Therapeutic Applications of Whey Protein - GSH Polska · 2015. 6. 19. · whey to a co-product in the manufacturing of cheese.2 Milk contains two primary sources of protein, the caseins

Alternative Medicine Review ◆ Volume 9, Number 2 ◆ 2004

Whey Protein Review

Copyright©2004 Thorne Research, Inc. All Rights Reserved. No Reprint Without Written Permission

Keri Marshall, MS, ND – 1996 Master of Science Socialand Preventive Medicine, State University of New York atBuffalo; 2001 graduate, National College of NaturopathicMedicine. Private practice, Sandpoint, Idaho.Correspondence address: 515 Pine Street, Suite H,Sandpoint, ID 83864Email: [email protected]

AbstractWhey, a protein complex derived from milk, isbeing touted as a functional food with a numberof health benefits. The biological componentsof whey, including lactoferrin, beta-lactoglobulin, alpha-lactalbumin,glycomacropeptide, and immunoglobulins,demonstrate a range of immune-enhancingproperties. In addition, whey has the ability toact as an antioxidant, antihypertensive,antitumor, hypolipidemic, antiviral,antibacterial, and chelating agent. The primarymechanism by which whey is thought to exertits effects is by intracellular conversion of theamino acid cysteine to glutathione, a potentintracellular antioxidant. A number of clinicaltrials have successfully been performed usingwhey in the treatment of cancer, HIV, hepatitisB, cardiovascular disease, osteoporosis, andas an antimicrobial agent. Whey protein hasalso exhibited benefit in the arena of exerciseperformance and enhancement.(Altern Med Rev 2004;9(2):136-156)

IntroductionIn recent years, milk constituents have

become recognized as functional foods, suggest-ing their use has a direct and measurable effect onhealth outcomes.1 Whey, a by-product of cheeseand curd manufacturing, was once considered awaste product. The discovery of whey as a func-tional food with nutritional applications elevatedwhey to a co-product in the manufacturing ofcheese.2 Milk contains two primary sources ofprotein, the caseins and whey. After processingoccurs, the caseins are the proteins responsible formaking curds, while whey remains in an aqueous

environment. The components of whey includebeta-lactoglobulin, alpha-lactalbumin, bovine se-rum albumin, lactoferrin, immunoglobulins, lac-toperoxidase enzymes, glycomacropeptides, lac-tose, and minerals.2 In additional, whey derivedfrom buttermilk versus cheese contains the lipidsphingomyelin.

Several cultures consider fermented foodspart of a healthful diet. Historically, whey wasconsidered a cure-all used to heal ailments rang-ing from gastrointestinal complaints to joint andligament problems. Nanna Rognvaldardottir, anIcelandic food expert, describes how whey, calledsyra by the Icelandic people, is fermented andstored in barrels. Syra is diluted with water andingested or used as a marinade or preservative formeat and other food. Syra was the most commonbeverage of Icelandic people and is thought to havereplaced ale, due to lack of grains in the region.3

Today, whey is a popular dietary proteinsupplement purported to provide antimicrobialactivity, immune modulation, improved musclestrength and body composition, and to preventcardiovascular disease and osteoporosis. Advancesin processing technology, including ultrafiltration,microfiltration, reverse osmosis, and ion-ex-change, have resulted in development of severaldifferent finished whey products. Whey proteinconcentrates (ranging from 80-95 percent protein),reduced lactose whey, whey protein isolate, de-mineralized whey, and hydrolyzed whey are now

Therapeutic Applicationsof Whey Protein

Keri Marshall, ND, MS


Alternative Medicine Review ◆ Volume 9, Number 2 ◆ 2004 Page 137

Review Whey Protein

available commercially. Each whey product var-ies in the amount of protein, carbohydrates, im-munoglobulins, lactose, minerals, and fat in thefinished product. These variables are importantfactors in the selection of whey fractions for spe-cific nutritional applications. Table 1 describes thevarious whey protein products available.

Whey Protein ManufacturingProtein from bovine whole milk consists

of approximately 20-percent whey protein. Whencasein is removed from whole milk, liquid wheyremains, having a protein concentration of about65 percent. The following is a summary of theOhio State University method of manufacturingwhey protein powder. Milk is high-temperature,short-time pasteurized (163 degrees F for 30 sec-onds) and held overnight at 40 degrees C. Thefollowing morning the mixture is cooled to 30degrees C, inoculated with a lactic acid culture,

and incubated for 30 minutes. Rennet extract isadded and the mixture is stirred, resulting in co-agulation of curd.

Rennet is derived from the abomasum(fourth stomach) of newly born calves. Chymosin,the active enzyme ingredient of rennet, aids in thecoagulation of milk by separating it into curds andwhey. In a newly born calf, chymosin aides in thedigestion and absorption of milk. Adult cows donot have this enzyme.

The liquid whey is drained through a stain-less steel screen and the remaining curd is cut andcooked at 30 degrees C. Whey liquid is then fil-tered at 45 degrees C and brought to a pH of 3 byadding citric acid. The liquid is filtered to one-fifth its original volume, resulting in whey con-centrate that is approximately 80-percent protein.This can be additionally micro-filtered to increaseprotein concentration to as high as 95 percent.

Table 1. Types of Commercially Available Whey Proteins

Product Description

Whey Protein Isolate

Whey Protein Concentrate

Hydrolyzed Whey Protein

Undenatured Whey Concentrate

Protein Concentration

90-95%

Ranges from 25-89%Most commonly available as 80%

VariableHydrolysis used to cleave peptide bonds Larger proteins become smaller peptide fractionsReduces allergic potential compared with non-hydrolyzed

VariableUsually ranges from 25-89%

Fat, Lactose, and Mineral Content

Little if any

Some fat, lactose, and mineralsAs protein concentration increases, fat, lactose, and mineral content decreases.

