Physicochemical Properties of Milk Fat Emulsions Stabilized with Bovine Milk Fat Globule Membrane

Milk fat globules (MFG) were reconstituted with milk fat globule membrane (MFGM) and milk fat (MF). Viscosity of the reconstituted MFG was highest at pH 5.0 and 4 min emulsifying, and rose with an increase of MFGM between 40–80 mg/g fat. Adsorbed protein/unit fat increased at acid pH with increase of MFGM. The composition of proteins adsorbed on the surface of MFG was not influenced by factors of reconstitution. The size and specific surface area of globules were influenced by emulsifying time, MFGM and MF concentrations, and pH. The size range of MFG prepared by standard method was 0.9–17 μm in diameter. Median diameter was 5 μm and specific surface area was 15,600 cm²/cm³ of emulsion.     

人乳与牛乳乳脂肪球膜蛋白质组的对比研究

人乳是婴幼儿生命初始阶段唯一能够摄入的食物,蛋白质是人乳中重要的营养成分,而牛乳作为代替人乳的常用原料已经被广泛的应用于婴幼儿食品中。本研究利用SDS-PAGE电泳和LC-MALDL-TOF蛋白组学方法将人乳与牛乳中乳脂肪球膜蛋白进行分离,能够发现人乳与牛乳脂肪球膜蛋白存在较大的差异。牛乳脂肪球膜中已鉴定出488种蛋白,人乳脂肪球膜中鉴定出的蛋白为1545种。牛乳脂肪球膜具有173种特异性蛋白,人乳脂肪球膜具有1230种特异性蛋白,在人乳与牛乳中存在315种同源蛋白。从蛋白质的GO(Gene Ontology)功能注释上来看,人乳脂肪球膜蛋白参与的生物过程有37%为代谢过程;具有的分子功能55%为结合作用;34%为参与细胞器构成。人乳脂肪球膜中有24种蛋白参与免疫相关的通路,主要为抗原加工和呈递。与牛乳相比,对人乳中乳脂肪球膜蛋白质在组成及功能上的研究,能够促进深入地了解人乳蛋白,并为以牛乳为原料的婴幼儿产品添加功能性蛋白提供参考。     

母乳和牛乳中乳脂肪球膜蛋白质的差异分析

为比较乳脂肪球膜（milk fat globule membrane,MFGM）蛋白在母乳和牛乳中的差异,采用有机试剂提取法,从母乳和牛乳中提取分离出MFGM蛋白,经皮尔斯TM去垢小柱去除十二烷基硫酸钠后,通过液相色谱-质谱联用技术检测分析,检测结果根据特异性肽段匹配数不小于1筛选后,在母乳和牛乳中分别检测到863 种和454 种蛋白,其中二者共有175 种,分别独有688 种和279 种。将母乳中差异表达的688 种蛋白质按PANTHER数据库进行蛋白质的Gene Ontology（GO）功能注释,这些蛋白主要是细胞部分、细胞器、细胞膜等组分,参与的生物途径有新陈代谢过程、生物调节、刺激性反应、免疫系统过程、再生作用等,具有催化、黏合、转运、酶调节等多种分子功能。另外,母乳中的这些差异蛋白参与氨酰tRNA生物合成、致病性大肠杆菌感染、脂肪酸代谢、丙酸代谢、甾体合成、酸降解等都是牛乳中的乳脂肪球膜蛋白不具有的功能。通过分析得出母乳和牛乳中的MFGM蛋白具有明显差异,虽然有部分组成和功能的重叠,但在丰度和代谢路径上,牛乳MFGM蛋白不能替代母乳MFGM蛋白。     

牛乳和双峰驼乳中乳脂球膜蛋白的差异分析

为阐明双峰驼乳中乳脂球膜蛋白（milk fat globule membrane proteins,MFGMP）的组成以及与牛乳MFGMP之间的差异,以牛乳和双峰驼乳中的MFGMP为研究对象,采用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳、双向电泳和液相色谱-串联质谱（liquid chromatograph-tandem mass spectrometry,LC-MS/MS）技术对其中的高丰度蛋白质进行表征,并对双峰驼乳MFGMP提取方法进行选择。结果显示,双峰驼乳和牛乳MFGMP中含量最高的蛋白组分依次为嗜乳脂蛋白、黄嘌呤氧化还原酶/脱氢酶、乳凝集素（MFG-E8或PAS6/7）、脂肪分化相关蛋白（ADRP或PLIN2）,3 种方法分析双峰驼乳和牛乳中MFGMP中高丰度蛋白组分的结果具有一致性。基于LC-MS/MS技术对双峰驼乳的MFGMP进行鉴定,并将对鉴定出的蛋白质进行基因本体功能注释和京都基因与基因组百科全书路径分析,发现这些蛋白主要是细胞膜、线粒体、内质网膜等组分,参与的生物途径有翻译和翻译的起始、氧化还原过程、细胞内蛋白质转运、蛋白质折叠、细胞氧化还原稳态等,具有结合活性和分子催化等分子功能。     

