Table 1 Comparison of growth parameters and metabolite profiles of Ruminococcus albus and Wolinella succinogenes in pure culture and co-culture

	Ruminococcus albus	Wolinella succinogenes	Co-culture
Growth parameters
Growth yield (OD₆₀₀)	0.79 ± 0.01	0.36 ± 0.01	0.93 ± 0.01
Specific growth rate (h⁻¹)	0.58 ± 0.19	0.33 ± 0.06	0.57 ± 0.34 (Ra) 0.54 ± 0.11 (Ws)
Concentration changes of extracellular metabolites
Hydrogen (%)	+5.3	−78.4	BDL
Fumarate (mM)	−5.5	−46.3	−43.1
Succinate (mM)	+2.2	+54.6	+55.4
Acetate (mM)	+21.8	0	+32.4
Ethanol (mM)	+8.7	0	+0.3
Formate (mM)	BDL	BDL	BDL
pH	−0.4	−0.4	−0.6

Growth of pure cultures and co-cultures of Ruminococcus albus 7 and Wolinella succinogenes DSM 1740 was monitored by qPCR. Values show means ± standard deviations of three biological replicates. Also shown is the change in extracellular pH, percentage hydrogen gas (measured by gas chromatography), and concentrations of fumarate, succinate, acetate, ethanol and formate (measured by ultra-fast liquid chromatography) between 0 and 12 h. Growth media was the same between the three conditions, except 80% H₂ was added for W. succinogenes growth, whereas no H₂ was added for the other conditions. Full liquid metabolite measurements are shown in Figure S8. BDL = below detection limit (1 mm for formate, 0.1% for H₂).

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