Varies with protein concentration

Some fat, lactose, and mineralsAs protein concentration increases, fat, lactose, and mineral content decreases. Processed to preserve native protein structures; typically have higher amounts of immunoglobulins and lactoferrin


Whey Protein Review


The final whey protein concentrate iswarmed and spray-dried to achieve whey proteinpowder. Whey protein concentrates can then beput through an ion-exchange process to remove

fat and lactose. In addition,some manufacturers hydro-lyze (cleaving peptide bondsvia enzymes or heat) the wheyto provide more peptides andfree amino acids in the finalproduct.4

The commercial suc-cess of whey protein has ledto the development of highquality whey protein supple-ments manufactured as pri-mary products and not as a by-product of cheese manufactur-ing. Manufacturers take spe-cial care to preserve the bio-logical activity, native proteinstructure, and protein-bound-fats in the finished product.Proteins are processed underlow temperatures and not ex-posed to fluctuating pHchanges to avoid denaturingthe native structures. In addi-tion, the source of milk and thehealth of the milking cows isthought to contribute to im-mune-enhancing activity ofwhey products.5,6

BiologicalComponentsAmino Acid Content

Collectively, wheyproteins have all the essentialamino acids and in higher con-centrations compared to vari-ous vegetable protein sourcessuch as soy, corn, and wheatgluten.2 In addition to havinga full spectrum of amino ac-

ids, the amino acids found in whey are efficientlyabsorbed and utilized, relative to free amino acidsolutions.7

Table 2. Amino Acid Residues in Bovine versus HumanLactoferrin

Alanine

Proline

Arginine

Lysine

Asparagine

Valine

Tryptophan

Cysteine

Threonine

Isoleucine

Serine

Glutamine

Glutamic acid

Phenylalanine

Methionine

Leucine

Glycine

Tyrosine

Aspartic Acid

Histidine

Total number of residues

67

30

39

54

29

47

13

34

36

15

45

29

40

27

4

65

48

22

36

9

689

63

35

43

46

33

48

10

32

31

16

50

27

42

30

5

58

54

21

38

9

691

Bovine Lactoferrin Human Lactoferrin

From: Pierce A, Colavizza D, Benaissa M, et al. Molecular cloning and sequence analysis of bovine lactoferrin. Eur J Biochem 1991;196:177-184.


Review Whey Protein


Relative to other protein sources, wheyhas a high concentration of branched-chain aminoacids (BCAAs) – leucine, isoleucine, and valine.BCAAs, particularly leucine, are important fac-tors in tissue growth and repair. Leucine has beenidentified as a key amino acid in protein metabo-lism during the translation-initiation pathway ofprotein synthesis.8

Whey proteins are also rich in the sulfur-containing amino acids cysteine and methionine.With a high concentration of these amino acids,immune function is enhanced through intracellu-lar conversion to glutathione.

LactoferrinLactoferrin, an iron-binding glycoprotein,

is a non-enzymatic antioxidant found in the wheyfraction of milk as well as in colostrums. Thelactoferrin component of whey consists of approxi-mately 689 amino acid residues, while humanlactoferrin consists of 691 residues.9 Wheylactoferrin is composed of a single polypeptidechain with two binding sites for ferric ions. Be-fore processing, bovine lactoferrin is only 15-20percent saturated with iron. Iron-depletedlactoferrin, defined as containing less than five-percent iron, is referred to as apolactoferrin. Hu-man breast milk contains apolactoferrin.10 Theconcentration of lactoferrin in human milk andcolostrums is approximately 2 mg/mL and 7 mg/mL, respectively, while in bovine milk and colos-trums it is approximately 0.2 mg/mL and 1.5 mg/mL, respectively.11 Lactoferrin is a dominant com-ponent of whey protein in human breast milk;however, the concentration in most commercialwhey protein powders is only 0.35-2.0 percent oftotal proteins. Table 2 illustrates the difference inamino acid profiles between bovine and humanlactoferrin.

ImmunoglobulinsAn immunoglobulin (Ig) is an antibody

or gamma-globulin. There are five classes ofantibodies – IgA, IgD, IgE, IgG, and IgM. IgGconstitutes approximately 75 percent of theantibodies in an adult. IgG is transferred frommother to child in utero via cord blood and by

breast-feeding, and serves as a child’s first line ofimmune defense – referred to as “passiveimmunity.” IgA is secreted in breast milk andultimately transferred to the digestive tract in thenewborn infant, providing better immunity than abottle-fed child.12 Colostrums contain significantlygreater concentrations of immunoglobulins thanmature milk. Immunoglobulins reach maximumconcentration the first 24-48 hours post-parturitionand decline in a time-dependent manner followingpeak concentration.13

Similarly, the whey fraction of milk ap-pears to contain a significant amount of immuno-globulins, approximately 10-15 percent of totalwhey proteins. An in vitro study demonstrated bo-vine milk-derived IgG suppresses human lympho-cyte proliferative response to T cells at levels aslow as 0.3 mg/mL of IgG. The authors further con-clude bovine milk IgG typically ranges between0.6-0.9 mg/mL and is therefore likely to conferimmunity that could be carried to humans.14 Stud-ies show raw milk from non-immunized cows con-tain specific antibodies to human rotavirus, as wellas antibodies to bacteria such as E. coli, Salmo-nella enteriditis, S. typhimurium, and Shigellaflexneri.15,16

beta-Lactoglobulinbeta-Lactoglobulin represents approxi-

mately half of the total protein in bovine whey,while human milk contains no beta-lactoglobu-lin. Besides being a source of essential andbranched chain amino acids, a retinol-binding pro-tein has been identified within the beta-lactoglo-bulin structure. This protein, a carrier of smallhydrophobic molecules including retinoic acid, hasthe potential to modulate lymphatic responses.17

alpha-Lactalbuminalpha-Lactalbumin is one of the main

proteins found in human and bovine milk. Itcomprises approximately 20-25 percent of wheyproteins and contains a wide variety of aminoacids, including a readily available supply ofessential and branched chain amino acids. Purifiedalpha-lactalbumin is most readily used in infant


Whey Protein Review


formula manufacturing, as it has the moststructurally similar protein profile compared tobreast milk. However, due to cost effectivemeasures, most dairy-based infant formulascontain ingredients such as demineralized wheywith higher levels of beta-lactoglobulin, makingthem less similar to human milk.

