Dataset Information

The proteome of Fibrobacter succinogenes S85 outer membrane vesicles

ABSTRACT: Here, we illustrate the proteomic composition and hydrolytic capacity of outer membrane vesicles (OMVs) produced by F. succinogenes during growth on Avicel cellulose and show that the OMVs are equipped with a highly versatile polysaccharide-degrading machinery. Proteomic assessment identified an enrichment of acidic proteins as well as carbohydrate-active enzymes of varying putative functions. Further, we show that the OMVs contain a membrane-bound multiprotein complex containing four proteins.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Fibrobacter Succinogenes Subsp. Succinogenes S85

SUBMITTER: Magnus Arntzen

LAB HEAD: Vincent Eijsink

PROVIDER: PXD005442 | Pride | 2017-06-30

REPOSITORIES: Pride

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Outer membrane vesicles from Fibrobacter succinogenes S85 contain an array of carbohydrate-active enzymes with versatile polysaccharide-degrading capacity.

Arntzen Magnus Ø MØ Várnai Anikó A Mackie Roderick I RI Eijsink Vincent G H VGH Pope Phillip B PB

Environmental microbiology 20170529 7

Fibrobacter succinogenes is an anaerobic bacterium naturally colonising the rumen and cecum of herbivores where it utilizes an enigmatic mechanism to deconstruct cellulose into cellobiose and glucose, which serve as carbon sources for growth. Here, we illustrate that outer membrane vesicles (OMVs) released by F. succinogenes are enriched with carbohydrate-active enzymes and that intact OMVs were able to depolymerize a broad range of linear and branched hemicelluloses and pectin, despite the inab ...[more]

PMID: 28447389

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Project description:Filamentous fungi are powerful producers of hydrolytic enzymes for the deconstruction of plant cell wall polysaccharides. However, the central question of how these sugars are perceived in the context of the complex cell wall matrix remains largely elusive. To address this question in a systematic fashion we performed an extensive comparative systems analysis of how the model filamentous fungus Neurospora crassa responds to the three main cell wall polysaccharides: pectin, hemicellulose and cellulose. We found the pectic response to be largely independent of the cellulolytic one with some overlap to hemicellulose, and in its extent surprisingly high, suggesting advantages for the fungus beyond being a mere carbon source. Our approach furthermore allowed us to identify carbon source-specific adaptations, such as the induction of the unfolded protein response on cellulose, and a commonly induced set of 29 genes likely involved in carbon scouting. Moreover, by hierarchical clustering we generated a co-expression matrix useful for the discovery of new components involved in polysaccharide utilization. This is exemplified by the identification of lat-1, which we demonstrate to encode for the physiologically relevant arabinose transporter in Neurospora. The analyses presented here are an important step towards understanding fungal degradation processes of complex biomass. Our data are based on carbon source transfer experiments. For this, N. crassa pregrown in sucrose for 16 hrs was washed in media without carbon and then transferred to either pectin, orange peel powder (OPP), xylan, Avicel (cellulose), sucrose, or media without carbon (NoC) for another 4 hrs. Biological triplicates (pectin, Avicel, sucrose, NoC) or duplicates (OPP) were used to identify differentially expressed genes in WT. For xylan only a single library was prepared, which was found to correlate well with previously published microarray data (Sun et al. 2012; doi: 10.1128/EC.05327-11)

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