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KMID : 1094720100150020314
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Biotechnology and Bioprocess Engineering
2010 Volume.15 No. 2 p.314 ~ p.323
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Production of Chum Salmon Cystatin from the Recombinant Saccharomyces cerevisiae Optimized using Response Surface Methodology
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Li De-Kun
Fu Xiao-Ting
Kim Sang-Moo
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Abstract
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Chum salmon cystatin was overexpressed on Saccharomyces cerevisiae YPH 499. At first, the culture condition for the production of recombinant chum salmon cystatin (RC) by S. cerevisiae YPH 499 was optimized in a shake flask using response surface methodology. Three independent variables; medium pH, inducing time, and the amount of inducing assistant, were analyzed to get the optimal condition for the production of RC. The results were fitted to a second-order polynomial equation, in which the determination coefficient (R2) was 0.904. The highest RC production in a shake flask, 0.57 U/mL was obtained at 5.7 of medium pH, 6.7 h of inducing time, and 5.6 g/L of inducing assistant. Based on the results of shake flask, the effects of agitation and aeration rates on the production of RC by S. cerevisiae YPH 499 were determined for scaleup in a fermentor. The highest production of RC in a fermentor, 0.56 U/mL was obtained at 350 rpm of agitation rate and 1.0 vvm of aeration rate. RC at 100 μg/g showed the highest inhibitory activity against the autolysis of Alaska pollock surimi based on the analysis of TCA-soluble peptides.
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KEYWORD
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Recombinant chum salmon cystatin, yeast, optimization, bioprocess design, scale-up, surimi
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