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.
Subject(s)
Chemical Fractionation/methods , Zea mays/chemistry , alpha-Amylases/isolation & purification , beta-Amylase/isolation & purification , Calcium Chloride/chemistry , Chemical Fractionation/instrumentation , Polyethylene Glycols/chemistry , Reproducibility of Results , alpha-Amylases/chemistry , beta-Amylase/chemistryABSTRACT
In this work the purification and biochemistry characterization of alpha-amylases from Aspergillus niger (FORILASE NTL) were studied. The effects of expansion degree of resin bed on enzyme purification by expanded bed adsorption (EBA) have also been studied. Residence time distributions (RTD) studies were done to achieve the optimal conditions of the amylases recovery on ion-exchange resin, and glucose solution was used as a new tracer. Results showed that height equivalent of the theoretical plates (HETP), axial dispersion and the Prandt number increased with bed height, bed voidage and linear velocity. The adsorption capacity of alpha-amylases, on the resin, increased with bed height and the best condition was at four-expansion degree. alpha-Amylase characterization showed that this enzyme has high affinity with soluble starch, good hydrolysis potential and molecular weight of 116 kDa.
