Infant formulas with increased concentrations of alpha-lactalbumin

Human and bovine milk differ substantially in the ratio of whey to casein protein (approximately 60:40 in human milk and approximately 20:80 in bovine milk) and in the proportions of specific proteins. Although current infant formulas closely mimic the ratio of total whey to casein inhuman milk, the concentration of a-lactalbumin (the dominant protein in human milk) is relatively low in formula, whereas beta-lactoglobulin, a protein not found in human milk, is the most dominant whey protein in formula. Because of the differences in the protein profiles of human milk and infant formula, amino acid profiles also differ. To meet all essential amino acid requirements of infants, formula concentrations of protein must be higher than those in human milk. Recently, whey sources with elevated concentrations of alpha-lactalbumin have become available, which permitted the development of formulas with increased concentrations of this protein and decreased concentrations of beta-lactoglobulin. alpha-Lactalbumin is rich in tryptophan, which is typically the limiting amino acid in formula, and as a result, formulas have been developed with lower protein but higher tryptophan concentrations. This type of formula may offer a number of advantages to the neonate, which include producing plasma tryptophan concentrations equal to those found in breastfed infants and obviating the need for the body to dispose of excess nitrogen loads.     

Substitution of casein by beta-casein or of whey protein isolate by alpha-lactalbumin does not affect mineral balance in growing rats

Bovine milk protein fractions that enable modification of the protein composition and amino acid profile of infant formulas to mimic those of human milk have recently become available. To determine the effects on protein quality and mineral bioavailability of replacing casein by beta-casein and of whey protein isolate by alpha-lactalbumin, 4 groups of growing rats were fed for 3 wk diets containing 10% protein as 1) casein (control); 2) beta-casein; 3) casein:whey (40:60); or 4) beta-casein:alpha-lactalbumin (40:60). Protein quality, determined as protein efficiency ratio (PER), net protein utilization (NPU), biological value (BV) and protein digestibility (PD), as well as body weight gain, were higher (P < 0.05) with consumption of the whey-adapted diets [casein:whey (40:60); beta-casein:alpha-lactalbumin (40:60)] compared with the casein diets (casein; beta-casein); however, there were no differences between the 2 casein diets or between the 2 whey-adapted diets. Apparent absorption of minerals (Ca, P, Fe, Zn) from the whey-adapted diets was higher than that from the casein diets (P < 0.05); but again, no differences were observed when casein or whey protein isolate were replaced by beta-casein or alpha-lactalbumin, respectively. Thus, substitution of casein by beta-casein or of whey protein isolate by alpha-lactalbumin does not affect protein quality or mineral bioavailability as determined in growing rats.     

Estimation of available methionine and cysteine in proteins of food products by in vivo and in vitro methods.

1. The available methionine and cysteine of proteins were determined by chemical methods after preliminary enzymic hydrolysis. 2. The values for the available methionine and cysteine contents of pure proteins (casein and bovine serum albumin) estimated by chemical methods were similar to those for the total content determined by the method of Moore, Spackman & Stein (1958). 3. Reductions of 15 and 11% respectively, when compared with unprocessed samples, were found in the available methionine contents of sweetened and unsweetened, condensed milks; of roller-dried milk and whey powders, and of mackerel sterilized at 126 degrees, the reductions were 22, 14 and 19% respectively. 4. The available cysteine content of sweetened, condensed milk was reduced by about 32%, whereas for mackerel sterilized at 115 and 126 degrees it was reduced by 64 and 75% respectively. 5. The contents of total sulphur amino acids for these food products did not differ from those for the unprocessed samples. 6. Values obtained for available S amino acid contents by rat bioassay confirmed the results of the in vitro estimations.     

Tryptophan intake influences infants' sleep latency.

