Study and improvement of the conditions for production of a novel alkali stable catalase

Abstract

Catalase (CAT) is an enzyme capable of catalyzing the conversion of H(2)O(2) to O(2) and H(2)O. It has recently acquired interest due to its attractive potential application in the textile industries. In a previous study, a bacterium with slight halophilic and alkaliphilic characteristics, Bacillus sp. F26, was isolated and found to produce high-level alkaline CAT. In the present study, the effects of culture conditions on the CAT production were investigated. The results showed that the highest activity of CAT (13.9 U/mg protein) was obtained when glucose (15 g/L) was used as carbon source. The utilization of the mixture of corn steep liquid and beef extract stimulated both bacterial growth and CAT synthesis. The highest biomass (4.5 g/L) and activity of CAT (16.5 U/mg protein) were found synchronously when 10 g/L corn steep liquid and 10 g/L beef extract were used as nitrogen source. The addition of H(2)O(2) as an oxidative stress was used to enhance CAT production in the flasks. It was found that the activity of CAT was increased by 51.3-22.8 U/mg protein compared with the control when 2 mmol/L H(2)O(2) was added at later exponential phases (16 h), although the cell growth was significantly inhibited. Based on the above, an exponential H(2)O(2) feed strategy was developed, in which the feed rate of H(2)O(2) was controlled according to specific cell growth rate (mu). In this way, the maximum CAT production (29.9 U/mL) was obtained, which was 92.8 and 20.7% higher than that in batch and constant rate fed-batch fermentation, respectively.