In a murine study, alpha-lactalbumin, inboth the native and hydrolyzed state, enhancedantibody response to systematic antigen stimula-tion.18 The same group proved alpha-lactalbuminhas a direct effect on B-lymphocyte function, aswell as suppressing T cell-dependent and -independentresponses.19

LactoperoxidaseWhey contains many types of enzymes,

including hydrolases, transferases, lyases, pro-teases, and lipases. Lactoperoxidase, an importantenzyme in the whey fraction of milk, is the mostabundant enzyme and the majority of it ends up inwhey following the curding process. Lactoperoxi-dase accounts for 0.25-0.5 percent of total proteinfound in whey. It has the ability to catalyze cer-tain molecules, including the reduction of hydro-gen peroxide.20 This enzyme system catalyzesperoxidation of thiocyanate and some halides(such as iodine and bromium), which ultimatelygenerates products that inhibit and/or kill a rangeof bacterial species.21 During the pasteurizationprocess, lactoperoxidase is not inactivated, sug-gesting its stability as a preservative.

Table 3. Components Found in Whey Protein

Whey Components

beta-Lactoglobulin

alpha-Lactalbumin

Immunoglobulins

Lactoferrin

Lactoperoxidase

Bovine Serum Albumin

Glycomacropeptide

% of Whey Protein

50-55%

20-25%

10-15%

1-2%

0.50%

5-10%

10-15%

Benefits

Source of essential and branched chain amino acids

Primary protein found in human breast milk Source of essential and branched chain amino acids

Primary protein found in colostrum Immune modulating benefits

Antioxidant Antibacterial, antiviral , and antifungal Promotes growth of beneficial bacteria Naturally occurs in breast milk, tears, saliva, bile, blood, and mucus

Inhibits growth of bacteria

Source of essential amino acids Large protein

Source of branched chain amino acids Lacks the aromatic amino acids phenylalanine, tryptophan, and tyrosine


Review Whey Protein


GlycomacropeptideGlycomacropeptide (GMP) is also re-

ferred to as casein macropeptide. GMP is a pro-tein present in whey at 10-15 percent, due to theaction of chymosin on casein during the cheese-making process. GMP is only present whenchymosin is used during processing; therefore,cheeses such as cottage cheese not made withchymosin do not produce GMP in the curding pro-cess.22 GMP is high in branched chain amino ac-ids and lacks the aromatic amino acids includingphenylalanine, tryptophan, and tyrosine. It is oneof the few naturally occurring proteins that lacksphenylalanine, making it safe for individuals withphenylketonuria (PKU).

Bovine SerumAlbumin

Bovine serumalbumin (BSA) is alarge protein that makesup approximately 10-15percent of total wheyprotein. BSA is a sourceof essential aminoacids, but there is verylittle available informa-tion regarding its poten-tial therapeutic activity.

Table 3 sum-marizes the compo-nents found in whey.

Mechanism ofAction

Whey has po-tent antioxidant activ-ity, likely by contribut-ing cysteine-rich pro-teins that aid in the synthesis of glutathione (GSH),a potent intracellular antioxidant.2 GSH is com-prised of glycine, glutamate, and cysteine (Figure1). Cysteine contains a thiol (sulfhydryl) groupthat serves as an active reducing agent in prevent-ing oxidation and tissue damage. As an antioxi-dant, glutathione is most effective in its reduced

form. Riboflavin, niacinamide, and glutathione re-ductase are essential cofactors in the reduction ofglutathione.23 As a result of the glutathione/anti-oxidant component of whey, it is being investi-gated as an anti-aging agent.24

As a detoxifying agent, glutathione per-oxidase (GSHPx), which is derived from seleniumand cysteine, is an endogenous antioxidant enzymewith the ability to convert lipid peroxides into lessharmful hydroxy acids. The peroxidases interactwith hydrogen peroxide to reduce it to water, ne-gating its oxidative potential. Both glutathioneperoxidase activity and selenium concentrationshave been shown to decrease as lactation contin-ues, peaking at approximately one month after

initiation. Practitioners use whey protein productsas a source of cysteine to increase intracellularglutathione levels25,26 and it has been reported thatGSHPx activity in cow’s milk, and presumablywhey, is the same as in human milk.27

Figure 1. Synthesis of Glutathione from Cysteine, Glutamate,and Glycine

g-Glutamylcysteinesynthetase

Glutathionesynthetase

Glutamic acid+

Cysteine+

ATP

g-Glutamylcysteine+

ADP+P

g-Glutamylcysteine+

Glycine+

ATP

Glutathione+

ADP+P

Step 1 Step 2


Whey Protein Review


Studies on lactoferrin have demonstratedits ability to activate natural killer (NK) cells andneutrophils, induce colony-stimulating factor ac-tivity, and enhance macrophage cytotoxicity.28-31

Lactoferrin also appears to have antivi-ral, antifungal, and antibacterial properties. Theantimicrobial effect is likely more potent in or-ganisms that require iron to replicate, as lactoferrinhas the unique ability to chelate iron in a way thatdeprives microorganisms of this essential nutri-ent for growth.32 In additional, lactoferrin has theability to release the outer membrane of gram-negative bacteria, the lipopolysaccharide compo-nent, thus acting as an antibiotic.33

