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  • Functional analysis of an anaerobic m-xylene-degrading enrichment culture using protein-based stable isotope probing.

Functional analysis of an anaerobic m-xylene-degrading enrichment culture using protein-based stable isotope probing.

FEMS microbiology ecology (2012-03-01)
Dragana Bozinovski, Steffi Herrmann, Hans-Hermann Richnow, Martin von Bergen, Jana Seifert, Carsten Vogt
ABSTRACT

A sulfate-reducing consortium maintained for several years in the laboratory with m-xylene as sole source of carbon and energy was characterized by terminal restriction fragment length polymorphism (T-RFLP) fingerprinting of PCR-amplified 16S rRNA genes and stable isotope probing of proteins (Protein-SIP). During growth upon m-xylene or methyl-labeled m-xylene (1,3-dimethyl-(13)C(2)-benzene), a phylotype affiliated to the family Desulfobacteriaceae became most abundant. A second dominant phylotype was affiliated to the phylum Epsilonproteobacteria. In cultures grown with methyl-labeled m-xylene, 331 proteins were identified by LC-MS/MS analysis. These proteins were either not (13)C-labeled (23%) or showed a (13)C-incorporation of 19-22 atom% (13)C (77%), the latter demonstrating that methyl groups of m-xylene were assimilated. (13)C-labeled proteins were involved in anaerobic m-xylene biodegradation, in sulfate reduction, in the Wood-Ljungdahl-pathway, and in general housekeeping functions. Thirty-eight percent of the labeled proteins were affiliated to Deltaproteobacteria. Probably due to a lack of sequence data from Epsilonproteobacteria, only 14 proteins were assigned to this phylum. Our data suggest that m-xylene is assimilated by the Desulfobacteriaceae phylotype, whereas the role of the Epsilonproteobacterium in the consortium remained unclear.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
m-Xylene, anhydrous, ≥99%
Supelco
m-Xylene, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
m-Xylene, ReagentPlus®, 99%
Supelco
m-Xylene, analytical standard
Sigma-Aldrich
m-Xylene, puriss. p.a., ≥99.0% (GC)