Summary
A central composite design (CCD) was used to evaluate, for the purpose of future process optimization, the influence of pH, yeast extract and ammonium chloride concentrations on the proportion of periplasmic hepatitisB pre-S2 antigen in the recombinant yeastHansenula polymorpha. Each factor was tested at five levels, and a second order polynomial model for the proportion of periplasmic antigen was fitted to the treatment combinations. pH showed the greatest effect: the proportion of periplasmic antigen was greatly increased at the higher pH levels. At the higher pH levels used, the proportion of periplasmic antigen was enhanced by a high concentration of ammonium chloride. Additional experiments have confirmed both the validity of the selected model and the optimal conditions found. A significant correlation was found between the proportion of periplasmic antigen and the total yield of antigen. These results indicated that is should be possible to modulate the distribution of the pre-S2 antigen between the periplasm and the cytoplasm of the yeast.
References
Blum, H.E., W. Gerok and G.N. Vyas. 1989. The molecular biology of hepatitisB virus. TIG 5: 154–158.
Box, G.E.P. and J.S. Hunter. 1957. Multifactor Experimental Designs for Exploring Response Surfaces. Ann. Math. Statist. 28: 195–242.
Draper, N.R. and H. Smith. 1981. Applied Regression Analysis. 2nd edn., John Wiley, New York.
Myers, R.H. 1976. Response Surface Methodology. Ann Arbor: Edwards Brothers (distributors).
De Roubin, M.R., L. Bastien, S.H. Shen and D. Groleau. 1991. Fermentation study for the production of hepatitis B virus pre-S2 antigen by the methylotrophic yeastHansenula polymorpha. J. Ind. Microbiol., 8: 147–156.
SAS/STAT User's Guide. 1989. Vol. 2, 4th edn., SAS Institute, NC, U.S.A.
Shen, H.-S., L. Bastien, T. Nguyen, M. Fung and S. Slilaty. 1989. Synthesis and secretion of hepatitis B middle surface antigen by the methylotrophic yeastHansenula polymorpha. Gene 84: 303–309.
Tsai, L.B., M. Mann, F. Morris, C. Rotgers and D. Fenton. 1987. The effect of organic nitrogen and glucose on the production of recombinant human insulin-like growth factor in high cell densityEscherichia coli fermentations. J. Ind. Microbiol. 2: 181–187.
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de Roubin, MR., Cailas, M.D., Shen, SH. et al. Some factors influencing the proportion of periplasmic hepatitisB virus pre-S2 antigen in the recombinant yeastHansenula polymorpha . Journal of Industrial Microbiology 9, 69–72 (1992). https://doi.org/10.1007/BF01576370
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DOI: https://doi.org/10.1007/BF01576370