Effects of α s1-casein (CSN1S1) and κ-casein (CSN3) genotypes on milk coagulation properties in Murciano-Granadina goats

The effects of the caprine α s1-casein (CSN1S1) polymorphisms on milk quality and cheese yield have been widely studied in French and Italian goat breeds. Much less is known about the consequences of κ-casein (CSN3) genotype on the technological and coagulation properties of goat milk. In the current study, we have performed an association analysis between polymorphisms at the goat CSN1S1 and CSN3 genes and milk coagulation (rennet coagulation time, curdling rate and curd firmness) and technological (time to cutting of curd and cheese yield) properties. In this analysis, we have included 193 records from 74 Murciano-Granadina goats (with genotypes constituted by different combinations of alleles B, E and F of the gene CSN1S1 and alleles A and B of the gene CSN3) distributed in three herds, which were collected bimonthly during a whole lactation. Data analysis, using a linear mixed model for repeated observations, revealed significant associations between CSN1S1 genotypes and the rate of the curdling process. In this way, milk from EE goats had a significantly higher curdling rate than milk from BB individuals (P<0?·05). Contrary to previous experiments performed in French breeds, cheese yield was not significantly different in BB, EE and EF goats. Moreover, we have shown that CSN3 genotype has a significant effect on the rennet coagulation time (BB>AB, P<0?·05) but not on cheese yield. No interaction between the CSN1S1 and CSN3 genotypes was observed.     

Genetic variation in the αS1-casein of Chinese yak (Bos grunniens)

The aim of this study was to investigate diversity of the αS1-casein gene in Maiwa yak. A polymorphism of αS1-casein gene has been identified in Chinese Maiwa yak by PCR single-strand conformation polymorphism protocol. Comparing the X59856 sequence, some non-coding and coding DNA variants of yak CSN1S1 gene were detected in this present study, including KJ397913 (promoter region, 9969A/G, 9984 A/G, 10153 C/T, 10175 A/C, 10261 G/C), KJ397914 (promoter region 10261 G/C), KJ397896 (intron III, 14862 T/C), KJ397897 (intron III, 14862 T/C, and deleted 14944-14946 CTT), KJ397899 (intron IV, 15297 A/C), KJ397901 (intron VIII, 17503 A/C, 17666 A/C), KJ397902 (exon IX, 18901C/A, intron IX, 18970 T/G, 19088 add A, 19120 add A, 19202 add T, 19296 G/T), KJ397903 (intron IX, 18970 T/G, 19088 add A, 19120 add A, 19202 add T, 19296 G/T), KJ397904 (intron IX, 19537 T/G, 19539T/G, 19599G/A, 19638 C/T), KJ397905 (intron XIV, 23402 T/G, 23417 C/T, 23480 T/C, 23511 G/A), and KJ397907 (intron XVI, 25976 G/T; exon XVII, 26181A/G). The polymorphism of αS1-casein gene may be helpful for future studies on genetic variation within and between yak populations or on associated traits.     

Genetic parameters for αS1-casein and αS2-casein phosphorylation isoforms in Dutch Holstein Friesian

Relative concentrations of α_S1-casein and α_S2-casein (α_S1-CN and α_S2-CN) phosphorylation isoforms vary considerably among milk of individual cows. We estimated heritabilities for α_S2-CN phosphorylation isoforms, determined by capillary zone electrophoresis from 1,857 morning milk samples, and genetic correlations among α_S2-CN phosphorylation isoforms in Dutch Holstein Friesian. To investigate if phosphorylation of α_S1-CN and α_S2-CN are due to the same genetic mechanism, we also estimated genetic correlations between α_S1-CN and α_S2-CN phosphorylation isoforms as well as the genetic correlations between the phosphorylation degrees (PD) of α_S1-CN and α_S2-CN defined as the proportion of isoforms with higher degrees of phosphorylation in total α_S1-CN and α_S2-CN, respectively. The intra-herd heritabilities for the relative concentrations of α_S2-CN phosphorylation isoforms were high and ranged from 0.54 for α_S2-CN-10P to 0.89 for α_S2-CN-12P. Furthermore, the high intra-herd heritabilities of α_S1-CN PD and α_S2-CN PD imply a strong genetic control of the phosphorylation process, which is independent of casein production. The genetic correlations between α_S2-CN phosphorylation isoforms are positive and moderate to high (0.33–0.90). Furthermore, the strong positive genetic correlation (0.94) between α_S1-CN PD and α_S2-CN PD suggests that the phosphorylation processes of α_S1-CN and α_S2-CN are related. This study shows the possibility of breeding for specific α_S1-CN and α_S2-CN phosphorylation isoforms, and relations between the phosphorylation degrees of α_S1-CN and α_S2-CN and technological properties of milk need to be further investigated to identify potential benefits for the dairy industry.     

