AU638441B2 - Biologically active whey protein composition, a method for producing it and use of the composition - Google Patents
Biologically active whey protein composition, a method for producing it and use of the composition Download PDFInfo
- Publication number
- AU638441B2 AU638441B2 AU47156/89A AU4715689A AU638441B2 AU 638441 B2 AU638441 B2 AU 638441B2 AU 47156/89 A AU47156/89 A AU 47156/89A AU 4715689 A AU4715689 A AU 4715689A AU 638441 B2 AU638441 B2 AU 638441B2
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- Prior art keywords
- whey protein
- whey
- composition according
- concentrate
- glutathione
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- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
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Links
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Classifications
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2250/00—Food ingredients
- A23V2250/54—Proteins
- A23V2250/542—Animal Protein
- A23V2250/5424—Dairy protein
- A23V2250/54244—Beta lactoglobulin
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2250/00—Food ingredients
- A23V2250/54—Proteins
- A23V2250/542—Animal Protein
- A23V2250/5434—Immunoglobulines
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2250/00—Food ingredients
- A23V2250/54—Proteins
- A23V2250/542—Animal Protein
- A23V2250/5436—Serum albumin [SAB]
Landscapes
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
Description
AUSTRALIA
Patents Act 638441 COMPLETE SPECIFICATION
(ORIGINAL)
Class Int. Class Application Number: Lodged: Complete Specification Lodged: Accepted: Published: Priority Related Art: Applicant(s): S Prof. Gustavo Bounous 1745 Cedar Avenue, Montreal, Quebec, CANADA Prof. Phil Gold 3225 The Boulevard, Westmount, Quebec, CANADA Address for Service is: PHILLIPS ORMONDE FITZPATRICK Patent and Trade Mark Attorneys 367 Collins Street Melbourne 3000 AUSTRALIA Complete Specification for the invention entitled: BIOLOGICALLY ACTIVE WHEY PROTEIN COMPOSITION, A METHOD FOR PRODUCING IT AND *:USE OF THE COMPOSITION Our Ref 156031 :POF Code: 1329/110829,110837
S
ow The following statement is a full description of this invention, including the best method of performing it known to applicant(s): -1- 6006 1 Biologically active whey protein composition, a method for producing it and use of the composition The present invention relates to a biologically active whey protein composition comprising substantive amounts of a whey protein concentrate comprising a whey protein isolate mixture having protein and/or peptide components containing glutamylcysteine groups wherein said components are present in an essentially undenatured state and wherein the biological activity of the whey protein concentrate is restricted to its undenatured conformation.
The present invention is also concerned with a method for producing said whey protein composition by ultrafiltration and subjecting the whey, the whey concentrate or a reconstituted whey protein concentrate powder to the actio. of an anion-exchange resine as well as with several application forms of the biologically active whey protein composition AS described in cl-aims 1 ou 24.
2t The invention also relates to food or food supplements, products for human or animal therapeutic nutrition, diet food, intensive care food, pharmaceutical compositions and anti-cancer therapeutic compositions comprising said biologically active whey composition and if necessary in combination with a neutral carrier (vehicle) suitable for oral feeding.
The present invention also relates to a method for increaso*,3p ing cellular levels of glutathione (GSH) by oral administra- S tion of substantive amounts of undenatured components S* of cow's whey protein mixture containing glutamylcysteine (Glu-Cys) groups i.e. beta-lactoglobulin and serum albumin S and possibly immunoglobulin.
Whey is a well-known dairy industry by-product. Its em- composition is approximately that of skim milk without 1 its casein. Acid whey is obtained by acidifying the milk either by adding an inorganic acid or by producing lactic acid (seeding the milk with lactic ferments) at a pH near the isoelectric point of the casein. The whey is recovered after separation of the curd. The addition of rennet to the milk also causes the flocculation or coagulation of the casein. The whey obtained after syneresis is called rennet whey. If the flocculation occurs at the pH of milk or at a slightly lower pH but above 5.8 to 6.0, the whey is called sweet whey.
