Supplier of Biologically Functional and Nutritional Food Ingredients

WHEY PROTEIN RESEARCH ARTICLES, from the peer reviewed technical papers.

Functional properties of whey, whey components, and essential amino acids: mechanisms underlying health benefits for active people (Review). Ewan Ha and B. Zemel. J. Nutr. Biochem. 14 (2003) 251-258. Whey proteins and amino acid supplements have a strong position in the sports nutrition market based on the purported quality of proteins and amino acids they provide. Recent studies employing stable isotope methodology demonstrate the ability of whey proteins or amino acid mixtures of similar composition to promote whole body and muscle protein synthesis. Other developing avenues of research explore health benefits of whey that extend beyond protein and basic nutrition. These functions are being investigated predominantly in tissue culture systems and animal models. The capacity of these compounds to modulate adiposity, and to enhance immune function and anti-oxidant activity presents new applications potentially suited to the needs of those individuals with active lifestyles. The paper will review the recent literature that describes functional properties of essential amino acids, whey proteins, whey-derived minerals and other compounds and the mechanisms by which they may confer benefits to active people in the context that exercise is a form of metabolic stress. The response to this stress can be positive, as with the accretion of more muscle and improved functionality or greater strength. However, overall benefits may be compromised if immune function or general health is challenged in response to the stress. From a mechanistic standpoint, whey proteins, their composite amino acids, and/or associated compounds may be able to provide substrate and bioactive components to extend the overall benefits of physical activity.

MILK, THE RICH SOURCE OF BIOACTIVE INGREDIENTS FOR FUNCTIONAL, NUTRACEUTICAL AND PHARMACEUTICAL INDUSTRY Biologically active components in the bovine milk, especially whey, have been well identified and their biological activities have been characterized. Their efficacies have been tested in vitro and in vivo but further human clinical test results are needed to support the bioactivity claims, especially for the pharmaceutical applications. As the industrial scale protein and peptide separation technologies such as membrane separation technology and chromatography are available, dairy industry should pay attention to the new business opportunities of producing the bioactive ingredients for the functional, nutraceutical and pharmaceutical industry, Ewan Ha, 2001 Dairy Ind & Technol. 1, 93-124, 2001.

Nutritional Quality of Whey Proteins (from Ewan Ha, 2001). Whey proteins are recognized as being of very high nutritional quality. They are rich in sulfur-containing amino acids such as cysteine and methionine and easily digested. The essential amino acid profile of whey proteins meets or exceeds all the nutritional requirements of the FAO/WHO (USDEC, 1997). The Protein Digestibility-Corrected Amino Acid Scoring (PDCAAS) method was adopted by FDA as the official method for determining protein quality in food products intended for children over one year of age and adults in 1993 (Morr and Ha, 1995). Using the PDCAAS method, whey protein rates the highest value of 1.0, because it is highly digestible and it meets or exceeds the recommended amount of each essential amino acid. Using another method to measure protein quality - the Protein Efficiency Ratio (PER) value - whey also ranks high on the scale. The higher the PER value, the better the protein. Casein, the reference protein, has a PER value of 2.5. Protein with a PER greater than 2.5 are considered to be quality proteins. Whey protein, with a PER greater than 3.0, is considered to be nutritionally excellent protein. Virtually every amino acid present in sweet-type whey exceeds Food Agriculture Organization/World Health Organization (FAO/WHO) nutritional intake recommendations, both for children aged 2-5 and for adults. In many cases, for adults, whey proteins offer more than double the minimum FAO/WHO standards. Whey proteins compare favorably to many common proteins that have a PER of less than 2.5, including soy (2.2), peanuts (1.8), corn (2.2) and wheat gluten (0.8). These products have limited concentrations of certain essential amino acids.

BIOLOGICALLY ACTIVE COMPONENTS IN MILK

Growth stimulating factors in milk
Insulin-like growth factors 1 and 2 (IGF-1 and IGF-2) together with a truncated form of IGF-1 were read.
Effects of bovine colostrum supplementation on serum IGF-I, IgG, hormone, and saliva IgA during training.Mero A et al.J Appl Physiol 1997 read
Colostrum is the first natural food produced by female mammals during the first 24-36h directly after giving birth. Chemically, colostrum is a very complex fluid rich in nutrients, antibodies and growth factors. In cows the antibodies...Pakkanen, R. / Aalto, J., International Dairy Journal, May 1997

