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。
With glsA1in BJ3-2 as the homologous sequence, glutaminase gene glsA2 with high activity was targeted andintegrated into the Bacillus subtilis BJ3-2 chromosome by constructing a single-exchange integrative vector. A recombinantstrain BJ3-2A2 was obtained with good genetic stability. The results of detection showed that glutaminase activity in therecombinant strain was 3.36 times higher than that in the original strain BJ3-2. The amino acid contents in douchi fermentedwith BJ3-2A2and the original BJ3-2 were measured, and the results showed that glutamate content in fermentation productsfrom the recombinant strain was increased by 12.8% as compared to that observed with the original strain. All the analysesshow that B. subtilis BJ3-2 is transformable as a RecE＋strain and that glutaminase gene glsA2can be highly expressed in theBJ3-2 chromosome, thus improving the flavour of douchi.
出处
《食品科学》
EI
CAS
CSCD
北大核心
2014年第1期141-144,共4页
Food Science
基金
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)
关键词
枯草芽孢杆菌
同源重组
单交换
谷氨酰胺酶基因
Bacillus subtilis
homologous recombinant
single-exchange
glutaminase-encoding gene