Whey is therefore defined with regard to the nature of the coagulation of the milk. The acid whey results mainly from the fabrication of fresh curds and from casein plants. The composition of the wheys may vary in rather wide ranges depending on the starting milk and the cheese processing employed. All wheys contain mineral, some fats, an amount of lactic acid, coagulant enzymes, the most interesting fraction obviously being the nitrogenous fraction. Indeed, it is the nitrogenous fraction which essentially comprises the soluble milk proteins possessing a high biological value.
The oldest technique for extracting proteins from whey 25 consists of making them insoluble by a denaturing heat treatment at a pH near their isoelectric point. The obvious drawback of such a technique is it denatures •S the proteins.
S3) As mentioned above, whey proteins are the group of milk proteins that remain soluble in "milk serum" or whey after precipitation of caseins at pH 4.6 and 20 0 C. The major whey proteins in cow's milk are beta-lactoglobulin alpha-lactalbumin immunoglobulin and serum albumin (SA) in order of decreasing amounts.
1 Whey and whey protein have been utilized from time immemorable for nutritional purposes. In addition, whey was recommended in folk and ancient medicine for the treatment of various diseases and, in one instance, lifetime feeding of hamsters with a whey protein diet has been shown to promote longevity with no explanation given.
All these conditions appear to be somehow related to changes in glutathione which is a ubiquitous element exerting a protective effect against superoxide radicals and other toxic agents.
Glutathione is a ubiquitous tripeptide thiol (L--glutamyl- L-cysteinylglycine) with a broad range of vital functions that include detoxification of xenobiotics and protection of cells against oxygen intermediates and free radicals, by-products of oxygen-requiring metabolism. Modulation of intracellular glutathione affects the proliferative immune response of lymphocytes which may be inhibited Q by oxidative injury. Glutathione protect the cells against radiation and alkylating agents. Age-related or experimentally induced glutathione depletion in the lens is associated with cataract formation. Oxidative DNA damage is rapidly and effectively repaired. The human body is continually repairing oxidized DNA. A small fraction of unrepaired lesions, however, could cause permanent changes in DNA and might be a major contributor to old age diseases and cancer. Indeed, several age associated diseases may be induced by free radicals. It appears that whereas data on age-related changes in tissue vitamin L and g* other antioxidants are, at best, contradictory, the tissue glutathione levels are more consistently reported to decline with old age in laboratory animals and man.
For these reasons there has been interest in the factors that influence intracellular glutathione synthesis and 4 1 especially in ways of increasing cellular levels of glutathione.
Glutathione is composed of three amino acids: glutamic acid, glycine and cysteine. Availability of cysteine is a limiting factor in the synthesis of glutathione.
Cysteine is derived from dietary protein and by transsulfuration from methionine in the liver. Various methods have been tried in order to increase cellular level of glutathione. Administration of free cysteine is not an ideal method because this amino acid is rapidly oxidized, toxic and may actually cause glutathione depletion.
Similar problems have been encountered with i.p. injection of N-acetyl-cysteine to rats, although oral administration of this compound appeared to prevent paracetamol-induced glutathione depletion. Administration of compounds that are transported and converted intracellularly into cysteine, such as L-2-oxothiazolidine-4-carboxylate are useful in increasing cellular glutathione acting as an intracellular delivery system for cysteine. Hepatic glutathione doubled four hous after injection, returned to normal 8 hours later but was below normal after 16 hours. Another approach for increasing tissue glutathione levels was found in s.c. injection of j^-glutamylcyst(e)ine in mice: glutathione increased in the kidney by about 55%, 40-60 minutes after injection, returning to near control values 2 hours later. The administered compound is transported intact and serves as a substrate for glutathione synthetase.
It was also reported that about 2 hours after i.p. administration of glutamyl-cysteinyl-glycyl-monomethyl (or monoethyl) ester to mice, the liver and kidney glutathione levels were doubled, with return to normal values after 8 hours. Similar increases in glutathione tissue levels were attained by Meister by administering an alkyl monoester of glutathione (US-A-4,784,685) to mice. Such esters are transported into tissue cells, and are de-esterified 1 within the cells, thus leading to increased cellular levels of glutathione. The kinetics of tissue glutathione increments attained with this method are similar to those described following i.p. injection of methyl or ethyl esters of glutathione. The effectiveness of these methods has been clearly demonstrated in acute experiments (US-A-4,784,685); in mice treated with L-2-oxothiazolidine- 4-carboxylate the expected drop in glutathione tissue level subsequent to acetaminophen injection, was replaced 110 by an actual increase in tissue glutathione values and survival. Other methods to increase tissue glutathione levels are based on the "overshoot" of glutathione concentration, following depletion by diethylmaleate or BSO.