Antimicrobial factors in milk
Bovine colostral antibodies against H. pylori read

Antiviral factors in milk
Antiviral effects of plasma and milk proteins: lactoferrin shows potent activity against both human immunodeficiency virus and human cytomegalovirus replication in vitro. Native and chemically derivatized proteins purified from serum and milk were assayed in vitro to assess their inhibiting capacity on the cytopathic effect of human immunodeficiency virus (HIV)-1 and human cytomegalovirus (HCMV) on MT4 cells and fibroblasts, respectively. read

WHEY PROTEINS (From Ewan Ha, 2001)
Whey is a co-product of cheese and casein manufacture and contains about 12% of protein as a solid basis. Whey proteins are universally defined as those proteins that remain in milk serum after coagulation of the caseins at pH 4.6 and 20°C (Morr and Ha, 1993). Whey proteins are compact globular proteins ranging from 14 to 1000 Kd. The percentage of each of the major whey proteins, that is, beta-lactoglobulin (beta-Lg), alpha-lactalbumin (alpha-La), bovine serum albumin (BSA), and the immunoglobulins (Ig) are in Table 2. beta-Lg exists as a dimer in solutions above its isoelectric point of pH 5.2. Between pH 3.5 and 5.2 the dimer polymerizes to an octomer. Below pH 3.5 and above pH 7.5, the dimer dissociates to a slightly expanded monomer. beta-Lg undergoes time- and temperature-dependent denaturation above 65°C. alpha-La was denatured at 62-63°C and that it was 90% renatured at pH 6.5 (de Wit et al., 1983; Morr and Ha, 1993). alpha-La binds Ca⁺², Zn⁺², and other metal ions. alpha-La undergoes a conformational transition at £pH 4, which is probably due to the replacement of this tightly bound Ca⁺² ion with a H⁺¹ ion at this pH. These physicochemical properties of individual whey proteins have been utilized in the whey protein fractionations (Bramaud et al., 1997; Gésan-Guziou et al. 1999; Pearce, 1983, 1987, 1992; Pierre and Fauquant, 1986; see Table 3). One of the key properties of the whey proteins is their abundance of sulfhydryl amino acid residues that allow them to form intermolecular covalent bonds during high-temperature processing of whey and whey protein concentrates solutions. Intermolecular disulfide bonding is essential for forming heat-induced whey protein gels and for stabilizing foam structures (Morr and Ha, 1993).

In addition to the major whey proteins, whey contains additional minor proteins and peptides. These minor components include: 1) b-casein fragments that are heat stable and soluble at pH 4.6, formerly known as proteose-peptone components (5, 8-slow, 8-fast), 2) the highly acidic casein glycomacropeptide (CMP also known as GMP) fragments of k-casein, 3) the highly basic proteins such as lactoferrin, lactoperoxidase, and growth-stimulating factors in whey, 4) phospholipoprotein complex (PLPC) consisting of skim milk membrane and milk fat globule membrane (MFGM) that are complexed with phospholipids and small milk fat globules (MFG), and 5) many other biologically active peptides.

Whey protein fractionation technologies have been reported using ion-exchange chromatography, affinity chromatography, selective precipitation and membrane fractionation. Spherosil-QMA anion exchange resin was used to preferentially adsorb b-lactoglobulin at pH 6.63 with minimal adsorption of the positively charged immunoglobulins, lactoferrins, and lactoperoxidase (Skudder, 1985). b-Lactoglobulin (Amundson et al., 1982) and a-lactalbumin (Pearce, 1983, 1987, 1992) were thermally aggregated and selectively precipitated at pH 4.65 and pH4.2 at 65°C, respectively. Although these reported chromatographic techniques could provide effective protein purification, most of them are not acceptable for the large-scale production due to the cost of production (Pearce, 1992).

The Proteose Peptone Components, identified as Component 5, 8-slow, and 8-fast by gel electrophoresis, have been reclassified as b-casein fragments 1P, 4P and 5P. These protein fragments contain low concentration of aromatic and sulfur amino acid residues that are not precipitated by heating milk at 95°C for 20 min and subsequently adjusting to pH 4.6 with acid. Although present in milk, additional b-casein fragments are likely to be formed during the cheese and whey manufacturing processes (Morr and Ha, 1993).
The proteose peptone component 3 (PP3), also known as lactophorin, is heat stable, acid soluble polypeptide. The PP3 component is a hydrophobic phosphoglycoprotein made up of 135 amino acids and is found in low level in milk (300mg/L). Bovine PP3 component is not sulfated but contains variable levels of sialic acids and fucose and not a membrane protein (Guimont, 1997). Hybridoma proliferation with the PP3 component of bovine, sheep and goat milk as a fetal bovine serum (FBS) substitute in cell culture was reported. The surface hydrophobicity and the glucosidic nature of PP3 appear to be essential to the mitogenic activity. When sialic acids are cleaved by neuraminidase, the PP3 component lost its ability to stimulate hybridoma mitosis. The negative charge of the carboxyl groups may play a role in the stabilization of the active conformation and in the solubilization of the glycoprotein.



