拮抗农业致病菌放线菌G5发酵条件的优化

刘杨，王勇（等）

摘要：微生物的发酵过程十分复杂，培养基配方的组成以及培养条件的变化都会对菌体的代谢造成很大的影响。为了使抗生素的产量达到最大化，试验运用了Plackett-Burman试验设计、最陡爬坡试验、Box-Behnken试验设计对放线菌（Actinomycetes）G5进行了发酵条件优化。利用Plackett-Burman试验设计对影响放线菌G5发酵的诸多相关因素进行评估并筛选出3个主要因素，即碳源含量、培养温度和培养时间；之后根据3个主要因素的作用大小和方向进行了最陡爬坡试验；最后运用Box-Behnken试验设计确定最优值，得出其最优培养条件为淀粉含量21.66 g/L、培养温度27.70 ℃、培养时间7.75 d。在此条件下，抗生素对苹果轮纹病菌的抑菌圈直径为23.00 mm，与模型预测值基本一致。

关键词：放线菌（Actinomycetes）G5；Plackett-Burman试验设计；最陡爬坡试验；Box-Behnken试验设计；发酵条件优化

中图分类号：TQ458 文献标识码：A 文章编号：0439-8114（2014）07-1634-03

Optimization of Fermentation Conditions for Actinomycetes G5

LIU Yang， WANG Yong， KANG Jing-qin， ZHANG Liang

（University of Science and Technology Liaoning， Anshan 114044， Liaoning，China）

Abstract：Microbial fermentation process were very complex due to effects of culture medium composition and culture conditions on cell metabolism. Fermentation conditions for actinomycetes G5 were optimized with Plackett-Burman experimental design， steepest ascent experiments， and Box-Behnken design. Three main factors affecting the fermentation of actinomycetes G5 screened by Plackeet-Burman design were carbon content， temperature and culture days. Then the steepest ascent experiments were conducted based on the action degree and direction of the three main factors. The optimal conditions were finally determined by Box-Behnken design. The results showed that the optimal fermentation condition for actinomycetes G5 were starch content of 21.66 g/L， culture temperature of 27.70 ℃ and culture days of 7.75 d. Under these conditions， the bacterial inhibition diameter of the acquired antibiotics against physalospora piricola was 23.00 mm， equal to the model prediction.

Key words： Actinomycetes G5； Plackett-Burman design； steepest ascent experiment； Box-Behnken； optimization

放线菌（Actinomycetes）G5的次级代谢产物抗生素对苹果轮纹病菌（Physalospora piricola）、黄瓜炭疽病菌（Colletotrichum orbiculare）等多种农业致病菌有较强的抑制作用[1]。在对放线菌G5的发酵过程中，液体培养基的配方和培养条件对抗生素的产量都有明显的影响[2]，因而发酵条件的优化工作就显得尤为重要。本试验运用响应面法对放线菌G5的发酵条件进行全面优化，以确保抗生素的产量达到最大。响应面法是利用简洁精确的试验设计，采用多元二次回归方程拟合因素和响应值之间的函数关系，通过对回归方程的分析来寻求最优条件参数，解决多变量问题的统计方法[3]。本试验运用响应面法中的Plackett-Burman设计、最陡爬坡试验、Box-Behnken设计来优化放线菌G5的培养基配方和培养条件。……