Enhanced catabolism of glycine betaine and derivatives provides improved osmotic stress protection in Methylorubrum extorquens PA1.

Autor:	Bruger EL; Department of Plant and Microbial Biology, University of Minnesota-Twin Cities, St. Paul, Minnesota, USA.; Biotechnology Institute, University of Minnesota-Twin Cities, St. Paul, Minnesota, USA., Hying ZT; Department of Plant and Microbial Biology, University of Minnesota-Twin Cities, St. Paul, Minnesota, USA.; Biotechnology Institute, University of Minnesota-Twin Cities, St. Paul, Minnesota, USA., Singla D; Department of Plant and Microbial Biology, University of Minnesota-Twin Cities, St. Paul, Minnesota, USA.; Biotechnology Institute, University of Minnesota-Twin Cities, St. Paul, Minnesota, USA., Márquez Reyes NL; Department of Plant and Microbial Biology, University of Minnesota-Twin Cities, St. Paul, Minnesota, USA.; Biotechnology Institute, University of Minnesota-Twin Cities, St. Paul, Minnesota, USA., Pandey SK; Department of Chemical and Biological Engineering, University of Idaho, Moscow, Idaho, USA., Patel JS; Department of Chemical and Biological Engineering, University of Idaho, Moscow, Idaho, USA., Bazurto JV; Department of Plant and Microbial Biology, University of Minnesota-Twin Cities, St. Paul, Minnesota, USA.; Biotechnology Institute, University of Minnesota-Twin Cities, St. Paul, Minnesota, USA.
Jazyk:	angličtina
Zdroj:	Applied and environmental microbiology [Appl Environ Microbiol] 2024 Jul 24; Vol. 90 (7), pp. e0031024. Date of Electronic Publication: 2024 Jun 27.
DOI:	10.1128/aem.00310-24
Abstrakt:	Integration of metabolites into the overall metabolic network of a cell requires careful coordination dependent upon the ultimate usage of the metabolite. Different stoichiometric needs, and thus pathway fluxes, must exist for compounds destined for diverse uses, such as carbon sources, nitrogen sources, or stress-protective agents. Herein, we expand upon our previous work that highlighted the nature of glycine betaine (GB) metabolism in Methylobacteria to examine the utilization of GB-derivative compounds dimethylglycine (DMG) and sarcosine into Methylorubrum extorquens in different metabolic capacities, including as sole nitrogen and/or carbon sources. We isolated gain-of-function mutations that allowed M. extorquens PA1 to utilize dimethylglycine as a carbon source and dimethylglycine and sarcosine as nitrogen source. Characterization of mutants demonstrated selection for variants of the AraC-like regulator Mext_3735 that confer constitutive expression of the GB metabolic gene cluster, allowing direct utilization of the downstream GB derivatives. Finally, among the distinct isolates examined, we found that catabolism of the osmoprotectant used for selection (GB or dimethylglycine) enhanced osmotic stress resistance provided in the presence of that particular osmolyte. Thus, access to the carbon and nitrogen and osmoprotective effects of GB and DMG are made readily accessible through adaptive mutations. In M. extorquens PA1, the limitations to exploiting this group of compounds appear to exist predominantly at the levels of gene regulation and functional activity, rather than being constrained by transport or toxicity.IMPORTANCEOsmotic stress is a common challenge for bacteria colonizing the phyllosphere, where glycine betaine (GB) can be found as a prevalent osmoprotectant. Though Methylorubrum extorquens PA1 cannot use GB or its demethylation products, dimethylglycine (DMG) and sarcosine, as a sole carbon source, utilization is highly selectable via single nucleotide changes for both GB and DMG growth. The innate inability to use these compounds is due to limited flux through steps in the pathway and regulatory constraints. Herein, the characterization of the transcriptional regulator, Mext_3735 (GbdR), expands our understanding of the various roles in which GB derivatives can be used in M. extorquens PA1. Interestingly, increased catabolism of GB and derivatives does not interfere with, but rather improves, the ability of cells to thrive under increased salt stress conditions, suggesting that metabolic flux improves stress tolerance rather than providing a distinct tension between uses. Competing Interests: The authors declare no conflict of interest.
Databáze:	MEDLINE
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