摘要
构建重组菌(JM109/pET-20b-Cel12B)生产极耐热内切葡聚糖酶,研究了不同表达宿主、IPTG诱导时间、IPTG浓度、重组菌生长不同阶段添加IPTG对重组菌生产极耐热内切葡聚糖酶的影响。结果表明:最佳诱导时间是OD值为1.0,IPTG浓度从0.5到3.0对酶活表达影响不大,诱导8h后极耐热内切葡聚糖酶表达量基本不变,优化后重组菌(E.coliBL21-CodonPlus(DE3)-RIL/pET-20b-Cel12B)表达酶活比原来菌株(E.coliJM109(DE3)/pET-20b-Cel12B)提高了53.9%,经过热处理和亲和层析得到电泳纯的蛋白条带。
Several factors influencing thermostable endoglucanase production with recombination E coli（JM109/ pET-20b-Ce112B）were investigated,The results showed that the production of thermostable endoglucanase did not improve after 8 hours of IPTG induction,Concentration of IPTG from 0.5 to 3mmol/L had almost the same effect on erpression of thermostable endoglucanase.The best induced time was OD600 at about 1.0. Enzyme activity produced by E.coli BL21-CodonPlus （DE3）-RIL/pET-20b-CeI12B was 1,539 times higher than that by E,coli JM109（DE3）/pET-20b-CeI12B. The recombinant protein was purified by heating and immobilizing metal affinity chromatography.
出处
《食品科技》
CAS
北大核心
2006年第10期27-30,共4页
Food Science and Technology
基金
江苏省教育厅自然科学研究项目(05KJB180006)
关键词
重组菌
诱导
内切葡聚糖酶
re-combination E coli
induction
endoglucanase
