Optimization of temperature-glycerol-pH conditions for a fed-batch fermentation process for recombinant hookworm (Ancylostoma caninum) anticoagulant peptide (AcAP-5) production by Pichia pastoris

Mehmet Inan, Vijay Chiruvolu, Kent M. Eskridge, George P. Vlasuk, Kenneth Dickerson, Scott Brown, Michael M. Meagher

Research output: Contribution to journalArticle

57 Scopus citations

Abstract

This study was undertaken to determine the optimum pH, temperature and glycerol feed rate for the production of recombinant hookworm (Ancylostoma caninum) anticoagulant peptide (rAcAP-5) by Pichia pastoris using response surface methodology (RSM). A central composite design was used as an experimental design for allocation of treatment combinations in three blocks. The variables selected for study were pH, temperature and glycerol feed rate. pH was the most important variable affecting yield, specific yield and specific activity of rAcAP-5. Glycerol feed rate had a significant effect on the specific activity of rAcAP-5 (% of total secreted protein) while temperature did not have a significant effect on the responses. The data showed a trend that gave maximum responses and there was no blocking effect on the responses. The RSM formulated three second order polynomial empirical models relating to the responses. From these models it was possible to determine the optimum conditions variables for maximum yield of rAcAP-5 (1.2 g l-1), the maximum specific yield of rAcAP-5 (11.5 mg g-1 dry cell) and the maximum specific activity of rAcAP-5 (96% of total secreted protein). Copyright (C) 1999 Elsevier Science Inc. All rights reserved.

Original languageEnglish (US)
Pages (from-to)438-445
Number of pages8
JournalEnzyme and Microbial Technology
Volume24
Issue number7
DOIs
StatePublished - May 15 1999

Keywords

  • Ancylostoma caninum anticoagulant peptide (rAcAP-5)
  • Fed-batch fermentation
  • Pichia pastoris
  • Recombinant protein
  • Response surface methodology

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Biochemistry
  • Applied Microbiology and Biotechnology

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