The optimization of fermentation conditions for Pichia pastoris GS115 producing recombinant xylanase

Abstract

Xylanase is a member of an important family of enzymes that has been used in many biotechnological processes. However, the overall cost of enzyme production has been the main problem in the industrial application of enzymes. To obtain maximum xylanase production, statistical approaches based on the Plackett–Burman design and response surface methodology were employed. The results of the statistical analyses demonstrated that the optimal conditions for increased xylanase production were the following: inoculum size, 3.8%; maize meal, 4.5%; histidine, 0.6%; methanol, 1%; culture volume, 20%; bean pulp, 30 g L⁻¹; and Tween‐80, 0.8%; and pH 5.0. Verification of the optimization demonstrated that 3273 U mL⁻¹ xylanase was observed under the optimal conditions in shake flask experiments. SDS–PAGE results showed that the size of xylanase protein was about 23 kDa. The results showed that the xylanase produced by fermentation came from Aspergillus Niger by MALDI‐TOF‐MS. The optimized medium resulted in 2.1‐ and 1.4‐fold higher the activity of xylanase compared with the unoptimized medium (the main nutrients are maize meal and bean pulp) and laboratory medium (the main nutrients are yeast extract and peptone), respectively. The optimization of fermentation conditions is an effective means to reduce production cost and improve xylanase activity.