响应面法优化链霉菌产支链蒽醌发酵培养基  

Optimization of Streptomyces sp.Fermentation Medium for Production of Alkylated Anthraquinone by Response Surface Methodology

作　　者：刘芳 权婷婷 张菲 石丽桥 刘曼莉 吴兆圆 LIU Fang;QUAN Tingting;ZHANG Fei;SHI Liqiao;LIU Manli;WU Zhaoyuan(Hubei Biopesticide Engineering Research Centre,Wuhan 430064,China;College of Plant Science and Technology,Huazhong Agricultural University,Wuhan 430070,China)

机构地区：[1]湖北省生物农药工程研究中心,武汉430064 [2]华中农业大学植物科技学院,武汉430070

出　　处：《中国畜牧兽医》2024年第7期3142-3154,共13页China Animal Husbandry & Veterinary Medicine

基　　金：国家重点研发计划(2021YFD1800402);湖北省农业科技创新中心创新团队项目(2021-620-000-001-027);中央引导地方科技发展资金项目(2022BGE250);武汉市知识创新专项曙光计划项目(2022020801020348)。

摘　　要：【目的】通过响应面法优化链霉菌(Streptomyces sp.)WS-13394株发酵培养条件及培养基成分,以提高菌株产支链蒽醌的水平,为该化合物在畜牧业领域的应用提供技术支撑。【方法】采用单因素试验确定链霉菌WS-13394株产支链蒽醌的最佳摇瓶发酵条件及最适碳氮源和无机盐组合,利用Plackett-Burman试验筛选出影响摇瓶发酵产量的显著因子;结合最陡爬坡试验、响应面设计分析,拟合显著因子与支链蒽醌产量的非线性方程,从而获得优化的菌株发酵培养基配方。【结果】优化后的最佳配方为可溶性淀粉28.12 g/L、鱼粉10.00 g/L、酵母膏6.34 g/L、碳酸钙0.71 g/L、硫酸镁0.50 g/L、磷酸氢二钾0.50 g/L,此模型预测发酵液支链蒽醌产量为408.35 mg/L。应用此优化组合进行摇瓶发酵试验,支链蒽醌产量为411.73 mg/L,产量较优化前(89.70 mg/L)提高了3.59倍。【结论】通过响应面优化获得了显著提高链霉菌WS-13394株产支链蒽醌的摇瓶发酵培养基配方,试验结果为该类化合物的抗菌活性机制研究及其在动物模型中的效果评估提供了技术支撑。【Objective】This study was aimed to optimize the fermentation conditions and medium composition of Streptomyces sp.WS-13394 by response surface methodology,so as to improve the production level of alkylated anthraquinones produced by the strain,and provide technical support for the application of this compound in animal husbandry.【Method】The optimal shake-flask fermentation conditions and the proper combination of carbon and nitrogen sources and mineral salts for the yield of alkylated anthraquinone produced by Streptomyces sp.WS-13394 were achieved through single-factor experiments.The significant factors affecting the yield of alkylated anthraquinone were fixed through Plackett-Burman design.The optimized medium for the yield of alkylated anthraquinone was achieved through the steepest ascending experiment,response surface design by fitting the significant factors with the yield of alkylated anthraquinone.【Result】The composition of the optimized medium for the yield of alkylated anthraquinone produced by Streptomyces sp.WS-13394 was as followings:Soluble starch 28.12 g/L,fish meal 10.00 g/L,yeast extract 6.34 g/L,CaCO_(3)0.71 g/L,MgSO_(4)0.50 g/L,and K_(2)HPO_(4)0.50 g/L.The theoretical yield of alkylated anthraquinone based on the nonlinear regression formula was 408.35 mg/L.The optimal combination was applied to shake bottle fermentation test,and the yield of alkylated anthraquinone was 411.73 mg/L,which was 3.59 times higher than that before optimization(89.70 mg/L).【Conclusion】Response surface optimization was used to obtain the shaking flask fermentation medium formulation that significantly improved the yield of alkylated anthraquinone produced by Streptomyces sp.WS-13394.The results provided technical support for the studies of the antibacterial activity mechanism of this compound and the evaluation of its effect in animal models.

关 键 词：链霉菌 发酵培养基 优化 响应面法 支链蒽醌 

分 类 号：S816.7[农业科学—饲料科学]