\|Chelated Minerals\|Colostrum\|Galacto-oligosaccharides \|Phospholipids\|Whey Protein\|
	WHEY PROTEIN RESEARCH ARTICLES, from the peer reviewed technical papers. Functional properties of whey, whey components, and essential amino acids: mechanisms underlying health benefits for active people (Review). Ewan Ha and B. Zemel. J. Nutr. Biochem. 14 (2003) 251-258. Whey proteins and amino acid supplements have a strong position in the sports nutrition market based on the purported quality of proteins and amino acids they provide. Recent studies employing stable isotope methodology demonstrate the ability of whey proteins or amino acid mixtures of similar composition to promote whole body and muscle protein synthesis. Other developing avenues of research explore health benefits of whey that extend beyond protein and basic nutrition. These functions are being investigated predominantly in tissue culture systems and animal models. The capacity of these compounds to modulate adiposity, and to enhance immune function and anti-oxidant activity presents new applications potentially suited to the needs of those individuals with active lifestyles. The paper will review the recent literature that describes functional properties of essential amino acids, whey proteins, whey-derived minerals and other compounds and the mechanisms by which they may confer benefits to active people in the context that exercise is a form of metabolic stress. The response to this stress can be positive, as with the accretion of more muscle and improved functionality or greater strength. However, overall benefits may be compromised if immune function or general health is challenged in response to the stress. From a mechanistic standpoint, whey proteins, their composite amino acids, and/or associated compounds may be able to provide substrate and bioactive components to extend the overall benefits of physical activity. MILK, THE RICH SOURCE OF BIOACTIVE INGREDIENTS FOR FUNCTIONAL, NUTRACEUTICAL AND PHARMACEUTICAL INDUSTRY Biologically active components in the bovine milk, especially whey, have been well identified and their biological activities have been characterized. Their efficacies have been tested in vitro and in vivo but further human clinical test results are needed to support the bioactivity claims, especially for the pharmaceutical applications. As the industrial scale protein and peptide separation technologies such as membrane separation technology and chromatography are available, dairy industry should pay attention to the new business opportunities of producing the bioactive ingredients for the functional, nutraceutical and pharmaceutical industry, Ewan Ha, 2001 Dairy Ind & Technol. 1, 93-124, 2001. Nutritional Quality of Whey Proteins (from Ewan Ha, 2001). Whey proteins are recognized as being of very high nutritional quality. They are rich in sulfur-containing amino acids such as cysteine and methionine and easily digested. The essential amino acid profile of whey proteins meets or exceeds all the nutritional requirements of the FAO/WHO (USDEC, 1997). The Protein Digestibility-Corrected Amino Acid Scoring (PDCAAS) method was adopted by FDA as the official method for determining protein quality in food products intended for children over one year of age and adults in 1993 (Morr and Ha, 1995). Using the PDCAAS method, whey protein rates the highest value of 1.0, because it is highly digestible and it meets or exceeds the recommended amount of each essential amino acid. Using another method to measure protein quality - the Protein Efficiency Ratio (PER) value - whey also ranks high on the scale. The higher the PER value, the better the protein. Casein, the reference protein, has a PER value of 2.5. Protein with a PER greater than 2.5 are considered to be quality proteins. Whey protein, with a PER greater than 3.0, is considered to be nutritionally excellent protein. Virtually every amino acid present in sweet-type whey exceeds Food Agriculture Organization/World Health Organization (FAO/WHO) nutritional intake recommendations, both for children aged 2-5 and for adults. In many cases, for adults, whey proteins offer more than double the minimum FAO/WHO standards. Whey proteins compare favorably to many common proteins that have a PER of less than 2.5, including soy (2.2), peanuts (1.8), corn (2.2) and wheat gluten (0.8). These products have limited concentrations of certain essential amino acids. BIOLOGICALLY ACTIVE COMPONENTS IN MILK Growth stimulating