Biochemical and molecular characterization of polymorphisms of αs1-casein in Sudanese camel (Camelus dromedarius) milk

This work was designed to detect occurrence of biochemical polymorphism of α_s1-casein in two ecotypes of Sudanese camel (Camelus dromedarius) and to characterize these variants on molecular level. Milk samples were screened for α_s1-casein variability by isoelectric focusing, using skimmed milk, as well as isolated α_s1-casein. Two protein patterns, named α_s1-casein A and C, were identified, whereas the major allele A revealed frequencies of 0.8214 and 0.8615 in the two ecotypes. CSN1S1*A and CSN1S1*C are both characterized by missing of exon 16 on mRNA-level compared with the previously described CSN1S1*B. However, the sequence of exon 16 occurs on DNA-level in both. Therefore, this exon seems to be skipped out during mRNA-processing. Furthermore, CSN1S1*C shows a single G > T nucleotide substitution in exon 5, leading to a non-synonymous amino acid exchange (p.Glu30 > Asp30; GenBank ID: JF429138). A polymerase chain reaction-restriction fragment length polymorphism-method was established as a DNA-based test for this nucleotide substitution.     

Immunoreactive properties of α-casein and κ-casein: Ex vivo and in vivo studies

The aim of this study was to evaluate the ex vivo and in vivo studies immune potential of α- and κ-casein. Ex vivo, naïve mouse splenocytes were stimulated with α- or κ-casein. After 120 h of culture, the proliferation index (PI), determined by 3-(4,5 dimethylthiazol-2yl)-2,5-diphenyl tetrazolium bromide (MTT) assay and carboxyfluorescein diacetate N-succinimidyl ester (CFSE) staining, did not vary for either antigen, suggesting similar ex vivo immunogenic potential of both casein fractions. In vivo, BALB/ccmdb mice were sensitized with α- or κ-casein and then gavaged with primary antigen. Mice immunized with α-casein had higher levels of IgG (2^16.33) and IgA (2^10.22) in serum at the end of the experiment compared with mice immunized with κ-casein (2¹⁵ and 2^9.3 for IgG and IgA, respectively). The use of α-casein for mouse immunization and ex vivo lymphocyte stimulation resulted in higher concentrations of secreted cytokines (IL-4, IL-10) compared with κ-casein stimulation. This is consistent with increasing regulatory T cell (Treg) lymphocyte populations, independent of the antigen used for stimulation. In summary, the immunogenic potential of α- and κ-casein was similar. Humoral and cellular immune responses confirmed their strong, independent potential to induce B and T cells. We propose that the lymphocyte proliferation index be used as an initial screening for protein immunogenicity.     

Genome-wide association study for αS1- and αS2-casein phosphorylation in Dutch Holstein Friesian

Phosphorylation of caseins (CN) is a crucial post-translational modification that allows caseins to form colloid particles known as casein micelles. Both α_S1- and α_S2-CN show varying degrees of phosphorylation (isoforms) in cow milk and were suggested to be more relevant for stabilizing internal micellar structure than β- and κ-CN. However, little is known about the genetic background of individual α_S2-CN phosphorylation isoforms and the phosphorylation degrees of α_S1- and α_S2-CN (α_S1-CN PD and α_S2-CN PD), defined as the proportion of isoforms with higher degrees of phosphorylation in total α_S1- and α_S2-CN, respectively. We aimed to identify genomic regions associated with these traits using 50K single nucleotide polymorphisms for 1,857 Dutch Holstein Friesian cows. A total of 10 quantitative trait loci (QTL) regions were identified for all studied traits on 10 Bos taurus autosomes (BTA1, 2, 6, 9, 11, 14, 15, 18, 24, and 28). Regions associated with multiple traits were found on BTA1, 6, 11, and 14. We showed 2 QTL regions on BTA1, one affecting α_S2-CN production and the other harboring the SLC37A1 gene, which encodes a phosphorus antiporter and affects α_S1- and α_S2-CN PD. The QTL on BTA6 harbors the casein gene cluster and affects individual α_S2-CN phosphorylation isoforms. The QTL on BTA11 harbors the PAEP gene that encodes for β-lactoglobulin and affects relative concentrations of α_S2-CN-10P and α_S2-CN-11P as well as α_S1-CN PD and α_S2-CN PD. The QTL on BTA14 harbors the DGAT1 gene and affects relative concentrations of α_S2-CN-10P and α_S2-CN-11P as well as α_S1-CN PD and α_S2-CN PD. Our results suggest that effects of identified genomic regions on phosphorylation of α_S1- and α_S2-CN are related to changes in milk synthesis and phosphorus secretion in milk. The actual roles of SLC37A1, PAEP, and DGAT1 in α_S1- and α_S2-CN phosphorylation in Dutch Holstein Friesian require further investigation.     

Allergic responses induced by goat milk αS1-casein in a murine model of gastrointestinal atopy

Up to 3% of young children develop milk allergy and this may influence the development of immune-mediated diseases in later life. One protein that has been associated with allergic reactions to ruminant milk is α(S1)-casein (CN). Studies suggest that goat milk with low levels of α(S1)-CN may reduce allergenicity of milk, but the dose response to α(S1)-CN has not been confirmed. In this study, we examined the immune response to varying levels of goat α(S1)-CN in a mouse model of gastrointestinal allergy. BALB/c mice (aged 5 wk) were given intraperitoneal injections with α(S1)-CN and aluminum as adjuvant at 1 and 3 wk to sensitize mice to the antigen. In wk 5, groups of fasting mice (n=8/group) were challenged 4 times on alternate days by intragastric gavage with saline or 2, 10, or 20mg of α(S1)-CN. Serum levels of specific IgE, IgG(1), and IgG(2a) antibodies and mouse mast cell protease-I were determined. Interleukin-4, IL-10, and IFN-γ responses to 48-h activation with antigen were measured in cultured splenocytes. We determined that mice sensitized with α(S1)-CN had higher titers of specific IgG(1) and IgE antibodies compared with controls; however, groups challenged with differing doses of α(S1)-CN did not differ. The group challenged with the highest dose of α(S1)-CN had a 10-fold increase in mouse mast cell protease-I compared with the group challenged with saline. Both IL-4 and IL-10 were produced in a dose-dependent manner by cultured splenocytes incubated with α(S1)-CN. Overall, α(S1)-CN stimulated the production of cytokines associated with allergic disease in a dose-dependent manner. Thus, milk with lower levels of α(S1)-CN should contribute to a lesser antigenic burden.     

Tryptic hydrolysis of bovine αS2-casein: identification and release kinetics of peptides

Bovine α_S2-casein was purified by anionic exchange and hydrophobic interaction chromatographies. The purified α_S2-casein was hydrolysed with trypsin; the resulting peptides were identified by amino acids composition and on line reversed-phase high performance liquid chromatography coupled with electrospray mass spectrometry. A study on the kinetics of peptide release enabled discrimination between different protein areas according to their surface-accessibility. Data from the kinetic study were reconciled with secondary structure predictions and a hypothetical model of the structural organisation of purified bovine α_S2-casein in solution was proposed.     

