Analysis of culture phase based on proteome analysis and control of extracellular enzyme production

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The aim of this study was to analyze culture phase based on informations obtained by intracelluar and extracelluar proteome analysis using a two dimensional electrophoresis (2 DE) and to control extracellular enzyme productions. Bacillus amyloliqucfaciens (ATCC 23350) was employed for the proteome analysis. Intracellular and extracelluar protein samples were taken from batch cultures at different physiological states including growth phase and a-amylase production phase. A novel 2 DE image analyzer was employed to analyze dynamic processes of 2 DE maps stained with silver nitrate. Using the novel image analyzer, overlapped protein spots on the 2 DE maps could be recognized as individual spots. The number of protein spots detected by the novel image analyzer was increased as compared with the conventional image analysis method. The detected protein spots were identified by use of the database of Bacillus subtilis proteins analyzed by a peptide mass fingerprinting. At the growth phase, proteins concerning glycolysis and TCA cycle were clearly identified, while extracellular proteins were not observed. However, at a-amylase production phase, many extracellular proteins were identified. Therefore, the culture phase was effectively analyzed at a protein level by proteome analysis using 2 DE.