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Controlled fed‐batch fermentation of recombinant Saccharomyces cerevisiae to produce hepatitis B surface antigen

1988; Wiley; Volume: 32; Issue: 3 Linguagem: Inglês

10.1002/bit.260320310

1097-0290

Jih‐Han Hsieh, Kun‐Yu Shih, Hsiang‐Fu Kung, Ming Shiang, Lan‐Ying Lee, Menghsiao Meng, Ching‐Chuan Chang, Hui‐Min Lin, Shu‐Ching Shih, Shu‐Ying Lee, Teh‐Yuan Chow, Teng‐Yung Feng, Tsong‐Teh Kuo, Kong‐Bung Choo,

Protein purification and stability

We have performed controlled fed-batch fermentation experiments to compare the production level of hepatitis B surface antigen (HBsAg) by recombinant yeast Saccharomyces cerevisiae strains (YNN27/pYBH-1, YNN27/ p2micro-S11, YNN27/pDCB-S2) containing plasmid vector with alcohol dehydrogenase (ADH1), acid phosphatase (PHO5), and glyceraldehyde-3-phosphate dehydrogenase (GPD) promoter, respectively. Yeast cell concentrations of 15-35 g dry cell weight/L were obtained. By limiting phosphorous concentration, HBsAg expression level for the YNN27/p2micro-S11 strain with inducible PHO5 promoter reached 0.2-0.3 mg/L. By controlling nutrient addition rate and dissolved oxygen concentration, HBsAg concentrations of 3-10 mg/L were achieved in 60-70 h fermentation using the YNN27/pDCB-S2 strain with the constitutive GPD promoter.