Department of Food Science, Faculty of Agricultural Sciences, Aarhus University, 8830 Tjele, Denmark.
J Dairy Sci. 2011 Oct;94(10):4787-99. doi: 10.3168/jds.2011-4343.
The aim of the present investigation was to study the underlying causes of noncoagulating (NC) milk. Based on an initial screening in a herd of 53 Danish Holstein-Friesians, 20 individual Holstein-Friesian cows were selected for good and poor chymosin-induced coagulation properties; that is, the 10 cows producing milk with the poorest and best coagulating properties, respectively. These 20 selected cows were followed and resampled on several occasions to evaluate possible changes in coagulation properties. In the follow-up study, we found that among the 10 cows with the poorest coagulating properties, 4 cows consistently produced poorly coagulating (PC) or NC milk, corresponding to a frequency of 7%. Noncoagulating milk was defined as milk that failed to form a coagulum, defined as increase in the storage modulus (G') in oscillatory rheometry, within 45min after addition of chymosin. Poorly coagulating milk was characterized by forming a weak coagulum of low G'. Milk proteomic profiling and contents of different casein variants, ionic contents of Ca, P and Mg, κ-casein (CN) genotypes, casein micelle size, and coagulation properties of the 4 NC or PC samples were compared with milk samples of 4 cows producing milk with good coagulation properties. The studies included determination of production of caseinomacropeptide to ascertain whether noncoagulation could be ascribed to the first or second phase of chymosin-induced coagulation. Caseinomacropeptide was formed in all 8 milk samples after addition of chymosin, indicating that the first step (cleavage of κ-CN) was not the cause of inability to coagulate. Furthermore, the effect of mixing noncoagulating and well-coagulating milk was studied. By gradually blending NC with well-coagulating milk, the coagulation properties of the well-coagulating samples were compromised in a manner similar to titration. Milk samples from cows that consistently produced NC milk were further studied at the udder quarter level. The coagulation properties of the quarter milk samples were not significantly different from those of the composite milk sample, showing that poor coagulation traits and noncoagulation traits of the composite milk were not caused by the milk quality of a single quarter. The milk samples exhibiting PC or NC properties were all of the κ-CN variant AA genotype, and contained casein micelles with a larger mean diameter and a lower fraction of κ-CN relative to total CN than milk with good coagulation properties. Interestingly, the relative proportions of different phosphorylation forms of α-CN differed between well-coagulating milk and PC or NC milk samples. The PC and NC milk samples contained a lower proportion of the 2 less-phosphorylated variants of α-CN (α(S1)-CN-8P and α(S2)-CN-11P) compared with samples of milk that coagulated well.
本研究旨在探讨非凝固(NC)牛奶的潜在原因。在对 53 头丹麦荷斯坦-弗里生牛进行初步筛选后,选择了 20 头个体荷斯坦-弗里生牛,以评估其凝乳酶诱导的良好和不良凝固特性;即,分别产生凝固特性最差和最好的牛奶的 10 头奶牛。对这 20 头选定的奶牛进行了多次随访和重新采样,以评估凝固特性的可能变化。在后续研究中,我们发现,在凝固特性最差的 10 头奶牛中,有 4 头奶牛始终产生活性差或非凝固(NC)牛奶,频率为 7%。非凝固牛奶是指在添加凝乳酶后 45 分钟内,未能形成凝乳的牛奶,这通过振荡流变仪中存储模量(G')的增加来定义。产生活性差凝乳的牛奶的特征是形成弱凝乳,其 G'低。将 4 份 NC 或 PC 样品的牛奶蛋白质组学分析和不同酪蛋白变体的含量、Ca、P 和 Mg 的离子含量、κ-酪蛋白(CN)基因型、酪蛋白胶束大小以及 4 份 NC 或 PC 样品的凝固特性与 4 份产生活性好凝乳的牛奶样品进行了比较。这些研究包括测定酪蛋白巨肽的产生,以确定非凝固是否归因于凝乳酶诱导凝固的第一阶段或第二阶段。在添加凝乳酶后,所有 8 份牛奶样品中都形成了酪蛋白巨肽,表明第一阶段(κ-CN 的切割)不是不能凝固的原因。此外,还研究了混合非凝固和凝固良好的牛奶的效果。通过逐渐将 NC 与凝固良好的牛奶混合,凝固良好的牛奶样品的凝固特性以类似于滴定的方式受到损害。持续产生 NC 牛奶的奶牛的牛奶样品在乳房象限水平进一步研究。乳房象限牛奶样品的凝固特性与复合牛奶样品的凝固特性没有显著差异,表明复合牛奶的凝固不良特性和非凝固特性不是由单个象限的牛奶质量引起的。表现出 PC 或 NC 特性的牛奶样品均为 κ-CN 变体 AA 基因型,并且与凝固特性良好的牛奶相比,其酪蛋白胶束的平均直径较大,κ-CN 相对于总 CN 的比例较低。有趣的是,与凝固良好的牛奶和 PC 或 NC 牛奶样品相比,不同磷酸化形式的 α-CN 的相对比例不同。与凝固良好的牛奶相比,PC 和 NC 牛奶样品中 2 种磷酸化程度较低的 α-CN 变体(α(S1)-CN-8P 和 α(S2)-CN-11P)的比例较低。