Standardized reaction times used to describe the mechanism of enzyme-induced gelation in whey protein systems

• Published in: Journal of dairy research Vol. 67; no. 3; p. 403
• Main Authors: Ipsen, R, Otte, J, Lomholt, S B, Qvist, K B
• Format: Journal Article
• Language: English
• Published: England 01.08.2000
• Subjects: Bacillus - enzymology; Gels; Hydrolysis; Milk Proteins - chemistry; Milk Proteins - metabolism; Peptides - metabolism; Protein Denaturation; Serine Endopeptidases - metabolism; Sodium Chloride; Temperature; Time Factors; Whey Proteins
• ISSN: 0022-0299
• DOI: 10.1017/S0022029900004337

Whey protein isolate (WPI), either untreated or pretreated at 80 degrees C for 30 min, was incubated with a proteinase from Bacillus licheniformis until a gel was formed. Standardized reaction times, directly linked to the degree of hydrolysis, were obtained from plots of the relative amount of peptides released v. reaction time obtained under different conditions (enzyme concentration, temperature, pH, NaCl addition). This provided a connection between the gelation profile and the degree of hydrolysis. In the case of untreated WPI, gelation occurred at lower degrees of proteolysis when the enzyme concentration was decreased, demonstrating that a rate-limiting aggregation process occurred at the same time as the proteolysis in a manner similar to the renneting of milk. This was not the case for preheated WPI, when gelation was found to take place at a constant degree of proteolysis, independent of the enzyme concentration. In this case, the mechanism could be described by assuming the thermally induced aggregates present in this substrate had progressively more stabilizing peptide segments shaved off, resulting in increased attraction between individual aggregates that ultimately led to gelation. Results obtained at 40-60 degrees C supported this, as we found no effect of temperature on the degree of proteolysis at gelation for the untreated WPI, whereas the degree of proteolysis decreased with increasing temperature when heated WPI was hydrolysed. The effect of pH and NaCl addition on the process was to reduce repulsion between the aggregating species so that gelation was induced at a decreased degree of proteolysis.