loading page

Fluorescent Labeling of Dicysteine-Tagged Peptide for Monitoring and Optimization of Protein Bio-production in Bacteria
  • +11
  • Shiyue Zheng,
  • Wenming Shao,
  • Sheng Lu,
  • Yahui Chen,
  • Lupeng Cui,
  • Long Jiang,
  • Hao Gu,
  • Yuanbo Wang,
  • Junqian Peng,
  • Yuying Wang,
  • Xin Yan,
  • Fang Wang,
  • Bin Wu,
  • Xiaoqiang Chen
Shiyue Zheng
Nanjing Tech University
Author Profile
Wenming Shao
Nanjing Tech University
Author Profile
Sheng Lu
Nanjing Tech University
Author Profile
Yahui Chen
Nanjing Tech University
Author Profile
Lupeng Cui
Nanjing Tech University
Author Profile
Long Jiang
Nanjing Tech University
Author Profile
Hao Gu
Nanjing Tech University
Author Profile
Yuanbo Wang
Nanjing Tech University
Author Profile
Junqian Peng
Nanjing Tech University
Author Profile
Yuying Wang
Nanjing Agricultural University
Author Profile
Xin Yan
Nanjing Agricultural University
Author Profile
Fang Wang
Nanjing Tech University
Author Profile
Bin Wu
Nanjing Tech University
Author Profile
Xiaoqiang Chen
Nanjing Tech University (formerly Nanjing University of Technology)
Author Profile

Abstract

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-Micheal 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 E. 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.
15 Aug 2022Submitted to AIChE Journal
16 Aug 2022Submission Checks Completed
16 Aug 2022Assigned to Editor
17 Aug 2022Reviewer(s) Assigned
24 Aug 2022Editorial Decision: Revise Minor
02 Sep 20221st Revision Received
02 Sep 2022Submission Checks Completed
02 Sep 2022Assigned to Editor
06 Sep 2022Reviewer(s) Assigned
13 Sep 2022Editorial Decision: Accept