Formula-fed infants have depressed plasma tryptophan concentration relative to human milk-fed infants. Because tryptophan alters sleep-waking patterns in adults, a study was designed to determine whether additional dietary tryptophan could elevate plasma tryptophan concentrations of formula-fed infants to concentrations characteristic of human milk feeding and whether differences in plasma tryptophan or the ratio of plama tryptophan to the sum of the other plasma large neutral amino acids (tryptophan:LNAA) were of behavioral significance. Infants were fed a formula (13 g protein/L; whey:casein, 34:66) containing either 0, 294, 588 or 882 mumol/L of added tryptophan. Infants fed human milk or commercial formula (15 g protein/L; whey:casein, 18:82) were included for comparison. In formula-fed groups, plasma tryptophan was directly related to tryptophan intake (r = 0.46, P less than 0.0005). Infants fed commercial formula or the formula without added tryptophan had lower (P less than 0.001) plasma tryptophan compared with infants fed human milk. Only the infants fed the highest tryptophan formula had significantly higher plasma tryptophan:LNAA ratios than the other experimental groups, and these ratios were similar to those of infants fed human milk. The plasma tryptophan:LNAA ratios, not plasma tryptophan concentrations, were predictive of differences in the infants' sleep latency; infants fed formula containing the highest tryptophan had sleep latencies of 18.7 min, significantly shorter (P less than 0.05) than those of infants fed formulas containing less added tryptophan (27.7 min). Feeding infants formulas differing in tryptophan concentration produced differences in sleep latency, which could influence neurobehavioral developments.(ABSTRACT TRUNCATED AT 250 WORDS)     

Casein content of human milk

Three methods for estimating the casein content of human milk were tested; isoelectric precipitation with washing and correction for co-precipitating proteins, sedimentation by ultracentrifugation, and indirect analysis (ie analyzing for the content of the major whey proteins and subtracting these from the total protein content). Gel electrophoresis and amino acid analysis were used to confirm some of the results. The casein content (mg/ml) of mature human milk (n = 9) was 2.33 +/- 1.69 by isoelectric precipitation, 1.80 +/- 0.48 by sedimentation and 2.96 +/- 1.08 by the indirect approach. A probable partition of nitrogen in breast milk would be casein N: whey protein N: non-protein N of 20:50:30; ie the correct ratio of casein nitrogen: whey nitrogen is approximately 20:80. Analysis of trace elements and minerals demonstrates that of total Ca 10%, Mg 5%, Zn 28%, Cu 17%, and Fe 27% is bound to casein when prepared by ultracentrifugation while isoelectric precipitation causes a redistribution of some of these elements. Since the protein ratio of human milk is considered a guideline when manufacturing infant formulas, these findings should be considered with regard to infant nutrition.     

Reduction in the antigenicity of whey proteins by heat treatment: a possible strategy for producing a hypoallergenic infant milk formula

Residual antigenic protein in heat-denatured cow's milk whey and in two commercial infant milk formulas was determined using enzyme-linked immunosorbent assays specific for beta-lactoglobulin, alpha-lactalbumin, bovine serum albumin, bovine IgG1 and alpha-casein. This immunochemical assessment of antigenicity was related to the capacity of the preparations to sensitize immunologically when fed to guinea-pigs for 2 weeks. Antibody production was measured and the susceptibility of the animals to systemic anaphylaxis was assessed by injecting them intravenously with heated or unheated milk proteins. Whey protein that had been heated at 100 degrees or 115 degrees for 30 min was extensively denatured and, in contrast to pasteurized whey, failed to sensitize guinea-pigs for anaphylaxis. Antibody production was undetected or very low. The proteins in SMA powder and SMA Gold Cap liquid concentrate were less denatured and animals given these formulas prepared according to the maker's instructions produced relatively high levels of antibodies to beta-lactoglobulin and alpha-casein and a majority developed anaphylaxis when injected intravenously with these products. As well as failing to sensitize, whey that had received severe heat treatment did not, in most cases, elicit anaphylaxis when injected into animals that had been sensitized with unheated milk. Discrimination between antibodies of the IgG1 and IgG2 subclasses specific for beta-lactoglobulin showed that IgG1, the principal anaphylactic antibody in guinea-pigs, was preferentially depressed in animals drinking heat-denatured milk preparations. The results suggest that heat denaturation of whey protein may be a logical and simple strategy for producing a hypoallergenic baby milk. Nevertheless, the value of experiments in guinea-pigs for predicting results in man is uncertain and the proposal awaits assessment in clinical trials.     

