酶标仪法测定微孔板发酵液中林可霉素

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英文篇名：Determination of lincomycin in the microplate fermentation broth by microplate reader 作者：余飞 ; 秦艳飞 ; 王洲 ; 孙俊峰 ; 张敏 ; 薛正莲 英文作者：Yu Fei;Qin Yan-fei;Wang Zhou;Sun Jun-feng;Zhang Min;Xue Zheng-lian;Institute of Biologic & Chemical Engineering of Anhui Polytechnic University,Microorganism Fermentation Engineering and Technology Research Center of Anhui Province;Xinyu Pharmaceutical Co.,Lid; 关键词：林可霉素 ; 测定 ; 分光光度计 ; 酶标仪 ; 优化 英文关键词：Lincomycin;;Determination;;Spectrophotometer;;Microplate reader;;Optimization 中文刊名：中国抗生素杂志 英文刊名：Chinese Journal of Antibiotics 机构：安徽工程大学生物与化学工程学院微生物发酵安徽省工程技术研究中心;新宇药业股份有限公司; 出版日期：2019-02-26 15:41 出版单位：中国抗生素杂志 年：2019 期：01 基金：安徽省科技攻关项目(No.07010302177) 语种：中文; 页：51-55 页数：5 CN：51-1126/R ISSN：1001-8689 分类号：O657.3;TQ927

摘要

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。
        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 0.02 mol/L palladium chloride solution was 150μL in the reaction system for the determination of lincomycin, and the appropriate concentration of hydrochloric acid was 1 mol/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 lincolnensis.

引文

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