Thermobifida fusca葡萄糖异构酶在枯草杆菌中的表达

Expression of glucose isomerase from Thermobifida fusca in Bacillus subtilis

为研究葡萄糖异构酶(GIase)在枯草杆菌中的表达效果,通过PCR扩增Thermobifida fusca GIase基因xyl A,分别克隆该基因序列到不同诱导表达方式的穿梭载体:p HCMC04、p MA09、p AL12,并转化枯草杆菌WB600。结果表明,含重组质粒p MA09-xyl A的枯草杆菌产酶最优,达到1.8 U/m L。对该重组枯草杆菌发酵产酶条件进行优化,以TB为发酵培养基,初始p H 7.0,温度为30℃时,摇瓶培养24 h后,GIase的产量达到5.6 U/m L。

For reasching expressive effect of GIase in Bacillus subtilis,Thermobifida fusca GIase gene xyl A was amplified by PCR,and cloned the gene sequences into shuttle carrieres with different induced expressionpattern： p HCMC04,p MA09,p AL12,and then transformed to Bacillus subtilis WB600. The results showed that enzyme production of Bacillus subtilis containing recombinant plasmid p MA09-xyl A was optimal,reached 1. 8 U / m L. Fermentation conditions of the recombinant Bacillus subtilis were further optimized,selecting fermentation culture medium for TB,initial p H 7. 0,the temperature of 30 ℃,shake flask culture after 24 h,the yield of GIase reached 5. 6 U / m L.