Reactions of proteins with oxidizing lipids. 2. Influence on protein quality and on the bioavailability of lysine, methionine, cyst(e)ine and tryptophan as measured in rat assays

The consequences of reactions between protein and oxidizing lipids on the nutritional quality of food proteins have been investigated using a whey protein-methyl linolenate-water model system. In rat assays, significant reductions were observed in protein efficiency ratio, net protein ratio, net protein utilization, biological value and true nitrogen digestibility, especially when the reaction had taken place at high moisture content, high temperature and in the presence of excess oxygen. The losses of bioavailable lysine and tryptophan as measured by rat assays followed a similar pattern. The chemical value of each amino acid multiplied by the true N digestibility closely resembled the rat assay value. In general, the reaction products of lysine and tryptophan formed during lipid oxidation were biologically unavailable. The bioavailabilities of methionine and of 'methionine plus cyst(e)ine' were determined in separate assays. Cyst(e)ine was calculated as 'methionine plus cyst(e)ine' minus methionine. In whey protein which had reacted with oxidizing methyl linolenate, the bioavailable methionine content was not significantly reduced even though 82% of the methionine residues were present as methionine sulphoxide. In hydrogen peroxide-treated casein in which all methionine residues were oxidized to the sulphoxide, methionine sulphoxide was found to be 96% as utilizable as a methionine source to the rat. Free methionine sulphoxide was 87% utilizable. Cyst(e)ine appeared to be as sensitive as lysine to reactions with lipid oxidation products. In whey protein which had reacted with oxidizing methyl linolenate, the bioavailabilities of cyst(e)ine, lysine, tryptophan and methionine were reduced by 28, 24, 11 and 8% respectively and true N digestibility by 9%. These results are discussed in relation to food products.     

Free amino acid content in standard infant formulas: comparison with human milk

OBJECTIVE: To compare the concentration of non-protein nitrogen (NPN) and free amino acids (FAA) in powdered and liquid commercial formulas with that in human milk. METHODS: The non-protein nitrogen and FAAs in pooled breast milk was compared with that in 11 protein-modified starting infant formulas (seven powdered, four liquid whey-predominant formulas) and one powdered soy-formula. Human milk was collected at the end of each feeding (hindmilk) over 24 hours in a group of 40 healthy lactating women after delivery of full-term infants at age one month. RESULTS: In human milk glutamic acid plus glutamine and taurine were the prevalent amino acids, accounting for around 50% total FAA. In the analysed formulas the total FAA fraction was 10% or even less than in human milk, mostly represented by taurine, while methionine was high in soy formula. The sum of glutamic acid and glutamine in all the formulas was much lower than in human milk. CONCLUSIONS: Breastfed infants are supplied with FAA, mainly glutamic acid and glutamine, compared to formula-fed counterparts. The different FAA intake might be the origin of some functional differences at the enteral level between breast- and formula-fed infants.     

Mediterranean milk and milk products

Milk and dairy products are part of a healthy Mediterranean diet which, besides cow's milk, also consists of sheep's, goat's and buffalo's milk--alone or as a mixture---as raw material. The fat and protein composition of the milk of the various animal species differs only slightly, but in every case it has a high priority in human nutrition. The milk proteins are characterized by a high content of essential amino acids. Beyond that macromolecules,which have various biological functions, are available or may be formed by proteolysis in milk. Taking this into consideration, the technology of different well-known Italian and German cheese types is presented and the differences as well as correspondences regarding nutrition are discussed. Especially Ricotta and Mascarpone are discussed in detail. Ricotta represents a special feature as this cheese is traditionally made of whey and cream. Thus the highly valuable whey proteins which contain a higher amount of the amino acids lysine, methionine and cysteic acid in comparison to casein and, additionally, to soy protein, are made usable for human nutrition. Finally, it is pointed out on the basis of individual examples that technologies to enrich whey proteins in cheese are already available and in use. Thus, the flavor of low fat cheese is improved and the nutritional value is increased.     

Free Amino Acid Content in Standard Infant Formulas: Comparison with Human Milk

Objective: To compare the concentration of non-protein nitrogen (NPN) and free amino acids (FAA) in powdered and liquid commercial formulas with that in human milk.

