磷酸甘露酶基因的阻断对刺糖多孢菌的形态及其多杀菌素合成的影响

发布时间：2018-08-14 16:24

【摘要】：磷酸甘露酶(PMM)是由manB基因编码的广泛存在于真核生物中的蛋白酶,在糖代谢过程中磷酸-6-甘露糖可在磷酸甘露酶的作用下转化为磷酸-1-甘露糖,与细胞的初级代谢过程及次级代谢产物的合成过程密切相关,可通过对刺糖多孢菌中与初级代谢途径相关的基因进行分析改造,促使多杀菌素的合成显著提高。本研究在刺糖多孢菌基因组中克隆了磷酸甘露酶编码基因manB的部分片段,将其与穿梭载体pOJ260连接,构建了阻断型载体pOJ260-manB。通过接合转移将其导入刺糖多孢菌野生菌中,获得刺糖多孢菌突变菌株S.spinosa-△manB。将突变菌株接种至不同培养基,观察突变菌株与野生菌株的孢子形成情况,发现突变菌株的孢子萌发有所提前,且孢子产生量比野生菌略多;在HPLC上检测多杀菌素的产量发现突变菌株的产量相对于野生菌株提高了180%,这说明manB基因对刺糖多孢菌的生长发育有一定的影响,并对调控多杀菌素的合成具有重要作用。
[Abstract]:Phosphomannase (PMM) is a protease encoded by manB gene in eukaryotes. In the process of glucose metabolism, phosphoric acid -6-mannose can be transformed into phosphate-1-mannose by the action of phosphomanase. It is closely related to the primary metabolic process of cells and the synthesis of secondary metabolites, which can be improved significantly by the analysis and modification of the genes related to primary metabolic pathway in polyspora spp. In this study, a partial fragment of phosphomannase encoding gene manB was cloned from the genome of Polysporium spp., and ligated with the shuttle vector pOJ260, the blocking vector pOJ260-man Bwas constructed. The mutant S.spinosa- manb was obtained by conjugation transfer. The spore formation of mutant strain and wild strain was observed by inoculating the mutant strain into different culture medium. It was found that the spore germination of mutant strain was earlier than that of wild strain, and the spore production of mutant strain was a little more than that of wild strain. The yield of the mutant strain was detected on HPLC. The yield of the mutant strain was increased by 180% compared with that of the wild strain, which indicated that the manB gene had a certain effect on the growth and development of polysporaria spp, and played an important role in regulating the synthesis of the bactericosporin.
【作者单位】：湖南师范大学生命科学学院/微生物分子生物学国家重点实验室培育基地;
【基金】：国家国际合作重点项目(2011DFA32610) 国家“973”计划(2011CB7221301) 国家“863”计划(2011AA10A203) 湖南省协同创新中心项目(20134486)
【分类号】：TQ920.1;TQ450.1

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