A2 Β-casein Research Articles

UHT A2 milks marketed in Brazil: Physicochemical and electrical characterization, rheological behavior and fatty acid profile

UHT A2 Milk offers a consumption alternative for consumers experiencing intestinal discomfort from consuming dairy products. A2 milk, derived from cows with the A2A2 genotype, does not promote the formation of betacasomorphin-7 (BCM-7). However, little is known about the comprehensive composition of this product, as well as its impact on physicochemical and electrical parameters. This study aimed to investigate potential physicochemical and chemical differences between UHT milk with the A2 β-casein genotype and conventional milk without A2 certification, sourced from brands available in the Brazilian market. Therefore, the physicochemical parameters, rheological behavior, electrical impedance, and fatty acid composition were evaluated. The results indicated that A2 milk had higher density, fat, protein, and total solids content compared to UHT milk without A2 certification. Furthermore, it obtained a higher electrical impedance modulus and lower admittance within the analyzed frequency range (1 kHz a 5 MHz). The UHT milk samples showed Newtonian fluid behavior at both temperatures analyzed (10 and 25 °C). Furthermore, the type of UHT milk (with and without A2 certification) did not significantly influence the absolute viscosity of the samples (p > 0.05) across both commercial brands and temperature. Overall, both types of milk had the same composition of fatty acids, with palmitic acid being the major compound. However, C12:0 and C18:1 cis-7 were more abundant in A2 milk. In this sense, the analytical methods used in this study were able to identify differences in macromolecules and compounds within the lipid fraction in the samples investigated. These analytes are suggested as potential identity markers to evaluate the authenticity of UHT bovine containing only beta-casein of the A2 genotype in its composition.

An A2 β-casein infant formula with high sn-2 palmitate and casein phosphopeptides supports adequate growth, improved stool consistency, and bone strength in healthy, term Chinese infants: a randomized, double-blind, controlled clinical trial.

The aim of this randomized, double-blind, controlled trial was to examine the effects of infant formula on the growth, stool consistency, and bone strength of infants (n = 120) over a period of 4 months. The investigational group was fed an A2 β-casein cow's milk infant formula containing casein phosphopeptides (CPP) and high sn-2 palmitate (54% of total palmitate at sn-2). The control group was fed a standard cow's milk formula without CPP and with low sn-2 palmitate (29% of total palmitate at sn-2). The third group was fed human milk (HM) (n = 60). All three groups had similar baseline characteristics, and maintained similar BMI, sleep habits, and growth rates in body weight and length throughout the study. However, compared to the control group, infants in the investigational and human milk groups had significantly: (i) greater body length at 90, 120, and 150 days of age; (ii) greater growth rate in head circumference from 30 to 60 days of age, with larger head circumference at 60 days of age; (iii) larger daily stool frequency at 60, 90, and 120 days of age; (iv) softer stool at 60, 90, and 120 days of age; (v) higher bone quality index and bone speed of sound at 150 days of age; (vi) fewer hours of crying at 60 and 90 days of age; (vii) less abdominal distention, burp, and flatus at 60, 90, and 120 days of age; and (viii) less constipation at 90 days of age. At other time points, no significant differences were observed between the three groups. No serious adverse events (AEs) related to the study products were reported, and significantly fewer infants in the investigational and HM groups experienced at least one AE compared to the control group. The study suggests that the A2 β-casein formula with high sn-2 palmitate and CPP supports adequate growth, is well tolerated, and may have beneficial effects on stool consistency, gastrointestinal comfort, crying duration, and bone density, comparable to HM. Clinical trial registration: https://clinicaltrials.gov/, NCT04749290.

Genotypic profiling of A1 and A2 β-casein alleles in Punganur cattle using PCR-RFLP

Milk is regarded as nature’s perfect food due to the presence of essential nutrients required for the growth and development of mammals. The consumption of milk with a specific β casein variant either A1 or A2 had a relationship with the risk of diseases has been identified. Considering the importance of β casein variants with the health significance, the present study was undertaken in Punganur cattle (N=175) by using PCR-RFLP with Dde I restriction enzyme that showed the distribution of A1A2 (121 bp, 86 bp, and 35 bp) in 5 animals, A2A2 (86 bp and 35 bp) in 170 animals with no evidence of A1A1 genotype. The genotypic frequency of the A1A2 heterozygous β-casein variant of exon7 of the CSN2 gene was 0.029, whereas, for the A2A2 homozygous β-casein variant was 0.971 and the A1A1 homozygous β-casein variant is absent in Punganur cattle. The A1 allelic frequency of exon7 of the CSN2 gene was 0.014, whereas, the A2 gene frequency is 0.986 in Punganur cattle. The study indicates that there is a minimum genetic variability in the tested population of the Punganur cattle breed suggesting less or lower level of crossbreeding activities with exotic animals.

