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Details	EV-TRACK ID	Experiment nr.	Species	Sample type	Separation protocol	First author	Year	EV-METRIC
	EV230776	1/1	Fibrobacter succinogenes	F. succinogenes S85 (ATCC 19169)	(d)(U)C DG Filtration UF	Arntzen MØ	2017	50%
Study summary Full title Outer membrane vesicles from Fibrobacter succinogenes S85 contain an array of carbohydrate-active enzymes with versatile polysaccharide-degrading capacity. All authors Arntzen MØ, Várnai A, Mackie RI, Eijsink VGH, Pope PB Journal Environ Microbiol Abstract Fibrobacter succinogenes is an anaerobic bacterium naturally colonising the rumen and cecum of herbi (show more...)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 inability of F. succinogenes to utilize non-cellulosic (pentose) sugars for growth. We hypothesize that the degradative versatility of F. succinogenes OMVs is used to prime hydrolysis by destabilising the tight networks of polysaccharides intertwining cellulose in the plant cell wall, thus increasing accessibility of the target substrate for the host cell. This is supported by observations that OMV-pretreatment of the natural complex substrate switchgrass increased the catalytic efficiency of a commercial cellulose-degrading enzyme cocktail by 2.4-fold. We also show that the OMVs contain a putative multiprotein complex, including the fibro-slime protein previously found to be important in binding to crystalline cellulose. We hypothesize that this complex has a function in plant cell wall degradation, either by catalysing polysaccharide degradation itself, or by targeting the vesicles to plant biomass. (hide) EV-METRIC 50% (87th percentile of all experiments on the same sample type) Reported Not reported Not applicable EV-enriched proteins Protein analysis: analysis of three or more EV-enriched proteins non EV-enriched protein Protein analysis: assessment of a non-EV-enriched protein qualitative and quantitative analysis Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected. electron microscopy images Particle analysis: inclusion of a widefield and close-up electron microscopy image density gradient Separation method: density gradient, at least as validation of results attributed to EVs EV density Separation method: reporting of obtained EV density ultracentrifugation specifics Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps antibody specifics Protein analysis: antibody clone/reference number and dilution lysate preparation Protein analysis: lysis buffer composition Study data Sample type Cell culture supernatant Sample origin Control condition Focus vesicles outer membrane vesicle Separation protocol Separation protocol Gives a short, non-chronological overview of the different steps of the separation protocol. dUC = (Differential) (ultra)centrifugation DG = density gradient UF = ultrafiltration SEC = size-exclusion chromatography IAF = immuno-affinity capture (Differential) (ultra)centrifugation Density gradient Filtration Ultrafiltration Protein markers EV: None non-EV: None Proteomics yes EV density (g/ml) not specified Show all info Study aim Identification of content (omics approaches) Sample Species Fibrobacter succinogenes Sample Type Cell culture supernatant EV-producing cells F. succinogenes S85 (ATCC 19169) EV-harvesting Medium Serum free medium Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Between 800 g and 10,000 g Between 10,000 g and 50,000 g Pelleting performed No Density gradient Type Discontinuous Number of initial discontinuous layers not specified if it was continuous or not Lowest density fraction not specified Highest density fraction not specified Total gradient volume, incl. sample (mL) not specified Sample volume (mL) not spec Speed (g) 200000 Duration (min) 120 Fraction volume (mL) not specified Fraction processing Centrifugation Pelleting: volume per fraction 12 ml Pelleting: speed (g) 100000 Filtration steps Between 0.22 and 0.45 Î¼m Ultra filtration Cut-off size (kDa) 10 en 10 Membrane type NS Characterization: Protein analysis Protein Concentration Method Bradford Proteomics database ProteomeXchange consortium Characterization: Lipid analysis No Characterization: Particle analysis DLS Report type Mean Reported size (nm) 49 EM EM-type TransmissionÂ-EM Image type Close-up

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EV-TRACK ID	EV230776
species	Fibrobacter succinogenes
sample type	Cell culture
cell type	F. succinogenes S85 (ATCC 19169)
condition	Control condition
separation protocol	dUC/ Density gradient/ Filtration/ Ultrafiltration
Exp. nr.	1
EV-METRIC %	50