武夷菌素高产基因工程菌发酵培养基的优化  

Optimization of Fermentation Medium for High Producing Strain of Wuyiencin by Genetically Engineered Bacterium

作　　者：葛蓓孛[1] 王家旺[1] 刘彦彦[1] 刘炳花 PARK Kyungseok PARK Yongsoon 张克诚[1] GE Beibei WANG Jiawang LIU Yanyan LIU Binghua PARK Kyungseok PARK Yongsoon ZHANG Kecheng(Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China Division of Agricultural Microbiology, National Academy of Agricultural Science, Rural Development Administration, Jeonju 560500, Republic of Korea Division of Biotechnology, Chonbuk National University, Jeonbuk 54596, Republic of Korea)

机构地区：[1]中国农业科学院植物保护研究所,北京100193 [2]韩国农村振兴厅国家农业科学院农业微生物研究所,全州560500 [3]韩国全北国立大学生物技术学院,全北54596

出　　处：《中国生物防治学报》2016年第5期642-649,共8页Chinese Journal of Biological Control

基　　金：国家公益性行业(农业)科研专项(201303025);国家自然科学基金(31371985;31401796)

摘　　要：为进一步提高武夷菌素的产量,以基因工程菌Streptomyces ahygroscopicus var.wuyiensis W273为试验菌株,采用单因子试验确定发酵培养基的最适6种营养成分为玉米粉、葡萄糖、黄豆面、氯化铵、碳酸钙和氯化镁;应用SAS软件中的Plackett-Burman设计法,对影响菌株发酵生产武夷菌素的6个因素进行了筛选,确定葡萄糖和碳酸钙为影响摇瓶发酵生产武夷菌素的主要因素。利用响应面分析法对2个主要因素的最佳水平及其交互作用进行研究,建立了二次回归方程,Y=218.18＋270.28X1＋1274.74X2―3.28X12―5.4X1X2―200.98X22,并最终确定最佳培养基配方为玉米粉20 g/L、葡萄糖39 g/L、黄豆面20 g/L、氯化铵4 g/L、碳酸钙2.65 g/L、氯化镁0.4 g/L。以新配方在28℃、220 r/min进行摇瓶发酵60 h后,武夷菌素效价平均值为7215μg/m L,较原始培养基发酵后武夷菌素效价提高了26.5%。To improving fermentation production of wuyiencin, the method of one-dimensional search was used to select suitable fermentation medium using the genetic engineering bacterium Streptomyces ahygroscopicus var. wuyiensis W273 as experimental material. Six kinds of nutrients were suitable for fermentation, which was corn starch, glucose, soybean flour, ammonium chloride, calcium carbonate and magnesium chloride, respectively. Plackett-Burman method of statistical anslysis system（SAS） was applied to screen the six factors which affected fermentation production of wuyiencin. By using the Plackett-Burman method, the concentrations of glucose and calcium carbonate were found to significantly influence the production of wuyiencin. Then, these two factors were optimized using central composite design of ＂response surface methodology＂. The quadratic regression equation was Y=218.18＋270.28X1＋1274.74X2―3.28X12―5.4X1X2―200.98X22,which was studied on optimal levels of two major factors and their interactions. Finally, the optimized fermentation medium was composed as follows（g/L）： corn starch 20, glucose 39, soybean flour 20, smmonium chloride 4, calcium carbonate 2.65 and magnesium chloride 0.4. The verifiable titer was 7215 μg/m L in the medium with shake-flask culture at 28 ℃ and 220 r/min for 60 h, increased 26.5% over the original medium of wuyiencin.

关 键 词：武夷菌素 基因工程菌 发酵培养基 响应面法 

分 类 号：S476[农业科学—农业昆虫与害虫防治]