Effects of Different Cow-Milk Beta-Caseins on the Gut-Brain Axis: A Narrative Review of Preclinical, Animal, and Human Studies.

The gut and brain communicate through bidirectional neural, endocrine, and immune signals to coordinate central nervous system activity with gastrointestinal function. Dysregulated inflammation can promote immune cell activation and increase entero-endocrine signaling and intestinal permeability; hence, a functional gut-brain axis is necessary for a healthy digestive system. The consumption of milk products can lead to gut discomfort via effects on gastrointestinal tract function and the inflammatory state, which, in turn, affect the brain. A1 β-casein and A2 β-casein are major components of bovine-milk protein, and their digestion may result in different physiological effects following the consumption of milk products. Peptides derived from A1 β-casein, such as β-casomorphins, may increase gut dysfunction and inflammation, thereby modulating the availability of bioactive metabolites in the bloodstream and contribute to changes in cognitive function. This narrative review examines the functional interrelationships between the consumption of cow-milk-derived β-caseins and their effect on the brain, immune system, and the gut, which together comprise the gut-brain axis.

The Effect of A2 Milk on Gastrointestinal Symptoms in Comparison to A1/A2 Milk: A Single-center, Randomized, Double-blind, Cross-over Study.

β-Casein, a major protein in cow's milk, is divided into the A1 and A2 type variants. Digestion of A1 β-casein yields the peptide β-casomorphin-7 which could cause gastrointestinal (GI) discomfort but A2 milk containing only A2 β-casein might be more beneficial than A1/A2 (regular) milk. The aim of this study was to evaluate the differences in GI discomfort after ingestion of A2 milk and A1/A2 milk. A randomized, double-blind, cross-over human trial was performed with 40 subjects who experienced GI discomfort following milk consumption. For each intervention period, either A2 milk first (A2→A1/A2) or A1/A2 milk was first consumed for 2 weeks (A1/A2→A2) following a 2-week washout period. GI symptom rating scale (GSRS) scores, questionnaire for digestive symptoms, and laboratory tests including fecal calprotectin were evaluated. For symptom analysis, generalized estimating equations gamma model was used. A2 milk increased bloating (P = 0.041) and loose stools (P = 0.026) compared to A1/A2 milk in GSRS. However, A2 milk caused less abdominal pain (P = 0.050), fecal urgency (P < 0.001) and borborygmus (P = 0.007) compared to A1/A2 milk in questionnaire for digestive symptoms. In addition, fecal calprotectin also decreased or less increased after consumption of A2 milk compared to A1/A2 milk (P = 0.030), and this change was more pronounced in males (P = 0.005) than in females. There were no significant adverse reactions during the trial. A2 milk alleviated digestive discomfort in Koreans following A2 milk consumption (ClinicalTrials.gov NCT06252636 and CRIS KCT0009301).

Prolonged Consumption of A2 β-Casein Milk Reduces Symptoms Compared to A1 and A2 β-Casein Milk in Lactose Maldigesters: A Two-Week Adaptation Study.

Approximately 30% of milk protein is β-casein. We aimed to determine whether lactose maldigesters who chronically consumed two cups of A1/A2 milk (containing 75% A1 β-casein and 25% A2 β-casein) would adapt to have fewer intolerance symptoms, lower serum inflammatory markers, and/or altered glutathione levels similar to those consuming A2 milk (containing 100% A2 β-casein). A double-blinded, randomized, crossover trial was conducted. Sixteen confirmed lactose maldigesters consumed 250 mL of A1/A2 milk and A2 milk twice daily with meals for two weeks. At the end of the adaptation period on day 15, lactose maldigestion was measured after a challenge with the same milk used for adaptation (0.5 g of lactose per kg of body weight) with a hydrogen breath test. Fecal urgency was higher during the two-week consumption of A1/A2 milk compared to A2 milk (p = 0.04, n = 16). Bloating (p = 0.03, n = 16) and flatulence (p = 0.02, n = 16) were also higher on the 15th day with A1/A2 milk compared to A2 milk challenge. However, day-to-day symptoms, hydrogen, serum inflammatory markers, and antioxidant concentrations were not different after A1/A2 and A2 milk consumption adaptation periods. Adaptation over two weeks did not improve lactose digestion or tolerance of A1/A2 milk to match that of A2 milk.

