泰乐菌素基因工程菌的构建Construction of a gene-engineered strain for the production of tylosin
范亮,杜宝华,陈光,田威,何建勇
FAN Liang,DU Bao-hua,CHEN Guang,TIAN Wei,HE Jian-yong(School of Life Science and Biopharmaceutics
摘要(Abstract)：
目的构建具有双拷贝tylF基因的泰乐菌素基因工程菌,以解决泰乐菌素基因生物合成的限速环节,提高泰乐菌素的发酵产量。方法通过SOE-PCR获得PermE-tylF基因,构建随机整合型重组质粒pBH05,通过接合转移将PermE-tylF基因及其载体随机整合到弗氏链霉菌的基因组中,并通过抗性筛选获得重组菌株D-120。将重组菌株D-120进行摇瓶发酵,采用生物效价测定的二剂量法测定泰乐菌素的发酵单位。结果在相同的发酵条件下,重组菌株泰乐菌素的发酵单位较出发菌株提高32.7%。结论该基因工程菌的构建改善了泰乐菌素生物合成的限速环节,大幅度提高了泰乐菌素的发酵效价。
Objective To construct a gene-engineered strain which producing tylosin by introducing PermE-tylF gene in the genomic DNA.Methods PermE-tylF gene was obtained by SOE-PCR.An E.coli-Streptomyces shuttle plasmid pBH05 containing PermE-tylF gene was constructed and was introduced into Streptomyces fradiae Ty-2010 through conjugal transfer.Tylosin fermentation was conducted by shaking flask method and tylosin titer was determined by bacterial inhibition tests.Results The tylosin productivity of recombinant strain D-120 was increased by 32.7% than the original strain.Conclusions The productivity of tylosin is obviously increased by the gene-engineered strain,in which the copy number of tylF gene is increased by introducing PermE-tylF gene in the genomic DNA of Streptomyces fradiae.
关键词(KeyWords)：
弗氏链霉菌;tylF基因;泰乐菌素;基因工程菌
Streptomyces fradiae;tylF gene;tylosin;gene-engineered strain
基金项目(Foundation):
作者(Author):
范亮,杜宝华,陈光,田威,何建勇
FAN Liang,DU Bao-hua,CHEN Guang,TIAN Wei,HE Jian-yong(School of Life Science and Biopharmaceutics
DOI: 10.14066/j.cnki.cn21-1349/r.2012.05.017
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