文献类型：期刊文献

英文题名：Penicillin G acylase purification with the aid of high-throughput screening approach

作者：Zhou, Jin[1];Wang, Yong-Hong[1];Chu, Ju[1];Gou, Bing-Quan[1];Zhuang, Ying-Ping[1];Zhang, Si-Liang[1];Yuan, Zhong-Yi[1]

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

年份：2008

卷号：39

期号：3

起止页码：195

外文期刊名：JOURNAL OF THE CHINESE INSTITUTE OF CHEMICAL ENGINEERS

收录：;EI(收录号：20085011775854);WOS:【SCI-EXPANDED(收录号:WOS:000255889400002)】；

语种：英文

外文关键词：96-well microtiter plate; high-throughput screening; penicillin G acylase; purification

摘要：Penicillin G acylase (PGA) is one of the most important enzymes for the production of semi-synthetic beta-lactam antibiotics and their key intermediates. Purification of penicillin G acylase from fermentation broth with the aid of high-throughput screening (HTS) process has been examined in this study. We used a microtiter-plate based on screening method to find appropriate purification conditions for the target protein. The screening method is based on a 96-well plate format, and different matrices and conditions (pH, salt concentration and type) were tested. Through analyses of all pooled fractions (flow-through and elution) we gained appropriate information to choose the best performing matrix and buffer conditions for upscaling. After an upscaled purification step the second unit operation is screened in the similar way and parameters for this operation can be chosen. The purification parameter of purified PGA at the small-screen and upscaling levels were measured, respectively. The results indicate that high-throughput progress based on a 96-well plate is a flexible and efficient paradigm for recombinant protein purification. (c) 2008 Published by Elsevier B.V. on behalf of Taiwan Institute of Chemical Engineers.