Assessing the iron chelation capacity of goat casein digest isolates

We isolated goat phosphopeptides via calcium and ethanol precipitation from a caseinate digest and investigated their feasibility as an iron-fortification ingredient in nutritional foods. Goat tryptic-digested phosphopeptides could bind 54.37 ± 0.50 mg of Fe/g of protein compared with goat milk, which could bind 3.83 ± 0.01 mg of Fe/g of protein, indicating that isolation did increase iron binding. However, the >13-fold increase in iron binding was only partly explained by the increased concentration of phosphoserine-rich residues in the isolated fraction: we observed a 77% increase in serine residue content and a 5.9-fold increase in phosphorus in the goat peptide isolate compared with the starting caseinate material. We investigated the effect of potential industrial processing conditions (including heating, cooling, holding time, and processing order) on iron binding by the tryptic-digested phosphopeptides. In addition, we tested the effect of ionic strength and the addition of peptides to a milk system to understand how food formulations could affect iron binding.     

Short communication: casein haplotype variability in sicilian dairy goat breeds

In the Mediterranean region, goat milk production is an important economic activity. In the present study, 4 casein genes were genotyped in 5 Sicilian goat breeds to 1) identify casein haplotypes present in the Argentata dell’Etna, Girgentana, Messinese, Derivata di Siria, and Maltese goat breeds; and 2) describe the structure of the Sicilian goat breeds based on casein haplotypes and allele frequencies. In a sample of 540 dairy goats, 67 different haplotypes with frequency ≥0.01 and 27 with frequency ≥0.03 were observed. The most common CSN1S1-CSN2-CSN1S2-CSN3 haplotype for Derivata di Siria and Maltese was FCFB (0.17 and 0.22, respectively), whereas for Argentata dell’Etna, Girgentana and Messinese was ACAB (0.06, 0.23, and 0.10, respectively). According to the haplotype reconstruction, Argentata dell’Etna, Girgentana, and Messinese breeds presented the most favorable haplotype for cheese production, because the casein concentration in milk of these breeds might be greater than that in Derivata di Siria and Maltese breeds. Based on a cluster analysis, the breeds formed 2 main groups: Derivata di Siria, and Maltese in one group, and Argentata dell’Etna and Messinese in the other; the Girgentana breed was between these groups but closer to the latter.     

Short communication: Polymorphism of casein cluster genes in Czech local goat breeds

The 4 casein loci were evaluated as haplotypes in 2 dairy goat breeds kept in the Czech Republic. Analysis of 41 White Shorthaired (WSH) trio families and 44 Brown Shorthaired (BSH) trio families revealed 14 and 20 haplotypes, respectively. Various genomic techniques were used to type the casein loci. Twenty-two different combinations of these alleles (casein haplotypes, in the order CSN1S1-CSN2-CSN1S2-CSN3) were found. Only 5 haplotypes in the WSH breed and 6 haplotypes in the BSH breed occurred at frequencies >0.05. For the WSH breed, the most common haplotype was FCFB (0.260), whereas for the BSH breed, the most common haplotype was FCFA (0.217). The information on the haplotype variability in both breeds could be used in breeding programs aimed at preserving biodiversity or selecting animals for specific protein production and cheesemaking.     

Comparative study on the microstructure and functional properties of casein in goat milk processed by different methods

This work compared the thermal stability, microstructure and functional properties of casein (CN) in goat milk processed by different methods, including homogenisation, low-temperature long-time (LTLT), high-temperature short-time (HTST), ultrapasteurisation (UP) and ultrahigh temperature (UHT) treatment. The greatest development of CN-whey protein complexes was observed in LTLT goat milk and the least in HTST goat milk. The CN micelles in LTLT, UP and UHT milk were more obviously changed than the CN micelles in HTST milk, and the random coil content increased significantly after the homogenisation, UP and UHT treatments. The homogenisation had a significant effect on the functional properties of CN. In addition, the most obvious changes in the properties of CN were observed in LTLT and UHT goat milk. The results suggested that homogenisation, pasteurisation and UHT can cause effective changes in the functional properties of CN in goat milk that facilitate its application in the milk industry.     

