title: Proteomic comparison of equine and bovine milks on renneting.
abstract: Rennet-induced coagulation of bovine milk is a complex mechanism in which chymosin specifically hydrolyzes κ-casein, the protein responsible for the stability of the casein micelle. In equine milk, this mechanism is still unclear, and the protein targets of chymosin are unknown. To reveal the proteins involved, the rennetability of equine milk by calf chymosin was examined using gel-free and gel-based proteomic analysis and compared to bovine milk. RP-HPLC analysis of bovine and equine milks showed the release of several peptides following chymosin incubation. The hydrolyses of equine and bovine casein by chymosin were different, and the major peptides produced from equine milk were identified by mass spectrometry as fragments of β-casein. Using two-dimensional electrophoresis, equine β-casein was confirmed as the main target of calf chymosin over 24 h at 30 °C and pH 6.5. The gel-based analysis of equine milk discriminated between the different individual proteins and provided information on the range of isoforms of each protein as a result of post-translational modifications, as well as positively identified for the first time several isoforms of κ-casein. In comparison to bovine milk, κ-casein isoforms in equine milk were not involved in chymosin-induced coagulation. The intensity of equine β-casein spots decreased following chymosin addition, but at a slower rate than bovine κ-casein.
authors: School of Food and Nutritional Sciences, University College , Cork, Ireland.
authors: Therese
authors: T
authors: Uniacke-Lowe
authors: François
authors: F
authors: Chevalier
authors: Sonia
authors: S
authors: Hem
authors: Patrick F
authors: PF
authors: Fox
authors: Daniel M
authors: DM
authors: Mulvihill
mesh_terms: D000818:Animals; D055162:Biocatalysis; D002364:Caseins / Q000737:chemistry; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D012085:Chymosin / Q000737:chemistry*; D015180:Electrophoresis, Gel, Two-Dimensional; D006736:Horses; D008892:Milk / Q000737:chemistry*; D008894:Milk Proteins / Q000737:chemistry*; D040901:Proteomics
chemical_list: D002364:Caseins; D008894:Milk Proteins; C025350:rennet; D012085:Chymosin