摘要
为了提高目的蛋白的产量和活性,将克隆自Bifidobacterium longum JCM1217的乳二糖磷酸化酶(LNBP)的基因构建到大肠杆菌BL21中,优化其表达条件。结果显示,诱导时间、诱导温度及IPTG浓度对LNBP的表达水平影响显著。经正交试验,优化出的重组菌表达LNBP的最佳诱导条件为诱导温度30℃, IPTG浓度0.5 mmol/L,诱导时间18 h。在此条件下获得的LNBP比活力达到15.18 U/mg。
To increase the yield and activity of the target protein, the gene cloned from Bifidobacterium longum JCM1217 lactobiose phosphorylase(LNBP) was constructed into E. coli BL21, and its expression conditions were optimized. The results showed that induction time, induction temperature and IPTG concentration had a significant effect on the expression of LNBP. The optimal conditions for LNBP expression by orthogonal test were induction temperature of 30 ℃, IPTG concentration of 0.5 mmol/L, and induction time of 18 h, and the specific activity reached 15.18 U/mg.
作者
佟超男
李苏红
孙晓
李文杰
Motomitsu Kitaoka
李拖平
TONG Chaonan;LI Suhong;SUN Xiao;LI Wenjie;Motomitsu Kitaoka;LI Tuoping(College of Food Science,Shenyang Agricultural University,Shenyang 110000;National Food Research Institute,Tsukuba 305-8642;Shenyang Women's and Children's Hospital,Shenyang 110000)
出处
《食品工业》
CAS
北大核心
2018年第11期172-175,共4页
The Food Industry
基金
沈阳市国际科技合作项目(F16-219-6-00)
沈阳市科技局科技创新平台建设计划(F-17-158-1-00)
关键词
乳二糖磷酸化酶
表达优化
纯化
lactobiose phosphorylase
expression optimization
purification