Α-lactalbumin Research Articles

Enhanced extracellular production of alpha-lactalbumin from Bacillus subtilis through signal peptide and promoter screening

Alpha-lactalbumin (α-LA) is a major whey protein found in breast milk and plays a crucial role in the growth and development of infants. In this study, Bacillus subtilis RIK1285 harboring AprE signal peptide (SP) was selected as the original strain for the production of α-LA. It was found that α-LA was identified in the pellet after ultrasonic disruption and centrifugation instead of in the fermentation supernatant. The original strain most likely only produced α-LA intracellular, but not extracellular. To improve the expression and secretion of α-LA in RIK1285, a library of 173 homologous SPs from the B. subtilis 168 genome was fused with target LALBA gene in the pBE-S vector and expressed extracellularly in RIK1285. SP YjcN was determined to be the best signal peptide. Bands in supernatant were observed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and purified by nickel column to calculate the highest yield signal peptide. In addition, different promoters (PaprE, P43, and Pglv) were compared and applied. The results indicated that the strain RIK1285-pBE-Pglv-YjcN-LALBA had the highest α-LA yield, reaching 122.04 μg/mL. This study demonstrates successful expression and secretion of human α-LA in B. subtilis and establishes a foundation for simulating breast milk for infant formulas and developing bioengineered milk.

New perspective into the interaction behavior explore of Nano-berberine with alpha-lactalbumin in the presence of beta-lactoglobulin: Multi-spectroscopic and molecular dynamic investigations

The present study was focus on the interaction of alpha lactalbumin (α-LA) and Nano-berberine (Nano-Ber) in the absence and presence of beta lactoglobulin (β-LG) as binary and ternary systems by the exertion of different spectroscopic techniques, conductometry, β-galactosidase activity, and molecular simulation measurements. The outcomes of fluorescence quenching clarified the dominance of static quenching mechanism throughout the interaction of α-LA-Nano-Ber and (α-LA-β-LG) Nano-Ber complexes and also determined the KSV values to be (4.68±0.05) × 104 and (5.18±0.06) × 104 M−1, respectively. The measured thermodynamic parameters exhibited the participation of electrostatic forces in the binding interaction of α-LA and α-LA-β-LG complex with Nano-Ber. Moreover, the negative value of ΔG0 represents the spontaneous mode of this binding process in both systems. Synchronous fluorescence and resonance light scattering data demonstrated the impact of β-LG attendance on altering the interaction behavior of α-LA and Nano-Ber. The changes in the secondary structure of both proteins were affirmed by circular dichroism spectroscopy (CD) technique. According to the displayed β-galactosidase activity in the Michaelis-Menten plot of both binary and ternary systems, a greater Vmax refers to the stronger affinity of lactose and lactose hydrolysis that can be effective on the lactose intolerance regarding dairy products. Additionally, molecular dynamic simulations were employed to simulate the atomic-level interactions and explore the stability and conformational changes of protein-Nano Ber complexes over time. The results of these comprehensive biophysical investigations shed light on the binding characteristics, structural alterations, and dynamic behavior of Nano-Ber upon interaction with α-lactalbumin in the absence and presence of β-lactoglobulin as the binary and ternary systems. This study reveals the Nano-Ber role in the protein–protein interaction behavior and provides valuable insights into the potential applications of α-lactalbumin in the design of functional food formulations and drug delivery systems