Role of Milk Fat Globule Membrane in Autoxidation of Milk Fat

The influence of the milk fat globule membrane (MFGM) on oxidation of milk fat under various conditions was monitored by fatty acid analysis and oxygen consumption at 50°C. The rate of copper-catalyzed oxidation of cream containing the membrane was faster than when the membrane was removed. In the dry state, the isolated MFGM, membrane lipids (ML) and the nonlipid membrane solids (NLMS) inhibited oxidation at 50°C and 95°C. In the presence of water, however, all membrane components enhanced oxidation at 50°C. The balance between the accelerating effects of membrane components and the inhibiting influence of other factors appeared to depend on processing conditions.     

牛乳脂肪球膜组分分离鉴定及其功能性研究进展

乳脂肪球膜（milk fat globule membrane,MFGM）是牛乳中脂肪球外表面包裹着的一层膜,在牛乳中的含量非常微小。但由于其具有良好的生理活性以及潜在的食品加工应用潜力,关于其展开的基础研究和应用基础研究受到广泛关注。本综述介绍MFGM的分离制备技术、结构、组分鉴定分析、生理活性及其应用特性的研究进展,以期为MFGM在食品工业尤其是在功能性食品中的应用提供一定的理论支持。     

乳脂肪球膜的特性、开发及在模拟母乳脂肪球结构中的应用

乳脂肪球膜（milk fat globule membrane,MFGM）是包裹在天然乳脂肪球外部的3 层膜状结构,然而牛乳基和大豆基婴儿配方奶粉缺少MFGM,脂肪球结构与母乳存在较大差异,因此添加外源MFGM以及制备与母乳脂肪球结构接近的婴儿配方奶粉成为了近期的研究焦点。本文综述了MFGM的相关特性和生产开发途径,以及牛乳MFGM在仿母乳脂肪球结构乳液和婴儿配方奶粉中的应用。体外模拟婴儿胃肠道消化实验以及啮齿动物体内实验结果表明,仿母乳脂肪球结构乳液和婴儿配方奶粉能够促进婴儿脂肪消化并且改善脂质代谢过程。     

乳脂肪球膜及其配料的科学共识

目的:富含乳脂肪球膜的乳清蛋白粉是婴幼儿配方乳粉的配料之一。有关专家在综合分析乳脂肪球膜对婴幼儿营养健康作用的研究文献基础上,形成《乳脂肪球膜及其配料的科学共识》。方法:组织科技界和产业界的相关专家,通过文献检索分析与研讨的形式开展研究。结果:乳脂肪球膜是包裹在乳脂肪液滴表面,由极性脂质、胆固醇和蛋白质等组成的复杂的3层磷脂蛋白膜,而鞘磷脂和神经节苷脂为乳脂肪球膜配料的特征性指标,其安全性和耐受性在临床试验中得到证实。现有研究表明在婴幼儿配方乳粉中添加“富含乳脂肪球膜的乳清蛋白粉”,可促进婴儿大脑认知发育,增强婴儿免疫力。目前乳脂肪球膜原料生产企业尚无统一的质量标准体系,乳脂肪球膜相关配料的组分复杂,且因获得方法不同而存有较大差异。对原料和终产品的检测方法,特别是对膜蛋白的检测方法尚不成熟,应加强乳脂肪球膜配料及主要特征成分检测方法、产品质量控制、营养功能与临床研究。政府有关部门、食品科技界及产业界也应加强对乳脂肪球膜及其配料的认知,助推我国婴幼儿配方食品产业健康发展。     

Effect of Milk Fat Globule Membrane in Reversing Heme-Stimulated Bacterial Growth in Whey

Growth of mastitic pathogenic bacteria was followed by turbidometry in hemin-enriched bovine whey. Hemin clearly promoted growth of bacteria. The milk fat globule membrane, when added to the hemin-enriched media, abolished much of the growth-enhancing effect offered by hemin. A Scatchard plot of hemin-binding showed that hemin was bound to the milk fat globule membrane by a specific mechanism as well as nonspecifically. Heme-binding to the milk fat globule membrane should be considered an important antibacterial mechanism in milk.     