factors in milk Insulin-like growth factors 1 and 2 (IGF-1 and IGF-2) together with a truncated form of IGF-1 were read. Effects of bovine colostrum supplementation on serum IGF-I, IgG, hormone, and saliva IgA during training.Mero A et al.J Appl Physiol 1997 read Colostrum is the first natural food produced by female mammals during the first 24-36h directly after giving birth. Chemically, colostrum is a very complex fluid rich in nutrients, antibodies and growth factors. In cows the antibodies...Pakkanen, R. / Aalto, J., International Dairy Journal, May 1997 Antimicrobial factors in milk Bovine colostral antibodies against H. pylori read Antiviral factors in milk Antiviral effects of plasma and milk proteins: lactoferrin shows potent activity against both human immunodeficiency virus and human cytomegalovirus replication in vitro. Native and chemically derivatized proteins purified from serum and milk were assayed in vitro to assess their inhibiting capacity on the cytopathic effect of human immunodeficiency virus (HIV)-1 and human cytomegalovirus (HCMV) on MT4 cells and fibroblasts, respectively. read Immune Milk The immunoglobulins of bovine colostrum provide the major antimicrobial protection against microbial infections and confer a passive immunity to the newborn calf until its own immune system matures. The concentration in colostrum of specific antibodies against pathogens can be raised by immunising cows with these pathogens or their antigens. Immune milk products are preparations made of such hyperimmune colostrum or antibodies enriched from it. read Probiotics and Whey Protein Concentrates. Yogurt is the most preferred vehicle to incorporate the health promoting live probiotic cultures into dairy foods. read RESEARCH ON THE EFFECT OF WHEY PROTEINS ON BODY BUILDING, LEAN TISSUE MASS AND MUSCLE STRENGTH, AND ATHLETIC PERFORMANCE. A pre-exercise alpha-lactalbumin-enriched whey protein meal preserves lipid oxidation and decreases adiposity in rats. Am J Physiol Endocrinol Metab. 2002 Sep;283(3):E565-72. read Effects of whey protein and resistance exercise on body cell mass, muscle strength, and quality of life in women with HIV. AIDS. 2001 Dec 7;15(18):2431-40 read The effect of whey protein supplementation with and without creatine monohydrate combined with resistance training on lean tissue mass and muscle strength. Int J Sport Nutr Exerc Metab. 2001 Sep;11(3):349-64. Males that supplemented with a combination of whey protein and creatine had greater increases in lean tissue mass and bench press than those who supplemented with only whey protein or placebo. However, not all strength measures were improved with supplementation, since subjects who supplemented with creatine and/or whey protein had similar increases in squat strength and knee flexion peak torque compared to subjects who received placebo. read Extraction from cheese whey by cation-exchange chromatography of factors that stimulate the growth of mammalian cells. J Dairy Sci. 1995 Jun;78(6):1209-18. read Effect of supplementation with a cysteine donor on muscular performance. J Appl Physiol. Lands LC, Grey VL, Smountas AA. 1999 Oct;87(4):1381-5. Oxidative stress contributes to muscular fatigue. Glutathione (GSH) is the major intracellular antioxidant, the biosynthesis of which is dependent on cysteine availability. We hypothesized that supplementation with a whey-based cysteine donor [Immunocal (HMS90)] designed to augment intracellular GSH would enhance performance. Twenty healthy young adults (10 men, 10 women) were studied presupplementation and 3 mo postsupplementation with either Immunocal (20 g/day) or casein placebo. Muscular performance was assessed by whole leg isokinetic cycle testing, measuring peak power and 30-s work capacity. Lymphocyte GSH was used as a marker of tissue GSH. There were no baseline differences (age, ht, wt, %ideal wt, peak power, 30-s work capacity). Follow-up data on 18 subjects (9 Immunocal, 9 placebo) were analyzed. Both peak power [13 +/- 3.5 (SE) %, P < 0.02] and 30-s work capacity (13 +/- 3.7%, P < 0.03) increased significantly in the Immunocal group, with no change (2 +/- 9.0 and 1 +/- 9.3%) in the placebo group. Lymphocyte GSH also increased significantly in the Immunocal group (35.5 +/- 11.04%, P < 0.02), with no change in the placebo group (-0.9 +/- 9.6%). This is the first study to demonstrate that prolonged supplementation with a product designed to augment antioxidant defenses resulted in improved volitional performance

