Abstract
An unstructured model based on mass balance equations for a recombinant methylotrophic yeast Pichia pastoris MutS (Methanol Utilization Slow) strain expressing the mini-proinsulin (MPI), was successfully established in quasi-steady state fed-batch fermentations with varying total quantity of biomass in a 7 l fermenter. The model describes the relationships between the total biomass and induction time, both in the batch and fed-batch phases. In addition, good correlations were obtained when the total quantity of MPI was correlated with the total biomass, demonstrating that the product of interest is associated with growth in the methanol phase. The parameters of the fermentation model obtained are similar to those reported by other researchers.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Brierley RA, Bussineau C, Kosson R, Melton A, Siegel RS (1990) Fermentation development of recombinant Pichia pastoris expressing the heterologous gene: bovine lysozyme. Ann. N.Y. Acad. Sci. 589: 350–362.
Cregg JM, Cereghino JL, Shi J, Higgins DR (2000). Recombinant protein expression in Pichia pastoris. Mol. Biotechnol. 16: 23–52.
D'Anjou MC, Daugulis AJ (2000) Mixed-feed exponential feeding for fed-batch culture of recombinant methylotrophic yeast. Biotechnol. Lett. 22: 341–346.
Goeddel DV, Kleid DG, Bolivar F, Heyneker HL, Yansura DG (1979) Expression in Escherichia coli of chemically synthesized genes for human insulin. Proc. Natl. Acad. Sci. USA 76: 106–110.
Kjeldsen T, Pattersson AF, Hach M (1999) Secretory expression and characterization of insulin in Pichia pastoris. Biotechnol. Appl. Biochem. 29: 79–86.
Markussen J, Damgaard U, Diers I, Fiil N, Hansen MT, Larsen P, Norris F, Norris K, Schou O, Snel L (1986) In: Theodoropoulos D, ed. Peptides. Berlin: Walter de Gruyter &; Co., pp. 189–194.
Pirt SJ (1975) Principles of Microbe and Cell Cultivation. Oxford: Blackwell Scientific Publications.
Roels JA (1983) Energetics and Kinetics in Biotechnology. Amsterdam: Elsevier Biomedical Press.
Thim L, Hansen MT, Norris K, Hoegh I, Boel E, Forstrom J, Ammerer G, Fiil NP (1986a) Secretion and processing of insulin precursors in yeast. Proc. Natl. Acad. Sci. USA 83: 6766–6770.
Thim L, Norris K, Hansen MT (1986b) Insulin precursors, process for their preparation and process for the preparation of human insulin. Eur. Pat. Applic. 86302133.3.
Wang Y, Liang ZH, Zhang YS, Yao SY, Xu YG, Tang YH, Zhu SQ, Cui DF, Feng YM (2001) Human insulin from a precursor overexpressed in the methylotrophic yeast Pichia pastoris and a simple procedure for purifying the expression product. Biotechnol. Bioeng. 73: 74–79.
Zhang W, Inam M, Meagher MM (2000a) Fermentation strategies for recombinant protein expression in the methylotrophic yeast Pichia pastoris. Biotech. Bioproc. Eng. 5: 275–287.
Zhang W, Bevins MA, Plantz BA, Smith LA, Meagher MM (2000b) Modeling Pichia pastoris growth on methanol and optimizing the production of a recombinant protein, the heavy-chain fragment C of botulinum neurotoxin, serotype A. Biotechnol. Bioeng. 70: 1–8.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
País, J.M., Varas, L., Valdés, J. et al. Modeling of mini-proinsulin production in Pichia pastoris using the AOX promoter. Biotechnology Letters 25, 251–255 (2003). https://doi.org/10.1023/A:1022398917213
Issue Date:
DOI: https://doi.org/10.1023/A:1022398917213