Continuous extraction of alpha- and beta-amylases from Zea mays malt in a PEG4000/CaCl2 ATPS

J Chromatogr B Analyt Technol Biomed Life Sci. 2007 Oct 15;858(1-2):227-33. doi: 10.1016/j.jchromb.2007.08.042. Epub 2007 Sep 8.

Abstract

In the present work, alpha- and beta-amylase enzymes from Zea mays malt were recovered by continuous extraction in a PEG/CaCl2 aqueous two-phase system (ATPS). The influences of the flux rate (RQ), free area of vane (A(free)) and vane rotation (RV) on enzyme recovery were studied by optimization using response surface methodology (RSM). The protein content and enzyme activity were measured from time to time in the extract and refined fluxes. RSM curves showed a squared dependence of recovery index with the RQ, A(free) and RV. The best system for recovering the maize malt enzymes was with low vane rotation and flux rate and high free area of vane. Alpha- and beta-amylases were purified 130-fold in the salt-rich phase.

MeSH terms

  • Calcium Chloride / chemistry
  • Chemical Fractionation / instrumentation
  • Chemical Fractionation / methods*
  • Polyethylene Glycols / chemistry
  • Reproducibility of Results
  • Zea mays / chemistry*
  • alpha-Amylases / chemistry
  • alpha-Amylases / isolation & purification*
  • beta-Amylase / chemistry
  • beta-Amylase / isolation & purification*

Substances

  • Polyethylene Glycols
  • alpha-Amylases
  • beta-Amylase
  • Calcium Chloride