Details zur Publikation |
| Kategorie | Textpublikation |
| Referenztyp | Zeitschriften |
| DOI | 10.1128/aem.00978-24 |
| Volltext | Akzeptiertes Manuskript |
| Titel (primär) | MecE, MecB, and MecC proteins orchestrate methyl group transfer during dichloromethane fermentation |
| Autor | Soder-Walz, J.M.; Deobald, D.; Vicent, T.; Marco-Urrea, E.; Adrian, L. |
| Quelle | Applied and Environmental Microbiology |
| Erscheinungsjahr | 2024 |
| Department | MEB |
| Band/Volume | 90 |
| Heft | 10 |
| Seite von | e00978-24 |
| Sprache | englisch |
| Topic | T7 Bioeconomy |
| Supplements | https://journals.asm.org/doi/suppl/10.1128/aem.00978-24/suppl_file/aem.00978-24-s0001.pdf https://journals.asm.org/doi/suppl/10.1128/aem.00978-24/suppl_file/aem.00978-24-s0002.xlsx |
| Keywords | in vitro activity test; anaerobic fermentation; heterologous expression; methyltransferase; metalloprotein; Dehalobacterium |
| Abstract | Dichloromethane (DCM), a common hazardous industrial chemical, is anaerobically metabolized by four bacterial genera: Dehalobacter, Dehalobacterium, Ca. Dichloromethanomonas, and Ca. Formimonas. However, the pivotal methyltransferases responsible for DCM transformation have remained elusive. In this study, we investigated the DCM catabolism of Dehalobacterium formicoaceticum strain EZ94, contained in an enriched culture, using a combination of biochemical approaches. Initially, enzymatic assays were conducted with cell-free protein extracts, after protein separation by blue native polyacrylamide gel electrophoresis. In the slices with the highest DCM transformation activity, a high absolute abundance of the methyltransferase MecC was revealed by mass spectrometry. Enzymatic activity assays with heterologously expressed MecB, MecC, and MecE from strain EZ94 showed complete DCM transformation only when all three enzymes were present. Our experimental results, coupled with the computational analysis of MecB, MecC, and MecE sequences, enabled us to assign specific roles in DCM transformation to each of the proteins. Our findings reveal that both MecE and MecC are zinc-dependent methyltransferases responsible for DCM demethylation and re-methylation of a product, respectively. MecB functions as a cobalamin-dependent shuttle protein transferring the methyl group between MecE and MecC. This study provides the first biochemical evidence of the enzymes involved in the anaerobic metabolism of DCM. |
| dauerhafte UFZ-Verlinkung | https://www.ufz.de/index.php?en=20939&ufzPublicationIdentifier=29750 |
| Soder-Walz, J.M., Deobald, D., Vicent, T., Marco-Urrea, E., Adrian, L. (2024): MecE, MecB, and MecC proteins orchestrate methyl group transfer during dichloromethane fermentation Appl. Environ. Microb. 90 (10), e00978-24 10.1128/aem.00978-24 |
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