The objective of this study was to optimize the submerged cultured conditions of carbonic anhydrase (CA) production from the strain of Bacillus mucilaginosus K02 by mono-factor tests with blank control and orthogonal design methodology. When the bacteria was cultured under the condition of monofactor tests with blank control and orthogonal designs, and conventional mono-factor test and orthogonal designs, respectively, the CA productions increased from 5.32 μU/L under the basic medium to 34.53 and 26.12 μU/L. The kinetic analysis of 5 L-fermentor showed that the specific production rate of CA in the optimized medium was almost equaled to the one obtained in the basic and optimized medium with conventional methods. The result demonstrated that CA synthesis was positively correlated with cell growth, and the mono-factor tests with blank control applied in the optimizing composition of the media could achieve better results in contrast to the conventional method.Key words: Bacillus mucilaginosus, carbonic anhydrase, specific production rate, kinetic analysis, orthogonal design
