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保加利亚乳杆菌异源表达纳豆激酶的性质研究 被引量:3

Studies on the Character of Heterogeneous Expressing Nattokinase in Lactobacillus delbrueckii subsp. bulgaricus
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摘要 目的:研究保加利亚乳杆菌中异源表达来自纳豆芽孢杆菌的纳豆激酶酶学性质及其体外溶栓作用。方法:从纳豆激酶的最适温度、热稳定性、最适pH值、pH值稳定性和金属离子及化学试剂等方面研究该酶的酶学性质,通过体外试验研究其溶血栓作用,并通过灭活纤溶酶原的方法分析其溶解纤维蛋白的作用方式。结果:保加利亚乳杆菌异源表达纳豆激酶的最适温度为40℃,在低于40℃时热稳定性较好,最适pH 7.0,pH值在6.0~8.0范围内较稳定。Ca^(2+)、Mn^(2+)、Fe^(2+)促进酶活的效果随浓度的变化而不同,其中Cu^(2+)、Hg^(2+)对该酶起明显的抑制作用,Ag+随浓度的增加抑制效果逐渐显著,而蛋白抑制剂PSMF对纳豆激酶具有明显的抑制作用,说明纳豆激酶为丝氨酸蛋白酶。体外试验表明:该酶溶解纤维蛋白的方式是直接溶解,而非通过激活纤溶酶原,且该酶具有明显的体外溶栓作用。结论:为纳豆激酶的应用及开发新型功能性乳制品提供了理论基础。 Objective: Studies on enzyme property and thrombolytic effect in vitro of heterogeneous expressing nattok- inase in L(wtobacilluz delbrueckii subsp, bulgaricus which was cloned from Bacillus natto. Methods: The enzyme property of nattokinase have been researched with the optimum temperature, thermal stability, optimum pH, pH stability, metal ions and the effects of chemicals. The enzyme thrombolytic effect in vitro was researched and dissolving fibrin ways was analyzed through the method of inactivated fibrinolytic. Results: The optimum reaction temperature of heterogeneous ex- pressing nattokinase in Lactobacillus delbrueckii subsp, bulgaricus was 40 ℃, optimum pH value was 7, it was more stable when its temperature belowed 40 ℃ and pH 6.0-8.0. The promoting effects of Ca2., Mn2+, Fe2+ on enzyme activity changed with different concentrations, Cu2+, Hg2+ inhibited the enzyme activity, and the inhibitive effect was more signifi- cant with the higher concentration of Ag+. Meanwhile, the inhibitor PSMF had significant inhibitory effect on nattokinase, which indicated that nattokinase was a serine protease. It showed the way of the enzyme dissolving fibrin was dissolution directly by inactivating the enzyme plasminogen, and the enzyme showed good thrombolytic activity with experiments in vitro. Conclusions: This study provides the theoretical basis for the application of nattokinase and the development of new functional dairy products.
出处 《中国食品学报》 EI CAS CSCD 北大核心 2017年第4期51-57,共7页 Journal of Chinese Institute Of Food Science and Technology
基金 河北省高等学校科学技术研究项目(QN2014065) 国家"863"计划项目(2006AA10Z317)
关键词 保加利亚乳杆菌 纳豆激酶 酶学性质 体外溶栓 Lactobacillus delbrueckii subsp. Bulgaricus nattokinase characterization thrombolytic
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