Fluorescent labeling of dicysteine‐tagged peptide for monitoring and optimization of protein bio‐production in bacteria

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-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.

AIEE Active Azomethine-Based Rhodamine Derivative For Ultrasensitive Multichannel Detection of Au3+ Through a Fluorimetrically, Electrochemically, and RGB-Based Sensing Assay

“…In addition to their remarkable catalytic feature, researchers have shown a significant degree of interest in gold species owing to their high bioactivity at appropriate levels . Several disorders, such as rheumatoid arthritis, cancer, asthma, and HIV, have been commonly treated with gold-based medications. − Moreover, the deposition of larger amounts of gold species (particularly its ionic form Au 3+ ) may cause health risks in creatures like fish, water fleas, fairy shrimp, flukes, and green algae because of their potential interactions with biomolecules like enzymes and DNA. , There are multiple reports of 200 μM AuCl 3 ingestion causing harm to major human organs, including the kidney, liver, and central nervous system. − …”