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发明公开
CN106754505A 高产乙酰氨基葡萄糖的重组枯草芽孢杆菌及其构建方法 审中-实审

高产乙酰氨基葡萄糖的重组枯草芽孢杆菌及其构建方法
摘要：
本发明涉及一种高产乙酰氨基葡萄糖的重组枯草芽孢杆菌，通过敲除枯草芽孢杆菌中调控氨基葡萄糖合成酶表达的glmS核酶，并插入终止子和启动子序列而得到。本发明还公开了一种高产乙酰氨基葡萄糖的重组枯草芽孢杆菌的构建方法，包括以下步骤：构建glmS核酶编码基因敲除框，所述敲除框依次包括glmS核酶编码基因上游同源臂、终止子、抗性基因、组成型启动子和glmS核酶编码基因下游同源臂序列；将得到的glmS核酶编码基因敲除框转化枯草芽孢杆菌，得到重组枯草芽孢杆菌。通过同源重组敲除氨基葡萄糖合成酶基因(glmS)核酶，阻断了宿主菌胞内6‑磷酸氨基葡萄糖反馈抑制氨基葡萄糖合成酶基因(glmS)的表达，促进乙酰氨基葡萄糖积累。
摘要（英）：

The invention relates to a recombinant bacillus subtilis of high-yield acetylglucosamine. The recombinant bacillus subtilis is obtained by knocking out glmS ribozyme expressed by regulatory glucosamine synthetase in bacillus subtilis and inserting terminator and promoter sequences. The invention further discloses a construction method of the recombinant bacillus subtilis of the high-yield acetylglucosamine. The construction method comprises the following steps that a glmS ribozyme encoding gene knockout frame is established, wherein the knockout frame sequentially comprises a glmS ribozyme encoding gene upstream homologous arm, a terminator, a resistant gene, a constitutional promoter and a glmS ribozyme encoding gene downstream homologous arm sequence; the obtained glmS ribozyme encoding gene knockout fram is converted into bacillus subtilis, and the recombinant bacillus subtilis is obtained. Expression of a 6-phosphoric glucosamine feedback inhibition glucosamine synthetase gene (glmS) in a host bacterial cell is blocked by homologous recombination and knockout of the glucosamine synthetase gene (glmS) ribozyme, and acetylglucosamine accumulation is promoted.

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