Lactoferrin demonstrates anti-inflamma-tory properties. A mouse study revealed lactoferrinhad the ability to regulate levels of tumor necro-sis factor (TNF) and interleukin 6 (IL-6), thusdecreasing inflammation and, ultimately, mortal-ity.34

In addition to the above-mentioned prop-erties, alpha-lactalbumin can chelate heavy met-als.35 It reduces oxidative stress because of its iron-chelating properties.36

Whey has been recently touted as a health-ful dietary supplement to reduce blood pressure.Antihypertensive peptides have been isolated inthe primary sequence of bovine beta-lactoglobu-lin.37 These peptides give whey significant angio-tensin I converting enzyme (ACE) inhibitory ac-tivity, which blocks the conversion of angiotensinI to angiotensin II, a highly potent vasoconstric-tor molecule.38 beta-Lactoglobulin has been de-scribed by Nagaoka et al as a cholesterol-lower-ing agent. In animal studies, beta-lactoglobulininhibited cholesterol absorption by changing mi-cellar cholesterol solubility in the intestine.39

Absorption and Digestion of MilkProteins: Casein versus Whey andLactoferrin

The physical manifestations of milk in thegut vary, based on the type of milk protein. Wheycomprises approximately 20 percent of milk pro-tein, while 80 percent of protein in milk is casein.32

These two proteins possess quite different prop-erties. Casein proteins form curds in the stomach,

increasing hydrolysis and slowing entrance to thesmall intestine. Whey proteins, on the other hand,do not coagulate under acidic conditions. They areconsidered to be “fast proteins,” as they reach thejejunum quickly after entering the gastrointesti-nal tract.40 After reaching the small intestine, thehydrolysis of whey is slower than that of casein,allowing for greater absorption over the length ofthe small intestine.

A study by Arnal et al demonstrates thatwhey’s rapid absorption patterns are superior forpostprandial protein utilization and overall nitro-gen balance in elderly women.41 Most recently, arandomized, single-blind study examining proteinsatiety found whey protein to exhibit a higher post-prandial level of plasma amino acids compared tocasein.42

A study by Troost et al examined the ef-fects of an orally administered recombinant hu-man lactoferrin (rhLF) in the digestive tract. Thestudy population consisted of eight female ileo-stomy patients who consumed 5 g rhLF and col-lected full ileostomy output for 24 hours. In a 24-hour period, only 4 µg of lactoferrin were excreted.The study concluded dietary rhLF does not reachthe colon because it is digested in the stomach andsmall intestine.43 A similar study by the same groupwas conducted on 12 healthy volunteers. On threeseparate days, volunteers were instructed to con-sume 4.5 g lactoferrin. After conclusion of thestudy, results revealed bovine lactoferrin survivedpassage through the stomach, demonstrating theability of this protein to survive in an acidic envi-ronment.44

Clinical Indications for WheyCancer

Whey protein concentrates have been re-searched extensively in the prevention and treat-ment of cancer. Glutathione stimulation is thoughtto be the primary immune-modulating mechanism.In a review of whey protein concentrates in thetreatment of cancer, Bounous discusses the anti-tumor and anticarcinogenic potential. The aminoacid precursors to glutathione available in wheymight: (1) increase glutathione concentration inrelevant tissues, (2) stimulate immunity, and (3)


Review Whey Protein


detoxify potential carcinogens.45 Other authorsconclude the iron-binding capacity of whey mayalso contribute to anticancer potential, as iron mayact as a mutagenic agent causing oxidative dam-age to tissues.46

Many animal studies have examined theeffects of whey and its immune-enhancing com-ponents, including lactoferrin and beta-lactoglo-bulin.47-54 In animal studies where colon cancer wasinduced, whey demonstrated significantly lowerincidence of tumors, as well as fewer aberrantcrypts.47,49,52,54 When whey-based protein powderswere compared to soy-based protein powders,similar effects were also observed.48,52 Yoo et aldemonstrated lactoferrin has the ability to inhibitmetastasis of primary tumors in mice with can-cer.50 Bovine serum albumin (10-15 percent of totalwhey protein) has demonstrated inhibition ofgrowth in human breast cancer cells in vitro.55 Ahamster study demonstrated fractionated whey hasthe ability to prevent and treat 5-fluorouracil che-motherapy-induced oral mucositis.56 This protec-tion is thought to occur via induction of tumorgrowth factor-beta (TGF-β), which reduces basalepithelial cell proliferation.

A recent in vitro study by Kent et al dem-onstrated that an isolate of whey protein, whencompared to a casein-based protein, increased glu-tathione synthesis and protected human prostatecells against oxidant-induced cell death.57 An invitro study on a human hepatoma cell line wasconducted using a high lactoferrin whey concen-trate (Immunocal®), a baicalein medium, or a com-bination of the two.58 Baicalein, a potential anti-cancer drug, is a flavonoid extracted fromScutellaria revularis that is thought to have anti-tumor activity. Immunocal alone did not have asignificant impact on the hepatoma cell line. How-ever, when Immunocal was combined withbaicelein, cytotoxicity was enhanced by inducinga higher rate of apoptosis than in the group treatedwith baicalein alone.

To date, few clinical trials on whey andcancer have been conducted. It has been proposedthat GSH concentrations are high in tumor cells,giving them resistance to chemotherapeuticagents.59 In 1995, a small trial was conducted onfive patients with metastatic carcinoma of the

breast, one patient with pancreatic cancer, and onewith liver cancer.59 Patients were given 30 g wheyprotein concentrate (Immunocal) for six months.In six patients, blood lymphocyte GSH was el-evated initially, suggesting high tumor GSH lev-els. After completion of the study, two of the pa-tients showed signs of tumor regression and a re-turn of lymphocyte GSH levels to normal, whiletwo patients showed signs of tumor stabilizationwithout normalization of glutathione levels. Threepatients had progression of disease and lympho-cyte GSH levels increased from initial measure-ments. The conflicting results and small size ofthis study indicate the need for a larger clinicaltrial to investigate the potential of a whey proteinconcentrate singly and as an adjunct to chemo-therapy.