The Human Microbiome and Mental Health: the Latest Views

Mental health is the emotional, psychological, and social well-being of a person who feels protected, needed, realizes his abilities, and can overcome life’s stresses. Mental disorders (MD) are a serious health problem worldwide today, which lead to a significant medical burden and economic losses for both medical institutions and patients and their families. Today, MD are one of the priority directions in the daily practice of Ukrainian doctors. MD can include anxiety, depression, bipolar disorder, autism spectrum disorder, schizophrenia, eating disorders, etc. Recently, the attention of scientists and doctors has been paid to the interaction between the microbiome and mental health. The researches which are aimed at defining what is a healthy microbiome have revealed significant individual differences in its composition and diversity. It has been established that a certain composition of the gut microbiome (GM) is associated with the development of anxiety disorders. Research results show that patients and animal models (mice) with anxiety disorders have a dramatic decrease in microbial richness and diversity. Patients with anxiety disorder typically had reduced Firmicutes and increased Bacteroidetes and Fusobacteria. In addition, GM changes were associated with the onset and development of depressive disorder. Differences in the composition of fecal microbiota in patients with major depressive disorder were revealed. The main methods of MD treatment today are pharmacotherapy and psychotherapy, which have limited effectiveness. However, practitioners are looking for alternative ways to help such patients. The results of many studies indicate that prebiotics (e.g., dietary fiber and alpha lactalbumin) as well as postbiotics show a protective effect on mental health, especially when used in combination. At the same time, prebiotics may be a potential agent for alleviating the side effects of antipsychotics used in the treatment of MD. More experimental researches and high-quality clinical trials are needed to study the effects of dietary components on MD through the brain–gut–microbiome axis.

Genetic variability among and within domestic Old and New World camels at the α-lactalbumin gene (LALBA) reveals new alleles and polymorphisms responsible for differential expression

α-Lactalbumin (α-LA), which is encoded by the LALBA gene, is a major whey protein that binds to Ca2+ and facilitates lactose synthesis as a regulatory subunit of the synthase enzyme complex. In addition, it has been shown to play central roles in immune modulation, cell-growth regulation, and antimicrobial activity. In this study, a multitechnical approach was used to fully characterize the LALBA gene and its variants in both coding and regulatory regions for domestic camelids (dromedary, Bactrian camel, alpaca, and llama). The gene analysis revealed a conserved structure among the camelids, but a slight difference in size (2,012 bp on average) due to intronic variations. Promoters were characterized for the transcription factor binding sites (11 found in total). Intraspecies sequence comparison showed 36 SNPs in total (2 in the dromedary, none in the Bactrian camel, 22 in the alpaca, and 12 in the llama), whereas interspecies comparison showed 86 additional polymorphic sites. Eight SNPs were identified as trans-specific polymorphisms, and 2 of them (g.112A>G and g.1229A>G) were particularly interesting in the New World camels. The first creates a new binding site for transcription factor SP1. An enhancing effect of the g.112G variant on the expression was demonstrated by 3 independent pGL3 gene reporter assays. The latter is responsible for the p.78Ile>Val AA replacement and represents novel allelic variants (named LALBA A and B). A link to protein variants has been established by isoelectric focusing (IEF), and bioinformatics analysis revealed that carriers of valine (g.1229G) have a higher glycosylation rate. Genotyping methods based on restriction fragment length polymorphism (PCR-RFLP) were set up for both SNPs. Overall, adenine was more frequent (0.54 and 0.76) at both loci. Four haplotypes were found, and the AA and GA were the most common with a frequency of 0.403 and 0.365, respectively. Conversely, a putative biological gain characterizes the haplotype GG. Therefore, opportunities for rapid directional selection can be realized if this haplotype is associated with favorable milk protein properties. This study adds knowledge at the gene and protein level for α-LA (LALBA) in camelids and importantly contributes to a relatively unexplored research area in these species.

HAMLET in human milk is resistant to digestion and carries essential free long chain polyunsaturated fatty acids and oleic acid.

The oleic acid/alpha-lactalbumin complex HAMLET (human alpha-lactalbumin made lethal to tumors) is cytotoxic to various cancerous cell lines and is assembled from alpha-lactalbumin (ALA) and free oleic acid (OA). HAMLET is also cytotoxic to normal immature intestinal cells. It remains unclear if HAMLET, experimentally assembled with OA and heat, can spontaneously assemble in frozen human milk over time. To approach this issue, we used a set of timed proteolytic experiments to evaluate the digestibility of HAMLET and native ALA. The purity of HAMLET in human milk was confirmed by ultra high performance liquid chromatography coupled to tandem mass spectrometry and western blot to resolve the ALA and OA components. Timed proteolytic experiments were used to identify HAMLET in whole milk samples. Structural characterization of HAMLET was performed by Fournier transformed infrared spectroscopy and indicated a transformation of secondary structure with increased alpha-helical character of ALA upon binding to OA.

Effect of genetic polymorphisms in LALBA and Prolactin genes on milk traits in Barki ewes

This study investigated the genetic polymorphisms in Alpha-lactalbumin (LALBA) and Prolactin (PRL) genes in the Egyptian Barki sheep. The investigated genes were previously identified to be associated with milk and reproductive traits in several livestock species. Nevertheless, little is known about their contribution to milk traits in Egyptian sheep. Milk traits of daily milk yield (DMY), protein percentage (PP), fat percentage (FP), lactose percentage, and total solid percentage (TSP) were available for 256 and 233 Barki ewes for LALBA and PRL, respectively. The association analysis between the phenotypes of milk traits was carried out using the general linear model implemented in SAS software. The single strand conformation polymorphism identified 3 (A, B and C) and 2 (G and C) distinctive alleles in LALBA and PRL, respectively. Allele frequencies for LALBA variants were 0.73 (A), 0.16 (B) and 0.11 (C), and 0.63 (G), and 0.37 (C) for PRL variants. Results showed that age and parity of the ewe significantly (P < 0.05) influenced the studied milk traits. Additionally, a significant (P < 0.05) effect of LALBA variants was reported on milk composition but not DMY. Likewise, a significant (P < 0.05) effect of PRL variants was observed on DMY and TSP. Interestingly, Animals with BB genotypes of LALBA gene produced milk with high FP (5.65 ± 1.25 %), PP (5.77 ± 1.17 %), LP (7.4 ± 1.22 %) and lower TSP (13.06 ± 5.1 %) compared with animals that carry other genotypes. Regarding the PRL polymorphisms, the heterozygote ewes (GC) tended to significantly produce more milk (368.57 ± 20.39 g/day) with higher TSP (19.81 ± 1.57) compared with the homozygotes. This study reported the LALBA and PRL as candidate genes with significant effects on milk yield and composition, which may be appropriate for a wide range of selection programs with breeding objectives that focus on milk yield and composition in sheep. However, further investigations with larger sample size are recommended to confirm these findings with higher accuracy.

Abstract P2-11-22: Expression of milk whey protein genes in human breast carcinoma cells isolated by LCM impacts prediction of clinical outcomes

Abstract Background: Milk whey protein consumption has been associated with breast cancer prevention, therapy and treatment tolerance, and other studies detected these proteins or their mRNAs in breast cancer models or tissues. Our objectives are to ascertain expression of certain whey protein genes in human breast carcinoma cells isolated by Laser Capture Microdissection (LCM) and to determine if gene expression is related to risk of recurrence and overall survival in lesions with various ER/PR status. Genes selected were: LALBA (alpha-lactalbumin), LPO (lactoperoxidase), LTF (lactotransferrin), PAEP (progestogen associated endometrial protein or glycodelin) and SSP1 (secreted phosphoprotein 1 or osteopontin). This retrospective investigation is unique in that highly quantified ER and PR protein levels and ESR1 and PGR relative expression from pure populations of carcinoma cells were employed to stratify cancers into ER/PR subcategories to assess relationships of expression of candidate genes with clinical outcomes. Methods: This investigation of primary breast carcinomas from 170 postmenopausal and 77 premenopausal patients used a matchless deidentified dataset of quantified estrogen receptor (ER) and progestin receptor (PR) protein results with clinical follow-up. ER/PR protein levels, expressed as fmol/mg cytosol protein, were quantified earlier from each breast carcinoma with FDA-approved kits, either Radioligand-Binding Assay (LBA, NEN/DuPont) and/or enzyme immunoassay (EIA, Abbott Labs). Patients were treated with the standard of care at the time of diagnosis. The PixCell IleTM (ThermoFisher, Arcturus) LCM instrument was used to procure only carcinoma cells from tissue sections of de-identified, frozen biopsies, using protocols we employed earlier. Total RNA of each fixed specimen was extracted, purified and amplified to obtain relative expression levels of approximately 22,000 genes by microarray, providing an assessment of mRNA in carcinoma cells only. Relationships of relative gene expression, quantified ER/PR and other features of primary breast cancers were analyzed with clinical results using R software v4.2.0 (Vigorous Calisthenics) by Univariable Cox Regression, Kaplan Meier plots and various tests (i.e., Wilcoxon, Log Ranked, Spearman and/or Chi Square tests). Bidirectional elimination stepwise regression was also used to derive best predictors of patient outcomes. Results: Violin plots of expression distribution of each milk protein gene of 247 biopsies regardless of patient menopausal status revealed that LALBA and SSP1 were highly elevated (p &amp;lt; 0.001) in ER+ (n = 146) or PR+ (n = 146) breast cancer cells while PAEP was quite diminished (p &amp;lt; 0.001). No difference was observed in LPO expression in ER+ or PR+ cells and LTF was only elevated slightly (p &amp;lt; 0.05) in ER+ cells. Regardless of ER/PR status of breast cancer, LTF, LALBA, SSP1, AR and ESR1 gene expression levels were increased significantly in postmenopausal versus premenopausal patients while PAEP was decreased (p &amp;lt; 0.001). No difference was observed in LPO or PGR expression. Correlation matrices examined gene to gene and receptor protein to gene relationships. Using Kaplan Meier analyses of each gene alone with Log Ranked Tests of outcomes, only SSP1 expression was associated with clinical outcome, i.e., overexpression was associated with lower PFS and OS (p = &amp;lt; 0.05). Conclusions: Application of bidirectional elimination stepwise regression of gene expression of the five milk proteins collectively with ER/PR protein status of each carcinoma revealed that LALBA gene expression enhanced prediction of PFS in ER+ cancer while PAEP expression was associated with improved prediction of OS in ER+ carcinomas. We report that expression of LALBA and PAEP genes, largely associated with milk production in normal breast tissue, are also expressed in certain breast carcinomas and influence prediction of patient survival. Citation Format: James L. Wittliff, Michael W. Daniels. Expression of milk whey protein genes in human breast carcinoma cells isolated by LCM impacts prediction of clinical outcomes [abstract]. In: Proceedings of the 2022 San Antonio Breast Cancer Symposium; 2022 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2023;83(5 Suppl):Abstract nr P2-11-22.

Binding interaction and stability of alpha-lactalbumin and retinol: Effects of pre- or post-acidification

The binding mechanisms of alpha-lactalbumin (ALa) with retinol (VA) and storage stability of their complex after pre-or post-acidification (pH 3.0) were investigated and compared by using multi-spectroscopic approaches. Fluorescence quenching results showed that combination of ALa with VA presented a typically static quenching mode at pH 3.0 with pre- or post-acidification. And the binding constant of post-acidification ALa-VA was greater than that of pre-acidification ALa-VA. Moreover, the corresponding thermodynamic parameters revealed that the binding of VA to ALa was spontaneous complexation processes through a driving force of hydrophobic interaction. Furthermore, post-acidification induced α-helix and β-sheet into β-turn and random coil of ALa-VA, while pre-acidification resulted in its β-sheet convert to disordered β-turn and random coil. The surface hydrophobicity of ALa-VA was notably decreased with the gradual addition of VA after two acid treatments, which were contrary to neutral ALa-VA. Additionally, ALa could effectively prevent VA from thermal degradation, and the stabilizing effect of post-acidification was stronger than that of pre-acidification during VA storage. These findings manifest that two acid treatments for interaction and stability of ALa with VA could cause significant affinity differences, and ALa as a good food matrix for delivering VA in dairy products.

Alpha-lactalbumin amyloid-like fibrils for intestinal delivery: Formation, physiochemical characterization, and digestive fate of capsaicin-loaded fibrils

Engineering protein architectures offers numerous opportunities to deliver consumers with added values yet mandates careful screening of possible risks. This study explores the fabrication of bovine alpha-lactalbumin (ALA) amyloid-like fibrils (AF) for gastro-intestinal delivery of capsaicin (CAP), a pungent compound which possesses health beneficial virtues. Acidic incubation of ALA with CAP accelerates formation of ALA-AF, as confirmed by transmission electron microscope, with a size increase from 3.7 ± 0.4 nm to 175 ± 58 nm and a melting temperature shift from Tm = 61.8 °C to Tm = 89.9 °C for native ALA and ALA-AF entrapping 200 μM CAP, respectively. In addition to encapsulation efficiency and loading capacity measurements, this work provides evidence that fibrillation attenuates the in vitro digestive proteolysis of ALA and diminishes the levels of bioaccessible bioactive peptides by an order of magnitude compared to native ALA. This is accompanied by sustained release of 7.8 ± 1.7% to 55.7 ± 12.3% CAP under gastric and intestinal conditions. Overall, this work presents a new possible avenue for designing protein structures to entrap bioactive moieties along-side a call to interrogate the possible ramifications to protein digestion and consumer health.

Generation of curcumin-loaded albumin nanoparticles by using off-the-shelf microfluidics driven by gravity

The generation albumin-based nanocarriers by precipitation from solution has great interest in the formulation of advanced nutritional products. Microfluidic techniques enable the implementation of low energy and continuum processes, with fast mass transfer and homogeneous mixing at the microscale. Here we describe the microfluidic generation of curcumin-loaded alpha lactalbumin nanoparticles in a simple and inexpensive way, by using off-the-shelf devices designed to produce solvent-shifting nanoprecipitation in core-sheath flows driven by gravity, which has not been reported before. Nanoparticles were characterized by dynamic light scattering, electron microscopy, and infrared spectroscopy. The microfluidic operating conditions were defined by theory and experiments, and the critical parameters controlling the nanoparticles diameter were identified. The prepared nanoparticles resulted practically monodisperse, the curcumin entrapment efficiency was about 40 %, and almost 70 % of the bioactive was gradually delivered in release experiments. The proposed methodology is a promising route to scale up the microfluidic elaboration of nanoparticles for the entrapment of active ingredients.

Bovine alpha-lactalbumin particulates for controlled delivery: Impact of dietary fibers on stability, digestibility, and gastro-intestinal release of capsaicin

Consumer demand for functional foods rich in proteins, fiber and bioactives, such as capsaicin (CAP), is constantly growing. This study hypothesized that the electrostatic biopolymer interactions between bovine alpha-lactalbumin (ALA), and the ionic polysaccharides alginate or chitosan (ALG and CHI, respectively) can be harnessed to form fine particulates with improved stability, attenuated susceptibility to digestive proteolysis and controlled release of CAP. Quantitative analysis show that the addition of dietary fiber increases the encapsulation efficiency of CAP up to 10%. Various light scattering techniques affirm that interactions between ALA and the oppositely charged polysaccharides significantly enhances particulates physical and pH stability. Thirty-day shelf-life studies show that the biopolymer complexation helps retain 50% of the entrapped CAP and limits protein aggregation. Semi-dynamic in vitro digestions highlight ALA-ALG particles control ALA digestive proteolysis and CAP release under oral and gastric conditions. Further, LC-MS/MS proteomic analysis of gastric aspirates suggests polysaccharide addition does not have a marked effect on the liberation of anti-inflammatory, ACE- and DPP-IV inhibitory peptides. Moreover, predictive PeptideRank tool helps demonstrate the potential release of possible novel bioactive peptides whose bioactivity requires further investigation. Thus, this study adds another tier to the existing body of evidence supporting protein-polysaccharide complexation as a possible avenue to develop edible delivery systems.

Characteristics of the lines of the Sibiryachka cattle breed by genes &lt;i&gt;CSN3, BLG, LALBA, LEP&lt;/i&gt; and their relationship with dairy productivity

The results of studies on productivity and genotypic structure of cows of the leading lines of Sibiryachka cattle breed, associative links of CSN3, BLG, LALBA, LEP genotypes with economically important traits are presented. Comparative evaluation showed that Reflection Sovering bull cows had the highest milk yield of 6851 kg, fat content of 4.05% and protein content of 3.15%. The Siberian bull lines Frank 937, Uragan 27 and Kursa 1949 which are being shaped are inferior to them in milk yield, fat and protein content with values of 5246-5504 kg, 3.92-3.94%; 3.10-3.12% respectively. The genotypic structure of the herd and the leading lines is identified. The Vis Back Aydiala bull line is characterized by a higher frequency of CSN3AA and LEPTT genotypes by 18.2 and 11.0%, in comparison with the Reflection Sovering line. For other genotypes, the differences do not reach the confidence threshold. The average level of homozygosity for the genes studied varies from 51.2% to 73.4%. The highest homozygosity was found for the CSN3 gene in the Vis Back Aydiala line at 79.6%. The number of effectively acting alleles is 1.66-1.72; the degree of genetic variability is 40.2-42.7%. The cows with CSN3AB genotype had 544.0 kg higher milk yield than homozygous animals for the A allele (p &lt;0.05). The highest milk yield was observed in BLGAA animals - 6790.1 kg, which is 947.2 kg higher than in cows with the alternative BLGВВ genotype (p &lt; 0.01). Animals with the LEPCC genotype outperformed LEPTT cows in milk yield by 718.7 kg. No priority genotypes were identified for the LALBA gene. Also, no connection has been established between genotypes and the quality indicators of milk.

Identification and in silico characterization of structural and functional impacts of genetic variants in milk protein genes in the Zebu breeds Guzerat and Gyr.

Whole genome sequencing of bovine breeds has allowed identification of genetic variants in milk protein genes. However, functional repercussion of such variants at a molecular level has seldom been investigated. Here, the results of a multistep Bioinformatic analysis for functional characterization of recently identified genetic variants in Brazilian Gyr and Guzerat breeds is described, including predicted effects on the following: (i) evolutionary conserved nucleotide positions/regions; (ii) protein function, stability, and interactions; (iii) splicing, branching, and miRNA binding sites; (iv) promoters and transcription factor binding sites; and (v) collocation with QTL. Seventy-one genetic variants were identified in the caseins (CSN1S1, CSN2, CSN1S2, and CSN3), LALBA, LGB, and LTF genes. Eleven potentially regulatory variants and two missense mutations were identified. LALBA Ile60Val was predicted to affect protein stability and flexibility, by reducing the number the disulfide bonds established. LTF Thr546Asn is predicted to generate steric clashes, which could mildly affect iron coordination. In addition, LALBA Ile60Val and LTF Thr546Asn affect exonic splicing enhancers and silencers. Consequently, both mutations have the potential of affecting immune response at individual level, not only in the mammary gland. Although laborious, this multistep procedure for classifying variants allowed the identification of potentially functional variants for milk protein genes.

Infant Formulas: A Long Story

AbstractThe ideal feeding for infants is the breast milk because it has a balanced nutritional composition, which includes all essential nutrients, in addition to many bioactive factors that contribute to the growth and development of the child, as well as to the maturation of the gastrointestinal tract. Among them are immunological factors, antimicrobials and anti-inflammatory components, digestive enzymes, various types of hormones, and growth factors. If human milk is not available, there is an indication of infant formulas that should follow the recommendations of the Codex Alimentarius of the Food and Agriculture Organization/World Health Organization (WHO). In a century of history, infant formulas have gone from a simple combination of cow milk (evaporated or condensed) and water to highly sophisticated products, elaborated by very refined technological processes to produce lactose-free, antiregurgitation, based on soy protein, hydrolyzed protein in various grades, and only amino acids formulas. The major milestones in the modification of infant formulas were the incorporation of nutrients/ingredients such as: iron, nucleotides, alpha lactalbumin, long-chain polyunsaturated fatty acids, prebiotics, probiotics, postbiotics, oligosaccharides similar to human milk, lactoferrin, and milk fat globule membrane. Many of these ingredients have shown benefits on the immunological system. Despite the technological advances, breast milk remains irreplaceable, being the gold standard for infant feeding.

