Enzymatic hydrolysis of whey protein concentrates : peptide HPLC profiles

• Autor(a) principal: Mota, M. V. T.
• Data de Publicação: 2004
• Outros Autores: Ferreira, I. M. P. L. V. O., Oliveira, M. B. P., Rocha, Cristina M. R., Teixeira, J. A., Torres, D., Gonçalves, M. P.
• Tipo de documento: Artigo
• Idioma: eng
• Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
• Texto Completo: http://hdl.handle.net/1822/2613
• topic: Whey proteins; Enzymatic hydrolysis; RP-HPLC; Science & Technology

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.