为构建遗传稳定的ATP高产工程菌,利用PCR技术扩增酿酒酵母ATP合成关键酶基因APT1,并将其克隆至质粒pCB1004-Pgpd的相应位点,得到有强启动子Pgpd驱动的APT1基因超表达质粒pCB1004-Pgpd-APT1。在PEG-CaCl2介导下,超表达质粒转化雅致放射毛霉原生质体,获得ATP高产工程菌。其ATP产量及摩尔转化率比出发菌株提高44.04%。

To construct the engineering strain with genetic stability which can efficient to synthesize ATP,the key enzyme gene APT1 for ATP synthesis which from Saccharomyces cerevisiae was cloned by PCR.The sequence was cloned to corresponding site of pCB1004-Pgpd.Then the super expression plasmid pCB1004-Pgpd-APT1 which controlled by strong promoter Pgpd was obtained.PEG and CaCl2 mediated protoplast transformation of Actinomucor elegans with super expression plasmid was performed and ATP-overproducing transformants were obtained.The ATP yield and Moore conversion were increased 44.04% in comparison with those from original strain.