重组有机磷降解酶OpdA发酵条件的优化  

Optimization of fermentation condition for recombination organophosphate-degrading enzyme(OpdA)

作　　者：邱丹[1] 李宁[2] 贺玉姣[1] 冯启武 王发春[1] 谢惠春[1] 王生全 

机构地区：[1]青海师范大学生命与地理科学学院,青海西宁810008 [2]青海大学化工学院,青海西宁810008 [3]都兰县环境保护和林业局,青海都兰816100

出　　处：《广东农业科学》2012年第11期116-118,共3页Guangdong Agricultural Sciences

基　　金：青海省自然科学基金(2009-N-109)

摘　　要：OpdA是具有有机磷降解功效的基因,通过构建重组质粒pET30-opdA,并将其转化到大肠杆菌BL21(DE3)中,获得了工程菌BL21(DE3)/p ET30-opdA。同时探讨了其在摇瓶及20 L发酵罐中的发酵条件,并将其应用于中试,为有机磷水解酶工业化生产和应用提供了基础。研究发现,在生长诱导温度30℃、100μg/mL Amp、0.15 mmol/L IPTG、0.1 mmol/L CoCl2、3%乙醇、pH值7.0的条件下诱导4 h(补料方式与pH值联动),发酵样品于4℃冰箱中过夜后离心破菌,发酵液的酶活可以达到约230 IU/mL。opdA was a gene having the effect of organophosphate degradation. The opdA gene inserted into vector pET30 to construct a recombination plasmid pET30-opdA, which was subsequently transformed into E.coli BL21（DE3）, producing a genetic engineering strain BL21 （DE3）/pET30-opdA. The optimized conditions of the fermentation experiments conducted in flasks and 20 L fermentor was used in pilot scale test. The resuh showed that the optimum temperature was 30℃, pH value was 7.0, the concentration of IPTG, COC12 and ethanol was 0.15 mmol/L, 0.1 mmol/L and 3% respectively, fed-batch was with pH and induction period was 4 h. The final concentration of OpdA was 230 IU/mL.

关 键 词：有机磷降解:大肠杆菌:条件优化 

分 类 号：Q815[生物学—生物工程]