From 934f8539993b52db777372d31bec99b7214abb5d Mon Sep 17 00:00:00 2001 From: Sabrina Zander <sabrina.zander@uni-duesseldorf.de> Date: Tue, 18 Mar 2025 12:34:41 +0000 Subject: [PATCH] Edit README.md --- README.md | 1 - 1 file changed, 1 deletion(-) diff --git a/README.md b/README.md index ef69675..536afab 100644 --- a/README.md +++ b/README.md @@ -4,7 +4,6 @@ The iron-containing porphyrin heme is of high interest for the food industry for the production of artificial meat as well as for medical applications. Recently, the biotechnological platform strain Corynebacterium glutamicum has emerged as a promising host for animal-free heme production. Beyond engineering of complex heme biosynthetic pathways, improving heme export offers significant yet untapped potential for enhancing production strains. In this study, a growth-coupled biosensor was designed to impose a selection pressure on the increased expression of the hrtBA operon encoding an ABC-type heme exporter in C. glutamicum. For this purpose, the promoter region of the growth-regulating genes pfkA (phosphofructokinase) and aceE (pyruvate dehydrogenase) was replaced with that of PhrtB, creating biosensor strains with a selection pressure for hrtBA activation. Resulting sensor strains were used for plate-based selections and for a repetitive batch f(luorescent)ALE using a fully automated laboratory platform. Genome sequencing of isolated clones featuring increased hrtBA expression revealed three distinct mutational hotspots: (i) chrS, (ii) chrA, and (iii) cydD. Mutations in the genes of the ChrSA two-component system, which regulates hrtBA in response to heme levels, were identified as a promising target to enhance export activity. Furthermore, causal mutations within cydD, encoding an ABC-transporter essential for cytochrome bd oxidase assembly, were confirmed by the construction of a deletion mutant. Reversely engineered strains showed strongly increased hrtBA expression as well as increased cellular heme levels. These results further support the proposed role of CydDC as a heme transporter in bacteria. Mutations identified in this study therefore underline the potential of biosensor-based growth coupling and provide promising engineering targets to improve microbial heme production. ## ARC Citation - Krüger, A., Göddecke, J., Osthege, M., Navratil, L., Weber, U., Oldiges, M., & Frunzke, J. (2024). Biosensor-based growth-coupling as an evolutionary strategy towards elevated hrtB expression in Corynebacterium glutamicum. DataPLANT. [](https://doi.org/10.60534/wr9ps-fmv73) -- GitLab