Author(s):
Mota, M. V. T. 
; Ferreira, I. M. P. L. V. O. ; Oliveira, M. B. P. ; Rocha, Cristina ; Teixeira, J. A. ; Torres, D. ; Gonçalves, M. P.
Date: 2004
Persistent ID: http://hdl.handle.net/1822/2613
Origin: RepositóriUM - Universidade do Minho
Subject(s): Whey proteins; Enzymatic hydrolysis; RP-HPLC
Description
Hydrolysis of whey protein concentrates (WPCs) at different temperatures
and pHs, using three enzymes: pepsin, trypsin, and Alcalase®, was
monitored during more than 5 hr by reversed phase HPLC/UV, using a column containing a polystyrene-divinylbenzene copolymer-based
packing, and an elution gradient from 8% to 80% acetonitrile containing
0.1% TFA. Peptides were separated according to their polarity and size,
and degradation of α-lactalbumin (α-la) and β-lactoglobulin (β-lg)
was evaluated. The three proteolytic enzymes (pepsin, trypsin, and
Alcalase®) employed for hydrolysis of WPCs led to different kinetics
of degradation of β-lg. α-la degradation after 15 min was almost complete
for the three enzymes. The hydrolysis catalysed by each enzyme resulted
in different peptide profiles by HPLC/UV. Hydrolysates produced by
pepsin (HP) were resolved into three main fractions of high retention
times, while tripsin hydrolysates (HT) were resolved into nine major
peaks and Alcalase® hydrolysates (HA) were resolved into 12 major
peaks, presenting a wide range of polarities and sizes. Although, with
different β-lg hydrolysis extension, chromatographic profiles of the
degradation and formation of peptides can be used as a finger print of
the type of enzyme used, because peptide profile is not affected either
by temperature or pH.
