Medium composition and culture conditions for the acid protease production byAspergillus nigerI1 were optimized by response surface methodology (RSM). A significant influence of temperature, KH2PO4, and initial pH on the protease production was evaluated by Plackett-Burman design (PBD). These factors were further optimized using Box-Behnken design and RSM. Under the proposed optimized conditions, the experimental protease production (183.13 U mL−1) closely matched the yield predicted by the statistical model (172.57 U mL−1) withR2=0.914. Compared with the initial M1 medium on which protease production was 43.13 U mL−1, a successful and significant improvement by 4.25 folds was achieved in the optimized medium containing (g/L): hulled grain of wheat (HGW) 5.0; KH2PO41.0; NaCl 0.3; MgSO4(7H2O) 0.5; CaCl2(7H2O) 0.4; ZnSO40.1; Na2HPO41.6; shrimp peptone (SP) 1.0. The pH was adjusted at 5 and the temperature at30°C. More interestingly, the optimization was accomplished using two cheap and local fermentation substrates, HGW and SP, which may result in a significant reduction in the cost of medium constituents.

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