# Expanding the Toolbox for Inducible Protein Expression With Automation‐enabled Generation of Glycomimetics

## TL;DR

Researchers developed new inducers for protein expression by adding fucose, allowing control through fucosidase activity and using automated synthesis methods.

## Contribution

The novel contribution is the design and automated synthesis of fucose-containing inducers that enable fucosidase-dependent protein expression.

## Key findings

- Automated continuous-flow processes were developed for synthesizing 2′F-IPTG and 2′F-IBCG.
- Fucosylated inducers enabled fucosidase-dependent expression of a reporter protein.

## Abstract

Inducible protein expression is a cornerstone of many aspects of industrial and molecular biotechnological processes. However, limited availability of inducible transcription factors can reduce our ability to control expression at a population level. The design and synthesis of a powerful inducer containing a fucose is demonstrated to induce protein expression through the lac operon only in cells with the ability to selectively de‐fucosylate them. Batch and automated continuous‐flow processes are reported for the syntheses of both 2′‐fucosyl isopropyl‐β‐D‐thiogalactopyranose (2′F‐IPTG) and isobutyl‐C‐galactoside (2′F‐IBCG) mimics. Fucosylation of the inducer allowed for fucosidase‐dependent expression of a reporter protein, providing an additional layer of control over inducible gene expression.

Inducible protein expression is a cornerstone of many aspects of industrial and molecular biotechnological processes. 2′‐fucosyl isopropyl‐β‐D‐thiogalactopyranose (2′F‐IPTG) and 2′‐fucosyl isobutyl‐C‐galactoside (2′F‐IBCG) mimics were explored for protein expression studies. Herein, we report the automated continuous flow syntheses of the mimics and their utility in protein expression.