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Cell Co-culture Patterning Using Aqueous Two-phase Systems
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Continuous laccase concentration in an aqueous two-phase system.

Michał Blatkiewicz1, Anna Antecka1, Andrzej Górak1,2

  • 11Faculty of Process and Environmental Engineering, Lodz University of Technology, ul. Wólczańska 213, 90-942 Lodz, Poland.

Chemicke Zvesti
|March 24, 2018
PubMed
Summary

A new method efficiently separates Cerrena unicolor laccase using a PEG 6000-phosphate system. This approach enables continuous, one-step extraction with high recovery and concentration, validated by experimental data.

Keywords:
ATPSDownstream processingLaccaseModel

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Area of Science:

  • Biochemical Engineering
  • Enzyme Technology
  • Separation Science

Background:

  • Laccase enzymes are valuable biocatalysts with diverse industrial applications.
  • Efficient separation and purification of laccase from microbial cultures are crucial for its economic viability.
  • Aqueous two-phase systems (ATPS) offer a promising, scalable method for biomolecule partitioning.

Purpose of the Study:

  • To develop and validate a continuous partitioning approach for Cerrena unicolor laccase.
  • To establish an empirical short-cut method for predicting ATPS performance in laccase extraction.
  • To assess the efficiency of a mixer-settler unit for one-step laccase concentration and recovery.

Main Methods:

  • Batch experiments were conducted to determine phase equilibrium and laccase activity in a PEG 6000-phosphate ATPS.
  • An empirical short-cut method was developed for calculating key partitioning parameters (mass/volume fractions, concentration factor, recovery).
  • Continuous extraction experiments were performed using a mixer-settler unit, with automatic substrate feed and phase collection.

Main Results:

  • The developed short-cut method accurately predicted phase compositions, volume fractions, concentration factors, and enzymatic yields within a 10% error range.
  • A one-step extraction process in the mixer-settler unit successfully achieved the required laccase concentration.
  • Continuous operation demonstrated good agreement between predicted and experimentally measured partitioning parameters at steady state.

Conclusions:

  • The proposed approach provides an effective and predictable method for continuous laccase partitioning in ATPS.
  • The empirical short-cut method simplifies the design and operation of ATPS for enzyme purification.
  • The mixer-settler unit is suitable for scalable, one-step extraction of concentrated laccase.