【摘要】 研究了三磷酸腺苷(adenosine triphosphate,ATP)调节变构菌素(tautomycetin,TMC)生物合成的机制。以分子生物学手段对Streptomyces griseochromogenes DM9562(WT)中的tmcN基因进行敲除、互补和ATPase结构域点突变;分析各突变菌株tmcN转录水平、TMC产量及胞内ATP水平;以外源添加的方法观察ATP对WT、基因敲除菌株、互补菌株和点突变菌株TMC生物合成的影响。结果显示:tmcN基因敲除和点突变显著影响TMC的生物合成及胞内ATP水平;外源添加5μmol/L ATP可显著上调WT中TMC产量,但对tmcN和点突变菌株TMC生物合成无影响。本研究提示调节蛋白TmcN可能介导了ATP对TMC生物合成的调节过程。更多还原

【Abstract】 The mechanism of tautomycetin biosynthesis regulated by adenosine triphosphate(ATP)was investigated.The tmcN gene in Streptomyces griseochromogenes DM9562(WT)was knockout through molecular biotechnologies,and the tmcNor sitedirect mutated tmcN were re-introduced to get the complementary and ATPase domain disrupted strains.The transcription level of tmcN,TMC yield and intracellular ATP in different strains were analyzed.The effects of ATP on TMC production of WT and different mutant strains were evaluated.The results show that tmcNgene knockout and site-direct mutation in ATPase domain of TmcN significantly affect the biosynthesis of TMC as well as the intracellular ATP levels.Addition of 5μmol/L ATP increases the TMC production of WT train,but there is no effect on tmcN and point mutant strain TMC biosynthesis.The results in this study suggest that the regulatory protein TmcN may mediate the ATP regulation of TMC biosynthesis.更多还原