Publication Details

Category Text Publication
Reference Category Journals
DOI 10.1128/aem.00978-24
Title (Primary) MecE, MecB, and MecC proteins orchestrate methyl group transfer during dichloromethane fermentation
Author Soder-Walz, J.M.; Deobald, D. ORCID logo ; Vicent, T.; Marco-Urrea, E.; Adrian, L. ORCID logo
Source Titel Applied and Environmental Microbiology
Year 2024
Department MEB
Volume 90
Issue 10
Page From e00978-24
Language 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.
Persistent UFZ Identifier 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