Biology subjects

Drepper, T.

Publications and source records attributed to Drepper, T..

3 recordsLinked to original sources

bioRxiv2026

A multichromatic UV-RGB optogenetic toolbox for control of gene expression in Pseudomonas putida

Optogenetics uses light to provide precise, reversible, and non-invasive control over bacterial functions including gene expression with high spatiotemporal resolution. Although many optogenetic systems have been developed for Escherichia coli, only a limited number is available for other prokaryotes, such as pseudomonads. Here, we establish a toolbox of genomically integrated optogenetic gene cassettes for light-responsive regulation of target gene expression in Pseudomonas putida with UV-A, blue, green, and red light. Using transposon Tn7-mediated chromosomal integration, we implemented four optogenetic systems: the photocaged IPTG/Ptac-LacI system, the LOV-based Dusk switch, the cyanobacteriochrome system CcaS/R, and different bacteriophytochrome-based REDusk variants. Benchmarking with the mCherry reporter demonstrated high dynamic ranges of up to [~]270-fold, low basal expression, and largely homogeneous population responses in P. putida. Spatial illumination further enabled patterned single- and dual-color gene expression. As a proof of concept, we applied the toolbox for light-controlled regulation of pyoverdine (PVD) biosynthesis in P. putida. The expression of the alternative sigma factor PfrI, which upregulates the production of the siderophore during iron-limitation, was placed under optogenetic control in a {Delta}pfrI background. The red-light responsive switches resulted in the strongest induction of PVD synthesis and enabled spatial control of siderophore-mediated microbial interactions. To demonstrate transferability, light-dependent pyoverdine production was further established in the human pathogen Pseudomonas aeruginosa PAO1. Together, this optogenetic plug-and-play toolbox enables non-invasive, spatiotemporal reprogramming of gene expression and cellular processes in pseudomonads and expands the available optogenetic repertoire beyond established model organisms.

synthetic biology↗

bioRxiv2026

Context-dependent siderophore exploitability shapes microbial community structure

1)Siderophores are classically viewed as shared iron-scavenging public goods, yet their ecological roles in multispecies communities remain poorly defined. Here, we establish a synthetic microbial community to dissect how different siderophores, their uptake compatibility and spatial structure shape iron competition. Using Corynebacterium glutamicum as a model, we show that this siderophore non-producer accesses diverse xenosiderophores, including enterobactin secreted by Escherichia coli. However, exploitation was constrained and co-cultures converged to stable compositions. Dose-response experiments combined with mathematical modelling indicated that the producer retains more effective access to enterobactin than the exploiter. Presence of Pseudomonas putida altered this interaction, as it exploited enterobactin while producing pyoverdine, a siderophore inaccessible to the other community members that restricted their iron access. Across different cultivation scales, community dynamics was strongly influenced by spatial organization and initial composition. These findings identify siderophores as context-dependent iron-allocation agents that can promote microbial coexistence or exclusion.

microbiology↗

bioRxiv2025

A novel biosensor for ferrous iron developed via CoBiSe:A computational method for rapid biosensor design

Genetically encoded biosensors enable monitoring of metabolite dynamics in living organisms. We present CoBiSe, a computational approach using Constraint Network Analysis to identify optimal insertion sites for reporter modules in molecular recognition elements (MREs). Applied to the iron-binding protein DtxR from Corynebacterium glutamicum, CoBiSe identified a flexible connective loop (residues 138-150) for inserting the reporter module, resulting in IronSenseR, a novel ratiometric biosensor for ferrous iron (Fe{superscript 2}). IronSenseR demonstrates high specificity for Fe{superscript 2} with dissociation constants of 1.55 {+/-} 0.08 {micro}M (FeSO4) and 2.44 {+/-} 0.28 {micro}M (FeCl2), while showing no binding to Fe3 and other divalent cations. In vivo assessment in Escherichia coli, Pseudomonas putida and Corynebacterium glutamicum confirmed IronSenseRs capability to detect changes in the intracellular iron pool. The creation of IronSenseR underlines that, by reducing search space and eliminating labor-intensive screening, CoBiSe streamlines biosensor development and enables precise creation of next-generation biosensors for diverse metabolites.

bioengineering↗