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Food Chemistry
Vol. 199, 2016, Pages: 409–415

pH-stat vs. free-fall pH techniques in the enzymatic hydrolysis of whey proteins

Ayoa Fernández, Phil Kelly

Teagasc and Food for Health Ireland (FHI), Moorepark Food Research Centre, Fermoy, Co. Cork, Ireland.


Enzymatic hydrolysis of a commercial whey protein isolate (WPI) using either trypsin or Protamex® was compared using controlled (pH-stat) and uncontrolled (free-fall) pH conditions. pH-stat control at the enzyme’s optimum value led to a more rapid rate of WPI hydrolysis by trypsin, while the opposite was the case when Protamex® was used. Furthermore, the choice of alkaline solution used to maintain constant pH during pH-stat experiments appeared to affect the reaction rate, being higher when KOH is added to the reaction mixture instead of NaOH. It would appear that potassium may play a role as co-factor or activator for the activity of this particular protease preparation. Although pH-stat techniques are usually considered to yield better hydrolysis kinetics, these findings suggest that the response of proteolytic enzyme preparations to static or free-fall pH control should be checked in advance, particularly when undertaking large scale production of WPI hydrolysates.

Keywords: Enzymatic hydrolysis; Whey proteins; Protease; pH-stat; Free-fall pH.

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