In a recent clinical trial, 20 patients withstage IV malignancies (one bladder, five breast,two prostate, one neuroblastoma, one ovarian, onegastric, three colon, one mesothelioma, two lym-phoma, two non-small cell lung, and one osteosar-coma) received a combination of 40 g/day non-denatured whey protein concentrate, 50-100 g/dayintravenous Transfer Factor Plus® (a supplementcontaining several immunoactive components), 1-2 g/day oral ascorbic acid, Agaricus blazei tea, amultiple vitamin/mineral complex, 500 mgAndrographis paniculata twice daily, and a soyextract for six months.60 After six months therewere 16 survivors, all of whom had significantlyhigher NK function and higher mean hemoglobinand hematocrit levels. All patients noted havingan improved quality of life during the course ofthe study. No comparison was made between thiscombination therapy and whey protein alone.

HepatitisWhey protein supplementation demon-

strates variable effects in patients infected withHepatitis B or C. Initially it was found that bo-vine lactoferrin prevented hepatitis C virus (HCV)infection in vitro in a human hepatocyte line.61

These results prompted further clinical trials.A pilot study was conducted on 11 patients

with chronic HCV. Each patient received either1.8 or 3.6 g bovine lactoferrin daily for eight


Whey Protein Review


weeks. In patients with low pre-treatment viral loads of HCV, de-creases in HCV RNA and serumalanine transaminase were ob-served. In patients with higherpretreatment HCV viral loads,levels did not change signifi-cantly.62

A dose-response trial of45 individuals with HCV wasconducted at doses of 1.8, 3.6,and 7.2 g lactoferrin daily foreight weeks.63 A virological re-sponse was observed in only fourpatients, although HCV RNAwas still detectable. Eight pa-tients had a virological response– a 50 percent or greater decreasein HCV RNA – eight weeks af-ter the treatment ended. Therewere no significant variations indose-dependant responses. Thistrial left many unanswered ques-tions for future studies regardingwhey supplementation and HCV,including optimum dose, dura-tion, and the potential effects ofcombining supplementation withconventional treatments such asinterferon therapy.

In an open study on 25patients diagnosed with eitherHepatitis B or C, patients weregiven 12 g whey (Immunocal)twice daily for 12 weeks.64 Priorto the start of treatment withwhey, patients were given 12 gcasein protein daily for twoweeks. Patients were also givencasein following Immunocal fora four-week period. In the 17 pa-tients with HCV, no significantchanges were noted. In the groupwith hepatitis B virus (HBV),serum lipid peroxidase levelsdecreased, while IL-2 and NKactivity increased. In six of eightHBV patients serum alanine

Table 4. Comparison of Whey Products Immunocaland Protectamin

Amino Acid

Aspartic

Glutamic

Serine

Glycine

Histidine

Arginine

Threonine

Alanine

Proline

Tyrosine

Valine

Methionine

Isoleucine

Leucine

Phenylalanine

Lysine

Tryptophan

Cysteine/Cystine

Minerals

Sodium

Potassium

Calcium

Chloride

Phosphate

Magnesium

Immunocal (g/100 g)

9.80

16.37

3.56

1.64

2.04

2.30

4.63

4.80

3.76

3.76

4.53

2.13

6.41

12.56

4.00

10.68

2.86

4.17

0.3

0.5

0.4

0.4

0.3

0.3

Protectamin (g/100 g)

10.50

17.48

5.58

1.84

1.73

2.24

6.97

4.87

6.01

1.97

6.06

2.27

6.32

9.96

3.15

9.19

1.53

2.28

0.1

0.5

0.3

0.1

0.3

0.1

From: Micke P, Beeh KM, Buhl R. Effects of long-term supplementation with whey proteins on plasma glutathione levels of HIV-infected patients. Eur J Nutr 2002;41:12-18.


Review Whey Protein


transferase levels were reduced, while plasma glu-tathione levels increased in five of the same eight.This trial shows promise for the use of whey pro-tein in the treatment of HBV.

Human Immunodeficiency Virus(HIV)

Glutathione deficiency is a common prob-lem for individuals infected with HIV. In an effortto increase cysteine, and ultimately glutathione,several studies have been conducted on the use ofwhey proteins in HIV-positive individuals.65,66 Ina study by Micke et al, 30 subjects with HIV wererandomized to receive a daily dose of 45 g wheyprotein from one of two sources – Protectamin®

or Immunocal. The two products have differentamino acid profiles and Immunocal is producedat a lower isolation temperature (


Whey Protein Review


against Pseudomonas aeruginosa, Listeriamonocytogenes, and E. coli.32,80 In addition to theabove-mentioned bacteria, lactoferrin has alsodemonstrated antifungal activity toward Candidaalbicans.81

Cardiovascular DiseaseIn recent years, studies have linked a high-

fat diet to an increased risk of cardiovascular dis-ease (CVD). Because CVD is linked to a numberof other factors, including increased age, genet-ics, obesity, sedentary lifestyle, and alcohol intake,quality of dietary fat must be taken into consider-ation. Milk is composed of more than 12 differenttypes of fat, including sphingolipids, free sterols,cholesterol, and oleic acid.82 Several studies havefound milk intake and milk products lower bloodpressure and reduce the risk of hypertension.83-85

A study was conducted on a group of 20healthy adult males to investigate whether a fer-mented milk supplement with an added whey pro-tein concentrate would affect serum lipids andblood pressure.86 The fermented milk containedboth Lactobacillus casei and Streptococcusthermophilus. During the course of eight weeks,volunteers consumed 200 mL of fermented milkwith whey protein concentrate or a placebo in themorning and evening. The placebo consisted of anon-fermented milk product without the additionof whey protein concentrate. After eight weeks,the fermented-milk group showed significantlyhigher HDLs and lower triglycerides and systolicblood pressure than did the placebo group. Whiletotal cholesterol and LDL levels were lower in thefermented-milk group, the difference was not sta-tistically significant.