The effect of single-nucleotide polymorphism in the promoter region of bovinealpha-lactalbumin(LALBA) gene onLALBAexpression in milk cells and milk traits of cows

Polymorphisms of milk protein genes have been proposed as candidate markers for dairy production traits in cattle. In the present study, a polymorphism was detected in the 5'-flanking (promoter) region of the bovine alpha-lactalbumin (LALBA) gene, a T/C transition located at nucleotide -1,001 relative to the transcription start site g.-1001T > C (NC_037332.1:g.31183170T > C), which is recognizable with PstI restriction endonuclease. In silico analyses showed that this mutation created novel retinoid X receptor alpha and vitamin D receptor transcription factor binding sites. Real-time PCR found that cows with different genetic variants of the promoter demonstrated different levels of expression of LALBA mRNA in milk somatic cells (MSCs). The TT genotype cows demonstrated low expression, whereas those with CT demonstrated much higher expression (P < 0.05). ELISA analysis found milk LALBA protein levels also differed between the TT and CT cows (P < 0.05) and that these levels were not correlated with the mRNA abundance in MSC. Association analysis found that the g.-1001T > C polymorphism in the promoter region of the LALBA gene influenced milk production traits in Polish Holstein-Friesian cows. High daily milk yield and dry matter yield, and high lactose yield and concentration were associated with the TT genotype. The TT genotype cows also had a lower number of somatic cells in the milk, considered as an indicator of udder health status. Therefore, the TT genotype could be more desirable from the breeder's perspective.

Detection of genomic regions underlying milk production traits in Valle del Belice dairy sheep using regional heritability mapping.

The aim of this study was to identify genomic regions underlying milk production traits in the Valle del Belice dairy sheep using regional heritability mapping (RHM). Repeated measurements for milk yield (MY), fat percentage and yield (F% and FY) and protein percentage and yield (P% and PY), collected over a period of 6 years (2006–2012) on 481 Valle del Belice ewes, were used for the analysis. Animals were genotyped with the Illumina 50k SNP chip. Variance components, heritabilities and repeatabilities within and across lactations were estimated, fitting parity, litter size, season of lambing and fortnights in milk, as fixed; and additive genetic, permanent environment within and across lactations, flock by test‐day interaction and residual as random effects. For the RHM analysis, the model included the same fixed and random effects as before, plus an additional regional genomic additive effect (specific for the region being tested) as random. While the whole genomic additive effect was estimated using the genomic relationship matrix (GRM) constructed from all SNPs, the regional genomic additive effect was estimated from a GRM matrix constructed from the SNPs within each region. Heritability estimates ranged between 0.06 and 0.15, with repeatabilities being between 0.14 and 0.24 across lactations and between 0.23 and 0.39 within lactation for all milk production traits. A substantial effect of flock‐test‐day on milk production traits was also estimated. Significant genomic regions at either genome‐wide (p < .05) or suggestive (i.e., one false positive per genome scan) level were identified on chromosome (OAR) 2, 3 and 20 for F% and on OAR3 for P%, with the regions on OAR3 in common between the two traits. Our results confirmed the role of LALBA and AQP genes, on OAR3, as candidate genes for milk production traits in sheep.

Identification of Goats’ and Cows’ Milk Protein Profile in Banyumas Regency by Sodium Dedocyl Sulphate Gel Electrophoresis (Sds-Page)

