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发明公开
CN110387345A 一种基于CRISPR-Cas9技术的氨糖合酶产生菌、构建方法及其应用 无效 - 驳回

一种基于CRISPR-Cas9技术的氨糖合酶产生菌、构建方法及其应用
摘要：
本发明公开了一种基于CRISPR-Cas9技术的氨糖合酶产生菌、构建方法及其应用，该氨糖合酶产生菌是将氨基葡萄糖乙酰化酶基因gnal和氨基葡萄糖合酶基因glms连接到载体pET-28a上，再将重组质粒转入CRISPR-Cas9基因编辑技术敲除E.coli BL21(DE3)基因组氨基葡萄糖代谢基因naggE和mannX得到的双基因缺失宿主菌中获得的。该重组菌应用于发酵培养，在摇瓶上可以累积氨基葡萄糖7.51g/L，乙酰氨基葡萄糖3.73g/L，总产量为11.24g/L；在5L发酵罐上可以累积氨基葡萄糖7.30g/L，乙酰氨基葡萄糖10.53g/L，总产量为17.83g/L。
摘要（英）：

The invention discloses a glucosamine synthase producing bacterium based on a CRISPR-Cas9 technology and a construction method and application of the glucosamine synthase producing bacterium. The glucosamine synthase producing bacterium is obtained in the modes that a glucosamine acetylase gene gnal and a glucosamine synthase gene glms are connected to a carrier pET-28a, and then a recombinant plasmid is transferred into a double-gene deletion host bacterium obtained by knocking out a glucosamine metabolism gene naggE and a glucosamine metabolism gene mannX of an E.coli BL21(DE3) genome through the CRISPR-Cas9 gene editing technology. The recombinant bacterium is applied to fermentation culture, on a shake flask, 7.51 g/L of glucosamine can be accumulated, 3.73 g/L of acetylglucosamine canbe accumulated, and the total output is 11.24 g/L; and on a fermentor of 5 L, 7.30 g/L of the glucosamine can be accumulated, 10.53 g/L of the acetylglucosamine can be accumulated, and the total output is 17.83 g/L.

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