摘要
用鸟枪法把嗜热脂肪芽孢杆菌313-1(供体菌)染色体上的蛋白酶基因克隆到质粒pPL603上,并在枯草杆菌中得到表达。此基因位于6.6kb的EcoRI酶切片段上,带有重组质粒的枯草杆菌所产生的蛋白酶活性比供体菌株约提高30倍左右。该酶的最适反应温度为75℃,最适pH为7.0左右,是一个中性蛋白酶,在80℃作用30min后仍保留85%以上的酶活。
The structural gene for a thermostable protease from Bacillus stea-rothermophilus (313-1) was cloned in plasmid pPL603 and was expressedin Bacillus subtilis. The gene for protease is coded on a 6.6kb EcoRIfragment. B.subtilis carrying the recombinant plasmid produced about29-fold more protease than the wildtype strain of B. stearothermophilus(313-1) did. Some properties of proteases partially purified from thetransformants of B.subtilis were examined. The protease is charecteri-zed as neutral one in pH, has an optimum reaction temperature of 75℃and retains above 85% of its activity even after treatment of 80℃ for30 min.
出处
《生物工程学报》
CAS
CSCD
北大核心
1991年第3期207-212,共6页
Chinese Journal of Biotechnology
关键词
基因克隆
耐热蛋白酶
基因
Shot-gun
gene clone
thermostable neutral protease gene
