Applied Biochemistry and Biotechnology

, Volume 108, Issue 1, pp 787–797

Partition behavior and partial purification of hexokinase in aqueous two-phase polyethylene glycol/citrate systems

  • George G. G. Oliveira
  • Daniel P. Silva
  • Ines Conceição Roberto
  • Michele Vitolo
  • Adalberto PessoaJr.
Article

DOI: 10.1385/ABAB:108:1-3:787

Cite this article as:
Oliveira, G.G.G., Silva, D.P., Roberto, I.C. et al. Appl Biochem Biotechnol (2003) 108: 787. doi:10.1385/ABAB:108:1-3:787

Abstract

This study dealt with the partition behavior and partial purification of hexokinase (HK) from baker’s yeast by liquid-liquid extraction using aqueous two-phase polyethylene glycol (PEG)/citrate systems. First, we investigated the effect of agitation type (vortex and 8 rpm rotation) on the stability of the system, and then the effects of sodium citrate concentration, PEG concentration, and molar mass of PEG on the partition coefficient of this enzyme by using a 25 factorial experimental design. The results of this factorial experiment showed the possibility of a partial purification of HK by using two extraction steps, since the enzyme preferentially migrated to the top phase and the total proteins (mainly contaminants) remained in the bottom phase. The purification factor (PurTOP) of the enzyme in the top phase was 1.87, and the partition coefficient of the total proteins (KProt) was 0.47.

Index Entries

Hexokinase liquid-liquid extraction aqueous two-phase systems Saccharomyces cerevisiae 

Copyright information

© Humana Press Inc. 2003

Authors and Affiliations

  • George G. G. Oliveira
    • 1
  • Daniel P. Silva
    • 1
  • Ines Conceição Roberto
    • 2
  • Michele Vitolo
    • 1
  • Adalberto PessoaJr.
    • 1
  1. 1.Department of Biochemical and Pharmaceutical Technology, Faculty of PharmacyUniversity of São PauloSão Paulo, SPBrazil
  2. 2.DEBIQ-FAENQUILLorena, SPBrazil

Personalised recommendations