ExerciseWhey protein supplements, including pu-

rified alpha-lactalbumin sports drinks, have beenreadily utilized in the consumer market becauseof the high protein quality score and high percent-age of BCAAs.87 Whey proteins can contain up to26-percent branched chain amino acids,2,88 whichare efficient substrates for synthesizing new pro-teins. For example, the BCAA leucine acts as a

signaling molecule for initiation of protein syn-thesis.8,89 It has been speculated that the quality ofa particular protein for enhancing muscle hyper-trophy and strength is related to its leucine con-tent.36 The amino acid profile in dietary proteinsinfluences their nitrogen utilization, and poor qual-ity dietary proteins have been shown to increasenitrogen losses and limit protein synthesis.90,91

Human studies documenting the benefi-cial effect of whey protein supplementation onmuscle size and strength are limited. Burke et aldemonstrated that men engaged in resistance train-ing programs while supplementing with whey pro-tein concentrates showed greater improvementsin strength than men using resistance trainingalone.92 Forty-two men, ages 18-31, familiar withweight training, followed the same high volume,heavy load, and free-weight resistance-trainingprogram for 12 weeks. The participants were di-vided into three groups in double-blind fashion:whey protein (1.2 g/kg body wt/day), a multi-in-gredient whey protein sports supplement (1.3 g/kg body wt/day), or placebo (1.2 g/kg body wt/day of maltodextrin ). At baseline there were nosignificant differences among the groups in leantissue mass or strength as determined by fourmeasurements, including bench press, squat,isokinetic knee extension, and isokinetic knee flex-ion. After 12 weeks men who received whey pro-tein or the multi-ingredient whey protein sportssupplement in combination with resistance train-ing showed greater improvements in one of fourmuscle strength measurements. In addition, wheysupplemented groups showed greater improve-ments in lean tissue mass over the placebo group.While this study does not demonstrate wheyprotein’s superiority over other protein sources, itdoes show improvement in resistance training withthe addition of protein supplementation.

Lands et al showed the effect of threemonths whey protein supplementation (10 g twicedaily) versus the same amount of casein as placeboon muscular performance in 18 men.93 At baselinethere were no significant differences in age, height,weight, percent ideal body weight, whole legisokinetic cycle testing, peak power, or 30-secondwork capacity. Peak power and 30-second work


Review Whey Protein


capacity improved significantly in the treatmentgroup with no change in the placebo group. Bodyweight remained unchanged in both groups, butthe supplemented group had a decrease in percentbody fat.

While moderate exercise enhances immu-nity,94 intense athletic training has been shown tostress the immune system.95-97 Free radical produc-tion and increased inflammatory activity arethought to contribute to impaired immune activ-ity in over-trained athletes. In highly trained indi-viduals muscular performance and recovery canbe hindered by oxidative stress.98-100 The availabil-ity of glutathione has been shown to reduce oxi-dative stress.101

Acting as a cysteine donor, whey has beenshown to raise intracellular GSH levels in vitro.102

Lands et al proposed enhanced biosynthesis ofintracellular glutathione, shown by an increase oflymphocyte GSH levels in test subjects, is themechanism responsible for the improved muscu-lar performance observed in their study.93

Whey’s amino acid profile makes it idealfor body composition and to support protein syn-thesis and muscle growth. Other bioactive com-ponents found in whey might benefit additionalaspects of health in active people and trained ath-letes by improving immune function and gas-trointestinal health and exhibiting anti-inflamma-tory activity. Whey components, such as IgA,glutamine, and lactoferrin, can beneficially impactcommon complaints among athletes, includingrepeated infections and gastrointestinal distur-bances.

Lower levels of sIgA and glutamine103,104

have been found after intensive exercise and inover-trained individuals, and have been correlatedwith increased frequency of infection.105 In addi-tion, glutamine deficiency has been proposed tocontribute to gastrointestinal complaints experi-enced by highly trained individuals.106 Free radi-cal damage is thought to delay muscle recoveryand impair performance.107 Whey prevents freeradical damage through supporting intracellularglutathione levels and supplying lactoferrin foradditional antioxidant activity.

ObesityObesity has reached epidemic proportions

in the United States. Low-fat, high-carbohydratedietary trends are being eschewed in favor ofhigher-protein, lower-carbohydrate diets. For in-dividuals eating high-protein diets, whey is an at-tractive source of dietary protein. Whey proteinisolates can be as high as 95-percent protein, afterthe removal of fat and lactose, and contain valu-able minerals and vitamins. Whey has made a sig-nificant commercial impact in the weight-loss in-dustry for its protein content alone. The essentialand non-essential amino acids in whey act as sub-strates for protein synthesis and may improve bodymass index in individuals participating in exer-cise programs.92

Calcium is thought to influence energymetabolism because intracellular calcium regu-lates adipocyte lipid metabolism and triglyceridestorage.108 Zemel et al demonstrated a greater ef-fect of dairy versus nondairy sources of calciumfor improving body composition. Calcium-forti-fied cereal was compared to calcium-fortified ce-real plus nonfat dried milk for accelerating weightand fat loss in mice. The addition of nonfat driedmilk resulted in substantial amplification of theseeffects. The mechanism responsible for increasedfat loss found with dairy-based calcium versusnondairy calcium has not yet been elucidated. Theresearchers speculate, “this additional activity re-sides in the whey fraction of milk.”108 Thebioactive components in whey are thought to actsynergistically with calcium to attenuate lipogen-esis, accelerate lipolysis, and effect nutrient parti-tioning between adipose tissue and skeletalmuscle.108

While there is increasing evidence dairyintake and whey protein may play a role in theprevention and/or attenuation of the rising epi-demic of obesity, there is minimal data to suggestdairy, calcium from dairy, or whey can impact in-dividuals with established obesity.


Whey Protein Review


Infant Formula and Infantile ColicCreating a substitute for mother’s milk has

proved to be challenging. It is estimated a nursinginfant ingests about 3 g lactoferrin daily during thefirst week of life,109 whereas a calf drinking two li-ters of milk a day ingests about 2 g lactoferrin daily.It is well accepted that nursing infants have a muchricher gut flora than do bottle-fed infants, particu-larly with Bifidobacteria and Lactobacilli.2 Such florais normally associated with an increased resistanceto colonization of the digestive tract with pathogenicbacteria.110 In a study by Roberts et al it was deter-mined that the addition of lactoferrin to a feedingformula increased levels of Bifidobacteria in bottle-fed babies. The levels of Bifidobacteria in formula-fed babies that were supplemented with lactoferrinwere not as high as those found in breast-fed babies.In addition, Bifidobacteria in formula-fed babies tookup to three months to develop, while Bifidobacteriadeveloped more rapidly in nursing infants.111

In a double-blind, German study, 102healthy infants less than two-weeks old were ran-domized to receive either a standard cow’s milk for-mula or an infant formula containing partially hy-drolyzed whey protein.112 Results of this study indi-cate the whey-protein fed infants had significantlymore Bifidobacteria in their stools, ultimately afford-ing improved gastrointestinal immunity. It has beenobserved in previous studies that higher levels ofBifidobacteria in the digestive system decrease thepotential for developing atopic disease for at-riskinfants with family history.113

A randomized, double-blind, placebo-con-trolled study of 43 infants with diagnosed infantilecolic was conducted to determine whether ahypoallergenic, hydrolyzed whey formula was su-perior to a standard cow’s milk formula.114 Infantilecolic is described as at least three hours per day ofcrying for at least three days a week for a minimumof three weeks.115 Parents kept a 24-hour diary fortwo weeks during the study. After a one-week quali-fication period, infants in the study were random-ized to receive either whey or cow’s milk formulafor one week. A clinically relevant result was ob-served in the whey formula group, with crying timereduced to less that one hour per day – a one-hourgreater reduction than found in the cow’s milkformula group.

OsteoporosisMilk has been proposed as a nutritional

food that aids in the prevention of osteoporosisdue to its bioavailable calcium content.116 Re-searchers have begun to examine the differentcomponents of milk to determine if a particularisolate is responsible for the bone-protective ef-fects. Initially, in vitro and animal studies deter-mined milk basic protein (MBP), a component ofwhey, has the ability to stimulate proliferation anddifferentiation of osteoblastic cells as well as sup-press bone resorption.117-119 MBP is prepared fromfractionated whey through cation exchange resin.The total protein concentration of MBP is 98 per-cent, containing lactoferrin, lactoperoxidase, andother minor components. Several in vivo studieson rats determined that both whey protein and frac-tionated whey protein had the ability to increasefemoral bone strength in young ovariectomizedrats.120-122

To date three clinical trials have been con-ducted to determine the effects of MBP in bonemetabolism.123-125 In a trial by Toba et al 30 healthyadult men were given a beverage containing 300mg MBP.123 Subjects were instructed to drink onebeverage daily for 16 days while maintaining anormal diet. After 16 days, serum calcium andurinary calcium excretion were unchanged amongall subjects. Both serum osteocalcin andprocollagen I carboxy-terminal propeptide (PICP)levels increased after 16 days, indicating increasedbone formation. Osteocalcin and PICP are bio-chemical markers released from osteoblasts toassess bone formation.

A similar study was conducted on a groupof 33 healthy adult women to assess bone metabo-lism.124 Women in the study were randomized toreceive either a placebo or a 40 mg MBP bever-age daily. Baseline left calcaneus bone-mineraldensity tests were performed on all women, andwere repeated again at the completion of the study.Results indicated bone mineral density was sig-nificantly increased in the MBP group comparedto placebo. Biochemical indices also revealed theMBP group exhibited a significant decrease inurinary cross-linked N-teleopeptides, indicating aninhibition of osteoclast-mediated bone resorption.


Review Whey Protein


The authors state 400-800 mL of milk is equiva-lent to the 40 mg dose of MBP. More recently,Yamura et al conducted a similar double-blindstudy examining 30 women over a six-month pe-riod. Results indicate a daily dose of 40 mg perday of MBP significantly increases radial bonedensity.125

Gastrointestinal SupportA small, randomized, double-blind, cross-

over study was conducted on 10 children withshort bowel syndrome to examine the effect of ahydrolyzed versus a non-hydrolyzed whey proteinon growth and development.126 Energy intake, ni-trogen balance, intestinal permeability, and weightgain were similar among all children, indicatingthe particular form of whey was not an essentialcomponent. In children experiencing bowel resec-tion, food introduction and promotion of normalgrowth and development is of utmost importance.The increased transit time of whey proteins in thesmall intestine makes it an ideal protein sourcefor this small subset of children.

