Investigating the synthesis and regulation mechanism of γ-PGA in Bacillus licheniformis WX-02 using

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  地衣芽孢杆菌(Bacillus licheniformis) WX-02作为一种重要的工业生产菌株,广泛地应用于农业及工业领域,其生产的代谢产物γ-聚谷氨酸(Poly-γ-glutamic acid,γ-PGA)能够作为重金属吸附剂、保湿剂、肥料吸收促进剂和饲料营养助剂等应用于农业、食品、饲料、化妆品、医药和污水处理领域.地衣芽孢杆菌WX-02菌株在正常条件下可以合成γ-PGA,而在高盐胁迫条件下却可以高产γ-PGA.本研究首先完整地解析了地衣芽孢杆菌WX-02的基因组信息,其基因组由长度为4,286,821bp的一条环状染色体组成,含有4512个蛋白编码基因,24个rRNA基因及79个tRNA基因,GC含量为46.1%.基于地衣芽孢杆菌WX-02完整的基因组信息,本研究结合转录组、蛋白相互作用网络和代谢网络等系统生物学方法系统地研究了γ-PGA的合成调控及其在盐胁迫条件下高产的分子机理.通过分析正常和高盐胁迫(6% NaCl)条件下的转录组,发现高浓度盐促进了谷氨酸合成相关基因(g#ABC,gltP及ycgMNO)的表达,从而使得γ-PGA在盐胁迫环境下能够高产.分析γ-PGA相关的蛋白相互作用子网络,发现转录调控因子CcpA与多数γ-PGA合成及调控相关的蛋白(PgsB,GltA,GltB,ProB,ProJ,YcgM及双信号转导系统ComP-ComA和DegS-DegU)具有直接或间接的互作关系,推断CcpA可能在γ-PGA合成及调控中起到一个非常至关重要的作用.为进一步研究地衣芽孢杆菌WX-02的基本代谢情况及γ-PGA合成调控相关的重要代谢物及路径,本研究构建了地衣芽孢杆菌WX-02的代谢网络模型iWX1009,并发现从网络中移除5个反应(rxn00800,rxn00097,rxn05164,rxn05299及rxn00047)可以使得γ-PGA的产量最大化.另外通过网络模型分析推导出了γ-PGA合成的相关代谢物及代谢路径.本研究应用系统生物学方法探究了调控γ-PGA合成的潜在因子、代谢物及代谢路径,初步阐明了γ-PGA的代谢调控机理,为构建γ-PGA高效规模化生产的基因工程菌株奠定了理论基础.
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