基于抑菌活性的纳他霉素微孔板生物检测法

为快速测定大量纳他霉素样品含量,基于其抑菌活性建立了一种新型的微孔板生物检测法。结果表明,裂殖酵母AS2.637为最适敏感指示菌,优化的敏感指示菌初始浓度为105～106cfu/mL,纳他霉素样品加入时间为0 h,培养时间为8 h。微孔板中纳他霉素浓度在0.95～2.14 mg/L与抑菌率呈较显著的线性关系,微孔板生物检测法与HPLC法测定的结果无显著差异,该方法具有处理量大、操作简单、平行性好等特点,可用于发酵过程中纳他霉素产量的定量和突变菌的高通量快速筛选工作。

A Microplate Bioassay to Determine Natamycin Based on Antimicrobial Activity

Based on the antimicrobial activity of natamycin,a novel microplate bioassay was established for rapid quantification of natamycin production from large numbers of fermentation samples.The results showed that Schizosaccharomyces octosporus AS2.637 was the optimum test organism,the optimal concentration of sensitive microorganism was 105～106 cfu/mL,the adding time of natamycin sample was 0h,the incubation time was 8h.The natamycin concentration in microplate was significantly linear with the inhibition ratio in the range of 0.95～2.14 mg/L.The results by microplate bioassay and HPLC method showed no considerable difference.It indicates that the microplate bioassay can be used for determination of natamycin in fermentation process and rapidly screening large numbers of mutants because of high throughput,simple operation and good parallelism.