洋葱伯克霍尔德氏菌Lu10-1产生抗菌活性物质的发酵培养基和发酵条件优化  

Optimization of Medium Components and Cultural Conditions for Producing Antibacterial Substances FromBurkholderia cepacia Lu10-1

作　　者：查传勇[1] 董法宝[1] 杨悦[1] 冀宪领[1] 牟志美[1] 

机构地区：[1]山东农业大学林学院蚕学系,山东泰安271018

出　　处：《蚕业科学》2009年第2期229-235,共7页ACTA SERICOLOGICA SINICA

基　　金：山东省自然科学基金项目(编号2007ZRB01872)

摘　　要：洋葱伯克霍尔德氏菌(Burkholderia cepacia)Lu10-1是从桑叶中分离得到的一株具有抗菌及促进植物生长等多种生物学功能的内生细菌。利用基于统计学的响应面法(response surface methodology,RSM)对影响该菌产生抗细菌活性物质的发酵培养基组成和发酵培养条件进行了优化。部分重复因子试验表明,酵母浸粉和氯化钠是培养基组分中的主要影响因子,其中酵母浸粉为正效应,氯化钠为负影响;结合最陡爬坡路径逼近最大响应区域和中心组合设计及响应面分析,确定了培养基中主要配方的最佳质量浓度为蔗糖17.0 g/L、酵母浸粉5.855 g/L、氯化钠4.519 g/L、磷酸二氢钾0.2 g/L。通过PB(plackeet-burman)试验发现接种量和发酵温度是该菌株产生抗菌活性物质发酵条件中的主要影响因子,经中心组合设计法优化的最佳发酵条件为:接种量0.027 7 mL/mL,摇瓶装液量100 mL,发酵温度30.29℃,培养基初始pH6.2,培养时间42 h。Burkholderia cepacia Lu10-1 is an endogenous bacterium isolated from mulberry leaves. It produces antibacterial substances and other plant growth regulators which have various biological functions. In present study, a statistics-based experimental design method, termed response surface methodology （RSM）, was employed to optimize the medium components and cultural conditions for producing antibacterial substance from Burkholderia cepacia Lu10-1. In the optimization of medium, the influences of sucrose, yeast extract powder, NaCl and KH2 PO4 on active substance production were first evaluated using a fractional facto- rial design. The results showed that the components of yeast extract powder and NaCl had significant effect on active substance production. The yeast extract powder content had a positive influence on active substance production, while the NaCI content affected active substance production negatively. The optimal region of the medium composition was approached using the path of steepest ascent method. The optimal content of medium components was determined by central composite design and response surface analysis. The components of optimum medium for antibacterial active substance production were as follows： sucrose 17 g/L, yeast extract powder 5.855 g/L, NaCl 4.519 g/L, and KH2PO4 0.2 g/L. The Plackeet-Burman design was used to optimize the fermentation condition. The results indicated that inoculation volume and fermentation time were the most significant factors, which optimal conditions were determined by central composite design and response surface analysis. The optimized fermentation conditions for antibacterial active substance production were as follows： inoculation volume 0.027 7 mL/mL, medium volume 100 mL, culture temperature 30.29 ℃, initial pH 6. 2, and fermentation time 42 h.

关 键 词：桑叶 内生细菌 洋葱伯克霍尔德氏菌Lu10-1菌株 抗细菌活性物质 响应面法 培养基 培养条件 

分 类 号：S476.1[农业科学—农业昆虫与害虫防治]