牛羊乳酪蛋白制备及电泳分析比较

采采用新鲜和复原的牛羊乳为原料,等电点沉淀,通过洗涤、干燥等步骤分别制得牛乳和羊乳酪蛋白,用凯氏定氮法对其进行定量检测,应用十二烷基硫酸钠聚丙烯酰胺凝胶电泳（SDS—PAGE）分析比较牛羊乳酪蛋白组分差异。结果表明：牛乳酪蛋白得率为86．75％,羊乳酪蛋白得率为90．55％,高含量的酪蛋白主要集中在电泳图谱的中分子量组,可分为ds,CN、as2-CN、β-CN和K—CN,牛羊乳as2-CN分子量羊乳大于牛乳,牛乳a-CN含量比较多,羊乳β—CN含量比较多,鲜乳与复原乳全蛋白主要组分在电泳图谱中除酪蛋白差别外,上端的高分子量组清蛋白区羊乳IgG重链的分子量比牛乳小。应用蛋白质电泳分析技术可以区分羊乳和牛乳的蛋白质组分,等电点沉淀法制备酪蛋白的方法简单,易于操作。     

Physicochemical and functional properties of goat milk whey protein and casein obtained during different lactation stages

During lactation, goat milk contains colostrum, transitional milk, mature milk, and end milk. The protein present in goat milk during different lactation periods has different characteristics. This study aimed to characterize the protein profile of goat milk samples obtained at different lactation stages and to identify changes in the physicochemical and functional properties of whey protein and casein from goat milk collected at 1, 3, 15, 100, and 200 d after calving. The results demonstrated that the lactation period had a great influence on the physicochemical and functional properties of goat milk whey protein and casein, especially the protein properties of colostrum on the first day after delivery. The denaturation temperature, hydrophobicity, and turbidity of whey protein were significantly higher on the first day postpartum than at other lactation periods. Correspondingly, the colostrum whey protein also had better functional properties, such as emulsification, oil holding capacity, and foaming properties on the first day postpartum than at other lactation periods. For casein, the turbidity, particle size, water holding capacity, and foaming properties on the first day after delivery were significantly higher than those at other lactation periods, whereas the denaturation temperature, oil holding capacity, and emulsification followed the opposite trend. For both whey protein and casein, the 2 indicators of emulsifying properties, namely, emulsifying activity index and the emulsion stability, also followed an opposite trend relative to lactation stage, whereas the changes in foaming capacity with the lactation period were completely consistent with the change of foaming stability. These findings could provide useful information for the use of goat milk whey protein and casein obtained during different lactation stages in the dairy industry.     

Developmental changes in proteins of casein micelles in goat milk using data-independent acquisition-based proteomics methods during the lactation cycle

Casein micelles (CM) play an important role in milk secretion, stability, and processing. The composition and content of milk proteins are affected by physiological factors, which have been widely investigated. However, the variation in CM proteins in goat milk throughout the lactation cycle has yet to be fully clarified. In the current study, milk samples were collected at d 1, 3, 30, 90, 150, and 240 of lactation from 15 dairy goats. The size of CM was determined using laser light scattering, and CM proteins were separated, digested, and identified using data-independent acquisition (DIA) and data-dependent acquisition (DDA)-based proteomics approaches. According to clustering and principal component analysis, protein profiles identified using DIA were similar to those identified using the DDA approach. Significant differences in the abundance of 115 proteins during the lactation cycle were identified using the DIA approach. Developmental changes in the CM proteome corresponding to lactation stages were revealed: levels of lecithin cholesterol acyltransferase, folate receptor α, and prominin 2 increased from 1 to 240 d, whereas levels of growth/differentiation factor 8, peptidoglycan-recognition protein, and 45 kDa calcium-binding protein decreased in the same period. In addition, lipoprotein lipase, glycoprotein IIIb, and α-lactalbumin levels increased from 1 to 90 d and then decreased to 240 d, which is consistent with the change in CM size. Protein–protein interaction analysis showed that fibronectin, albumin, and apolipoprotein E interacted more with other proteins at the central node. These findings indicate that changes in the CM proteome during lactation could be related to requirements of newborn development, as well as mammary gland development, and may thus contribute to elucidating the physical and chemical properties of CM.     

