Optimization and stabilization of antifungal protein from Bacillus amyloliquefaciens strain HRH317
-
摘要: 本文以抑菌圈直径为指标,采用单因素实验和响应面法对解淀粉芽孢杆菌HRH317菌株产抗菌蛋白的发酵条件进行优化。得到最佳发酵条件为:初始p H为7.0的NB培养液,接种量4%,37℃下振荡培养24 h,转速为200 r/min。在此条件下抗菌蛋白的抑菌圈直径达21.85 mm,结果表明该抗菌蛋白具有良好的抑菌效果。稳定性研究结果表明:蛋白在40~80℃、p H2~11稳定,对蛋白酶K有一定耐受性。Abstract: This paper is concerned with the antifungal protein extraction from the strains Bacillus amyloliquefaciens HRH317.The inhibitive circle diameter as evaluating indicator, the fermentation condition were optimized by single factor experiments and response surface methodology ( RSM) .The results showed the optimized extraction parameters were that the initial p H value of nutrient broth medium ( NB) was 7, inoculation volume of 4%, 37 ℃, and the shaker rotation speed 200 r/min.The inhibitive circle diameter was up to 21.85 mm.The results indicated that the antifungal protein was proved to have strong inhibition effect.The antifungal protein was tested to have strong tolerant to proteinase K, which could be steady with 40~80 ℃ and p H2~11.
-
[1] 梁泉峰.间型假比酵母菌对果蔬采后腐败霉菌拮抗作用的研究[D].泰安:山东大学, 2002. [2] Zheng Y, Chen YS, Lu HS.Screening for antibacterial and antifungal activities in some marine algaefrom the Fu-jian coast of China[J].China Oceanol Limnol, 2001, 19 (4) :326-331.
[3] 黄春萍, 张宏, 张晓喻.植物抗真菌蛋白的分类及抗菌机制研究进展[J].国外医药抗生素分册, 2007, 28 (4) :167-171. [4] Yuan Bo, Wang Zhe, Qin Sheng, et al.Study of the antisapstain fungus activity of Bacillus amyloliquefaciens CGMCC5569 associated with Ginkgo biloba and identification of its active components[J].Bioresource Technology, 2012, 114:536-541.
[5] Mohamed S O, Dharini S, Lise K.生物抑制剂解淀粉芽孢杆菌结合1-甲基环丙烯处理在果实采后病害控制及品质保持中的作用效果[J].保鲜与加工, 2011 (2) :56-56. [6] 郝卫宁, 李辉, 胡美英, 等.柑桔绿霉病拮抗细菌的筛选、鉴定及其抑制效果[J].中国生物防治学报, 2011, 27 (2) :284-288. [7] 韩玉竹, 邓钊, 张宝, 等.解淀粉芽孢杆菌H15产抗菌肽的发酵条件优化和提取方法比较研究[J].食品科学, 2015, 36 (15) :135-141. [8] Zhao Zhenzhen, Wang Qiushuo, Wang Kaimei, et al.Study of the antifungal activity of Bacillus vallismortis ZZ185 in vitro and identification of its antifungal components[J].Bioresource Technology, 2010, 101:292-297.
[9] Slavokhotova A A, Rogozhin E A, Musolyamov A K, et al.Novel antifungal a-hairpinin peptide from Stellaria media seeds:structure, biosynthesis, gene structure and evolution[J].Plant Molecular Biology, 2014, 84:189-202.
[10] 胡剑, 林心怡, 张九一, 等.拮抗菌BS-98分泌抗菌蛋白的条件及其发酵液特性[J].微生物学通报, 1996, 23 (6) :323-326. [11] 洪鹏, 安国栋, 胡美英, 等.解淀粉芽抱杆菌HF-01发酵条件优化[J].中国生物防治学报, 2013, 29 (4) :569-578. [12] 杨桥, 韩文菊, 张文俊, 等.产Maerolactin A抗生素海洋解淀粉芽孢杆菌的鉴定及发酵条件优化[J].药物生物技术, 2009, 16 (4) :311-315 [13] 辛媛媛, 侯美玲, 郝志敏, 等.一株拮抗细菌HJ5高产抗菌物质发酵条件优化[J].河北农业大学学报, 2012, 35 (4) :70-74. [14] 周映华, 李秋云, 曾艳, 等.益生菌蜡状芽孢杆菌发酵条件及培养基的优化[J].饲料研究, 2007 (10) :56-58. [15] 黄海蝉, 裘娟萍.枯草芽孢杆菌防治植物病害的研究进展[J].浙江农业科学, 2005 (3) :213-215. [16] 汪燕, 罗水忠, 蔡静, 等.响应面法优化茂源链霉菌产谷氨酰胺转氨酶发酵条件[J].食品工业科技, 2015, 36 (16) :203-207. [17] 郝秋娟, 李树立, 陈颖.温度对淀粉液化芽孢杆菌5582产蛋白酶的影响[J].中国酿造, 2008 (11) :37-39. [18] 方传记.陆兆新, 孙力军.淀粉液化芽孢杆菌抗菌脂肽发酵培养基及发酵条件的优化[J].中国农业科学, 2008, 41 (2) :533-539. [19] Claude P, Selitemnikoff.Antifungal Proteins[J].Applied and Environmental Microbiology, 2001, 67 (7) :283-289.
[20] 高学文, 姚仕义, Huongpham, 等.基因工程菌枯草芽抱杆菌GEB3产生的脂肽类抗生素及其生物活性研究[J].中国农业科学, 2003, 36 (12) :1496-1501. [21] SL Wang, YH Yen, GC Tzeng, et al.Production of antifungal material by bioconversion of shellfish wastes fermented by Pseudomonas fluorescens K-188[J].Enzyme and Microbial Technology, 2005, 36:49-56.
[22] Ongena M, Jacques P.Bacillus lipoeptides:versatile weapons for plant disease biocontrol[J].Trends in Microbiology, 2008, 6 (3) :115-125.
[23] 刘永峰.枯草芽孢杆菌 (Bacillus subtilis) Bs-916胞外抗菌蛋白质的纯化及其鉴定[D].南京:南京农业大学, 2006. [24] 张宝俊, 张家榕, 韩巨才, 等.内生解淀粉芽孢杆菌LP-5抗菌蛋白的分离纯化及特性[J].植物保护学报, 2010, 37 (2) :143-147. [25] 王军华, 权春善, 徐洪涛, 等.解淀粉芽孢杆菌Q-12抗真菌特性的研究[J].食品与发酵工业, 2006, 32 (6) :47-50.
计量
- 文章访问数: 212
- HTML全文浏览量: 21
- PDF下载量: 165
下载:
