Development of the milk fat microstructure during the manufacture and ripening of Emmental cheese observed by confocal laser scanning microscopy

Changes in the physico-chemical properties and microstructure of milk fat globules were investigated during the manufacture and ripening of Emmental cheese. The measurement of fat globule size and apparent zeta-potential showed that they were slightly affected during cheese milk preparation, i.e. storage of cheese milk overnight at 4 °C and pasteurisation. After rennet-induced coagulation and heating of curd grains, coalescence caused the formation of large fat globules (i.e.>10 μm). The structure of fat in Emmental cheese was characterised in situ using confocal laser scanning microscopy (CLSM). The rennet-induced coagulation lead to the formation of a continuous network of casein strands in which fat globules of various sizes were entrapped. Heating of curd grains induced the formation of fat globule aggregates. Pressing of the curd grains resulted in the greatest disruption of milk fat globules, their coalescence, the formation of non-globular fat (free fat) and the release of the milk fat globule membrane (MFGM) material. This study showed that milk fat exists in three main forms in ripened Emmental cheese: (i) small fat globules enveloped by the MFGM; (ii) aggregates of partially disrupted fat globules and (iii) free fat, resulting from the disruption of the MFGM and allowing free triacylglycerols to fill voids in the protein matrix. The curd grain junctions formed in Emmental cheese were also characterised using CLSM: they are compact structures, rich in protein and devoid of fat globules.