Casein haplotype structure in five Italian goat breeds

The aim of this work was to investigate the genetic structure of the casein gene cluster in 5 Italian goat breeds and to evaluate the haplotype variability within and among populations. A total of 430 goats from Vallesana, Roccaverano, Jonica, Garganica, and Maltese breeds were genotyped at alphas1-casein (CSN1S1), alphas2-casein, (CSN1S2), beta-casein (CSN2), and kappa-casein (CSN3) loci using several genomic techniques and milk protein analysis. Casein haplotype frequencies were estimated for each breed. Principal component analysis was carried out to highlight the relationship among breeds. Allele and haplotype distributions indicated considerable differences among breeds. The haplotype CSN1S1*F- CSN1S2*F-CSN3*D occurred in all breeds with frequencies >0.100 and was the most common haplotype in the Southern breeds. A high frequency of CSN1S1*0-CSN1S2*C-CSN3*A haplotype was found in Vallesana population (0.162). Principal component analysis clearly separated the Northern and Southern breeds by the first component. The variability of the caprine casein loci and variety of resulting haplotypes should be exploited in the future using specific breeding programs aiming to preserve biodiversity and to select goat genetic lines for specific protein production.     

Short communication: Genetic variability in the predicted microRNA target sites of caprine casein genes

The main goal of the current work was to identify single nucleotide polymorphisms (SNP) that might create or disrupt microRNA (miRNA) target sites in the caprine casein genes. The 3′ untranslated regions of the goat α_S1-, α_S2-, β-, and κ-casein genes (CSN1S1, CSN1S2, CSN2, and CSN3, respectively) were resequenced in 25 individuals of the Murciano-Granadina, Cashmere, Canarian, Saanen, and Sahelian breeds. Five SNP were identified through this strategy: c.*175C > T at CSN1S1; c.*109T > C, c.*139G > C, and c.*160T > C at CSN1S2; and c.*216C > T at CSN2. Analysis with the Patrocles Finder tool predicted that all of these SNP are located within regions complementary to the seed of diverse miRNA sequences. These in silico results suggest that polymorphism at miRNA target sites might have some effect on casein expression. We explored this issue by genotyping the c.*175C > T SNP (CSN1S1) in 85 Murciano-Granadina goats with records for milk CSN1S1 concentrations. This substitution destroys a putative target site for miR-101, a miRNA known to be expressed in the bovine mammary gland. Although TT goats had higher levels (6.25 g/L) of CSN1S1 than their CT (6.05 g/L) and CC (6.04 g/L) counterparts, these differences were not significant. Experimental confirmation of the miRNA target sites predicted in the current work and performance of additional association analyses in other goat populations will be an essential step to find out if polymorphic miRNA target sites constitute an important source of variation in casein expression.     

牛羊乳蛋白组分比较研究

羊乳被国际营养学界誉为"奶中之王",逐渐被人们列为日常生活的营养保健佳品。该文对羊乳和牛乳的蛋白组分(主要是酪蛋白和乳清蛋白)含量、氨基酸组成及变异体等方面的差异进行了综述,并且对两者酪蛋白胶束的差异进行了比较,为羊乳检验和加工提供理论依据。     

酪蛋白磷酸肽-钙络合物对酸乳贮藏特性的影响

为探究酪蛋白磷酸肽-钙（CPP-Ca）络合物对酸乳贮藏特性的影响,研究CPP-Ca络合物不同添加量对酸乳21 d贮藏期内乳酸菌总数、pH值、滴定酸度、乳清析出率及黏度的影响。与对照组比较,当CPP-Ca络合物添加量为0.15 g/100 mL时,酸乳中乳酸菌存活率从32.92%提高至47.15%（P＜0.05）;滴定酸度增长率从9.96%降低至7.06%（P＜0.05）;乳清析出率降低了0.4%（P＜0.05）,黏度比提高了5 729 mPa·s （P＜0.05）。结果表明,CPP-Ca络合物的添加能增加酸乳贮藏期内的乳酸菌总数,延缓酸度积累,降低乳清析出率,增加黏度,改善酸乳贮藏期内的产品品质。     

A first step toward genomic selection in the multi-breed French dairy goat population

The objectives of this study were to describe, using the goat SNP50 BeadChip (Illumina Inc., San Diego, CA), molecular data for the French dairy goat population and compare the effect of using genomic information on breeding value accuracy in different reference populations. Several multi-breed (Alpine and Saanen) reference population sizes, including or excluding female genotypes (from 67 males to 677 males, and 1,985 females), were used. Genomic evaluations were performed using genomic best linear unbiased predictor for milk production traits, somatic cell score, and some udder type traits. At a marker distance of 50 kb, the average r² (squared correlation coefficient) value of linkage disequilibrium was 0.14, and persistence of linkage disequilibrium as correlation of r-values among Saanen and Alpine breeds was 0.56. Genomic evaluation accuracies obtained from cross validation ranged from 36 to 53%. Biases of these estimations assessed by regression coefficients (from 0.73 to 0.98) of phenotypes on genomic breeding values were higher for traits such as protein yield than for udder type traits. Using the reference population that included all males and females, accuracies of genomic breeding values derived from prediction error variances (model accuracy) obtained for young buck candidates without phenotypes ranged from 52 to 56%. This was lower than the average pedigree-derived breeding value accuracies obtained at birth for these males from the official genetic evaluation (62%). Adding females to the reference population of 677 males improved accuracy by 5 to 9% depending on the trait considered. Gains in model accuracies of genomic breeding values ranged from 1 to 7%, lower than reported in other studies. The gains in breeding value accuracy obtained using genomic information were not as good as expected because of the limited size (at most 677 males and 1,985 females) and the structure of the reference population.     

Comparative analysis of caseins in Saanen goat milk from 3 different regions of China using quantitative proteomics

A quantitative proteomic technique based on data-independent acquisition (DIA) was used to analyze differentially expressed caseins of Saanen goat milk samples collected from 3 regions in China (Guangdong, GD; Inner Mongolia, IM; Shaanxi, SX). A total of 345 proteins were quantified in each sample. Gene Ontology (GO) analysis showed that proteins were mainly involved in cellular process and cell and binding functions. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis indicated that proteins were mainly involved in metabolic pathways. Differentially expressed proteins (DEP) between goat milk from 3 comparison groups composed of paired regions were compared and analyzed. The number of DEP was 114, 69, and 79 for GD versus IM, GD versus SX, and IM versus SX, respectively. The GO enrichment analysis of the 3 comparison groups showed that differences were mainly related to the regulation of biological quality, biological regulation, and response to stimulus in terms of biological process; extracellular region for cellular component; and binding function for molecular function. Pathways in which DEP of GD versus IM, GD versus SX, and IM versus SX were mostly protein processing in endoplasmic reticulum for the first 2 groups and metabolic pathways for the last. Protein-protein interaction network analysis demonstrated that the most prominent DEP was heat shock protein 90 β family member 1 for both the GD versus IM and the GD versus SX groups, and haptoglobin for the IM versus SX group. Data from this study may offer useful information for further investigation of the protein composition of Saanen goat milk and its application in the dairy industry.     

Effects of breed and casein genetic variants on protein profile in milk from Swedish Red,Danish Holstein,and Danish Jersey cows

In selecting cows for higher milk yields and milk quality, it is important to understand how these traits are affected by the bovine genome. The major milk proteins exhibit genetic polymorphism and these genetic variants can serve as markers for milk composition, milk production traits, and technological properties of milk. The aim of this study was to investigate the relationships between casein (CN) genetic variants and detailed protein composition in Swedish and Danish dairy milk. Milk and DNA samples were collected from approximately 400 individual cows each of 3 Scandinavian dairy breeds: Swedish Red (SR), Danish Holstein (DH), and Danish Jersey (DJ). The protein profile with relative concentrations of α-lactalbumin, β-lactoglobulin, and α_S1-, α_S2-, κ-, and β-CN was determined for each milk sample using capillary zone electrophoresis. The genetic variants of the α_S1- (CSN1S1), β- (CSN2), and κ-CN (CSN3) genes for each cow were determined using TaqMan SNP genotyping assays (Applied Biosystems, Foster City, CA). Univariate statistical models were used to evaluate the effects of composite genetic variants, α_S1-β-κ-CN, on the protein profile. The 3 studied Scandinavian breeds differed from each other regarding CN genotypes, with DH and SR having similar genotype frequencies, whereas the genotype frequencies in DJ differed from the other 2 breeds. The similarities in genotype frequencies of SR and DH and differences compared with DJ were also seen in milk production traits, gross milk composition, and protein profile. Frequencies of the most common composite α_S1-β-κ-CN genotype BB/A²A²/AA were 30% in DH and 15% in SR, and cows that had this genotype gave milk with lower relative concentrations of κ- and β-CN and higher relative concentrations of α_S-CN, than the majority of the other composite genotypes in SR and DH. The effect of composite genotypes on relative concentrations of the milk proteins was not as pronounced in DJ. The present work suggests that a higher frequency of BB/A¹A²/AB, together with a decrease in BB/A²A²/AA, could have positive effects on DH and SR milk regarding, for example, the processing of cheese.     

Preparation and characteristics of yak casein hydrolysate–iron complex

The effect of mass ratio between yak casein hydrolysate and FeSO₄, reaction temperature, pH and holding time on ferrous‐binding capacity of yak casein hydrolysate was investigated. Iron‐releasing percentage and structural characteristics of the formed yak casein hydrolysate–iron complex were also examined. Results showed that casein hydrolysates with different hydrolysis degrees (DH) possessed different ferrous‐binding capacities with the highest of 7‐h hydrolysate. The optimal binding conditions between yak casein hydrolysate and iron are mass ratio of casein hydrolysate to FeSO₄ of 15:1 for 20 min at 40 °C and pH 6.0. The principal binding site for Fe²⁺ in yak casein hydrolysate consists of the carboxylate groups and amide groups. Compared with ferrous sulphate, yak casein hydrolysate–iron complex has higher iron‐releasing percentage at basic conditions, such as pH 7.0, 7.5 and 8.0. Ferrous‐iron enriched casein hydrolysate, i.e. yak casein hydrolysate–iron complex, may have great potential as an effective delivery vehicle of bioavailable iron.     

Grazing season and forage type influence goat milk composition and rennet coagulation properties

Two different types of pasture (cultivated and rangeland) and 2 different hay qualities (high and low quality) were examined for their effects on goat milk composition and rennet coagulation properties. Furthermore, the effect of dietary treatments in both the early and late grazing season was studied. As lactation stage is known to influence milk composition, the goats in the early and late grazing season were in the same lactation stage at the start of the experiment. The milk composition was influenced both by dietary treatment and season. Milk from goats on pasture was superior to those on hay by containing a higher content of protein and casein, and the goats on cultivated pasture had the highest milk yield. Casein composition was significantly influenced by forage treatment. Goats grazing on cultivated pasture had higher contents of α_s1-casein and also of κ-casein compared with the other treatments, whereas goats grazing on rangeland had the highest content of β-casein. Factors such as milk yield, casein micelle size, α_s2-casein, and calcium content were reduced in late compared with early season. More favorable rennet coagulation properties were achieved in milk from the early grazing season, with shorter firming time and higher curd firmness compared with milk from the late grazing season, but the firming time and curd firmness were not prominently influenced by forage treatment. The content of α_s2-casein and calcium in the milk affected the firming time and the curd firmness positively. The influence of season and forage treatment on especially milk yield, casein content, and rennet coagulation properties is of economic importance for both the dairy industry and goat milk farmers.     

