Title: Fermentation Process of Pichia pastoris Expressing Recombinant Human Soluble TRAIL
Abstract: Objective To optimize the fermentation process of Pichia pastoris expressing recombinant human soluble TRAIL(rhsTRAIL) in a 5 L fermentor.Methods Study the influence of fermentation parameters,such as medium,seed cell density,glycerol content,pH value,the concentration and flow rate of methanol,as well as the time for induction,on the growth of Pichia pastoris and expression of target protein.Determine the activity of rhTRAIL in inducing the aoptosis of tumor cells by electron microscopy.Results After inorganic salt medium was inoculated with the seed cells at a density(A_ 600 ) of 8-12 and a m.o.i of 10% and incubated for 24 hours,the density of Pichia pastoris(A_ 600 ) reached 76.When the glycerol content was 1%,the Pichia pastoris grew rapidly(A_ 600 =160).When Pichia pastoris was incubated in the medium at a pH value of 5.0 and a final methanol concentration of 1% for 96 hours,the rhsTRAIL reached a maximum expression level of 120 mg/L,and showed the activity in inducing the apoptosis of tumor cells.Conclusion The fermentation process of Pichia pastoris expressing rhsTRAIL was optimized as follows:To inorganic salt medium(pH 5.0),inoculate the seed cells of Pichia pastoris at a density(A_ 600 ) of 8-12 and a m.o.i of 10%,add glycerol and methanol to a final concentration of 1% separately and induce for 96 hours.
Publication Year: 2005
Publication Date: 2005-01-01
Language: en
Type: article
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