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In order to investigate the effect of different pH adjustment methods on Coenzyme Q10 fermentation of Rhodobacter sphaeroides, two batch cultures in different pH adjustment were carried out with NH3.H2O and CaCO3 respectively in a 10 L stirred fermentor.The results showed two fermentation processes had little different effect on the cell biosynthesis.But it could reduce the rate of glucose consumption and increase the translation rate of Coenzyme Q10 to glucose and CoQ10 concentration with CaCO3 for pH adjustment.The metabolic flux distribution analyses for two fermentation processes of 42h were determined.It showed different methods of pH adjustment had serious effect on the metabolic flux distribution of chorismate、fructose-6-phosphate and glucose-6-phosphate nodes.The pH adjustment with CaCO3 could enhance hexose monophosphate pathway, provide more NADPH for Coenzyme Q10 biosynthesis pathway and improve the ability of Coenzyme Q10 biosynthesis.