IPTG与乳糖联合诱导重组大肠杆菌右旋糖酐蔗糖酶表达

doi:10.7506/spkx1002-6630-201401036

食品科学 ›› 2014, Vol. 35 ›› Issue (1): 185-188.doi: 10.7506/spkx1002-6630-201401036

IPTG与乳糖联合诱导重组大肠杆菌右旋糖酐蔗糖酶表达

李攀峰，张洪斌*，胡雪芹

合肥工业大学医学工程学院制药工程系，安徽合肥 230009

收稿日期:2013-04-06 修回日期:2013-12-30 出版日期:2014-01-15 发布日期:2014-01-22
通讯作者: 张洪斌 E-mail:zhb5678@163.com
基金资助:
安徽省长三角科技攻关基金项目（10140702001)；安徽省自主创新专项（2013AKKG0391)

Combinatorial Induction of Recombinant Dextransucrase Expression in E. coli by IPTG and Lactose

LI Pan-feng, ZHANG Hong-bin*, HU Xue-qin

Department of Pharmaceutical Engineering, School of Medical Engineering, Hefei University of Technology, Hefei 230009, China

Received:2013-04-06 Revised:2013-12-30 Online:2014-01-15 Published:2014-01-22
Contact: zhang hong-bin E-mail:zhb5678@163.com
Supported by:
Yangtze River Science and Technology Joint Research Projects

摘要/Abstract

摘要：

研究异丙基硫代-β-D-呋喃半乳糖苷（isopropyl-β-D-1-thiogalactopyranoside，IPTG）与乳糖联合诱导重组大肠杆菌右旋糖酐蔗糖酶表达的效果。在利用IPTG和乳糖分别作为诱导剂对右旋糖酐蔗糖酶工程菌Escherichia coliBL21（DE3）/pET28-dexYG进行诱导表达的基础上，尝试将此两种诱导剂联合使用，在降低成本的同时获得较好的表达效果。在获得最佳培养基的基础上，考察菌体IPTG与乳糖的联合加入量、菌体浓度、诱导时间对右旋糖酐蔗糖酶表达的影响。在菌浓（OD600 nm）达到3.0时，加入0.1 mmol/L IPTG 95 μL＋2.5 g/L乳糖，25 ℃混合诱导培养4 h，酶活力最高，达到40.44 U/mL。IPTG与乳糖联合诱导重组大肠杆菌右旋糖酐蔗糖酶表达可行。

关键词: 异丙基-&beta, -D-硫代吡喃半乳糖苷；乳糖；右旋糖酐蔗糖酶；表达；联合诱导

Abstract:

In order to explore the combinatorial induction of recombinant dextransurase experssion in E. coli by isopropyl
β-D-1-thiogalactopyranoside (IPTG) and lactose, a mixture of IPTG and lactose was added to the cell culture to induce the
expression of recombinant dextransurase. Based on the optimal culture medium, IPTG concentration and the amount of
lactose added, bacteria concentration and induction time revealed significant impacts on the enzyme activity. The results
showed that the optimal concentrations of IPTG and lactose were 0.1 mmol/L and 2.5 g/L, respectively, and optimal
induction time was 4 h at 25 ℃. Under these culture conditions, the activity of dextransucrase could reach up to 40.44 U/mL. The
combinatorial induction of recombinant E. coli dextransurase by IPTG and lactose was feasible, greatly reducing the dosage
of IPTG and industrial production costs.

Key words: isopropyl-β-D-1-thiogalactopyranoside (IPTG), lactose, dextransucrase, expression, combinatorial induction

中图分类号:

Q814

李攀峰，张洪斌*，胡雪芹. IPTG与乳糖联合诱导重组大肠杆菌右旋糖酐蔗糖酶表达[J]. 食品科学, 2014, 35(1): 185-188.

LI Pan-feng, ZHANG Hong-bin*, HU Xue-qin. Combinatorial Induction of Recombinant Dextransucrase Expression in E. coli by IPTG and Lactose[J]. FOOD SCIENCE, 2014, 35(1): 185-188.

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IPTG与乳糖联合诱导重组大肠杆菌右旋糖酐蔗糖酶表达

Combinatorial Induction of Recombinant Dextransucrase Expression in E. coli by IPTG and Lactose

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