Medium optimization for endochitinase production by recombinant Pichia pastoris ZJGSU02 using response surface methodology
Plackett-Burman design and response surface methodology were employed to optimize the medium components for endochitinase production by Pichia pastoris. A Plackett- Burman design of seven factors with 12 runs was applied to evaluate the effects of different medium components. Yeast extract, oleic acid and Tween-80 were found to have significant influence on endochitinase production. The optimal concentrations of three factors were investigated by the response surface methodology using Box- Behnken design. The optimal medium components obtained for achieving the maximum activity of the endochitinase were as follows: Yeast extract 24.36 g/l, tryptone 20 g/l, YNB 5.0 g/l, potassium phosphate 100 mM, methanol 5 ml/l, oleic acid 1.758 ml/l, Tween-80 6.2 ml/l, Pichia trace metals (PTM1) 4.0 ml/ l and biotin 4.00 × 10-4 g/l. Under these conditions, endochitinase activity was up to 88.26 μ/ml, which was about 1.14-fold higher than using the original medium (77.62 μ/ml). This work will be very helpful for large-scale production of endochitinase for future industrial application.
Keywords: Recombinant endochitinase, Plackett-Burman design, response surface methodology, fermentation