This study evaluated the effect of 8 mg/L sulfide on the speciation and bioavailability of 10 mg/L iron (Fe2+), with and without a biodegradable chelating agent, Ethylenediamine-N, N'-disuccinic acid (EDDS). These tests were carried out in the presence of sulfide, in both batch assays, and in a continuous submerged anaerobic membrane bioreactor (SAMBR). The EDDS was added at 1:1 M ratio to the Fe2+ (10 mg/L), either simultaneously, or as Fe-EDDS complex. Results showed that adding 8 mg/L of sulfide limited the bioavailability of Fe2+ by shifting its speciation toward less bioavailable fractions based on the Bureau Communautaire de Reference (BCR) extraction analysis, i.e. organic matter/sulfide and residual, resulting in a decrease in methane yields and an increase in VFA levels. An Fe-EDDS complex was found to be more effective controlling the change in sulfide levels in the SAMBR as it helped to reverse the shift in Fe2+ speciation, and increased methane yields by 9.46%.