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多杀菌素发酵培养基的研究
李能威,张晓琳,郭伟群,熊犍,宋炜,宋渊
作者单位
李能威 国家粮食局 科学研究院北京 100037 
张晓琳 国家粮食局 科学研究院北京 100037 
郭伟群 国家粮食局 科学研究院北京 100037 
熊犍 华南理工大学 轻工与食品学院广东 广州 510640 
宋炜 国家粮食局 科学研究院北京 100037 
宋渊 中国农业大学 生物学院北京 100094 
摘要:
【目的】 以刺糖多孢菌C4-10-8B为试验菌株,对多杀菌素发酵培养基进行优化,以提高刺糖多孢菌的多杀菌素发酵产量。【方法】 采用单因素和多因素试验,分别考察了不同碳源、氮源、前体、无机盐和微量元素对多杀菌素生物合成的影响,最后通过正交试验综合考察发酵培养基中各组分对多杀菌素产量的影响。【结果】 葡萄糖是较优碳源,最佳质量浓度为40.0 g/L,当葡萄糖与糊精(质量比5∶2)复配时,多杀菌素产量较对照培养基提高18.35%;棉籽蛋白为最优氮源,25.0 g/L棉籽蛋白与15.0 g/L玉米蛋白胨或0.6 g/L硫酸铵复配时,多杀菌素产量分别较对照提高40.0%和45.3%;乙酸钠和谷氨酰胺质量浓度为1.0 g/L时,均能使多杀菌素产量较对照提高21.5%;K2HPO4对多杀菌素合成有明显的促进作用,在其质量浓度为2.5 g/L时,多杀菌素产量较对照提高24.5%;ZnCl2对多杀菌素合成也有促进作用,而MgSO4和CoCl2则显著抑制多杀菌素合成;通过2次正交试验获得最优发酵培养基,利用该培养基时多杀菌素产量可达395.54 μg/mL,比对照提高了62.5%。【结论】 多杀菌素生物合成的最优发酵培养基为:葡萄糖50.0 g/L,棉籽蛋白25.0 g/L,玉米蛋白胨20.0 g/L,(NH4)2SO4 0.8 g/L,谷氨酰胺1.25 g/L,乙酸钠0.8 g/L,NaCl 3.0 g/L,K2HPO4 2.5 g/L,FeSO4 50.0 mg/L,ZnCl2 25.0 mg/L,CaCO3 1.0 g/L。
关键词:  刺糖多孢菌  多杀菌素  发酵培养基
DOI:
分类号:
基金项目:国家“十一五”科技支撑计划项目子课题(No.2006BAD08B02)“高效生物储粮防护剂-多杀菌素的研制开发”
Study on the fermentation media for spinosad production
LI Neng-wei,ZHANG Xiao-lin,GUO Wei-qun,XIONG Jian,SONG Wei,SONG Yuan
Abstract:
【Objective】 The fermentation media of Saccharopolyspora spinosa C4-10-8B were optimized for better spinosad production.【Method】 One-factor and multifactor experimental designs to optimize carbon sources,nitrogen sources,precursors,minerals and trace elements for the spinosad production were performed,and each selected element was discussed comprehensively with orthogonal designs for better spinosad-producing fermentation media.【Result】 The results showed that glucose was the optimum carbon source,and the optimal mass concentration was 40.0 g/L,and additional 18.35% spinosad yield was achieved with the combination of 5 parts(m) glucose and 2 parts(m) dextrin;cottonseed protein was the preferred nitrogen source,and with the mass concentration of 15.0 g/L corn peptone or 0.6 g/L (NH4)2SO4 compounded with 25.0 g/L cottonseed protein,the spinosad yield increased respectively by 40.0% and 45.3%;with the mass concentration of 1.0 g/L L-glutamine or sodium acetate,the spinosad yield increased by 21.5%;K2HPO4 was positive for the spinosad production,and additional 24.5% spinosad yield was achieved when its mass concentration was 2.5 g/L,ZnCl2 also had the positive effect;the spinosad production significantly reduced in the presence of CoCl2 or MgSO4.With orthogonal designs,the spinosad yield in the optimized fermentation medium increased by 62.5%,up to 395.54 μg/mL.【Conclusion】 The optimal fermentation medium for spinosad production was glucose 50.0 g/L,cottonseed protein 25.0 g/L,corn peptone 20.0 g/L,(NH4)2SO4 0.8 g/L,L-glutamine 1.25 g/L,sodium acetate 0.8 g/L,NaCl 3.0 g/L,K2HPO4 2.5 g/L,FeSO4 50.0 mg/L,ZnCl2 25.0 mg/L,CaCO3 1.0 g/L.
Key words:  Saccharopolyspora spinosa  spinosad  fermentation medium