Production of Bacillus licheniformis ATCC 21415 alkaline protease in batch, repeated batch and continuous culture

ABSTRACT

Bacillus licheniformis ATCC 21415 cells were immobilized on different carriers using different methods of immobilization including physical adsorption, covalent binding, ionic binding and entrapment. The immobilized cells were prepared by covalent binding on wool (as a new carrier) through 1% glutaraldehyde had the highest enzyme activity (9.0 U/mL) with the highest specific productivity (6.17 U/g wet cells/h). Alkaline protease production and the stability of biocatalyst were investigated in both free and immobilized cells. The results showed that the immobilized cells were more efficient for enzyme production by repeated batch fermentation (5 cycles, 480 h) with 57% residual activity whereas the free cells retained 35% after 2 cycles. In continuous production the highest enzyme activity (9.9 U/mL) was obtained at a dilution rate of 0.1/h while the highest enzyme yield (763.6 U/h) and the highest reactor productivity (3.32 U/mL/h) were attained at a dilution rate of 0.4/h. Packed-bed bioreactor was a successful method for continuous production of alkaline protease for a long time (168 h) with 53% relative activity. The bioreactor affected the highest specific productivity (118.2 U/g wet cells/h) which was 12-24 times higher than other systems of enzyme production.