Species identification of ruminant milk by genotyping of the κ-casein gene

The development of molecular genetic and bioinformatic systems for identifying the species of milk and the raw material composition of dairy products is of great scientific and practical importance with the purpose of introducing developments in the system for controlling the turnover of falsified products. The aim of the research is to develop a method of PCR-RFLP analysis for species identification of milk and dairy products from agricultural ruminant animals by the κ-casein gene (CSN3) with the possibility of qualitative and relative quantitative assessment of species-specific DNA of the tested biomaterial. The objects of research were samples of raw milk and milk powder, pasteurized cream, and hard and semi-hard cheeses. The developed method of species identification of milk and dairy products includes sample preparation of the studied samples, nucleic acid extraction, combined PCR-RFLP technique, detection of obtained results by the method of horizontal electrophoresis in agarose gel and their analysis, including using the developed mathematical algorithms and software. The synergistic effect established in combined operation of 2 restriction enzymes ensured their application in a mix with increased performance in an ergonomic way in the context of DNA authentication of cow, goat, and sheep milk and dairy products based on them. The specificity and sensitivity of the proposed method is potentially suitable for implementing the development of a system to control the turnover of falsified and counterfeit goods.     

Effect of temperature and pH on the solubility of caseins: environmental influences on the dissociation of α(S)- and β-casein

Selective precipitation is a common method for the isolation of β-casein, using the different calcium sensitivities of the individual caseins and the selective solubility of β-casein at a low temperature. In previous studies, it has been indicated that the β-casein yield depends on the physicochemical characteristics of the casein raw material used for fractionation. The objective of this study was to evaluate and compare the solubility of α(S)- and β-casein in solutions of micellar casein, sodium caseinate, and calcium caseinate as a function of pH and temperature. Additionally, the solubility of isolated α(S)- and β-casein fractions in demineralized water, ultrafiltration permeate, and a calcium-depleted milk salt solution was investigated depending on the pH and temperature. Furthermore, micellar casein, sodium caseinate, and calcium caseinate were subjected to a calcium chloride-precipitation process to determine the solubility of α(S)- and β-casein in calcium chloride precipitate, which is produced during selective precipitation, as a function of temperature and pH. Generally, the temperature had only a marginal influence on the α(S)-casein solubility compared with the β-casein solubility, whereas the solubility was shown to be strongly influenced by the pH. Our results suggest that the yield of β-casein obtained during isolation by means of selective precipitation may be a result of the solubility characteristics of α(S)- and β-casein in calcium chloride precipitate. Manufacturers may consider a simple solubility experiment before the β-casein isolation process by means of selective precipitation to predict β-casein yield.     

Effects of diet on casein and fatty acid profiles of milk from goats differing in genotype for αS1-casein synthesis

This study investigated the interactions between nutrition and the genotype at α_S1-CN loci (CSN1S1) in goats, evaluating the impact of fresh forage-based diets and an energy supplement on the casein and fatty acid (FA) profiles of milk from Girgentana goats. Twelve goats were selected for having the same genotype at the α_S2-CN, β-CN, and κ-CN loci and differing in the CSN1S1 genotype: homozygous for strong alleles (AA) or heterozygous for strong and weak alleles (AF). Goats of each genotype were divided into three groups and, according to a 3 × 3 Latin square design, fed ad libitum three diets: sulla fresh forage (SFF), SFF plus 800 g/day of barley (SFB), and mixed hay plus 800 g/day of barley (MHB). The SFB diet led to higher-energy intake and milk yield. The energy-supplemented diets (SFB, MHB) reduced milk fat and urea and increased coagulation time. The fresh forage diets (SFF, SFB) increased dry matter (DM) and crude protein (CP) intake and milk β-CN. Diet had a more pronounced effect than CSN1S1 genotype on milk FA profile, which was healthier from goats fed the SFF diet, due to the higher content of rumenic acid, polyunsaturated, and omega-3 FAs. The AA milk had longer coagulation time and higher curd firmness, higher short- and medium-chain FAs (SMFA), and lower oleic acid than AF milk. Significant diet by genotype interactions indicated the higher milk yield of AA goats than AF goats with the higher-energy SFB diet and the lower synthesis of SMFA in AF than in AA goats with the SFF diet.     