Effects of A1 Milk, A2 Milk and the Opioid-like Peptide β-Casomorphin-7 on the Proliferation of Human Peripheral Blood Mononuclear Cells.

Special attention is given to cow's milk and its variants, with ongoing discussions about health-related impacts primarily focusing on the A1 variant in contrast to the A2 variant. The difference between these variants lies in a single amino acid alteration at position 67 of β-casein. This alteration is presumed to make the A1 variant more susceptible to enzymatic breakdown during milk digestion, leading to an increased release of the peptide β-casomorphin-7 (BCM-7). BCM-7 is hypothesized to interact with µ-opioid receptors on immune cells in humans. Although BCM-7 has demonstrated both immunosuppressive and inflammatory effects, its direct impact on the immune system remains unclear. Thus, we examined the influence of A1 and A2 milk on Concanavalin A (ConA)-stimulated human peripheral blood mononuclear cells (PBMCs), as well as the effect of experimentally digested A1 and A2 milk, containing different amounts of free BCM-7 from β-casein cleavage. Additionally, we evaluated the effects of pure BCM-7 on the proliferation of ConA-stimulated PBMCs and purified CD4+ T cells. Milk fundamentally inhibited PBMC proliferation, independent of the β-casein variant. In contrast, experimentally digested milk of both variants and pure BCM-7 showed no influence on the proliferation of PBMCs or isolated CD4+ T cells. Our results indicate that milk exerts an anti-inflammatory effect on PBMCs, regardless of the A1 or A2 β-casein variant, which is nullified after in vitro digestion. Consequently, we deem BCM-7 unsuitable as a biomarker for food-induced inflammation.

The Immunomodulatory Effects of A2 β-Casein on Immunosuppressed Mice by Regulating Immune Responses and the Gut Microbiota.

The aim of this study was to investigate the immunomodulatory effects of A2 β-casein (β-CN) in cyclophosphamide-induced immunosuppressed BALB/c mice. Experiments conducted in vitro revealed that A2 β-CN digestive products have potent immunostimulatory activities. Animal studies demonstrated that A2 β-CN improved the immunological organ index reduction trend caused by cyclophosphamide, reduced the pathological damage to the spleen tissue in immunosuppressed mice, increased the release of IL-17A, IgG, and IgA, and reduced the production of IL-4. By regulating the relative abundance of advantageous bacteria like Oscillospira, Lactobacillus, and Bifidobacteria and harmful bacteria like Coprococcus and Desulfovibrionaceae, A2 β-CN improved gut microbiota disorders in immunosuppressed mice. Moreover, A2 β-CN promoted the production of short-chain fatty acids and increased the diversity of the gut microbiota. Therefore, ingestion of A2 β-CN is beneficial to the host's immune system and gut health. These findings provide insights for the future application of A2 β-CN-related dairy products.

A discussion on A1-free milk: Nuances and comments beyond implications to the health.

This chapter provides an overarching view of the multifaceted aspects of milk β-casein, focusing on its genetic variants A1 and A2. The work examines the current landscape of A1-free milk versus regular milk, delving into health considerations, protein detection methods, technological impacts on dairy production, non-bovine protein, and potential avenues for future research. Firstly, it discussed ongoing debates surrounding categorizing milk based on A1 and A2 β-casein variants, highlighting challenges in establishing clear regulatory standards and quality control methods. The chapter also addressed the molecular distinction between A1 and A2 variants at position 67 of the amino acid chain. This trait affects protein conformation, casein micelle properties, and enzymatic susceptibility. Variations in β-casein across animal species are acknowledged, casting doubt on non-bovine claims of "A2-like" milk due to terminology and genetic differences. Lastly, this work explores the burgeoning field of biotechnology in milk production.

Determination of A1 and A2 β-Casein in Milk Using Characteristic Thermolytic Peptides via Liquid Chromatography-Mass Spectrometry

β-casein, a protein in milk and dairy products, has two main variant forms termed as A1 and A2. A1 β-casein may have adverse effects on humans. The fact that there is only one amino acid variation at the 67th position between A1 and A2 β-casein makes it difficult to distinguish between them. In this study, a novel method using characteristic thermolytic peptides is developed for the determination of A1 and A2 β-casein in milk. Firstly, caseins extracted from milk samples are thermolytic digested at 60 °C without any denaturing reagents required for unfolding proteins, which simplifies the sample pretreatment procedure. The characteristic thermolytic peptides (i.e., fragments 66–76 and 59–76 for A1 and A2 β-casein, respectively) selected to specifically distinguish A1 and A2 β-casein only have eleven or eighteen amino acid moieties. Compared with tryptic characteristic peptides with a length of 49 amino acid moieties, these shorter thermolytic characteristic peptides are more suitable for LC-MS analysis. This novel method, with the advantages of high specificity, high sensitivity, and high efficiency, was successfully applied for the analysis of six milk samples collected from a local supermarket. After further investigation, it is found that this method would contribute to the development of A2 dairy products for a company and the quality inspection of A2 dairy products for a government.

Effects of Feeding A1 and A2 Cow Milk-based Diet on Hematological Parameters in Diabetic Mice Model

Background: Milk is essential part of diet across the globe and is a rich source of protein and calcium. Major protein component of milk is casein with beta-casein (β-casein) as the second most prevalent protein in cow milk. β-casein has 15 different genetic variants and of these A1 and A2 have gained research focus. All livestock as well as well human have proline at amino acid position 67 of β-casein, which is referred as A2 variant, but in cattle breeds, other genetic variant called A1 with histidine at amino acid position 67 is also present. This A1 type variant of â-casein or A1 type milk has been implicated as a potential etiological factor in several pathologies. Methods: The objective of the present study was to evaluate the A1and A2 β-casein variants of cow milk as factors affecting different hematological parameters and other parameters like glucose and insulin in streptozotocin (STZ)-induced diabetic C57/BL6 mice after the induction of diabetes. Diabetes was induced by injecting STZ intraperitoneally at dose of 45mg per kg of body weight for consecutive 5 days. Milk powder prepared from milk with A1A1 and A2A2 genotypes was used for feeding for three months. Result: After 3 months of feeding trial, it was observed that diabetic mice fed with A1A1 milk (STZ+A1A1) exhibited significantly elevated levels of glucose and insulin. Similarly, the levels of white blood cells, lymphocytes and neutrophils showed significant changes in STZ+A1A1 group compared to control and STZ+A2A2 group indicating the probable association A1A1 milk with inflammatory reaction. However, no significant changes were observed in parameters like red blood cells, hemoglobin, hematocrit or mean cell volume. In the mice group fed with A2A2 milk powder-based diet, no significant change was observed in the observed parameters except lymphocyte percentage which was lower compared to control group. In summary, our results show that A1 form of cow milk might have a proinflammatory effect.

Evaluating the potential impact of selection for the A2 milk allele on inbreeding and performance in Australian Holstein cattle

Selection decisions are generally based on estimated breeding values (EBV) for a combination of traits that are polygenic (e.g. milk production). However, in some cases, there is additional intense selection for a single allele, or SNP, for a perceived benefit, such as selection for polled or A2 milk. Using a dataset where the A2 mutation was imputed using a reference population with whole genome sequence, we tested the hypothesis that intense selection in Australian Holstein cattle for the A2 allele in the β-casein gene may have resulted in increased inbreeding. We also estimated the average difference in performance between animals homozygous for the A1 or A2 allele for a range of traits. Using high-density genotypes we compared differences in genome-wide and regional inbreeding between Holstein cows homozygous for either the A1 or A2 β-casein alleles i.e. A1/A1 or A2/A2. This study shows that between the years 2000 to 2017, the frequency of the A2/A2 genotype increased by 20% in Holstein cows (from 32% to 52%). Our results suggest that selection for homozygosity at the β-casein A2 allele has increased inbreeding both across the genome and on chromosome 6 in A2/A2 Holstein cows. Animals that were A2/A2 were twice as likely to have a run of homozygosity of at least 1Mb long across the β-casein locus compared to animals that were A1/A1. Cows that are homozygous for the A2 allele had an average protein yield EBV advantage of 0.24 genetic standard deviations (SD) compared to A1/A1 homozygous cows. In contrast, A2/A2 homozygous animals were on average 0.2 genetic SD inferior on fertility EBV. As a result, the difference in the overall economic index (that includes traits contributing to profitability) there was only a small advantage of 0.05 SD for A2/A2 cows compared to A1/A1 cows. However, strong selection for the A2 allele has likely led to a higher level of regional and overall inbreeding which in the long term could harm genetic progress for some or all economic traits. Therefore, applying approaches that mitigate rapid inbreeding while selecting for preferred alleles and quantitative traits may be desirable.

Effectiveness of Growing-Up Milk Containing Only A2 β-Casein on Digestive Comfort in Toddlers: A Randomized Controlled Trial in China.

Emerging clinical evidence indicates the potential gastrointestinal (GI) benefits of milk containing only A2 β-casein, but data from randomized controlled trials is sparse among pediatric populations. We aimed to evaluate the effectiveness of growing-up milk (GUM) containing only A2 β-casein on GI tolerance in toddlers. A total of 387 toddlers aged 12-36 months were recruited in Beijing, China, and randomized in a 1:1:1 ratio to consume one of two commercially available A2 GUMs (combined in the analysis as A2 GUM) or continue their current feeding regimen of conventional milk for 14 days. The primary outcome was the total Gut Comfort Score (GCS) (range: 10-60; higher values indicate greater GI distress) derived from a 10-item (score range: 1-6 per item) parent-reported questionnaire, reflecting GI tolerance. The GCS (mean ± SD) was comparable between the A2 GUM and conventional milk groups on day 7 (14.7 ± 5.0 vs. 15.0 ± 6.1, p = 0.54) and day 14 (14.0 ± 4.5 vs. 14.3 ± 5.5, p = 0.51). Parents reported less constipation in those consuming A2 GUM vs. conventional milk on day 14 (1.3 ± 0.6 vs. 1.4 ± 0.9, p = 0.020). Among 124 participants with minor GI distress at baseline (GCS ≥ 17, top tertile range 17-35), GCS was significantly lower in those consuming A2 GUM on day 7 (18.2 ± 5.1 vs. 21.2 ± 6.8, p = 0.004) and day 14 (17.1 ± 5.3 vs. 19.6 ± 6.3, p = 0.026), as were individual GI symptoms (all p < 0.05). In the toddlers without GI issues at baseline (GCS < 17), a low GCS was maintained throughout the study period after switching to A2 GUM (mean values range 10-13). Growing-up milk containing only A2 β-casein were well-tolerated and associated with lower parent-reported constipation scores after two weeks when compared to conventional milks. In healthy toddlers with minor GI distress, A2 GUM improved overall digestive comfort and GI-related symptoms within one week.

Gastric Emptying of New-World Milk Containing A1 and A2 Β-Casein Is More Rapid as Compared to Milk Containing Only A2 Β-Casein in Lactose Maldigesters: A Randomized, Cross-Over Trial Using Magnetic Resonance Imaging.

Lactose maldigesters report an increase in abdominal pain due to the consumption of milk containing a mixture of A1 and A2 β-casein as compared to milk containing only A2 β-casein. Gastric transit affects gastrointestinal symptoms and rapid transit has been associated with an increase in abdominal pain. We conducted a double-blinded, randomized, crossover trial in 10 lactose maldigesters. Subjects consumed each of the two types of milk: conventional milk containing 75% A1 β-casein and 25% A2 β-casein and A2 milk containing 100% A2 β-casein. Magnetic resonance images were acquired, and abdominal pain was rated and recorded at 0, 10, 30, 60 and 120 min after milk consumption. The volume of milk in the stomach was calculated using FSL software. The volume of milk in the stomach after consuming milk with 75% A1 β-casein and 25% A2 β-casein was significantly lower at 30 (p = 0.01), 60 (p = 0.002) and 120 (p < 0.001) minutes as compared to milk with 100% A2 β-casein in the 10 lactose maldigesters. The transit of New-World milk containing A1 and A2 β-casein was more rapid as compared to Old-World milk containing only A2 β-casein. This difference in transit may mediate symptoms of lactose intolerance.

Modification of a previously patented method to unequivocally score A2-like and A1-like bovine β-casein variants

A growing interest in the production and commercialization of A2 cow's milk has been observed in many countries in the last few years due to the beneficial properties for human health attributed to A2 β-casein variant. Methods of varying complexity and different equipment requirements have been proposed for the determination of the β-casein genotype of individual cows. We proposed herein a modification of a previously patented method based on an amplification-created restriction site PCR followed by restriction fragment length polymorphism analysis. This method allows to identify and differentiate A2-like from A1-like β-casein variants, after differential endonuclease cleavage flanking the nucleotide that determines the amino acid at position 67 of β-casein. The advantages of this method are that it:• enables to unequivocally score A2-like as well as A1-like β-casein variants,• can be performed at low cost in simply equipped molecular biology laboratories, and• can be scaled up to analyze hundreds of samples per day.For these reasons, and based on the results obtained from the analysis carried out in this work, it showed to be a reliable method for the screening of herds to selective breeding of homozygous cows and bulls for A2 or A2-like alleles.

DNA extraction procedures and validation parameters of a real-time PCR method to control milk containing only A2 β-casein

Bovine milk mainly contains two types of β-casein: A1 and A2 variants. In recent years, a new variety of cows’ milk has emerged in dairy sector called “A2 milk”. This novel product is characterised by the absence of A1 β-casein, which has been associated with possible gastrointestinal discomfort due to β-casomorphin-7 (BCM-7) release during gastrointestinal digestion. In this context, methods to verify A1 allele absence in A2 milk are required as a quality control in the A2 milk commercialisation. Therefore, the aim of the present study was to develop a locked nucleic acid (LNA) probe-based duplex real-time PCR (qPCR) assay for A1 allele detection in A2 milk samples. Firstly, four DNA isolation methods from milk somatic cells were optimised and evaluated. Results suggests that the commercial kit NucleoSpin Tissue was the most suitable method in terms of DNA quality and amplificability for downstream applications. Then, optimisation and validation of the qPCR assay were carried out. For both A1 and A2 alleles, the absolute limits of detection of this qPCR assay were 7.3 DNA copies/reaction (2 x 10 −5 ng DNA) and 30.4 DNA copies/reaction (0.1 ng DNA) at a 95% confidence level with synthetic reference DNA samples and heterozygous genotyped DNA sample, respectively. The relative limits of detection were 2% (15 copies) and 5% (152 copies) for A1 allele in A2 samples at a 95% confidence with synthetic reference and genotyped DNA samples, respectively. The qPCR assay was robust, with intra- and inter-assay variability below 4.3%, and specific, differentiating between A1 and A2 alleles with 100% genotyping accuracy. In conclusion, this cost-effective and fast method could be used to discriminate A1 allele in A2 samples and, consequently, to verify A1 allele absence in “A2 milk” by screening commercial product on the market. • The LNA duplex qPCR assay developed could be used to verify A1 allele absence in “A2 milk”. • LNA probes reliably discriminated between A1 and A2 alleles. • The relative limits of detection reach up to 2% (15 copies) for A1 allele in A2 samples at a 95% confidence. • Milk is a challenging matrix for DNA isolation due to the presence of PCR inhibitors that leads to low DNA yield.

Impact of in vitro static digestion method on the release of β-casomorphin-7 from bovine milk and cheeses with A1 or A2 β-casein phenotypes

Beta-casomorphin-7 (BCM7) represents the fragment Val60-Ile66 of bovine β-casein (β-CN), and there is evidence that it is more easily released during gastrointestinal digestion (GID) of A1 β-CN variant, in comparison to the A2 variant. This study aimed at investigating the effect of type of enzymes and the protease/protein (P/S) ratio on BCM7 release during the intestinal step of in vitro static GID of bovine milk and cheeses with A1 or A2 β-CN phenotypes. BCM7 occurred in digests of both A1 and A2 samples, being the release more marked for A1 counterparts. Nonetheless, the BCM7 release depended on both the type of GID enzymes and the P/S ratio. These findings highlight the importance of GID conditions which may affect the outcomes for possible differences between A1 and A2 milk based on BCM7 release during in vitro GID.

