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KMID : 1094720180230010055
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Biotechnology and Bioprocess Engineering
2018 Volume.23 No. 1 p.55 ~ p.63
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Statistical Optimization of Medium and Fermentation Conditions of Recombinant Pichia pastoris for the Production of Xylanase
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Lee Nam-Keun
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Abstract
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Recombinant xylanase (rPcXynC) from Pichia pastoris was produced on large-scale by optimizing production-medium composition using statistical experimental methods. Production medium was optimized through the use of statistical methods such as one factor at a time (OFAT), Plackett-Burman design, fractional factorial design (FFD), steepest ascent method (SAM), and response surface methodology (RSM). The optimum medium composition was established to be (g/L); wheat bran 11.62, yeast extract 30, Tween 60.5, DL-β-Phenylalanine 0.5, Thiamine 0.5, FeSO4 0.01, KH2PO4 0.66, and KHSO4 0.09. The optimum medium composition yielded 3,051 mU/mL of xylanase activity which was three times higher than that obtained from the initial medium composition. Finally, fermentation conditions were examined using the optimized production medium in a laboratory bioreactor. The optimal fermentation conditions were found to be 25ºC, pH 6, 170 rpm and 1 vvm with intermittent feeding of methanol (67.5 mL) and the xylanase activity was 3,683 mU/mL. In repeated-batch fermentation using optimized production medium and fermentation condition, the xylanase activity was 3,680 mU/mL at the first cycle of 96 h harvesting time using 90% of the culture solution. The activity was similarly maintained until the last cycle of 264 h.
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KEYWORD
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Pichia pastoris, response surface methodology, optimum medium composition, xylanase
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