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| Expression of recombinant uricase in E.coli JM109(DE3)Induced by lactose |
| DENG Yong-kang1, WU Min-lu2, LIU Sheng-bang1, DU Lin-fang1, WU Li-li1, LI Man1, MENG Yan-fa1 |
| 1 College of Life Science,Sichuan University,Chengdu610064, China
2 Department of Lab Medicine,Chengdu Medical Colledge,Chengdu610083,China
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Abstract Abstract Using lactose as a succedaneous inducer of IPTG to induce the expression of Candida utilis uricase gene cloned in E.coli JM109(DE3) was deeply investigated for establishing a high-performance but low-cost method of recombinant uricase production. The adopted lactose concentration for induction, the time point of induction, the duration of induction and the dynamics of uricase expression were optimized and studied in detail by shake flask experiment. The results of optimization were verified by enlarged fermentation in 5 L fermentor and then lactose was used as the inducer in the high density fermentation. As showed by the experiments, the best inducing concentration of lactose was 5 g/L, the midanaphase of logarithmic phase was the best time for induction and the duration of induction were 9~10h; according to the optimum conditions, compared with IPTG, both in the shake flask and 5 L fermentor with batch culture, the better inducing effect could be obtained: the expression level of uricase was about 26% of the total bacterial protein and about 36% of the soluble protein, the final cell density(OD600) was over 40 in the 5 L fermentor and uricase expression level was about 25% with high density fermentation.
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Received: 02 February 2009
Published: 28 July 2009
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