Biology subjects

Jensen, S. I.

Publications and source records attributed to Jensen, S. I..

3 recordsLinked to original sources

bioRxiv2025

A Robust Isotope Ratio LC-MS/MS Workflow for High-Throughput Metabolic Profiling of Bacteria

Stable isotope dilution mass spectrometry (IDMS) has become a cornerstone of quantitative metabolomics, enabling accurate intracellular metabolite quantification across a range of biological systems. However, the broader adoption of IDMS in high-throughput studies remains limited by the high costs of commercially available 13C-labeled internal standards (IS), labor-intensive in-house IS production, and the narrow applicability of existing methods to different organisms. Here, we present a robust and scalable IDMS-based LC-MS/MS workflow for high-throughput metabolic profiling of diverse bacteria. The analytical method couples ion-pairing liquid chromatography with multiple reaction monitoring (MRM) to quantify 96 intracellular metabolites in under 16 minutes, with an average RSD of 21%. We developed a protocol for large-scale production of high-quality 13C-labeled IS, considerably lowering the cost for high-throughput IDMS studies. We then applied the workflow to 5 bacterial species in different cultivation conditions. This work provides a versatile platform for microbial metabolomics, supporting systems biology and data-driven metabolic engineering at scale. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=98 SRC="FIGDIR/small/669109v2_ufig1.gif" ALT="Figure 1"> View larger version (21K): org.highwire.dtl.DTLVardef@1f3e9e9org.highwire.dtl.DTLVardef@81eb0eorg.highwire.dtl.DTLVardef@16efe09org.highwire.dtl.DTLVardef@1e83a06_HPS_FORMAT_FIGEXP M_FIG C_FIG

systems biology↗

bioRxiv2024

Nanomotif: Identification and Exploitation of DNA Methylation Motifs in Metagenomes using Oxford Nanopore Sequencing

DNA methylation is found across all domains of life but is a rarely used feature in recovery of metagenome-assembled genomes (MAGs). Recently, Oxford Nanopore introduced all context methylation detection models. We leveraged this to develop Nanomotif, which identifies and exploits methylation motifs for enhanced MAG recovery. We demonstrate how Nanomotif enables database-independent contamination removal from high-quality MAGs and host association of plasmids directly from Nanopore sequencing data in complex metagenomes.

bioinformatics↗

bioRxiv2022

From empirical to data-driven host selection: a broad-host-range expression platform to facilitate chassis screening

Nature has provided a vast landscape of organisms through evolution, each with unique phenotypic traits adapted to varying environments. Nevertheless, host selection in biotechnological research is exceedingly dominated by empirical preference, where the endogenous physiology of the selected host is often not suited to the desired application. Considering that large parts of cellular regulation and metabolism remain obscure, empirical selection of a preferred model organism may lead to undue caveats in further engineering attempts, arising from intrinsic metabolism. One reason for the empirical host selection is the lack of engineering tools for screening novel organisms. In this study, we provide a modular, single vector-based expression platform, compatible with a wide range of prokaryotes. It centers around a tight and titratable promoter system, inducible by anhydrotetracyclin within an 84-fold dynamic range. It enables easy screening of recombinant proteins and pathways in both mesophilic and thermophilic Gram-negative and Gram-positive hosts. Overall, this platform enables simple screening of heterologous expression and production in a variety of hosts, including the exploration of previously unconsidered hosts thereby aiding the transition from empirical to data-driven host selection.

synthetic biology↗