研究报告

Bacillus cereus胶原蛋白酶功能基因分析与结构预测

  • 刘丽莉 ,
  • 代晓凝 ,
  • 陈珂 ,
  • 孟圆圆 ,
  • 郝威铭
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  • 1(河南科技大学 食品与生物工程学院,河南 洛阳,471023)
    2(食品加工与安全国家级教学示范中心(河南科技大学),河南 洛阳,471023)
博士,教授(本文通讯作者,E-mail:yangliuyilang@126.com)。

收稿日期: 2018-10-15

  修回日期: 2019-01-05

  网络出版日期: 2019-05-14

基金资助

国家自然科学基金(31401622);河南省重大专项(1611 00110900,161100110600-2,161100110700-2,161100110800-06)

Structure prediction and functional analysis of aBacillus cereus collagenase

  • LIU Lili ,
  • DAI Xiaoning ,
  • CHEN Ke ,
  • MENG Yuanyuan ,
  • HAO Weiming
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  • 1(College of Food and Bioengineering, Henan University of Science and Technology, Luoyang 471023, China)
    2(College of food and bioengineering, Henan University of Science and Technology, National Experimental Teaching Demonstration Center for Food Processing and Security, Luoyang 471023, China)

Received date: 2018-10-15

  Revised date: 2019-01-05

  Online published: 2019-05-14

摘要

以一株降解骨胶原蛋白的菌种蜡样芽孢杆菌(Bacillus cereus)MBL13-U作为出发菌种,通过对菌全基因组测序,针对胶原蛋白酶功能基因进行了定位和结构预测。结果表明,从全基因组序列中得到胶原蛋白酶的功能基因序列长度为 2 916 bp;该功能基因中α螺旋占总氨基酸的37.1%,β转角占12.8%,无规则卷曲占26.0%,且该酶的三维结构预测为镊子状结构。将胶原蛋白酶基因转入大肠杆菌宿主菌株BL21,构建出工程菌pET30a-ColM13/BL21。对工程菌所产的重组蛋白酶ColM13酶解Ⅰ型骨胶原蛋白的结构进行分析表明,酶解作用使骨胶原蛋白产生多肽和大量游离氨基酸,其微观结构发生显著变化,表明ColM13在工程菌中已成功表达。

本文引用格式

刘丽莉 , 代晓凝 , 陈珂 , 孟圆圆 , 郝威铭 . Bacillus cereus胶原蛋白酶功能基因分析与结构预测[J]. 食品与发酵工业, 2019 , 45(8) : 29 -35 . DOI: 10.13995/j.cnki.11-1802/ts.019054

Abstract

The genome of Bacillus cereus MBL13-U that degrades bone collagen was sequenced, and a putative gene, ColM13, which encodes a collagenase was closed and its activity was assayed. It consisted of 2 916 bp. Its deduced amino acids sequence contained α-helix, β-rotation, and irregular curl structures, which accounted for 37.1%, 12.8%, and 26.0% of total amino acids, respectively. The mimic three-dimensional structure of the enzyme was to be tweezers. It was cloned and transformed into Escherichia coli BL21 to obtain E. coli BL21/pET30a-ColM13. The recombinant ColM13 degraded type I bone collagen and formed polypeptides as well as a large amount of free amino acids. Besides, the microstructure of collagen changed significantly after treating with the recombinant enzyme. Based on these observations, it was concluded that ColM13 has been successfully expressed in E. coli.

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