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当前位置: 首页 > 文章 > 高活性谷氨酰胺酶基因glsA_2在枯草芽孢杆菌BJ3-2染色体上的定点整合 食品科学 2014,35 (1) 141-144
Position: Home > Articles > Targeted Integration of Glutaminase Gene glsA_2 with High Activity into the Bacillus subtilis BJ3-2 Chromosome FOOD SCIENCE 2014,35 (1) 141-144

高活性谷氨酰胺酶基因glsA_2在枯草芽孢杆菌BJ3-2染色体上的定点整合

作  者：
卢彪;吴拥军;刘艳敏;王亚娟;唐雪
单  位：
贵州大学生命科学学院
关键词：
枯草芽孢杆菌;同源重组;单交换;谷氨酰胺酶基因
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。
译  名：
Targeted Integration of Glutaminase Gene glsA_2 with High Activity into the Bacillus subtilis BJ3-2 Chromosome
作  者：
LU Biao;WU Yong-jun;LIU Yan-min;WANG Ya-juan;TANG Xue;College of Life Science,Guizhou University;
关键词：
Bacillus subtilis;;homologous recombinant;;single-exchange;;glutaminase-encoding gene
摘  要：
With glsA 1 in BJ3-2 as the homologous sequence, glutaminase gene glsA 2 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 glsA 2 can be highly expressed in the BJ3-2 chromosome, thus improving the flavour of douchi.

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