Rheological and structural properties of acid-induced milk gels as a function of β-casein phenotype

This study aimed to investigate if differences in β-casein polymorphic structure would affect the acid-induced (glucono-δ-lactone) gelation behaviour of corresponding skim milks. Gels obtained from skim milk containing A2/A2 β-casein had significantly lower elastic modulus, water holding capacity, and gel permeability compared to A1/A1 and A1/A2 gels. Microscopy images also showed a denser microstructure and smaller pore size in acid-induced gels prepared with A1/A1 and A1/A2 milks compared to A2/A2 milk. A number of reasons may account for these differences in gelation, specifically, gels with A1 β-casein contained greater amounts of α-helixes and aggregated β-sheets in their secondary structure compared to A2/A2 gel that was comprised mainly of random coils or polyproline II helixes. In addition, compositional differences such as greater total and micellar calcium and higher levels of total κ-casein existed in A1/A1 and A1/A2 milks compared to A2/A2 milk and may have contributed to the difference in gelation and gel structure. Although the role of β-CN phenotype on skim milk gelation may need further investigation, findings from this study clearly indicate that A2/A2 milk was associated with poor acid gelation properties. • Effects of acidification on milks with various β-caseins were studied. • Gels carrying A1 β-casein possessed greater water retention and lower permeability. • Increased β-sheets in gels carrying A1 β-casein related to higher storage modulus. • A2/A2 gel possessed a high amount of polyproline II structures. • Gels prepared of A1/A1 and A1/A2 milks possessed a denser microstructure.

Bovine milk with variant β-casein types on immunological mediated intestinal changes and gut health of mice.

Dietary proteins provide bioactive peptides, which are important for host gastrointestinal functions. We hypothesized that A2-type β-casein could provide gastrointestinal benefits and improve the immune and gut health. This study was conducted to investigate those effects and mechanisms. Thirty BALB-c mice (3–4 weeks old) were fed with either a control diet (control), a diet supplemented with bovine milk containing A1 and A2 type β-casein (A1A2, contains 63.62% A2 β-casein of total β-casein) or a diet containing A2 type β-casein (A2A2, contains 95.96% A2 β-casein of total β-casein) (10 ml/kg body weight) for 4 weeks. Immunoglobulin and inflammation factors were measured in serum, and histological variations were measured in duodenal and ileum, and stool 16S rRNA and short-chain fatty acids (SCFAs) contents were measured in fecal samples. Results showed that consumption of A2-type β-casein milk could improve proximal small intestine villus and crypt morphology (p < 0.05), increase IgG and IgE responses, and modulate the composition and diversity of gut microbiota by increase the relative abundance of phylum Proteobacteria, class Clostridia, family Ruminococcaceae and species Lactobacillus animalis (p < 0.05). There were also significant associations between gut microbes, immune response, and SCFAs, especially isobutyric acid (p < 0.05), which may potentially regulated gastrointestinal benefits. Moreover, intake of A2-type β-casein milk had no impact on inflammation. These findings explained potential benefits of consumption of A2-type β-casein milk on host immune system and gut health outcomes, and provide insights to the future application of nutritional modulation.

Worldwide Research Trends on Milk Containing Only A2 β-Casein: A Bibliometric Study.

Simple SummaryA1 β-casein has been correlated with adverse health outcomes, and, as a consequence, milk containing only A2 β-casein has emerged on the market. There has been a relevant increase in publications in this area since 2010. Food Science Technology and Agriculture were the main research areas of this topic. The term β-casomorphin was the most frequently used. The USA, New Zealand, and Australia were the most productive countries, though the most productive research institutions were, in absolute terms, from India, France, and Germany. The majority of the most cited studies that refer to A2 β-casein and health were reviews, and a few clinical trials have also been published.The protein fraction of β-casein may play a key role in the manifestation of a new intolerance: milk protein intolerance. The most common forms of β-casein among dairy cattle breeds are A1 and A2 β-casein. During gastrointestinal digestion of A1 β-casein, an opioid called peptide β-casomorphin-7 (BCM-7) is more frequently released, which can lead to adverse health outcomes. For that reason, novel products labelled as “A2 milk” or “A1-free dairy products” have appeared on the market. In this context, a bibliometric analysis on A2 β-casein research was carried out through the Web of Science (WoS) database. The main objective of this work was to provide an overview of the state of the art in the field of β-casein A2 by analyzing the number of publications per year, trends in thematic content, the most frequently used terms, and the most important institutions and countries in the field. This bibliometric study showed that a greater effort is needed to determine the possible implications of this novel product for human health and the market.