依赖于豆粕原材料的纳豆激酶制备和纯化工艺研究  

Study on Preparation and Purification Process of Nattokinase from Soybean Meal Raw Materials

作　　者：吴丹 杨苗苗 杜小平 祁蒙 杨水云[3] WU Dan;YANG Miao-miao;DU Xiao-ping;QI Meng;YANG Shui-yun(College of Life Sciences and Food Engineering,Shaanxi Xueqian Normal University,Xi'an 710100,China;Ankang R&D Center for Se-enriched Products,Ankang 725000,China;School of Life Science and Technology,Xi'an Jiaotong University,Xi'an 710049,China)

机构地区：[1]陕西学前师范学院生命科学与食品工程学院,西安710100 [2]安康市富硒产品研发中心,陕西安康725000 [3]西安交通大学生命科学与技术学院,西安710049

出　　处：《中国调味品》2023年第3期79-85,共7页China Condiment

基　　金：中国富硒产业研究院富硒专项研发计划项目(2021FXZX07-02);陕西省教育厅2021年度自然科学基金项目(21JK0586);陕西高校青年创新团队“秦岭药食同源生物资源保护与利用创新团队”资助;陕西学前师范学院食品科学与工程重点学科资助。

摘　　要：纳豆激酶(nattokinase,NK)是纳豆芽孢杆菌发酵产生的一种丝氨酸蛋白酶,有良好的溶解血栓作用。但目前市面上NK产品纯度较低,需要食用大量粗品才能达到预防血栓的作用,因此,提高NK产品的纯度显得非常必要和重要。对发酵培养基组分进行优化,优化后的天然培养基组成为豆粕3%、甘油0.5%、谷氨酸钠1%、硫酸镁0.5%,在此条件下,发酵NK产量可达到约1486.6 Fu/mL,满足扩大生产要求。在下游纯化技术研究方面,首先调高pH至10,然后采用十六烷基三甲基溴化铵(hexadecyl trimethyl ammonium bromide,CTAB)聚合沉淀NK的伴随产物γ-聚谷氨酸(γ-polyglutamic acid,γ-pGA),在此基础上,利用三项夹心法浓缩NK,透析冻干后获得效价高达(7.88±0.76)×10^(5) Fu/g的高纯NK产品。该技术为生产高品质NK产品提供了重要的技术思路。Nattokinase(NK)is a kind of serine protease produced by the fermentation of Bacillus subtilis natto,which has good thrombolytic effect.However,at present,the purity of NK products on the market is low,and a large amount of crude products are needed to prevent thrombosis.Therefore,it is very necessary and important to improve the purity of NK products.The components of fermentation medium are optimized,and the composition of natural medium after optimization is 3%soybean meal,0.5%glycerin,1%sodium glutamate,0.5%magnesium sulfate.Under these conditions,the yield of fermented NK can reach about 1486.6 Fu/mL,which can meet the requirements of expanded production.In terms of downstream purification technology,firstly,adjust the pH to 10,and then use hexadecyl trimethyl ammonium bromide(CTAB)to polymerize and precipitate the accompanying products of NKγ-polyglutamic acid(γ-PGA).Based on this,NK is concentrated by three-phase partitioning method,and the high-purity NK products with the titer as high as(7.88±0.76)×10^(5) Fu/g are obtained after dialysis and freeze-drying.This technology has provided an important theoretical idea for the production of high-quality NK products.

关 键 词：高纯纳豆激酶 碱性条件 沉淀伴随产物 培养基优化 

分 类 号：TS201.25[轻工技术与工程—食品科学]