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CN105755031A 高产细菌素的副干酪乳杆菌基因工程菌株的构建方法 无效 - 驳回

高产细菌素的副干酪乳杆菌基因工程菌株的构建方法
摘要：
高产细菌素的副干酪乳杆菌基因工程菌株的构建方法，涉及一种副干酪乳杆菌基因工程菌株的构建方法。本发明是要解决现有副干酪乳杆菌的细菌素产量低的问题。方法：一、从副干酪乳杆菌HD1.7中扩增prcK基因；二、构建质粒pMD18?T?prcK；三、构建重组过表达载体pRSFDuet?1?prcK；四、构建重组过表达载体pRR?tet；五、将过表达载体pRR?tet电转化副干酪乳杆菌HD1.7感受态细胞；六、prcK基因过表达突变株的筛选和鉴定。该基因工程菌株的抑菌能力比原始菌株提高了17.86％。传代后发酵液细菌素效价2373.46±46.27AU/mL，传代后效价比较稳定。本发明用于产细菌素。
摘要（英）：

The invention provides a construction method of a gene engineering strain of Lactobacillus paracasei with high bacteriocin yield, relates to a construction method of the gene engineering strain of the Lactobacillus paracasei and aims to solve the problem of low bacteriocin yield of the Lactobacillus paracasei at present. The method comprises steps as follows: step one, a prcK gene is amplified from Lactobacillus paracasei HD1.7; step two, a plasmid pMD18-T-prcK is constructed; step three, a recombinant overexpression vector pRSFDuet-1-prcK is constructed; step four, a recombinant overexpression vector pRR-tet is constructed; step five, the overexpression vector pRR-tet is used for performing electrotransformation on Lactobacillus paracasei HD1.7 competent cells; step six, overexpression mutant strains of the prcK gene are screened and identified. The bactericidal capacity of the gene engineering strain is increased by 17.86% compared with that of original strains; bacteriocin titer of fermentation broth after passage is 2372.46 plus or minus 46.27 AU/mL, and the titer after the passage is more stable; the method is applied to bacteriocin production.

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