基于响应面法优化拮抗大丽轮枝菌枯草芽孢杆菌BS-Z15发酵培养基  

Optimization of Fermentation Medium Against Verticillium Dahliae Bacillus Subtilis BS-Z15 Based on Response Surface Methodology

作　　者：宁焕宸 吴梦君 赵惠新[1] 刘会强[1] NING Huan-chen;WU Meng-jun;ZHAO Hui-xin;LIU Hui-qiang(Xinjiang Key Laboratory of Special Environmental Species Protection and Regulation Biology,College of life Sciences,Xinjiang Normal University,Urumqi,Xinjiang,830054,China;Union Hospital Affiliated to Tongji Medical College,Huazhong University of Science and Technology,Wuhan,Hubei,430032,China)

机构地区：[1]新疆特殊环境物种保护与调控生物学自治区重点实验室,新疆师范大学生命科学学院,新疆乌鲁木齐830054 [2]华中科技大学同济医学院附属协和医院,湖北武汉430032

出　　处：《新疆师范大学学报（自然科学版）》2022年第2期54-62,共9页Journal of Xinjiang Normal University(Natural Sciences Edition)

基　　金：新疆维吾尔自治区高校科研计划项目(XJEDU2021I023);国家自然科学基金项目(32160074);新疆维吾尔自治区重点实验室开放课题(2020D04010)。

摘　　要：前期从新疆和硕县的连作棉花根际土壤中分离得到了一株枯草芽孢杆菌(BS-Z15),该菌能发酵产生高效拮抗大丽轮枝菌、防治棉花黄萎病的代谢产物,有较好的应用前景。为了提高该活性物质的发酵产量,本研究以发酵菌液浑浊度为菌体生长速度指标,以等体积发酵液浓缩物拮抗大丽轮枝菌活性为该活性物质产量指标,通过单因素试验,明确了最优培养基中的碳源、氮源及无机盐分别为葡萄糖、酵母浸粉、Mn^(2+)(或Fe^(2+));进一步采用响应面法对培养基中最优碳源、氮源和无机盐的浓度进行优化,优化培养基的葡萄糖、酵母浸粉、MnSO_(4)·H_(2)O浓度分别为11.61g/L、11.22 g/L、0.99 g/L.本研究采用优化培养基发酵BS-Z15,通过HPLC法分析表明其抗菌活性物质产量较对照提高77%,且显著提高BS-Z15菌体生长速度,为高效制备枯草芽孢杆菌BS-Z15菌剂及发酵生产抗大丽轮枝菌等真菌的活性物质提供了原料基础。In the early stage,a strain of Bacillus subtilis(BS-Z15)was isolated from the rhizosphere soil of continuous cropping cotton in Heshuo area,Xinjiang.This strain can ferment and produce metabolites that can effec-tively antagonize Verticillium Dahliae and control cotton Verticillium wilt,which has a good application prospect.In order to improve the fermentation yield of the active substance,the turbidity of fermentation broth was taken as the index of cell growth rate,and the activity of equal volume fermentation broth concentrate against Verticillium Dahliae was taken as the index of the yield of the active substance.Through single factor test,it was determined that the carbon source,nitrogen source and inorganic salt in the optimal medium were glucose,yeast extract powder,Mn^(2+)(or Fe^(2+));Response surface methodology was further used to optimize the concentration of the optimal carbon source,nitrogen source and inorganic salt in the medium.The concentrations of glucose,yeast extract powder and MnSO_(4)·H_(2)O in the optimized medium were 11.61 g/L,11.22 g/L and 0.99 g/L respectively.BS-Z15 was fermented in the optimized medium,and the analysis of HPLC showed that the yield of antibacterial active substances was 77%higher than control group,and the growth rate of BS-Z15 was significantly increased.This study provides a raw material basis for the efficient preparation of Bacillus Subtilis BS-Z15 and the fermentation of active substances against Verticillium Dahliae and other fungi.

关 键 词：枯草芽孢杆菌BS-Z15 培养基优化 菌体生长 抗真菌活性物质 响应面法 

分 类 号：Q939.9[生物学—微生物学]