Effect of CSN1S1-CSN3 (α(S1)-κ-casein) composite genotype on milk production traits and milk coagulation properties in Mediterranean water buffalo

The aim of this study was to estimate effects of CSN1S1-CSN3 (α_S1-κ-casein) composite genotypes on milk production traits and milk coagulation properties (MCP) in Mediterranean water buffalo. Genotypes at CSN1S1 and CSN3 and coagulation properties [rennet clotting time (RCT), curd firming time (K₂₀), and curd firmness (A₃₀)] were assessed by reversed-phase HPLC and computerized renneting meter analysis, respectively, using single test-day milk samples of 536 animals. Alternative protein variants of α_S1-CN and κ-CN were detected by HPLC, and identification of the corresponding genetic variants was carried out by DNA analysis. Two genetic variants were detected at CSN1S1 (A and B variants) and 2 at CSN3 (X1 and X2 variants). Statistical inference was based on a linear model including the CSN1S1-CSN3 composite genotype effect (7 genotypes), the effects of herd-test-day (8 levels), and a combined days in milk (DIM)-parity class. Composite genotype AB-X2X2 was associated with decreased test-day milk yield [?0.21 standard deviation (SD) units of the trait] relative to genotype BB-X2X2. Genotypes did not affect milk protein content, but genotype AB-X1X1 was associated with increased fat content compared with genotype BB-X2X2 (+0.28 SD units of the trait) and AB-X1X1 (+0.43 SD units of the trait). For RCT, the largest difference (+1.91 min; i.e., 0.61 SD units of the trait) was observed between genotype AA-X1X2 and AB-X1X1. Direction of genotype effects on K₂₀ was consistent with that for RCT. The maximum variation in K₂₀ due to genotype effects (between AA-X1X2 and AB-X1X1 genotypes) was almost 0.9 SD units of the trait. Magnitude of genotype effects was smaller for A₃₀ than for RCT and K₂₀, with a maximum difference of 0.5 SD units of the trait between genotype AA-X1X2 and AA-X1X1. The B allele at CSN1S1 was associated with increased RCT and K₂₀ and with weaker curds compared with allele A. Allele X2 at CSN3 exerted opposite effects on MCP relative to CSN1S1 B. Because of linkage disequilibrium, allele B at CSN1S1 and allele X2 at CSN3 tend to be associated and this likely makes their effects cancel each other. This study indicates a role for casein genes in variation of MCP of buffalo milk. Further studies are necessary to estimate the effects of casein genetic variants on variation of cheese yield.     

In vitro gastrointestinal digestion of bovine αS1- and αS2-casein variants gives rise to different IgE-binding epitopes

The occurrence and differences of resistant regions containing IgE-binding epitopes of α_S1-casein (α_S1-CN) variants B and C, as well as α_S2-CN A and B, after in vitro gastrointestinal digestion was investigated using mass spectrometry. The amino acid substitutions characterising the genetic variants affected the peptide pattern arising from the caseins and thus modifications in their allergenic epitopes occurred. Peptides f174–193 in α_S1-CN B and f179–198 in α_S1-CN C correspond to the IgE-binding epitope f173–194, which has been reported as one of the major epitopes in α_S1-CN B. Within α_S2-CN, the two variant-specific peptides, f7–29 from variant A and f1–22 from variant B, contain the previously identified IgE-binding epitope f1–20. These peptides, and in consequence the protein variants, may exhibit different immunoreactions, which could be significant in the production of milk with improved nutritional properties, such as hypoallergenic quality, by selection and breeding of cows with particular milk protein genotypes.     

Additive and dominance effects of the αs1-casein locus on milk yield and composition traits in dairy goats