乳脂肪球膜的组成、营养及制备研究进展

乳中的脂肪以乳化的脂肪球形态分散于水相中,脂肪球被三层脂肪球膜包裹,近年来的研究表明脂肪球膜中的成分对婴幼儿的生长发育有至关重要的作用。本文介绍了乳脂肪球膜特别是其磷脂的组成及含量,添加入婴幼儿配方奶粉中乳脂肪球膜的营养作用,以及目前市售的富含脂肪球膜的产品及制备方式,为缩小配方奶与婴幼儿营养需求的差异提供科学依据。     

The Macrostructure of Milk Lipids: The Fat Globules

The aim of this review article is to summarize the information available related to milk fat globules (MFGs) in order to highlight their contribution to the nutritional and technological characteristics of milk and dairy products. The macrostructure of milk lipids is composed of globules made up of triglycerides with different melting points, enveloped by a biological membrane from the mammary epithelial cell. In milks of different animal species, there are different-sized MFGs, ranging from diameters of less than 0.2 μm to a maximum of 15 μm. The average diameter and the number of globules are affected by endogenous, physiological, and exogenous factors. The size of the globules in turn affects the qualitative characteristics of milk and cheese. In addition, the average diameter of the globules and their surface that is exposed to the digestive system affect fat digestibility in different ways. Finally, the components of the MFG membranes have been shown to take part in the secretion process of globules and to have a beneficial effect on human health. In conclusion, by acting on factors influencing the dimensions of the fat globules and by increasing the content of the milk membrane could help adapt milk production to specific consumer targets and improve milk nutritional properties.     

驴初乳与常乳乳脂球膜蛋白质组的对比分析

为阐明驴初乳与常乳中乳脂球膜蛋白的差异,利用蛋白质组学技术对二者进行蛋白质组差异分析。在驴初乳和驴常乳中分别鉴定到216 种和215 种乳脂球膜蛋白,并在其中筛选出40 种差异蛋白,包括15 种差异表达蛋白和25 种特异表达蛋白。将驴初乳与常乳中的差异蛋白进行生物信息学分析发现,差异蛋白主要参与的细胞组成为细胞外泌体、胞外囊泡、胞外细胞器等;主要参与的生物过程为对外部刺激的反应过程、细胞增殖过程、血管形态生成过程等;主要参与的分子功能为金属离子结合、阳离子结合、钙离子结合等。此外,这些差异蛋白主要参与补体和凝血级联,为生成IgA而形成的肠道免疫网络等代谢通路。利用蛋白质网络互作分析发现,差异蛋白中存在具有高连接度的关键乳脂球膜蛋白因子。本研究有助了解驴乳脂球膜蛋白的生物学特性,并为驴乳中乳脂球膜蛋白的营养学研究及相关配方乳粉的研发提供理论基础。     

Milk Fat Globule Membrane Substances Inhibit Mouse Intestinal β-Glucuronidase

The β-glucuronidase inhibitory action of milk fat globule membrane (MFGM) was studied. By increasing the MFGM concentration to 0.2%, nearly 90% of the β-glucuronidase activity was inhibited. The trypsin-derived digests of MFGM inhibited β-glucuronidase activity by ～ 42%. The inhibitory effect of MFGM on intestinal β-glucuronidase of mouse was studied. We administered MFGM (groups I: 5%; group II: 10%; group III: 20%) to mice and investigated the time course of changes in fecal β-glucuronidase activity. In group I and II, enzyme activity had decreased 15–20% on the 5th day of treatment, but in group III, a decrease in enzyme activity was seen with an inhibition rate of 50%, showing a marked intracolonic efficacy of MFGM. Thus MFGM may have potential in explaining, treating or preventing intestinal cancers.     

添加富含乳脂肪球膜乳清蛋白粉对发酵乳代谢组学的影响

目的:研究添加富含乳脂肪球膜(MFGM)乳清蛋白粉(M-WPI)前、后的保加利亚乳杆菌发酵乳中保加利亚乳杆菌存活率,发酵乳风味及抗氧化能力。采用代谢组学分析添加M-WPI前、后保加利亚乳杆菌发酵乳的显著差异代谢标志物及其作用通路。方法:通过样本主成分分析、偏最小二乘判别分析、热图分析和KEGG富集分析等方法确定加入M-WPI前、后发酵乳代谢过程中的差异性代谢产物,并以此分析M-WPI对发酵乳产生的影响。结果表明:在脱脂乳中加入M-WPI,可以提高保加利亚乳杆菌存活率,增加发酵乳的风味及抗氧化活性。采用主成分分析(PCA)和偏最小二乘判别分析(PLS-DA)共确定77种显著差异代谢物,涉及的主要代谢通路为19条。加入M-WPI后葡萄糖、半乳糖等,代谢物上调,提高了保加利亚乳杆菌存活率;癸酸、苹果酸等有机酸上调,能提高发酵乳的口感及改善发酵乳风味。保加利亚乳杆菌发酵乳中上调的代谢物以氨基酸类较多,其中丙氨酸、天冬氨酸和泛酸等参与β-丙氨酸代谢,与发酵乳抗氧化活性提高有关。本研究为加入M-WPI发酵乳营养评价以及相关加工技术和产品开发提供参考。     

