基于快速筛选的纳他霉素产生菌的微量培养

The Micro-cultivation of Natamycin-producing Strains for Rapid Screening

以纳他霉素产生菌——褐黄孢链霉菌(Streptomyces gilvosporeus)为研究对象,以96深孔板为载体,建立适合大规模菌株快速筛选的微量培养体系。首先用8层纱布外加有孔不锈钢盖子较好地解决了水分蒸发及交叉污染的问题;确定了深孔板装液量600μL、转速300r/min、振幅40mm的最佳培养条件,此时菌体生长和产物合成的变化趋势均与摇瓶培养过程非常类似。实验发现微量培养体系的最大板内和板间差异分别为1.93%和6.62%,通过统计学软件分析,两种不同培养体系下获得的菌株产量之间具有极显著的线性回归关系(F=39.303,P=0.00<0.01),产量分布的排序大致相同,这表明微孔板作为培养体系,具有标准化、平行化等优点,能很好地应用于大量菌株的快速筛选。

As natamycin-producing strain, Streptomyces giIvosporeus was studied to develop the micro-cultivation system in 96 deep-well microplates for rapid screening from a large number of isolates. Firstly, the stainless steel cover perforated with holes and dressed with eight gauzes was designed to solve water evaporation and cross-contamination in 96 deep-well microplates. The optimal conditions for micro-cultivation were as follows： culture volume 600 gL, shaking speed 300 r/min, shaking diameter 40 mm, and the pattern of microbial growth and natamycin biosynthesis with the above micro-cultivation mode was quite similar with that in shake flasks. It was found that the maximum intramicroplate and inter-microplate difference were 1.93% and 6.62%, respectively. Using the statistic software analysis, the productivity in 96 deep-well microplates were in good linear correlation with those in shake flasks （F = 39.303, P = 0.00 〈 0.01） and the rank of production distribution in two different cultivation systems was consistent. Therefore, the micro-cultivation in microplates was proved to be good standardization and parallelization, which was potential to be applied in rapid screening from a large number of strains.