›› 2009, Vol. 11 ›› Issue (1): 68-72.

• 863课题进展 • 上一篇    下一篇

谷氨酸棒杆菌麦芽寡糖基海藻糖合成酶基因的克隆与表达

张文德,乔宇,丁宏标   

  1. (中国农业科学院饲料研究所, 北京100081)
  • 收稿日期:2008-11-14 修回日期:2008-12-26 出版日期:2009-02-15 发布日期:2009-07-29
  • 通讯作者: 丁宏标,研究员,博士生导师,主要从事饲料生物技术研究。Tel:010-82106074; E-mail:dinghongbiao@mail.caas.net.cn
  • 作者简介:张文德,硕士研究生,主要从事饲料生物技术研究。Tel:010-62133407|E-mail:wendezhang2008@yahoo.com.cn。
  • 基金资助:

    国家863计划项目(2005AA246010);国际科技合作重点项目(2005DFA31070)资助。

Cloning and Expression of Gene Encoding Maltoologosyl Trehalose
Synthase from Corynebacterium glutamicum

ZHANG Wen-de, QIAO Yu, DING Hong-biao   

  1. ( Feed Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081,China)
  • Received:2008-11-14 Revised:2008-12-26 Online:2009-02-15 Published:2009-07-29

摘要:

采用PCR方法从谷氨酸棒杆菌(Corynebacterium glutamicum)基因组中分离得到2.4 kb的麦芽寡糖基

海藻糖合成酶基因(TreY),将其插入原核表达载体pET-30a中,转化大肠杆菌BL21(DE3)pLysS获得重

组基因工程菌。经IPTG诱导表达得到约90 kDa的目的蛋白,可溶性蛋白占细胞总蛋白的45.3%。重组酶

作用于麦芽糊精,可使其DE值降低,得到麦芽寡糖基海藻糖的混合物。

关键词: 麦芽寡糖基海藻糖合成酶;麦芽糊精;谷氨酸棒杆菌;重组表达

Abstract:

A 2.4 kb DNA fragment encoding maltooligosyl trehalose synthase was cloned from

Corynebacterium glutamicum by PCR. It was inserted into prokaryotic expression vector

pET-30a and then was introduced into the host Escherichia coli BL21 (DE3) pLysS. A high

yield of the recombinant enzyme at a molecular weight of 90 kDa was obtained by IPTG

induction. The soluble recombinant enzyme accounted for 45.3% of the total cell proteins

in supernatant. This recombinant enzyme was capable of decreasing the DE value of dextrin

and producing a maltooligosyl trehalose mixture.

Key words: maltoologosyl trehalose synthase, malt dextrin, Corynebacterium glutamicum, recombinant expression

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