Synthesis and Application of Photocaged Isopropyl β-D-1-Thiogalactopyranoside for Light-Mediated Control of Bacterial Gene Expression.

Photocaged compounds are chemical conjugates that are designed to release an active molecule upon exposure to light of a specific wavelength. In recent years, photocaged inducer molecules such as caged isopropyl β-D-1-thiogalactopyranoside (cIPTG) have been increasingly used as a powerful tool for light-driven gene expression in bacteria, allowing researchers to precisely and noninvasively tune the expression of specific target genes. In this chapter, we present a guideline for the synthesis of 6-nitropiperonyl photocaged IPTG (NP-cIPTG) as well as its in vivo application as an optochemical on-switch of gene transcription in Escherichia coli and other bacteria.

Analysis of protein secretion in Bacillus subtilis by combining a secretion stress biosensor strain with an in vivo split GFP assay.

Background: Bacillus subtilis is one of the workhorses in industrial biotechnology and well known for its secretion potential. Efficient secretion of recombinant proteins still requires extensive optimization campaigns and screening with activity-based methods. However, not every protein can be detected by activity-based screening.

Exploring engineered vesiculation by Pseudomonas putida KT2440 for natural product biosynthesis.

Pseudomonas species have become promising cell factories for the production of natural products due to their inherent robustness. Although these bacteria have naturally evolved strategies to cope with different kinds of stress, many biotechnological applications benefit from engineering of optimised chassis strains with specially adapted tolerance traits. Here, we explored the formation of outer membrane vesicles (OMV) of Pseudomonas putida KT2440.

Enabling oxygen-controlled microfluidic cultures for spatiotemporal microbial single-cell analysis.

Microfluidic cultivation devices that facilitate O control enable unique studies of the complex interplay between environmental O availability and microbial physiology at the single-cell level. Therefore, microbial single-cell analysis based on time-lapse microscopy is typically used to resolve microbial behavior at the single-cell level with spatiotemporal resolution. Time-lapse imaging then provides large image-data stacks that can be efficiently analyzed by deep learning analysis techniques, providing new insights into microbiology.

The modular pYT vector series employed for chromosomal gene integration and expression to produce carbazoles and glycolipids in .

The expression of biosynthetic genes in bacterial hosts can enable access to high-value compounds, for which appropriate molecular genetic tools are essential. Therefore, we developed a toolbox of modular vectors, which facilitate chromosomal gene integration and expression in KT2440. To this end, we designed an integrative sequence, allowing customisation regarding the modes of integration (random, at Tn7, or into the 16S rRNA gene), promoters, antibiotic resistance markers as well as fluorescent proteins and enzymes as transcription reporters.