Bioreactor technology for bioprocessing graywater solutions in microgravity is under development by NASA at Johnson Space Center and at major aerospace companies. Inoculum sources have been inconsistent. Startup and subsequent operation of ground-based bioreactors may have been adversely affected by this inconsistency and/or by inoculation procedures. The goal of the research reported in this paper is to develop an inoculum that will completely biodegrade Igepon T42 soap to carbon dioxide and water under anaerobic, denitrifying conditions and with process conditions set by bioreactor design requirements for microgravity operation. Potential inoculum sources from two habitats within the KSC-ALS breadboard project were developed for potential use. The effects of pH (7.2 vs. 9.0, buffered) on soap degradation by the two inocula was determined in a flask study. Nearly all of the soap was degraded at pH 7.2 while nearly none was degraded at pH 9.0. Both inocula behaved similarly. Stability and reliability of the flask culture enrichments was demonstrated continuous denitrification and soap degradation for 5 months with daily replacement of the liquid contents of the flasks was achieved. Bacterial isolates were obtained from the rootinoculated flask cultures and to determine if human pathogens were present.