文献类型：期刊文献

英文题名：High-Throughput System for Screening of Cephalosporin C High-Yield Strain by 48-Deep-Well Microtiter Plates

作者：Tan, Jun[1];Chu, Ju[1];Hao, Yuyou[1];Guo, Yuanxin[1];Zhuang, Yingping[1];Zhang, Siliang[1]

机构：[1]E China Univ Sci & Technol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China

年份：2013

卷号：169

期号：5

起止页码：1683

外文期刊名：APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY

收录：;EI(收录号：20140617268028);WOS:【SCI-EXPANDED(收录号:WOS:000316116500017)】；

基金：This work was financially supported by a grant from the Major State Basic Research Development Program of China (973 Program), No. 2012CB721006, the National High Technology Research and Development Program of China (863 Program), No. 2006AA020302, National Major Scientific Technological Special Project No. 2012YQ 15008709, and National High Technology Research and Development Program of China (863 Program), No. 2012AA021201

语种：英文

外文关键词：Cephalosporin C; High-throughput screening; High-throughput bioactivity assay; 48-deep MTP; Mixture culture

摘要：Improvement of microbial strains for the high-production of industrial products has been the hallmark of all commercial fermentation processes. Strain improvement has been conventionally achieved through mutation and selection. However, most of the screenings were performed in shake flasks, which made the screening procedure very complex, time-consuming, and inefficient. Most mutant spore suspension had no chance to be screened due to the low-throughput of shake flasks and had to be sacrificed. In this paper, in order to get a Cephalosporin C (CPC) high-yield stain, traditional mutagenesis was employed to obtain the mutant library and gave them the equal screening chance by a novel mixture culture method combined with high-throughput screening method. The good correlation of fermentation results between differing-scale cultivations confirmed the feasibility of utilizing the 48-deep microtiter plates as a scale-down tool instead of shake flasks for culturing high-aerobic microbes with long cultivation period. The microbioassay based on the antibacterial activity of CPC against Alcaligenes faecalis was used to select mutants. As a result, the high-yield strain W-6 was successfully screened out and the CPC titer was nearly 50 % higher than that of the parental strain in the shake flask. The CPC production of strain W-6 was further validated in 50 l bioreactor, and the CPC production reached 32.0 g/l, twofold higher than that of the wild strain.