AbstractAn AnSBBR (anaerobic sequencing batch reactor containing biomass immobilized on an inert support) with liquid phase recirculation, containing a 3.5 L working volume, treated 1.5 L of cheese whey wastewater in 3 and 4 h cycles at 30 ºC to produce biohydrogen. From startup the bioreactor presented process instability. To overcome this problem the following measures were taken, however without success: adaptation of the biomass with uncontaminated easily degradable substrates, pH control at very low levels, and a different form of inoculation (natural fermentation of the feed medium). The problem was solved by cooling the feed medium to 4 ºC to prevent acidification in the storage container, by eliminating nutrient supplementation to prevent possible formation of H2S by sulfate-reducing bacteria and by periodic washing of the support material to improve the food/microorganism ratio. Hence, stable hydrogen production could be achieved with minimal presence of methane (36% H2; 62% CO2; 2% CH4) and the AnSBBR fed with cheese whey (influent concentration of 4070 mgCOD.L-1 and 3240 mgCarbohydrate.L-1 and applied volumetric organic loading of 14.6 gCOD.L-1.d-1) presented improved productivity and yield indicators compared to pure lactose and other reactor configurations, reaching values of 420 NmLH2.L-1.d-1 and 0.60 molH2.molCarbohydrate-1 in the steady-state phase (conversions of carbohydrates and COD were 98% and 30%, respectively).