酶标仪法测定微孔板发酵液中林可霉素

目的 建立酶标仪法测定微孔板发酵液中林可霉素的含量。方法 对酶标仪法的主要影响因素进行优化，并验证此 方法的精密度；在此基础上，分别利用酶标仪法与分光光度计法测定 25 株突变株经 48 孔板发酵后林可霉素的含量并进行曲线 拟合，以验证两种方法的相关性。结果 确定了在测定林可霉素含量的适宜反应体系中 0.02mol/L 氯化钯溶液为 150μL，盐酸添 加浓度为 1mol/L，且该方法精密度良好；拟合曲线相关系数 R2 =0.9551，显示两种方法具有很好的相关性。结论 该实验结果表 明酶标仪法能准确、快速地测定微孔板发酵液中林可霉素的含量，为后续高通量选育林可霉素高产菌株奠定了基础。

Determination of lincomycin in the microplate fermentation broth by microplate reader

Objective To determine the content of lincomycin in the microplate fermentation broth by microplate reader. Methods The main influencing factors of the method were optimized and the precision test was implemented simultaneously. In order to verify the feasibility of this method, the concentration of lincomycin in 25 kinds of mutant strains after fermentation in 48-well plates were respectively measured by spectrophotometer and microplate reader. Results It was determined that the appropriate volume of 0.02mol/L palladium chloride solution was 150μL in the reaction system for the determination of lincomycin, and the appropriate concentration of hydrochloric acid was 1mol/L. Under this condition, the concentration of lincomycin could be detected by microplate reader. The data regression showed a strong correlation between these two methods, and the correlation coefficient was 0.9551. Conclusion The experimental results showed that the microplate reader could accurately, quickly, and easily determine the content of lincomycin in the microplate fermentation broth. It also laid the foundation for the highthroughput selecting and breeding of high-yield lincomycin from Streptomyces