当前位置: X-MOL 学术AlChE J. › 论文详情
Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)
Fluorescent labeling of dicysteine-tagged peptide for monitoring and optimization of protein bio-production in bacteria
AIChE Journal ( IF 3.7 ) Pub Date : 2022-09-19 , DOI: 10.1002/aic.17912
Shiyue Zheng 1 , Wenming Shao 2 , Sheng Lu 1 , Yahui Chen 1 , Lupeng Cui 2 , Long Jiang 1 , Hao Gu 1 , Yuanbo Wang 1 , Junqian Peng 1 , Yuying Wang 3 , Xin Yan 3 , Fang Wang 1 , Bin Wu 2 , Xiaoqiang Chen 1
Affiliation  

A rapid and convenient strategy to monitor the productivity of biomanufacturing is essential for the research in optimizing relevant bioprocesses. In this work, we have developed a fluorescein-derived probe (FL-DT) that reacts rapidly with thiol groups via 1,4-Michael addition reaction of the sulfhydryl to unsaturated ketone and releases fluorescence. FL-DT specifically forms fluorescent adduct with two adjacent thiols in a protein of interest (POI), making the probe a reliable tool for protein quantification. The production of xylanase fused with a short di-Cys tag was then successfully monitored and quantified with FL-DT in Escherichia coli system under different protein expression conditions, providing useful information for optimizing the bioprocess. Our work provides a convenient and efficient strategy for POI labeling and monitoring bioproduction.

中文翻译:

双半胱氨酸标记肽的荧光标记用于监测和优化细菌中的蛋白质生物生产

监测生物制造生产率的快速便捷策略对于优化相关生物过程的研究至关重要。在这项工作中,我们开发了一种荧光素衍生探针 (FL-DT),它通过巯基与不饱和酮的 1,4-迈克尔加成反应与硫醇基团快速反应并释放荧光。FL-DT 与感兴趣的蛋白质 (POI) 中的两个相邻硫醇特异性地形成荧光加合物,使探针成为蛋白质定量的可靠工具。然后用 FL-DT 在大肠杆菌中成功监测和量化与短 di-Cys 标签融合的木聚糖酶的产生系统在不同的蛋白质表达条件下,为优化生物过程提供有用的信息。我们的工作为 POI 标记和监测生物生产提供了一种方便有效的策略。
更新日期:2022-09-19
down
wechat
bug