These studies were done in vitro on murine cell lines.
Also preexposure of rats to hypoxia was found to increase lung glutathione.
The administration of glutathione itself is of little g o consequence on tissue glutathione levels, because it apparently cannot be transported intact across the cell membrane.
Some of the previously discussed methods of increasing intracellular levels of glutathione concentration are either toxic or dangerous owing to the risks related to the initial phase of glutathione depletion. The methods involving the use of -glutamylcyst(e)ine, athiazolidine I: or glutathione esters (US-A-4,784,685) offer an interesting o* possibility for short term intervention. However, their 3p long term effectiveness in producing sustained elevation of cellular glutathione has not been shown, nor has the possible toxicity of their long term use been disproved.
Indeed, glutathione and glutathione disulfide were found to be positive in the most commonly used short term tests for carcinogenicity and mutagenicity.
1 Accordingly, an object of the invention is to provide a novel whey protein composition which is biologically active and which can be used in the nutrition of human and animals and as drugs, the composition should contain a special whey protein fraction or fractions which increase host cellular glutathione, reducing the total amount required in comparison to that what is needed to obtain similar objectives using undenatured whey protein concentrate, the new product should be administrated either in capsule or as a powder to be easily dissolved in water or other liquids.
Another object of the invention is to provide a way to increase the intracellular levels of glutathione by oral administration of the cysteine precursor as contained in the undenatured whey protein concentrate of bovine origin. Similar amounts of cysteine given as free cysteine are ineffective. On the other hand, the same amounts of another protein such as egg white, .0 with an identical high cysteine content, were found to be ineffective in raising tissue glutathione levels.
S* Hence, the specific whey protein concentrate physico-chemical composition, in its undenatured form the position of cysteine in relation to the primary, secondary and tertiary structure of the protein), appears to be a crucial factor in the bioavailability of cysteine as a precursor for intracellular synthesis of glutathione.
Another object of the invention is to provide a method for increasing cellular levels of glutathione without exposure to the toxic effects associated with other known methods: the long term ingestion of bovine whey protein concentrate has been shown to be non-toxic in mice, hamsters and humans.
A further object of the invention is to provide a sustained A further object of the invention is to provide a sustained 1 elevation of tissue glutathione for any purpose for which a prophylactic long term elevation of cellular glutathione levels is desired in the prior art, such as for cellular protection against free radicals, foreign hazardous compounds, drug detoxification, radiation, immunodeficiency states etc.
It was surprisingly found upon technical changes in the industrial methods of preparation of whey from milk and of whey protein concentrate that w.p.c.
must be undenatured in order to be biologically active.
Accordingly, a new concept relating to the preparation of whey protein concentrate, with the specific aim of preserving the undenatured conformation upon which its newly discovered biological activity is dependant is proposed in the present invention. This naturally involves the two principal phases in the industrial production of whey proteins: 1 1. A process of separation of whey from milk which brings about minimal deviation of the whey protein conformation from the range normally accepted as native. Hence the process should exclude levels of heat treatment and shaking in the presence of air which could denature the whey protein.
•O 0 O 2. A process for whey protein concentrate production designed to recover from whey the proteins in their native conformation. Hence ultra-filtration and the 0: other stages of the process are very lenient excluding or limiting heating, pumping, aeration and other processing treatment which promotes protein denaturation.
*The above-meoni ed pr-b I ems were solved by the characterL '-i4-n-f-eatures of rCelaim 1 rpcti vcly 11
"Q
0 7A- According to one aspect of the invention, there is provided a method of producing a biologically active whey protein concentrate composition comprising a suitable concentration of undenatured whey protein concentrate comprising the following steps: immediately after milking cooling the milk to a temperature in the range of 2 0 C to 10 0
C,
after another cleaning of the milk, precipitation of the curd by reducing the pH to about 4.6 with lactic acid initially at 20 0
C,
addition of rennet and raising the temperature to about 30 0 C for 20 minutes to promote expulsion of whey from the curd, allowing the agitation in a vat to resolve at low speed, thermal treatment of the pasteurization type of the remaining product in the vat and agitating at high speed, irradiation and separation of the whey and ultrafiltration of the whey using a membrane having a molecular weight cut off of substantially 17,000 or less to retain B-lactoglobulin and serum albumin, said method being characterized in that the fraction S of whey protein concentrate is not heated and the material .'3D from which it is derived is slowly agitated to minimize protein denaturation and that said ultrafiltration is carried out in a production line comprising up to 20 frame-type S modules holding a large number of said membranes achieving a final undenatured protein concentrate in dry matter, wherein said ultrafiltration is carried out at a temperature in the range of 4 0 C to 20 0
C.
The retentate may then be subject to further ultrafiltration 4 with membranes having molecular weight cut offs of 100,000 '7 4!0 and/or 500,000.
Il 0 IT 1 7B- A further aspect of this invention provides biologically-active whey protein composition obtained by the above method comprising a whey protein concentrate containing a whey protein isolated mixture having protein and/or peptide components containing glutamylcysteine groups wherein said components are present in substantially undenatured state and wherein the biological activity of the whey protein concentrate is restricted to its undenatured conformation, and further wherein the protein isolate mixture contains beta-Lactoglobulin and/or serum albumin and/or immunoglubolin and still further wherein the alpha-Lactalbumin fraction is removed. The protein isolate mixture can then be concentrated and refined to medicinal purity. A whey protein composition is further provided wherein the whey protein concentrate has immunoenhancing proteins being heat-labile and insensitive to digestion and wherein the immunoenhancing properties depend upon the undenatured state of the peptides and proteins contained in the whey protein mixture.
Whilst the whey protein concentrate of the invention is not limited to that of any particular species, importantly it includes bovine and/or sheep and/or goat and/or human whey protein concentrate.
Whey protein compositions of this invention are provided :i wherein the whey protein concentrate is preferably present in an amount of 18 to 28g, more preferably 20g per 100g diet.
The whey protein composition according to the invention further comprises in combination whey protein composition together with vitamins B1 and B2 in amount in excess of minimum daily requirements preferably in amounts of 1.5 to 2.Omg B 1 and 1.5 to 2.0mg B 2 per 100g diet.
The invention further provides a food or food supplement including a milk or milk protein formulation comprising a whey protein composition as described above adapted for specific nutritional requirements and a neutral vehicle. It still further provides a method of treatment of humans and/or animals including colon cancer comprising administration of i fE^
(A
0 7Ctherapeutically or prophylactically effective amounts of whey protein composition with a neutral vehicle suitable for oral f eeding.
OV.
8 -oA'e invpntion i furthpr HpvplnnD hv t hel aim Claims 16 to 24 comprise veral application forms of the novel whey in composition.
IT advantaq offered by tha invntionarc mainl.y that: 1. Oral administration of substantive amounts of bovine whey protein concentrate enhances the glutathione content in the liver, heart and spleen of mice (see Fig. 1, 2, 3).
2. This change is moderate but sustained over time and biologically significant.
3. This property is restricted to the undenatured conformation of whey protein concentrate.
4. Glutathione is a tripeptide -glutamyl-L-cysteinyl- SO glycine).
The administration of either cysteine (limiting sub- 6oo strate) or glutathione itself or other protein sources o such as egg white are ineffective in raising cellular 25 levels of GSH. However, i.p. injection of -glutamylcysteine was found to enhance for a few hours the tissue GSH level of mice, 5. Whey protein concentrate contains substantial amounts of glutamylcysteine groups, unlike casein, which does not increase tissue GSH when fed to mice.
6. The glutamylcyteine groups are located in the beta-lactoglobulin and serum albumin fractions (see table 1).
35 The aminoacid sequence of bovine milk immunoglobulin is not totally known.
1 7. Pancreatic digestion does not split the disulfide bond of the whey proteins; instead heat denaturation is known to split this bond, thus unfolding the globular structure of the molecule to form a random coil conformation.
