食品科学 ›› 2014, Vol. 35 ›› Issue (1): 141-144.doi: 10.7506/spkx1002-6630-201401027

• 生物工程 • 上一篇    下一篇

高活性谷氨酰胺酶基因glsA2 在枯草芽孢杆菌BJ3-2染色体上的定点整合

卢 彪,吴拥军*,刘艳敏,王亚娟,唐 雪   

  1. 贵州大学生命科学学院,贵州 贵阳 550025
  • 收稿日期:2012-12-20 修回日期:2013-11-20 出版日期:2014-01-15 发布日期:2014-01-22
  • 通讯作者: 吴拥军 E-mail:wyjbio@163.com
  • 基金资助:

    国家自然科学基金项目(31260394);贵阳市科技计划项目(筑科工合同字[2010] 第1-68号);
    贵州省科技重大专项(黔科合重大专项字[2013]6013号)

Targeted Integration of Glutaminase Gene glsA2 with High Activity into the Bacillus subtilis BJ3-2 Chromosome

LU Biao, WU Yong-jun*, LIU Yan-min, WANG Ya-juan, TANG Xue   

  1. College of Life Science, Guizhou University, Guiyang 550025, China
  • Received:2012-12-20 Revised:2013-11-20 Online:2014-01-15 Published:2014-01-22
  • Contact: WU Yong-jun E-mail:wyjbio@163.com

摘要:

以枯草芽孢杆菌BJ3-2 的glsA1 基因为同源序列,通过构建单交换整合载体,将高活性谷氨酰胺酶基因glsA2 定点整合入BJ3-2 菌株染色体中,获得能够稳定遗传的重组菌株BJ3-2A2。经检测重组菌株谷氨酰胺酶活力为BJ3-2 菌株的3.36 倍。利用重组菌株BJ3-2A2 发酵豆豉,全氨基酸检测显示,重组菌株发酵的豆豉谷氨酸含量比出发菌株高12.8%。说明枯草芽孢杆菌BJ3-2 可以转化且为RecE+ 菌株,glsA2 基因在BJ3-2 菌株染色体上可实现较高活性表达,并提高发酵豆豉的鲜味。

关键词: 枯草芽孢杆菌, 同源重组, 单交换, 谷氨酰胺酶基因

Abstract:

With glsA1 in BJ3-2 as the homologous sequence, glutaminase gene glsA2 with high activity was targeted and
integrated into the Bacillus subtilis BJ3-2 chromosome by constructing a single-exchange integrative vector. A recombinant
strain BJ3-2A2 was obtained with good genetic stability. The results of detection showed that glutaminase activity in the
recombinant strain was 3.36 times higher than that in the original strain BJ3-2. The amino acid contents in douchi fermented
with BJ3-2A2 and the original BJ3-2 were measured, and the results showed that glutamate content in fermentation products
from the recombinant strain was increased by 12.8% as compared to that observed with the original strain. All the analyses
show that B. subtilis BJ3-2 is transformable as a RecE+ strain and that glutaminase gene glsA2 can be highly expressed in the
BJ3-2 chromosome, thus improving the flavour of douchi.

Key words: Bacillus subtilis, homologous recombinant, single-exchange, glutaminase-encoding gene

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