Production of chum salmon cystatin from the recombinant <Emphasis Type="Italic">Saccharomyces cerevisiae</Emphasis> optimized using response surface methodology |
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Authors: | Dekun Li Xiaoting Fu Sang Moo Kim |
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Institution: | (1) Department of Hotel and Restaurant Management, Chia-Nan University of Pharmacy and Science, 60 Erh-Jen Rd., Sect. 1, Jen-Te, Tainan, 71710, Taiwan;(2) Department of Biotechnology, Chia-Nan University of Pharmacy and Science, 60 Erh-Jen Rd., Sect. 1, Jen-Te, Tainan, 71710, Taiwan |
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Abstract: | 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 (R
2) 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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