响应面法优化发酵液中多拉菌素的脱色工艺

Study on decoloring process of doramectin in fermentation broth of Streptomyces avermitilis by response surface methodology

目的研究多拉菌素发酵提取液的脱色工艺,简化后续提取过程及提高多拉菌素成品的质量。方法用UV检测发酵液的脱色率,以HPLC检测多拉菌素的损失率,以脱色率和损失率考察活性炭对多拉菌素发酵提取液的脱色效果。先以单因素实验确定因素水平范围,然后再以响应面法优化脱色工艺参数(添加量,脱色温度及溶液pH)。结果在单因素实验基础上,确定脱色时间为40min后,通过响应面方法对其它脱色工艺参数加以优化,得到最佳工艺条件为:活性炭的添加量1.0%(W/V)、温度33℃、pH3,该条件下多拉菌素发酵提取液的脱色率为70.35%。验证试验中脱色率达到68.78%,无显著差异。结论该脱色工艺简单可靠,脱色效果好,适合于工业化生产。

Objective Response surface methodology was applied to study the decolorization of activated carbon for doramectin in fermentation broth of Streptomyces avermitilis. The aim was not only to simplify further extraction process but also to increase the quality of final product. Methods Based on decoloring rate by the detection of UV and loss of doramectin detected by HPLC, effect of activatied carbon to decoloration was investigated. The levels of factors were determined by single-factor experiments, and then optimal conditions were carried out by response surface methodology with additive amount of activated carbon, temperature and pH. Results The decoloring time was 40 min which was determined by the single-faetor experiments. The optimal technical conditions were as follows： 1.0% activated carbon（W/V）, temperature 33℃ and decoloration pH of 3. Under the optimal conditions, the decoloring rate was above 68.78% in further verified experiment and the difference was not significant with predicted value of 70.35%. Conclusion The method was effective, simple and reliable, also warranted for the commercial process.