Multi-stage laccase extraction and separation using aqueous two-phase systems: Experiment and model

Axel Prinz, Katharina Koch, Andrzej Górak, Tim Zeiner*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review


This work presents results of experimental and model investigation of continuous multi-stage enzyme extraction using aqueous two-phase systems for the first time. The aqueous two-phase system comprised polyethylene glycol 3000 and phosphate with additional sodium chloride buffered to pH 7. Two different laccases served as model enzymes. One of the laccases was directly taken from fungal culture supernatant, while the other laccase was solubilized lyophilisate. The modeling is based on an equilibrium stage approach. Equilibrium data were taken from single-stage experiments and approximated by different correlation equations. The model describes densities, phase equilibrium, enzyme activity partitioning between the phases. Moreover it allows to consider activity changes due to the aqueous two-phase system. Eight multi-stage mixer-settler experiments under varying operation conditions were performed to validate the proposed model; whereas the total throughput of all multi-stage extraction experiments was about 350 g h-1. The average relative deviation of modeled activities from experimentally measured activities was 23%. Therefore, the model is able to calculate the behavior of the phases as well as the partitioning of the two enzymes between the two phases for a multi-stage process based on single-stage data.

Original languageEnglish
Pages (from-to)1020-1031
Number of pages12
JournalProcess Biochemistry
Issue number6
Publication statusPublished - 2014


  • Aqueous two-phase extraction
  • Equilibrium-stage-modeling
  • Laccase
  • Multi-stage experiments
  • Scale-up

ASJC Scopus subject areas

  • Biochemistry
  • Applied Microbiology and Biotechnology
  • Bioengineering

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