The objective of this study was to evaluate the effects of the CSN1S1 locus polymorphism on 305-d records of milk, fat, protein, lactose and total solids yields, fat, protein, lactose and total solids contents in Mexican dairy goats. A total of 514 lactation records belonging to Alpine (n=60), Saanen (n=105) and Toggenburg (n=74) goats, born from 2003 to 2006 in three herds were used. Discrimination between alleles E, F, N, A* (CSN1S1 A, G, H, I, O1 and O2) and B* (CSN1S1 B1, B2, B3, B4, C and L) were made by amplification of fragments of the gene CSN1S1 and digestion with the restriction endonuclease XmnI. In order to estimate additive and dominance effects, data sets including (1) all genotypes, and (2) only homozygote genotypes, were analysed using linear mixed models. The allele A*, had significant additive effects for protein content (0·21±0·07%; P=0·002) and total solids content (0·66±0·23%; P=0·005) when compared with allele F. An unfavourable additive effect of allele A* on milk yield was found in the Alpine breed (-81·4±40·2; P=0·046) when compared with allele F. Favourable dominance effects were found for some genotypes (P<0·05) for milk yield (A*N and B*N), fat yield (A*N and B*E), protein yield (A*N and B*E), lactose yield (A*N) and total solids yield (A*N). Also, unfavourable dominance effects were found (P<0·05) for protein content (A*B* and A*N) and total solids content (A*B*, A*N, and A*F). Allele A* was the only one with a positive effect for protein content. Significant allele-year interaction effects were also observed. The presence of significant dominance effects, estimated between specific pairs of alleles, challenged the purely additive nature of the genetic effect at the CSN1S1 locus. Implications from use of CSN1S1 effects in goat breeding programmes are presented.     

Relation between αS1-casein,genotype, and quality traits of milk from Swedish dairy goats

Locally produced food is becoming popular among Swedish consumers. One product that has increased in popularity is artisan-manufactured goat cheese, and although the dairy goat industry in Sweden is small-scale, production is gradually increasing. In goats, the CSN1S1 gene regulates expression of the protein α_S1-casein (α_S1-CN), which has been found to be important for cheese yield. Over the years, breeding animals have been imported to Sweden from Norway. Historically, a high frequency of the Norwegian goat population carried a polymorphism at the CSN1S1 gene. This polymorphism, called the Norwegian null allele (D), leads to zero or significantly reduced expression of α_S1-CN. Using milk samples from 75 goats, this study investigated associations between expression of α_S1-CN and genotype at the CSN1S1 gene on milk quality traits from Swedish Landrace goats. Milk samples were grouped according to relative level of α_S1-CN (low: 0–6.9% of total protein; medium-high: 7–25% of total protein) and genotype (DD, DG, DA/AG/AA). While the D allele leads to extremely low expression of α_S1-CN, the G allele is low expressing and the A allele is highly expressing for this protein. Principal component analysis was used to explore the total variation in milk quality traits. To evaluate the effect of different allele groups on milk quality attributes, 1-way ANOVA and Tukey pairwise comparison tests were used. The majority (72%) of all goat milk samples investigated showed relative α_S1-CN content of 0% to 6.82% of total protein. The frequency of individuals homozygous for the Norwegian null allele (DD) was 59% in the population of sampled goats, and only 15% carried at least one A allele. A low relative concentration of α_S1-CN was associated with lower total protein, higher pH, and higher relative concentration of β-casein and levels of free fatty acids. Milk from goats homozygous for the null allele (DD) showed a similar pattern as milk with low relative concentration of α_S1-CN, but total protein was only numerically lower, and somatic cell count and α_S2-CN were higher than for the other genotypes. The associations between levels of α_S1-CN and the investigated genotype at the CSN1S1 gene indicate a need for a national breeding program for Swedish dairy goats.     

Effect of αS1-casein genotype on phenolic compounds and antioxidant activity in goat milk yogurt fortified with Rhus coriaria leaf powder

The aim of this work was to evaluate the phenolic compounds and antioxidant activity of goat milk yogurt characterized by different α_S1-casein genotypes and fortified with Rhus coriaria leaf powder. The α_S1-casein genotype was determined by isoelectric focusing, total phenol content was determined by the Folin–Ciocalteu method, phenolic compounds were identified and quantified by HPLC-UV analysis, and antioxidant activity was measured using 2,2′-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid and ferric-reducing antioxidant power. The statistical analysis showed a significant effect of the studied factors. Comparing different genotypes it emerged that yogurt from goats with weak alleles at CSN1S1 loci (FF) showed the lowest phenolic compounds and therefore a lower antioxidant activity compared with yogurt from goats with strong alleles at CSN1S1 loci (AA, BB, AB). Rhus coriaria-fortified yogurt showed a significant increase in total phenolic compounds and antioxidant activity in comparison with plain yogurt. The FF-fortified yogurt presented the lowest total phenol content and antioxidant activity. This could be due to a greater capacity of proteins and peptides in this yogurt to form stable complexes with phenols. The different total phenol content detected in R. coriaria-fortified yogurt indicates that the α_S1-casein genotype influenced the amount of added phenols that are bound to the caseins and, therefore, the part that remains free and that affects the biological capacity of the final product.     

