文献类型：期刊文献

英文题名：High-throughput system for screening of high L-lactic acid-productivity strains in deep-well microtiter plates

作者：Lv, Xiangyun[1];Song, Jiali[1];Yu, Bo[1];Liu, Huilan[1];Li, Chao[1,2,3];Zhuang, Yingping[1,2,3];Wang, Yonghong[1,2,3]

机构：[1]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, 130 Meilong Rd,POB 329, Shanghai 200237, Peoples R China;[2]Shanghai Inst Biomfg Technol, 130 Meilong Rd,POB 329, Shanghai 200237, Peoples R China;[3]Collaborat Innovat Ctr, 130 Meilong Rd,POB 329, Shanghai 200237, Peoples R China

年份：2016

卷号：39

期号：11

起止页码：1737

外文期刊名：BIOPROCESS AND BIOSYSTEMS ENGINEERING

收录：;EI(收录号：20163102650281);WOS:【SCI-EXPANDED(收录号:WOS:000384354100010)】；

基金：This work was financially supported by grants from Major State Basic Research Development Program of China (2013CB733600), the National High Technology Research and Development Program (2015AA021005) and the National Natural Science Foundation of China (Grant No. 31200024).

语种：英文

外文关键词：High-throughput screening; 24-well MTPs; L-lactic acid; Bromocresol green; L-lactate oxidase assay

摘要：For strain improvement, robust and scalable high-throughput cultivation systems as well as simple and rapid high-throughput detection methods are crucial. However, most of the screening methods for lactic acid bacteria (LAB) strains were conducted in shake flasks and detected by high-performance liquid chromatography (HPLC), making the screening program laborious, time-consuming and costly. In this study, an integrated strategy for high-throughput screening of high l-lactic acid-productivity strains by Bacillus coagulans in deep-well microtiter plates (MTPs) was developed. The good agreement of fermentation results obtained in the MTPs platform with shake flasks confirmed that 24-well U-bottom MTPs could well alternate shake flasks for cell cultivation as a scale-down tool. The high-throughput pH indicator (bromocresol green) and l-lactate oxidase (LOD) assays were subsequently developed to qualitatively and quantitatively analyze l-lactic acid concentration. Together with the color halos method, the pH indicator assay and LOD assay, the newly developed three-step screening strategy has greatly accelerated the screening process for LAB strains with low cost. As a result, two high l-lactic acid-productivity mutants, IH6 and IIIB5, were successfully screened out, which presented, respectively, 42.75 and 46.10 % higher productivities than that of the parent strain in a 5-L bioreactor.