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...
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| Format: | Article |
| Language: | English |
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Zhejiang University Press
2006-01-01
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| Series: | 浙江大学学报. 农业与生命科学版 |
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| Online Access: | https://www.academax.com/doi/10.3785/1008-9209.2006.01.0096 |
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| author | HUANG Jun MEI Le-he ZHONG Chun-long DENG Fu-huang YAO Shan-jing |
| author_facet | HUANG Jun MEI Le-he ZHONG Chun-long DENG Fu-huang YAO Shan-jing |
| author_sort | HUANG Jun |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-37b70ffa33a34ca2984a689a0da45928 |
| institution | Kabale University |
| issn | 1008-9209 2097-5155 |
| language | English |
| publishDate | 2006-01-01 |
| publisher | Zhejiang University Press |
| record_format | Article |
| series | 浙江大学学报. 农业与生命科学版 |
| spelling | doaj-art-37b70ffa33a34ca2984a689a0da459282025-08-20T03:58:11ZengZhejiang University Press浙江大学学报. 农业与生命科学版1008-92092097-51552006-01-01329610010.3785/1008-9209.2006.01.009610089209Study on the expression conditions and primary purification of recombinant P450 BM-3 monooxygenaseHUANG JunMEI Le-heZHONG Chun-longDENG Fu-huangYAO Shan-jingThe 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.https://www.academax.com/doi/10.3785/1008-9209.2006.01.0096P450 BM-3expressionpurificationion-exchange chromatography |
| spellingShingle | HUANG Jun MEI Le-he ZHONG Chun-long DENG Fu-huang YAO Shan-jing Study on the expression conditions and primary purification of recombinant P450 BM-3 monooxygenase 浙江大学学报. 农业与生命科学版 P450 BM-3 expression purification ion-exchange chromatography |
| title | Study on the expression conditions and primary purification of recombinant P450 BM-3 monooxygenase |
| title_full | Study on the expression conditions and primary purification of recombinant P450 BM-3 monooxygenase |
| title_fullStr | Study on the expression conditions and primary purification of recombinant P450 BM-3 monooxygenase |
| title_full_unstemmed | Study on the expression conditions and primary purification of recombinant P450 BM-3 monooxygenase |
| title_short | Study on the expression conditions and primary purification of recombinant P450 BM-3 monooxygenase |
| title_sort | study on the expression conditions and primary purification of recombinant p450 bm 3 monooxygenase |
| topic | P450 BM-3 expression purification ion-exchange chromatography |
| url | https://www.academax.com/doi/10.3785/1008-9209.2006.01.0096 |
| work_keys_str_mv | AT huangjun studyontheexpressionconditionsandprimarypurificationofrecombinantp450bm3monooxygenase AT meilehe studyontheexpressionconditionsandprimarypurificationofrecombinantp450bm3monooxygenase AT zhongchunlong studyontheexpressionconditionsandprimarypurificationofrecombinantp450bm3monooxygenase AT dengfuhuang studyontheexpressionconditionsandprimarypurificationofrecombinantp450bm3monooxygenase AT yaoshanjing studyontheexpressionconditionsandprimarypurificationofrecombinantp450bm3monooxygenase |