牛初乳与牛常乳乳脂肪球膜中丰度差异蛋白的表征与分析

为阐明不同泌乳期间牛乳乳脂肪球膜蛋白的差异,选定牛初乳和牛常乳中的乳脂肪球膜蛋白作为研究对象,利用定量蛋白质组学技术对其中的蛋白质进行表征,并对两组间的丰度差异蛋白进行筛选及生物信息学分析。本研究共鉴定到763 种蛋白,其中197 种为两组共有蛋白,进一步从其中筛选出80 种丰度差异蛋白,包括41 种上调蛋白和39 种下调蛋白（牛初乳/牛常乳）。对上述丰度差异蛋白进行生物信息学分析发现,这些蛋白参与的主要细胞组分为细胞外泌体、细胞外空间、细胞外区域等,参与的主要代谢通路为代谢途径、嘌呤代谢、苯丙氨酸代谢、酪氨酸代谢、丙酮酸代谢、糖酵解/糖异生等,并进一步筛选出31 种与其他蛋白存在相互作用的关键蛋白,包括结合珠蛋白、2-磷酸-D-甘油酸水解酶等。研究结果将有助于了解泌乳期间乳蛋白成分的差异及其功能性,为牛乳制品的精深加工奠定基础。     

不同均质工艺处理对巴氏杀菌乳脂肪球膜蛋白质组成的影响

乳脂肪球大小和膜蛋白组分的变化决定了脂肪球稳定性,从而影响巴氏杀菌乳在贮藏期内的品质稳定性。本研究通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和液相色谱-质谱联用法分析不同预热温度（50、60、70?℃）和不同均质压力（20、30、40、45?MPa）对脂肪球膜蛋白的影响,并通过稳定性分析仪探究脂肪球膜蛋白变化与稳定性的关系。结果表明,均质工艺对脂肪球膜蛋白种类无显著影响,但对脂肪球膜蛋白的含量有一定影响：黄嘌呤氧化还原酶（xanthine dehydrogenase/oxidase,XO）、嗜乳脂蛋白（butyrophilin,BTN）、脂肪滴结合蛋白（adipophilin,ADPH）在预热60 ℃时的含量最低,黏液素（mucin 1,MUC1）在预热60 ℃时的含量最高,脂肪酸结合蛋白（fatty acid binding protein,FABP）、组织糖蛋白高碘酸烯夫6/7（periodic acid Schiff 6/7,PAS6/7）的含量随着预热温度的升高而减少,组织糖蛋白高碘酸薛夫IV（cluster of differentiation 36,CD36）的含量随着预热温度的升高而增多;XO、PAS6/7、BTN、ADPH、FABP在均质压力40 MPa时含量最低,CD36的含量随均质压力的增加而减少;脂肪球膜蛋白构成中,牛血清白蛋白（bovine serum albumin,BSA）、αs1-酪蛋白（casein,CN）和β-CN含量增加,其中BSA含量增加最明显。综上,推测均质工艺导致αs1-CN、β-CN和BSA组分参与了脂肪球膜的构建,并对脂肪球稳定性产生了积极的影响。     

Interactive Effects of Milk Fat Globule and Casein Micelle Size on the Renneting Properties of Milk

The size of the casein micelles (CM) and the milk fat globules (MFG) vary depending on farming factors, such as seasonal variation and stage of lactation, and cow genetics. The MFG and CM size of milk can influence the renneting behavior and texture of manufactured dairy products. In this work, we investigated the combined effects of MFG and CM size on the onset of gelation, the maximum rate of gelation, the value for G′_60 min (the final storage modulus) and G″_60 min (the final loss modulus), and tan δ upon renneting. Fractionation of MFG on the basis of size was carried out using laboratory-based centrifugation, whereas milk of predominantly large (184–218 nm) or small (147–159 nm) CM was selected naturally on-farm. Casein micelle size had the dominant effect on curd firmness and gelation rates of milk, where small CM milk formed rennet gels earlier and resulted in a firmer gel than milk with large CM. However, MFG size also influenced the renneting properties. The strongest rennet gels were obtained when large MFG (3.88–5.78 μm) was combined with small CM (153–159 nm). Selecting milk on the basis of the microstructure of key milk components could be achieved by natural selection of dairy cows or via fractionation technologies. Selection may provide a useful tool for efficient manufacturing of different dairy products based on the desirable characteristics specific to each.