Ice-Structuring Peptides Derived from Bovine Collagen

Abstract

Antifreeze proteins belonging to structurally diverse families of genetically coded proteins from several living organisms have been isolated and characterized in the past. This paper reports that collagen peptides of a certain molecular size range derived from Alcalase hydrolysis of bovine gelatin are able to inhibit recrystallization of ice in frozen ice cream mix as well as in frozen sucrose solutions in a manner similar to natural antifreeze proteins. The optimum conditions for producing such ice-structuring peptides (ISP) were hydrolysis at pH 9.0 for 30 min at 45 degrees C and an Alcalase-to-gelatin ratio of 0.176 unit per gram of gelatin. The collagen peptides were fractionated on size exclusion (Sephadex G-50) and ion exchange (sulfopropyl-Sephadex C-25) columns, and the molecular mass distribution of the ice-structuring peptide fractions was determined by matrix-assisted laser desorption ionization-time-of-flight (MALDI-TOF) mass spectrometry. The collagen peptide fractions in the molecular mass range of 600-2700 Da inhibited ice recrystallization in a supercooled ice cream mix and in concentrated sucrose solutions. The cationic collagen peptides within this fraction with molecular mass in the range of 1600-2400 Da were more effective than the anionic peptides in inhibiting ice crystal growth.