文献类型：期刊文献

中文题名：紫外分光光度法快速测定庆大霉素C_(1a)含量在高通量筛选中的应用

英文题名：UV spectrophotometric determination of the concentration of gentamicin C_(1a) in the application of high throughput screening

作者：田江涛[1];李敏超[1];杭海峰[1];郭美锦[1];储炬[1];庄英萍[1]

机构：[1]华东理工大学生物反应器工程国家重点实验室,上海200237

年份：2019

卷号：44

期号：1

起止页码：59

中文期刊名：中国抗生素杂志

外文期刊名：Chinese Journal of Antibiotics

收录：CSTPCD;;Scopus;CSCD:【CSCD2019_2020】；

基金：科技部国家重大科学仪器设备开发专项(No.2012YQ15008706)

语种：中文

中文关键词：庆大霉素C1a;分光光度法;ARTP;高通量筛选

外文关键词：Gentamicin C1a;Spectrophotometry method;ARTP;High throughput screening

摘要：目的探索庆大霉素C_(1a)含量的快速检测方法,实现诱变菌株的高通量筛选。方法以紫外分光光度法作为检测庆大霉素C_(1a)含量的手段,对庆大霉素C_(1a)单组分生产菌株进行ARTP、LiCl、微波和ARTP+LiCl诱变,通过高通量筛选获得高产菌株,并在5L发酵罐中对高产稳定性菌株进行验证。结果紫外分光光度法和高效液相色谱法对发酵液中庆大霉素C_(1a)含量进行测定,最大相对误差为3.98%;筛选获得了10株高产菌株,其中AL324菌株与出发菌株相比,摇瓶效价提高了52%;通过稳定性传代实验,获得了4株高产稳定性菌株。5L发酵罐验证实验表明AL324与出发菌株相比效价提高了81.3%。结论紫外分光光度法可以快速检测发酵液中庆大霉素C_(1a)含量,该方法应用于高通量筛选中,获得了高产菌株,筛选结果表明ARTP+LiCl复合诱变方法的正突变率较高,是一种有效的庆大霉素C_(1a)高产菌株诱变方式。
Objective To explore the rapid detection method for the content of gentamicin C1a during high throughput screening.Methods In this paper,a rapid and simple detection method for GM C1a in the broth was established and integrated into the high throughput screening model.Four different mutation methods of atmospheric room temperature plasma (ARTP),microwave,LiCl,and ARTP+LiCl with above mentioned high throughput cultivating and detecting systems were used.Furthermore,the high yield strain AL324 was tested in the 5L fermentation.Results Comparing to the results of high performance liquid chromatography (HPLC)detection,a maximum deviation of 3.98% was found in this rapid detection method.Among them,a mutant of AL324 could enhance the titer by 52% in the shake flask,compared to the parent strain.After the following rescreening and genetic stability verification processes,only 4 mutants still had better fermentation performances.Moreover,the titer improvement further reached to 81.3% in the 5 L fermentation.Conclusion This rapid detection method can be used to detect the GM C1a titer and integrated into the high throughput screening model.The screening results illuminate the ARTP+LiCl is an effective method for producing high yield strains of GM C1a.