Percoll gradient centrifugation effectively separated Syntrophomonas wolfei cells from Methanospirillum hungatii cells, resulting in a 70- to 80-fold enrichment of S. wolfei cells relative to M. hungatii cells. The separated S. wolfei cells were viable. Gram quantities of cellular protein which was enzymatically active and had low levels of contamination by the methanogenic cofactor, factor(420), were obtained.