Methods: The non-protein nitrogen and FAAs in pooled breast milk was compared with that in 11 protein-modified starting infant formulas (seven powdered, four liquid whey-predominant formulas) and one powdered soy-formula. Human milk was collected at the end of each feeding (hindmilk) over 24 hours in a group of 40 healthy lactating women after delivery of full-term infants at age one month.

Results: In human milk glutamic acid plus glutamine and taurine were the prevalent amino acids, accounting for around 50% total FAA. In the analysed formulas the total FAA fraction was 10% or even less than in human milk, mostly represented by taurine, while methionine was high in soy formula. The sum of glutamic acid and glutamine in all the formulas was much lower than in human milk.

Conclusions: Breastfed infants are supplied with FAA, mainly glutamic acid and glutamine, compared to formula-fed counterparts. The different FAA intake might be the origin of some functional differences at the enteral level between breast- and formula-fed infants.     

母乳α-乳清蛋白、β-酪蛋白与婴幼儿健康的研究进展

母乳蛋白质分为乳清蛋白和酪蛋白两大类。α-乳清蛋白和β-酪蛋白分别是母乳乳清蛋白和酪蛋白中含量最高的蛋白质。本文将对α-乳清蛋白和β-酪蛋白对婴幼儿健康影响进行综述。[营养学报,2020,42(1):78-82]     

Human milk nonprotein nitrogen components: changing patterns of free amino acids and urea in the course of early lactation

Free amino acids and urea were analyzed in 78 human milk samples obtained during the first 5 wk of lactation from 10 mothers delivering at term. Significant differences (p less than 0.05) in the concentrations between colostral and mature milk were found for glutamic acid, glutamine, alanine, glycine, cystine, and phosphoethanolamine which increased, and with serine, phosphoserine, aspartic acid + asparagine, arginine, lysine, isoleucine, phenylalanine, proline, methionine, tryptophan, and beta-alanine which decreased. Some of these changes occurred within the first 5 days of lactation, so that differences between transitional and mature milk became negligible (glutamic acid, alanine, and serine, aspartic acid + asparagine, lysine, isoleucine, methionine, tryptophan, respectively). No significant differences between any of the three stages of lactation were found regarding the concentrations of total free amino acids, urea, taurine, threonine, valine, leucine, histidine, and tyrosine. Possible relevances for free amino acids, including nonprotein ones, in human milk are discussed.     

Effects of alpha-lactalbumin-enriched formula containing different concentrations of glycomacropeptide on infant nutrition

BACKGROUND: Formula-fed infants have growth and plasma amino acid patterns different from those of breastfed infants. OBJECTIVE: alpha-Lactalbumin is a major protein in human milk, and the addition of bovine alpha-lactalbumin to infant formula has been proposed to modify the plasma amino acid pattern of the recipient infant, possibly allowing a reduction in the protein content of the formula, which may affect growth. DESIGN: We compared breastfed infants and infants fed standard formula or alpha-lactalbumin-enriched formulas (25% of protein) with glycomacropeptide accounting for 15% or 10% of the protein. The protein content of each formula was 13.1 g/L. Ninety-six infants aged 6 +/- 2 wk were recruited. Anthropometric measures were recorded, and interviews were conducted at enrollment and monthly until 6 mo of age. Blood samples were collected at enrollment and at 4 and 6 mo. RESULTS: Formula intake did not differ between groups, and weight gain in the alpha-lactalbumin-enriched formula groups were similar to that of the breastfed infants. The standard formula group gained significantly more weight than did the breastfed infants. All formula-fed infants had significantly higher plasma concentrations of most essential amino acids at 4 and 6 mo than did the breastfed infants, and serum urea nitrogen was also higher in the formula-fed infants. Insulin and leptin concentrations did not differ between groups. CONCLUSIONS: Compared with standard formula-fed infants, infants fed formula with a modified protein composition had growth patterns more similar to those of breastfed infants. All formula-fed groups had plasma amino acid concentrations similar to or higher than those of breastfed infants. This indicates that the protein content of alpha-lactalbumin-enriched formula can be further reduced, which should be evaluated.     

Amino acid contents of infant foods

The protein quality of three milk–cereal-based infant foods (paps) was evaluated by determining their amino acid contents and calculating the amino acid score. Proteins were subjected to acid hydrolysis, prior to which cysteine and methionine were oxidized with performic acid. Amino acids were determined by reverse-phase high-performance liquid chromatography with fluorescence detection with a prior derivatization with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate. Tryptophan was determined by reverse-phase high-performance liquid chromatography with ultraviolet detection after basic hydrolysis. Glutamic acid, proline and leucine were the most abundant amino acids, whereas tryptophan and cysteine had the lowest contents. Tryptophan was the limiting amino acid in the analyzed infant foods. A pap serving (250 ml) contributes significantly to fulfillment of the recommended dietary allowances of essential and semi-essential amino acids for infants (7–12 months old) and young children (1–3 years old).     

Comparative proteomic exploration of whey proteins in human and bovine colostrum and mature milk using iTRAQ-coupled LC-MS/MS

Whey, an essential source of dietary nutrients, is widely used in dairy foods for infants. A total of 584 whey proteins in human and bovine colostrum and mature milk were identified and quantified by the isobaric tag for relative and absolute quantification (iTRAQ) proteomic method. The 424 differentially expressed whey proteins were identified and analyzed according to gene ontology (GO) annotation, Kyoto encyclopedia of genes and genomes (KEGG) pathway, and multivariate statistical analysis. Biological processes principally involved biological regulation and response to stimulus. Major cellular components were extracellular region part and extracellular space. The most prevalent molecular function was protein binding. Twenty immune-related proteins and 13 proteins related to enzyme regulatory activity were differentially expressed in human and bovine milk. Differentially expressed whey proteins participated in many KEGG pathways, including major complement and coagulation cascades and in phagosomes. Whey proteins show obvious differences in expression in human and bovine colostrum and mature milk, with consequences for biological function. The results here increase our understanding of different whey proteomes, which could provide useful information for the development and manufacture of dairy products and nutrient food for infants.

The advanced iTRAQ proteomic approach was used to analyze differentially expressed whey proteins in human and bovine colostrum and mature milk.     

重组人α-乳清蛋白奶粉的营养价值评价

目的 评价转人α-乳清蛋白奶粉的营养价值。方法 根据《转基因生物及其产品食用安全检测—蛋白质功效比实验方法》测定刚断乳（出生后20～23 d）SD大鼠1个月后的体重增长和进食量,计算蛋白质的功效比;测定重组α-乳清蛋白奶粉的氨基酸含量,计算氨基酸评分;测定重组α-乳清蛋白奶粉营养素含量,比较与普通奶粉差异。结果 重组人乳清蛋白奶粉雄性动物组总体蛋白功效比值无统计学差异（P > 0.05）。雌性动物重组人乳清蛋白奶粉组和其他对照组蛋白功效比值无统计学差异（P > 0.05）;重组人乳清蛋白奶粉的氨基酸含量除异亮氨基酸含量低于普通奶粉,其他氨基酸含量都高于普通奶粉;重组人乳清蛋白奶粉和普通奶粉的营养含量有差别。 结论 重组α-人乳清蛋白奶粉的蛋白功效和其他组相当,营养成分虽有点低,但氨基酸组成更合适。     

S-Carboxyethylcysteine (a constituent of Acacia seed) negatively affects casein protein utilization by rats

ObjectiveTwo rat bioassay experiments are reported. The first investigated the first limiting amino acid in Acacia colei and the second experiment investigated the effect of S-carboxyethylcysteine (CEC; a compound present in acacia seed) on protein use.MethodsIn the first experiment, Wistar rats were fed A. colei seed supplemented with three levels of methionine, cysteine, and tryptophan (0.1%, 0.2%, and 0.4%). In the second experiment, the Wistar rats were fed CEC-incorporated casein diets.ResultsSupplementation of A. colei with tryptophan had no significant effect on the protein efficiency ratio, cysteine showed the highest protein efficiency ratio value at the 0.4% level, and the protein efficiency ratio increased significantly with the increase in methionine content, making methionine the first limiting amino acid. The methionine-induced growth rate was suppressed by the incorporation of CEC, which also had a negative effect on the plasma amino acid levels.ConclusionThe results indicated that methionine is the first limiting amino acid in A. colei and that CEC could affect the seed’s protein use. Acacia colei seed can be used effectively as famine food only if it is complemented with other cereals known to be rich in sulfur amino acids.     

