title: Optimization of fermentation conditions for maximal recombinant hepatitis B surface antigen particle production in Pichia pastoris.
abstract: Hepatitis B virus (HBV) infection can cause the severe threat to the health of the people around the world. It depends upon the development of efficient diagnostic reagent and vaccine to prevent the prevalence of HB. In this study, we constructed the high expression recombinant Pichia pastoris and performed the screening tests in shake flasks to obtain the optimal values of several key fermentation parameters. Based on their effects on the growth and expression level of recombinant strains, FBS was the optimal industrial medium. The optimal values for the dissolve oxygen (DO), the final concentrations of methanol and the pH values were 50 mL, 1% (V/V) and 5.4-6.0, respectively. The optimal values of the parameters simulated in shake flasks were successfully scaled up to 10 L bioreactors to achieve high-throughput production: 310 OD600 in biomass and 27 mg/L in recombinant HBsAg. The expressed recombinant HBsAg in P. Pastoris was confirmed by SDS-PAGE and Western blotting. Electron microscopy examination showed that the purified protein could be self-assembled to 22 nm virus-like particles. The results provided a basis for industrial scale-up production of diagnostic reagent and vaccine of next generation against HB.
authors.affiliation: Colleges of Life Sciences, Hunan Normal University, Key Laboratory of Microbial Molecular of Hunan Province, Chnagsha 410081, China.
authors.forename: Rushi
authors.initials: R
authors.lastname: Liu
authors.forename: Qinlu
authors.initials: Q
authors.lastname: Lin
authors.forename: Yi
authors.initials: Y
authors.lastname: Sun
authors.forename: Yilan
authors.initials: Y
authors.lastname: Qiu
authors.forename: Tian
authors.initials: T
authors.lastname: Xu
authors.forename: Hai
authors.initials: H
authors.lastname: Ding
authors.forename: Xiangrong
authors.initials: X
authors.lastname: Guo
mesh_terms: D019149:Bioreactors; D005285:Fermentation; D006514:Hepatitis B Surface Antigens / Q000096:biosynthesis* / Q000235:genetics; D006801:Humans; D010843:Pichia / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D006514:Hepatitis B Surface Antigens; D011994:Recombinant Proteins