Growth inhibition by ammonia and use of a pH-controlled feeding strategy for the effective cultivation of Mycobacterium chlorophenolicum

The inhibitory effect of ammonia on the growth of the polychlorinated xenobiotic-degrading bacterium Mycobacterium chlorophenolicum was examined. The strain is inhibited by both the ionized and nonionized forms of ammonia, At pH 6.9, 50% reduction of the growth rate was observed at 6.8 g/l total ammonium. For 23 experiments performed in shake-flask culture at different pH values, and ammonium concentrations a growth model based on the extended Monod kinetic fits the data with a deviation of 5.3%. To overcome growth inhibition in bioreactors, a pH- controlled feeding strategy was developed for effective cultivation of M. chlorophenolicum at an ammonium level below 0.3 g/l. The ammonium addition was controlled online by the stoichiometric interdependence of ammonium consumption and pH decline. With this online control strategy, a biomass concentration as high as 26.2 g/l can be achieved within less than a week of cultivation. The yield is also increased from 0.32 g to 0.43 g biomass (per gram glucose). The strategy developed provides an effective method for the production of biomass o M. chlorophenolicum serving as the inoculum in remediation technologies.