具有溶栓活性重组乳酸乳球菌构建.pdfVIP

2011 No．1 ·100 · Serial No．226 China Brewing Research Report 具有溶栓活性的重组乳酸乳球菌的构建 余凤云，冯 浩，许 芳，闫达中* （武汉工业学院生物与制药工程学院，湖北 武汉 430023） 摘 要：利用PCR 方法从纳豆芽孢杆菌（Bacillus subtilis subsp. natto）基因组DNA 中扩增纳豆激酶原基因，构建了纳豆激酶原基因的 表达载体pFY002 ，转化Lactococcus lactis NZ9000 ，得到活性表达纳豆激酶的重组菌Lactococcus lactis NZ9000/ pFY002 。利用纤维蛋白 平板法检测其溶栓活性，并对诱导剂乳链菌肽（NisinA）的浓度、温度、诱导时间对重组菌产酶的影响进行探究。 关 键 词：纳豆激酶；乳酸乳球菌；克隆表达 中图分类号：Q933 文献标识码：A 文章编号：0254-5071 （2011）01-0100-04 Construction of recombinant Lactococcus lactis with fibrinolytic activity YU Fengyun, FENG Hao, XU Fang, YAN Dazhong* (School of Biology and Pharmaceutical Engineering, Wuhan Polytechnic University, Wuhan 430023, China) Abstract: Pro-nattokinase gene was amplified by PCR from Bacillus subtilis subsp. natto genomic DNA, which was cloned into pGT-19 vector and then sequenced. The recombinant plasmid pFY002 was constructed by inserting pro-nattokinase gene into expression vector pNZ8048 and was transformed into Lactococcus lactis NZ9000. The fibrinolytic activity was analyzed after the expression of gene was induced by nisin A. The optimum conditions including nisinA concentration, temperature and induction time in the engineering L. lactis were further investigated. Key words: nattokinase; Lactococcus lactis; expression 纳豆激酶（nattokinase NK）是从日本传统的保健食品 酸乳球菌，获得具有溶栓活性的乳酸菌，并进行相关表达 纳豆（natto）中提取的一种碱性丝氨酸蛋白酶，能显著溶解 产物生物活性分析；通过摇瓶培养探究诱导剂乳链菌肽 （ 体内外血栓，并能激活静脉内皮细胞产生纤维蛋白溶酶原 NisinA）的浓度、温度、诱导时间对重组菌产酶的影响，优 [1] 激活剂 。纳豆激酶作为一种很有潜力的新型溶栓药