不同投料比酪蛋白改性丙烯腈纤维的性能

 研究4种不同酪蛋白投料比的接枝改性丙烯腈纤维,从化学组成、成分含量、形态结构以及基本力学性能、摩擦性能、吸湿性能、染色性能等方面进行比较,探讨不同投料比的酪蛋白对改性丙烯腈纤维性能的影响。结果发现：酪蛋白改性丙烯腈纤维属于三元混合纤维,包括接枝上丙烯腈的酪蛋白,丙烯腈均聚物以及未反应的游离酪蛋白;随着酪蛋白含量的增加,纤维强度在一定范围内增大,吸湿、染色性能均有显著提高,但是过高含量的酪蛋白使得纤维的成型性能下降,纤维品质降低。同时对酪蛋白改性丙烯腈纤维的可加工性及开发利用提出了看法与建议。     

Short communication: Influence of composite casein genotypes on additive genetic variation of milk production traits and coagulation properties in Holstein-Friesian cows

The aim of the study was to quantify the effects of composite β- and κ-casein (CN) genotypes on genetic variation of milk coagulation properties (MCP); milk yield; fat, protein, and CN contents; somatic cell score; pH; and titratable acidity (TA) in 1,042 Italian Holstein-Friesian cows. Milk coagulation properties were defined as rennet coagulation time (RCT) and curd firmness (a₃₀). Variance components were estimated using 2 animal models: model 1 included herd, days in milk, and parity as fixed effects and animal and residual as random effects, and model 2 was model 1 with the addition of composite β- and κ-CN genotype as a fixed effect. Genetic correlations between RCT and a₃₀ and between these traits and milk production traits were obtained with bivariate analyses, based on the same models. The inclusion of casein genotypes led to a decrease of 47, 68, 18, and 23% in the genetic variance for RCT, a₃₀, pH, and TA, respectively, and less than 6% for other traits. Heritability of RCT and a₃₀ decreased from 0.248 to 0.143 and from 0.123 to 0.043, respectively. A moderate reduction was found for pH and TA, whereas negligible changes were detected for other milk traits. Estimates of genetic correlations were comparable between the 2 models. Results show that composite β- and κ-CN genotypes are important for RCT and a₃₀ but cannot replace the recording of MCP themselves.     

牦牛"曲拉"乳酸干酪素生产工艺研究

采用乳酸凝结剂、护色、离心分离等方法，通过正交试验筛选出活性炭最佳使用参数（质量分数2．0％，吸附温度50℃，吸附时间30min）。复合护色剂配合比例mNa2S2O3：mNa2SO3：mNaHSO3=1.0：2．0：2．5），以及离心工艺参数（转速8000r／min，时间10min，温度50℃）：确定了牦牛“曲拉”乳酸干酪素生产工艺；制品在感官、理化、微生物等方面符合QB／T3780-1999要求。     

牦牛与奶牛酪蛋白制备CPPs的酶工艺优化及其离子交换纯化研究

优化了以牦牛酪蛋白与奶牛酪蛋白为原料酶法制备酪蛋白磷酸肽（CPPs）的工艺条件，并评估了离子交换对CPPs粗制品的纯化效果。结果表明，牦牛酪蛋白与奶牛酪蛋白的产率最优化条件以及氮磷比r（N／P）最优化的酶解工艺条件均基本一致，产率最大的胰蛋白酶酶解工艺条件为：底物浓度13．1％，酶解反应时间104min，酶底物浓度比0．9％；r（N／P）最优（小）的酶解工艺条件为：底物浓度7．6％，酶解反应时间155min，酶底物浓度比1．2％。牦牛及奶牛CPPs粗制品经过阴离子交换树脂纯化后，所获得的精制CPPs其，（N／P）均明显降低，对于CPPs的产率最优样品其，（N／P）由原来的12．2～12．4降低到6．3～6．9；对于r（N／P）最优样品，其r（N／P）由原来的9．90～0．95进一步降低至5．6～5．8。研究表明：无论以产率或制品质量为优化指标，牦牛与奶牛酪蛋白的酶解工艺优化条件基本一致。而且利用阴离子交换树脂处理可以有效地降低CPPs粗制品的r（N／P）、提高制品的质量。