Whey protein rich in alpha-lactalbumin increases the ratio of plasma tryptophan to the sum of the other large neutral amino acids and improves cognitive performance in stress-vulnerable subjects

BACKGROUND: Cognitive performance often declines under chronic stress exposure. The negative effect of chronic stress on performance may be mediated by reduced brain serotonin function. The uptake of the serotonin precursor tryptophan into the brain depends on nutrients that influence the availability of tryptophan by changing the ratio of plasma tryptophan to the sum of the other large neutral amino acids (Trp-LNAA ratio). In addition, a diet-induced increase in tryptophan may increase brain serotonergic activity levels and improve cognitive performance, particularly in high stress-vulnerable subjects. OBJECTIVE: We tested whether alpha-lactalbumin, a whey protein with a high tryptophan content, would increase the plasma Trp-LNAA ratio and improve cognitive performance in high stress- vulnerable subjects. DESIGN: Twenty-three high stress-vulnerable subjects and 29 low stress-vulnerable subjects participated in a double-blind, placebo-controlled, crossover study. All subjects conducted a memory-scanning task after the intake of a diet enriched with either alpha-lactalbumin (alpha-lactalbumin diet) or sodium caseinate (control diet). Blood samples were taken to measure the effect of dietary manipulation on the plasma Trp-LNAA ratio. RESULTS: A significantly greater increase in the plasma Trp-LNAA ratio after consumption of the alpha-lactalbumin diet than after the control diet (P = 0.0001) was observed; memory scanning improved significantly only in the high stress-vulnerable subjects (P = 0.019). CONCLUSION: Because an increase in the plasma Trp-LNAA ratio is considered to be an indirect indication of increased brain serotonin function, the results suggest that dietary protein rich in alpha-lactalbumin improves cognitive performance in stress-vulnerable subjects via increased brain tryptophan and serotonin activities.     

Changes in food proteins reacted with nitrite at gastric pH

Casein and egg albumin were reacted with nitrite at pH 3. The reaction mixtures containing nitrite yellowed when the reaction began; after 24 hours of reaction with 7.5 X 10(-3) M nitrite, about 70% of added casein was sedimented as yellow precipitate. There was a considerable loss of lysine, arginine, tyrosine, and tryptophan residues. This was determined from the amino acid compositions of the proteins obtained by the reaction with 2.5 X 10(-2) M nitrite at pH 3 for 48 hours. An amino acid elution pattern of the treated casein showed five peaks due to new amino acids; three of these were provisionally identified to be cysteic acid, methionine sulfone, and 3-nitrotyrosine.     

Gelation of the heat-induced complex between kappa-casein and alpha-lactalbumin

Whey is a by-product of the manufacture of cheese from milk. The usual practice is to dispose of it, the usage of whey being not sufficiently developed, though it contains proteins of excellent quality such as beta-lactoglobulin and alpha-lactalbumin. Interaction between kappa-casein and whey protein was examined in order to form new food materials. Gelation of the heat-induced complex between kappa-casein and alpha-lactalbumin is described in this paper. alpha-Lactalbumin was easily separated from whey protein by the gel filtration technique on a Toyopearl HW-50 column at pH 6.0. Heat treatment facilitated the hydrolysis of a mixture of kappa-casein and alpha-lactalbumin by trypsin, alpha-chymotrypsin and pronase. Heat treatment at above 75 degrees C and a protein level of over 5% were needed to form gel in 35 mM phosphate buffer, pH 7.6, containing 0.4 M NaCl. The temperature was in agreement with that at which alpha-lactalbumin denatured and formed the complex with kappa-casein. Decrease of soluble protein concentration and increase of turbidity were induced with gelation. Gel was not formed when only kappa-casein or alpha-lactalbumin was heated under the appropriate conditions. It was considered that a kappa-casein-alpha-lactalbumin gel was formed from a complex of the two proteins by heat treatment. The breaking stress of kappa-casein-alpha-lactalbumin gel was less than that of kappa-casein-beta-lactoglobulin gel. If the pH was reduced to 5.8 after complex formation by the two proteins, gel was formed at a low protein concentration compared with that with no alteration of pH.