Protein is one of the nutrient components in milk that is related to product quality. The components of milk protein are divided into casein alpha-s1, beta, alpha-s2, kappa, and whey fractions such as alpha lactalbumin and beta lactoglobulin. There are no existing data of milk protein fraction in dairy cow and goats in Banyumas Regency. This study aimed to determine the profile in form of protein fractions of cow and goat milk in Banyumas. Milk sample from fifty cows and thirty dairy goats was taken by random sampling in some areas. The milk protein profile was identified by the technique of sodium dodecyl sulphate gel electrophoresis (SDS-PAGE) and protein quantity prediction by software. The data obtained were analyzed statistically by Mann-Whitney between cows and goats. The results were significantly different (p&lt;0.05) between cows and goats in molecular weight of protein alpha-S1 casein (29.66 vs 33.37 kDa), alpha-S2 (27.76 vs 29.49 kDa), beta (24 , 48 vs 25.59 kDa) and beta lactoglobulin (15.75 vs 15.97 kDa). The quantity of casein alpha-S1 (7.88 vs 4.16 g/l), alpha-S2 (1.31 vs. 4.02 g/l), beta (8.74 vs 14.24 g/l), kappa (2.41 vs. 4.28 g/l) and alpha lactalbumin (0.91 vs 0.7 g / l) was significantly different (p &lt;0.05) between cow's and goat's milk, respectively. In conclusion, milk protein profile of cows and goats in Banyumas Regency is different.

Comparison of carrying mechanism between three fat-soluble vitamins and alpha-lactalbumin: Effects on structure and physicochemical properties of alpha-lactalbumin

This study aims to explore the interaction mechanisms between alpha-lactalbumin (ALa) and three fat-soluble vitamins (VA, VD3 and VE) at pH 7.0 and their impacts on structure and functionality of ALa. Analysis of fluorescence spectroscopy showed that vitamins quenched intrinsic fluorescence of ALa in a static mode and cholecalciferol (VD3) had a higher binding affinity for ALa than Retinol (VA) and alpha-tocopherol (VE). Synchronous fluorescence and three-dimensional fluorescence spectra demonstrated the conformation changes of ALa. Meanwhile, fourier transform infrared spectroscopy manifested that three vitamins induced a transition from α-helix to β-sheet in ALa, and VD3 notably increased its random coil content. Furthermore, a combination of VE with ALa generated macromolecular aggregates, based on particle size results. The addition of VA and VD3 increased zeta potential and surface hydrophobicity of ALa, while VE triggered that of ALa initially increased and then decreased with vitamins ranging from 0 to 0.35 mmol/L. Additionally, molecular simulation suggested that VA and VE had a same binding site in the cleft of ALa, but VD3 bound in a hydrophobic pocket in protein, and their binding driving forces were hydrophobic interaction and hydrogen bond. This study reveals that ALa might be a potentially suitable carrier for delivering vitamins, providing a theoretical basis of interaction between proteins and small molecules.

Properties of hydrolyzed α-lactalbumin, β-lactoglobulin and bovine serum albumin by the alcalase and its immunemodulation activity in Raw 264.7 cell

Properties of hydrolyzed α-lactalbumin, β-lactoglobulin and bovine serum albumin by the alcalase and its immunemodulation activity in Raw 264.7 cell

Association of κ-casein, β-lactoglobulin, α-lactalbumin and leptin gene polymorphisms with bovine productivity traits in Western Siberia

The Simmental breed (n=182) and Siberian breed (n=131) cows were genotyped using the CSN3, BLG, LALBA, and LEP genes. In both populations, the frequency of homozygotes did not exceed 10 %, and the proportion of heterozygotes was at the level of 40-45%. The actual genotype distribution corresponded to the theoretically expected Hardy-Weinberg distribution. In terms of milk yield, cows of the «Sibiryachka» breed with the BLGAA genotype significantly outperformed cows with the BLGBB genotype (p&lt;0.05), and in the Simmental breed, cows with the BLGAA genotype had a higher milk yield than BLGAB (p&lt;0.01). According to the LALBA gene in the Simmental breed, cows with the LALBABB genotype were superior to cows with LALBAAA in fat content (p&lt;0.05), in the «Sibiryachka» breed, higher fat content was observed in cows with the LALBAAB genotype (p&lt;0.05). In both breeds, cows with the LEPCC genotype outperformed cows with the LEPTT genotype in fat content (p&lt;0.05). According to the reproduction indicators, cows with the LALBAAA and LEPCT genotypes had an earlier age of insemination in comparison with cows with the LALBABB and LEPCC genotypes (p&lt;0.05). Cows with the LEPCT genotype had a longer calving interval than cows with the LEPCC genotype (p&lt;0.05).