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Fluorescent Labeling of Dicysteine-Tagged Peptide for Monitoring and Optimization of Protein Bio-production in Bacteria
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  • 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

Corresponding Author:473167846@qq.com

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Wenming Shao
Nanjing Tech University
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Sheng Lu
Nanjing Tech University
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Yahui Chen
Nanjing Tech University
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Lupeng Cui
Nanjing Tech University
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Long Jiang
Nanjing Tech University
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Hao Gu
Nanjing Tech University
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Yuanbo Wang
Nanjing Tech University
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Junqian Peng
Nanjing Tech University
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Yuying Wang
Nanjing Agricultural University
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Xin Yan
Nanjing Agricultural University
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Fang Wang
Nanjing Tech University
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Bin Wu
Nanjing Tech University
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Xiaoqiang Chen
Nanjing Tech University (formerly Nanjing University of Technology)
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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
Dec 2022Published in AIChE Journal volume 68 issue 12. 10.1002/aic.17912