青霉素高产菌株高通量检测和筛选方法的研究

A high-throughput penicillin titer detecting method and its application in high-throughput screening of penicillin-producing strains

目的建立高通量筛选青霉素高产菌株的方法。方法采用青霉素酶水解青霉素形成青霉噻唑酸,产物与碘液反应,以淀粉为指示剂,青霉素含量与碘液用量呈现良好的线性关系。结果在室温条件下,不超过10min,可并行检测多个发酵液样品的青霉素含量,该方法简便、迅速、微量并且不受发酵液中其他杂质成分的干扰,也可在10min内对超过200个发酵液样品中的青霉素含量进行相对评估,淘汰90%以上的低效价菌株。结论提供了高通量筛选青霉素高产菌株的方法。

Objective To estabolish a rapid penicillin titer detecting method to facilitate the high-throughput screening of penicillin-producing strains. Methods Penicillinase was employed to hydrolysis penicillin to peniciUanic acid, which would react with iodine. Results The penicillin titer can be quantified within 10min at room temperature regardless of other by-products in the fermentation broth. In addition, and more than 200 samples can be simultaneously assessed by one operator within 10min. Conclusion This novel method provide a platform for high- throughput screening penicillin high producing strains.