		WHEY PROTEINS (From Ewan Ha, 2001) Whey is a co-product of cheese and casein manufacture and contains about 12% of protein as a solid basis. Whey proteins are universally defined as those proteins that remain in milk serum after coagulation of the caseins at pH 4.6 and 20°C (Morr and Ha, 1993). Whey proteins are compact globular proteins ranging from 14 to 1000 Kd. The percentage of each of the major whey proteins, that is, beta-lactoglobulin (beta-Lg), alpha-lactalbumin (alpha-La), bovine serum albumin (BSA), and the immunoglobulins (Ig) are in Table 2. beta-Lg exists as a dimer in solutions above its isoelectric point of pH 5.2. Between pH 3.5 and 5.2 the dimer polymerizes to an octomer. Below pH 3.5 and above pH 7.5, the dimer dissociates to a slightly expanded monomer. beta-Lg undergoes time- and temperature-dependent denaturation above 65°C. alpha-La was denatured at 62-63°C and that it was 90% renatured at pH 6.5 (de Wit et al., 1983; Morr and Ha, 1993). alpha-La binds Ca⁺², Zn⁺², and other metal ions. alpha-La undergoes a conformational transition at £pH 4, which is probably due to the replacement of this tightly bound Ca⁺² ion with a H⁺¹ ion at this pH. These physicochemical properties of individual whey proteins have been utilized in the whey protein fractionations (Bramaud et al., 1997; Gésan-Guziou et al. 1999; Pearce, 1983, 1987, 1992; Pierre and Fauquant, 1986; see Table 3). One of the key properties of the whey proteins is their abundance of sulfhydryl amino acid residues that allow them to form intermolecular covalent bonds during high-temperature processing of whey and whey protein concentrates solutions. Intermolecular disulfide bonding is essential for forming heat-induced whey protein gels and for stabilizing foam structures (Morr and Ha, 1993). In addition to the major whey proteins, whey contains additional minor proteins and peptides. These minor components include: 1) b-casein fragments that are heat stable and soluble at pH 4.6, formerly known as proteose-peptone components (5, 8-slow, 8-fast), 2) the highly acidic casein glycomacropeptide (CMP also known as GMP) fragments of k-casein, 3) the highly basic proteins such as lactoferrin, lactoperoxidase, and growth-stimulating factors in whey, 4) phospholipoprotein complex (PLPC) consisting of skim milk membrane and milk fat globule membrane (MFGM) that are complexed with phospholipids and small milk fat globules (MFG), and 5) many other biologically active peptides. Whey protein fractionation technologies have been reported using ion-exchange chromatography, affinity chromatography, selective precipitation and membrane fractionation. Spherosil-QMA anion exchange resin was used to preferentially adsorb b-lactoglobulin at pH 6.63 with minimal adsorption of the positively charged immunoglobulins, lactoferrins, and lactoperoxidase (Skudder, 1985). b-Lactoglobulin (Amundson et al., 1982) and a-lactalbumin (Pearce, 1983, 1987, 1992) were thermally aggregated and selectively precipitated at pH 4.65 and pH4.2 at 65°C, respectively. Although these reported chromatographic techniques could provide effective protein purification, most of them are not acceptable for the large-scale production due to the cost of production (Pearce, 1992). The Proteose Peptone Components, identified as Component 5, 8-slow, and 8-fast by gel electrophoresis, have been reclassified as b-casein fragments 1P, 4P and 5P. These protein fragments contain low concentration of aromatic and sulfur amino acid residues that are not precipitated by heating milk at 95°C for 20 min and subsequently adjusting to pH 4.6 with acid. Although present in milk, additional b-casein fragments are likely to be formed during the cheese and whey manufacturing processes (Morr and Ha, 1993). The proteose peptone component 3 (PP3), also known as lactophorin, is heat stable, acid soluble polypeptide. The PP3 component is a hydrophobic phosphoglycoprotein made up of 135 amino acids and is found in low level in milk (300mg/L). Bovine PP3 component is not sulfated but contains variable levels of sialic acids and fucose and not a membrane protein (Guimont, 1997). Hybridoma proliferation with the PP3 component of bovine, sheep and goat milk as a fetal bovine serum (FBS) substitute in cell culture was reported. The surface hydrophobicity and the glucosidic nature of PP3 appear to be essential to the mitogenic activity. When sialic acids are cleaved by neuraminidase, the PP3 component lost its ability to stimulate hybridoma mitosis. The negative charge of the carboxyl groups may play a role in the stabilization of the active conformation and in the solubilization of the glycoprotein.

BIOLOGICALLY ACTIVE COMPONENTS IN MILK

RESEARCH ON THE EFFECT OF WHEY PROTEINS ON BODY BUILDING, LEAN TISSUE MASS AND MUSCLE STRENGTH, AND ATHLETIC PERFORMANCE.