The effect of pH on phosphine formation during anaerobic cultivation of granular sludge was investigated. The sludge was taken from full-scale anaerobic reactors treating brewery wastewater. Acetate and phosphate were used as the carbon source and phosphorus source respectively. After 10 days cultivation in the dark, results showed that acidic conditions were more favorable for free phosphine production. At pH 5, the optimum concentration 86.42 ng PH3 m-3 of free phosphine was obtained. The level at pH 7 was reduced to 18.53 ng PH3 m-3, about 1/5 of the maximum. The maximum concentration of matrix-bound phosphine of 3.30 ng PH3 kg-1 wet sludge was achieved at pH 6. More than 83% of the total phosphine was matrix-bound phosphine, which accounted for 0.003-0.009 per thousand of the phosphate removal, while free phosphine comprised 0.00002-0.001 per thousand of the phosphate removal. Most of the phosphorus removal from solution was turned into chemical precipitation or was adsorbed by sludge. The mechanism of the phosphate reduction-step in the formation of phosphine production is still unknown. The promotion of phosphine formation by low pH is compatible with an acidic bio-corrosion mechanism of metal particles in the sludge or of metal phosphides which form phosphine at low pH.