang University Press
2006-01-01
|
| Series: | 浙江大学学报. 农业与生命科学版 |
| Subjects: | |
| Online Access: | https://www.academax.com/doi/10.3785/1008-9209.2006.01.0096 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| Summary: | The optimum culture conditions of P450 BM-3 expression by recombinant Escherchia coli DH 5α were studied in detail. Using inoculum quantity corresponding to 1% of the culture medium, adding of 0.1mg·L<sup>-1</sup> FeCl<sub>3</sub> of the culture solution and rapid temperature shift from 37℃ to 42℃ as soon as OD<sub>578</sub> reached about 1.0 and maintained for 5 hours, were proved to be the favorable conditions for growth of the bacteria and expression of the gene coding for P450 BM-3 monooxygenase. Purification of P450 BM-3 was investigated by two anion-exchange media DEAE-Sepharose FF, Resource Q. The results indicated that purification effectiveness of DEAE-Sepharose FF was inferior to that of Resource Q. The parameter of purification by Resource Q was optimized on ÄKTA explorer 100 system. By optimising the elution condition of P450 BM-3, the results showed that the purification factor was 2.12 and P450 BM-3 activity recovery was 30.1% through 0.3 mol· L<sup>-1</sup>, 0.5 mol· L<sup>-1</sup> two-step NaCl elution. |
|---|---|
| ISSN: | 1008-9209 2097-5155 |