满丽莉,向殿军

• 1:内蒙古民族大学生命科学学院
• 2:内蒙古民族大学农学院

摘要(Abstract)：

以传统发酵豆制品为供试材料,应用脱脂牛乳平板法和纤维蛋白平板法分离到1株纳豆激酶高产菌株MX-6,经菌落形态、菌体形态及16S rDNA基因同源性分析,确定其为枯草芽孢杆菌。通过单因素试验确定碳源、氮源、无机盐及表面活性剂对纳豆激酶活力影响的基础上,应用响应面法的Box-Behnken设计进行枯草芽孢杆菌MX-6产纳豆激酶发酵培养基的优化。当可溶性淀粉2.91%、大豆蛋白胨1.92%、氯化钙0.04%、硫酸镁0.04%、羧甲基纤维素0.39%时可获得最大的纳豆激酶溶圈直径33.87 mm,与优化前的溶圈直径相比提高了56.81%,表明该模型能较好地预测纳豆激酶的合成情况。

关键词(KeyWords)： 纳豆激酶;筛选鉴定;枯草芽孢杆菌;响应面法;培养基优化

Abstract:

Keywords:

基金项目(Foundation): 内蒙古自治区自然科学基金资助(2018MS03060);; 内蒙古民族大学博士科研启动基金资助(BS403);内蒙古民族大学实验室开放项目

作者(Author): 满丽莉,向殿军

参考文献(References)：

• [1]Mohanasrinivasan V,Mohanapriya A,Potdar S,et al.In vitro and in silico studies on fibrinolytic activity of nattokinase:a clot buster from Bacillus sp.[J].Frontiers in Biology,2017(3):219-225.
• [2]Furie B,Furie B C.Mechanisms of thrombus formation[J].New England Journal of Medicine,2008(9):938-949.
• [3]Tai M W,Sweet B V.Nattokinase for prevention of thrombosis[J].American Journal of Health-system Pharmacy,2006(12):1121-1123.
• [4]Jo H D,Kwon G H,Park J Y,et al.Cloning and overexpression of aprE3-17 encoding the major fibrinolytic protease of Bacillus licheniformis CH3-17[J].Biotechnology and Bioprocess Engineering,2011(16):352-359.
• [5]Kamiya S,Hagimori M,Ogasawara M,et al.In vivo evaluation method of the effect of nattokinase on carrageenan-induced tail thrombosis in a rat model[J].Acta Haematologica,2010(4):218-224.
• [6]Cai D,Zhu C,Chen S.Microbial production of nattokinase:current progress,challenge and prospect[J].World Journal of Microbiology and Biotechnology,2017(33):84.
• [7]Mahajan P M,Gokhale S V,Lele S S.Production of nattokinase using Bacillus natto NRRL 3666:media optimization,scale up,and kinetic modeling[J].Food Science and Biotechnology,2010(6):1593-1603.
• [8]Wang H,Sun X,Wang L,et al.Coproduction of menaquinone-7 and nattokinase by Bacillus subtilis using soybean curd residue as a renewable substrate combined with a dissolved oxygen control strategy[J].Annals of Microbiology,2018(10):655-665.
• [9]Zhang F,Zhang J,Linhardt R J.Interactions between nattokinase and heparin/GAGs[J].Glycoconjugate Journal,2015(9):695-702.
• [10]Dabbagh F,Negahdaripour M,Berenjian A,et al.Nattokinase:production and application[J].Applied Microbiology and Biotechnology,2014(22):9199-9206.
• [11]Yatagai C,Maruyama M,Kawahara T,et al.Nattokinase promoted tissue plasminogen activator release from human cells[J].Pathophysiology of Haemostasis and Thrombosis,2008(5):227-232.
• [12]Zhang X,Yun L,Peng L,et al.Optimization of douchi fibrinolytic enzyme production by statistical experimental methods[J].Journal of Huazhong University of Science and Technology,2013(1):153-158.
• [13]张超凤,严美婷,杜霞,等.响应面法优化纳豆激酶液态发酵条件[J].中国生物制品学杂志,2017(6):658-662.