Construction of High Throughput Screening Platform and Its Applications
作者单位：华东理工大学
学位级别：硕士
导师姓名：储炬
授予年度：2014年
学科分类：0710[理学-生物学] 1007[医学-药学(可授医学、理学学位)] 100705[医学-微生物与生化药学] 07[理学] 071005[理学-微生物学] 10[医学]
摘 要：The aim of this study is to establish an efficient platform for high throughput screening and apply it to the increasing the yield of cephalosporin C （CPC）, which was one of β-lactam antibiotics, through nitrogen source optimization as well as preliminary exploration on high throughput screening of high-yield isoleucine producer strains.. In this study, the following problems are to be resolved.（1） Selecting the type of microtiter plates to study. The types of mirotiter plates often used were24-well,48-well,96-well microtiter plates. Microtiter plates can be chosen based on the oxygen transfer coefficient （KLa） and liquid evaporation （%）.（2） Optimizing the filling volume of microtiter plates and testing the feasibility of microtiter plates for cultivation and for turbidimetric assay.（3） Investigating of parallelity among wells in24-well and48-well microtiter plates.（4） Optimizing cultural conditions such as inoculum age, inoculum volume and fermentation time. In addition, the effect of different concentration starch on production of CPC in24-well microtiter plates was studied.（5） Establishing the high throughput methods for optimization medium, especially nitrogen source.（6） Use of Genipin color reaction with the amino acid L-isoleucine for high throughput screening of high-yield isoleucine producer strains. Through the study, the platform for high throughput screening was successfully established. It was found that24-well microtiter plates and48-well microtiter plates were more appropriate for cultivate aerobic microorganism than widely used96-well microtiter plates. There were higher oxygen transfer coefficient and larger filling volume enough for analyzing of the broth after fermentation. In addition, the water evaporation ratio in24-well is smaller than in48-well and the water evaporation ratio in48-well is smaller than in96-well microtiter plates. Hence,24-well and48-well microtiter plates were suggested for fermentation of aerobic microorganism-Cephalosp
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