Characterization of new strain Lactobacillus paracasei I-N-10 with proteolytic activity: Potential role in decrease in β-casein immuno-reactivity

The proteolytic activity of thirty-three LAB isolates from Mongolian tarag was tested on skimmed milk. The strain displaying the highest proteolytic activity was purified and presented by 16S rDNA sequencing 99.9?% homology with Lactobacillus paracasei 1-4-2A. It was named L. paracasei I-N-10. Proteases of L. paracasei I-N-10 hydrolyze predominately ??-casein and in some level ??_S2-casein; hydrolysis of ??_S1-casein was not observed. Proteolytic activity was optimal at 42?°C and neutral pH. Proteases of L. paracasei I-N-10 were inhibited by serine- and metalloproteases inhibitors. PCR amplification revealed the presence of prtP gene, which was identical to prtP gene of L. paracasei genus. Mass spectrometry analysis of ??-casein hydrolysate allowed to characterize 7 peptides resulting from proteolysis by L. paracasei I-N-10. The isolated strain was able to cleave ??-casein in different sites including 2 of the major linear epitopes implicated in its allergenicity. Being sensitive to main antibiotics classes, L. paracasei I-N-10 could be considered as safe and used as starter culture with a potential role in decreasing ??-casein immuno-reactivity.     

Nutritional utilization in Malagueña dairy goats differing in genotypes for the content of alphaS1-casein in milk

A study was carried out with 20 goats of the Malagueña breed, half with a high (HG) and half with a low (LG) genetic capability for α_S1-casein (AS1-CN) synthesis, to determine whether the 2 different genotypes (that cause differences in goat milk composition) are related to differences in nutritional feed utilization. Among the 10 HG goats, 7 had BB and 3 had AB genotypes for AS1-CN, whereas there were 7 EF and 3 FF genotypes in the 10 LG goats. The goats were fed diets differing in crude protein content (13.6 vs. 17.7% dry matter for diets 1 and 2, respectively). For each genotype group, a balance trial was conducted with each of the 2 diets in a 2-period balanced changeover designed with half the animals consuming diet 1 and the other half diet 2, determining individual feed intake and the utilization of N and energy in the diets. Greater voluntary feed intake on a metabolic body weight basis among the HG goats was identified as the first possible cause of their milk production. The HG goats also had a greater level of feed utilization, on a metabolic body weight basis, for N and energy intake. Greater ratios of N balance/ digestible N, milk protein N/digestible N, milk energy/ digestible energy, and milk energy/ME were found for HG goats compared with LG. These effects appear to be dependent on the level of protein in the diet, indicating interactive effects. The greater N and energy utilization of HG versus LG goats may explain the differences in milk composition between the 2 genotype groups.     

Heat-induced aggregation of whey proteins in the presence of κ-casein or sodium caseinate

Aggregates were formed by heating mixtures of whey protein isolate (WPI) and pure κ-casein or sodium caseinate at pH 7 and 0.1 M NaCl. The aggregates were characterized by static and dynamic light scattering and size exclusion chromatography. After extensive heat-treatment at 80 °C for 24 h, almost all whey proteins and κ-casein formed mixed aggregates, but a large proportion of the sodium caseinate did not aggregate. At a given WPI concentration the size of the aggregates decreased with increasing κ-casein or sodium caseinate concentration, but the overall self-similar structure of the aggregates was the same. The presence of κ-casein or caseinate therefore inhibited growth of the heat-induced whey protein aggregates. The results were discussed relative to the reported chaperone-like activity of casein molecules towards heat aggregation of globular proteins.