Whey protein has demonstrated a protec-tive effect on the gastric mucosa. This effect isthought to be related to the sulfhydryl component,particularly cysteine and its link with glutamic acidin the production of glutathione.127,128 Several ani-mal studies have demonstrated this protective ef-fect.129,130 In a study by Rosaneli et al, rats fed awhey protein concentrate showed a 41-percentreduction in ulcerative lesions caused by ethanolingestion, while a 73-percent reduction rate wasobserved following repeat doses of whey.129 Astudy by Matsumoto et al demonstrated an iso-lated alpha-lactalbumin formula had a four-foldeffect on reducing ulcerative lesions compared toa whey protein formula containing only 25-per-cent alpha-lactalbumin.130

Other Uses of Whey ProteinThe wide range of essential and non-es-

sential amino acids, minerals, fats, and biologi-cally active proteins in whey provide extensiveapplication in clinical nutrition. Adequate proteinintake is essential for post-surgical wound heal-ing and protein depletion delays healing time.131

Whey provides protein necessary for wound heal-ing. Surgery stresses the body, altering naturaldefenses, leading to various post-surgical compli-cations. Zimecki et al demonstrated lactoferrinregulated the immune response and provided pro-tective measures in post-surgical patients.132

Human studies demonstrated whey pro-tein improved cognitive function and coping abil-ity in highly stressed individuals.133,134 A rise inserotonin is thought to improve adaptation tostress,135 and the authors proposed the tryptophanavailable in whey provides a substrate to increasebrain serotonin levels. The treatment groups forboth studies received alpha-lactalbumin-enrichedwhey protein because it has the highest tryptophanconcentration of whey protein fractions.

Table 5 summarizes the clinical trials us-ing whey.

ConclusionMilk is one of the oldest functional foods

available to mammals. From birth, mammals relyon mother’s milk for nutrition and immune pro-tection. Scientists are beginning to develop anunderstanding of the various components of milk,including whey, and how they may impact healthand disease. Considerable positive research re-garding whey and its biological components con-tinues to be published. It is noteworthy some ofthe research has been funded by commercial dairyorganizations, although a significant amount ofclinical data from Asia was not supported by dairy-aligned interest groups.

To date, no severe adverse reactions havebeen noted following administration of whey pro-tein products, although some patients note minorgastrointestinal disturbances. For individuals withfrank milk allergies, whey products may not besuitable, although many individuals sensitive todairy find that casein is the culprit and they cantolerate whey. Other dairy-sensitive individuals arelactose-intolerant. Most whey proteins are pro-cessed to remove lactose and finished whey prod-ucts only contain trace amounts. De-lactosedwhey, produced from crystallizing a majority ofthe lactose out and recovering the remaining whey,is appropriate for lactose-intolerant individuals.


Whey Protein Review


A challenge test with a small portion of a particu-lar whey supplement for an individual with dairysensitivities would be indicated before beginningtherapeutic amounts.

As researchers further investigate wheycomponents it can be expected that new functional

foods will develop. In addition to the isolatedcolostrum, lactoferrin, and alpha-lactalbuminproducts currently available, glycomacropeptide,lactoperoxidase, whey immunoglobulin, andbovine serum albumin isolated products may beinvestigated.

Table 5. Summary of Clinical Trials Using Whey Protein

Condition

Cancer

Hepatitis B

HIV

Cardiovascular Risk Factors

Exercise

Infant Formula

Dose of Whey Protein 30 g daily

40 g daily Stage 4 malignanciesUsed other natural therapies.

12 g daily

45 g daily

200 mL of fermented milk combined with liquid whey daily

1.2 g/kg body weight daily

10 g twice daily

Standard Cow Formula vsPartially Hydrolyzed Whey Formula(doses varied)

Study Duration

6 months

6 months

12 weeks

2 weeks and6 months

8 weeks

12 weeks

3 months

12 weeks

1 week

Results

2 of 5 patients had tumor regressionResults suggested an increase in glutathione levels in healthy cells and decreased glutathione levels in cancer cells

16/20 survivors at 6 monthsIncreased NK cell function Increased glutathione Increased hemoglobin and hematocritImproved quality of life

Decreased serum lipid peroxidase levelsIncreased IL-2 and NK activityDecreased serum alanine transferase activity Increased plasma glutathione levels

Increased glutathione levels in both trials

Increased HDL Decreased triglycerides Decreased systolic BP Decreased total cholesterol

Improved lean tissue mass Improvement in one of four muscle strength measurements

Significant improvements in peak powerSignificant increase of 30-second work capacityIncreased lymphocyte glutathione levels

Increased Bifidobacteria proportionIncreased gastrointestinal immunityDecreased potential for developing atopic diseases

Decreased incidence of infantile colic


Review Whey Protein


Currently the variety of whey proteinsavailable allows for tailoring their use for specificclinical indications. Hydrolyzed whey providesreadily available di- and tri-peptide fractions at-tractive to athletes and other individuals desiringa quickly absorbed, low allergenicity proteinsource. Undenatured whey provides the highestconcentrations of intact native proteins such aslactoferrin and immunoglobulins for immunemodulation. This is attractive to practitioners us-ing whey as a functional food for immune-com-promised patients and as an antimicrobial agent.Glycomacropeptide isolates do not contain theamino acids phenylalanine, tryptophan, or ty-rosine, providing a valuable protein source forindividuals with PKU.

Because of the wide variety of whey prod-ucts, practitioners using whey could benefit fromobtaining an analysis of whey proteins being rec-ommended to patients. Helpful information fordeciding whether a particular whey protein is ap-propriate for individuals includes total proteinconcentration and percent beta-lactoglobulin, al-pha-lactalbumin, glycomacropeptides, immuno-globulins, bovine serum albumin, and lactoferrin.In addition, the presence of fat, lactose, and min-erals should be considered.

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Therapeutic Applications of Whey Protein - GSH Polska · 2015. 6. 19. · whey to a co-product in the manufacturing of cheese.2 Milk contains two primary sources of protein, the caseins