The invention is now described with reference to the drawings and tables which show: Fig. 1 the spleen glutathione content of mice fed different diets, Fig. 2 and 3 liver and heart glutathione content of mice fed different diets, Fig. 4 enhancement of spleen cell immune response to SRBC, in mice fed a diet containing undenatured whey protein concentrate, Fig. 5 role of glutathione in the immuno-enhancing effect of dietary whey protein, Fig. 6 *Va table 1 the protein composition of cow and human milks, es Fig. 7 table 2 amino acid composition of whey protein concentrate and egg white protein.
S* The interaction of dietary protein, GSH and the host immune response was explored. It was investigated whether 30 a different protein source such as egg white, with the S same high level of cysteine as whey protein concentrate (Table had a similar effect in promoting higher GSH tissue content. It was found out that an egg white protein diet does not enhance the host immune response above average. Whereas the static GSH level in spleen was found unaltered by U-Lacp feeding for three weeks, the present studies in young adult C3H mice showed that 1 enhancement of spleen cell immune response to SRBC (Fig. 4) is associated with sustained elevation of splenic GSH during the antigen-driven clonal expansion of the lymphocytes in U-Lacp (undenatured whey protein)-fed mice in comparison to a pattern of decline observed in spleen GSH levels in mice fed either of the nutritionally equivalent D-Lacp (denatured whey protein), casein, cysteine enriched casein, or egg white protein diets (Fig. 1).
The latter four groups also exhibited a lower immune response (Fig. Administration of S-(n-butyl) homocyteine sulfoximine, which reduces the splenic glutathione level by half, produces a marked drop in the humoral immune response of whey protein (U-Lacp) diet-fed mice. This is further evidence of the important role of glutathione in the immunoenhancing effect of dietary whey protein (Fig. Tissue Glutathione Assay: Ninety milligrams of mouse heart or liver were homogenized in 0 acid Homogenates are centrifuged for 5 minutes in a microfuge at 10,000 x g. The assay is carried out using the supernatants on the same day according to the method of Anderson. Values are expressed as pmol/g wet tissue (Fig. 2 and 3).
After three months on either diet initiated at age 17 months, GSH content was found to be higher in the liver o* 0 and heart of U-Lacp (undenatured whey protein) fed mice compared to the D-Lacp (denatured whey protein), casein, 30 egg white protein or Purina diet-fed counterparts (Fig. 2 and The GSH values in heart and liver of mice fed Purina laboratory chow was similar at age 10 weeks, 17, 20, 21 months. The U-Lacp diet appears to enhance Sthe GSH content of heart and liver above "normal" values 85 after 3 and 4 months of continuous feeding (Fig. 2 and 3).
11 1 In conclusion, after three weeks on the U-Lacp diet, spleen GSH content is increased during the antigen driven clonal expansion of the lymphocytes in young adult C3H/HeN mice as compared to a decline in controls fed D-Lacp, casein or egg white protein diets (Fig. In old C57BL/6N1A mice, long term feeding of U-Lacp diet results in a moderate but sustained increase in liver and heart GSH levels (Fig. 2 and The GSH enhancing activity of WPC (whey protein concentrate) is restricted to its undenatured form (U-lacp). This property is not solely due to the high cysteine content of WPC because another protein source with similar cysteine content (egg white) does not exhibit this biological activity. This property of U-Lacp does not depent specifically on its nutritional efficiency as evaluated by body weight, serum proteins, and food consumption, but appears to depend on the primary, secondary and tertiary structure of the protein in its native form.
6e '0 Data in Fig. 2 and 3 show that the concentration of liver and heart glutathione in control Purina fed mice remains very constant over time. On the other hand a moderate but sustained elevation of tissue GSH was noted in mice fed the nutritionally equivalent undenatured whey protein (U-Lacp) diet. Only minuscule quantities of glutathione and no breakdown products that can be readily attributed to glutathione are excreted in urine.
The magnitude of change in cellular glutathione concentration that can be achieved may be quite limited, perhaps 3. reflecting the critical importance of this molecule and the attendent tight regulatory control. Glutathione itself serves as a negative feedback on the GSH synthetic enzymes, which obviously limits cellular capacity to increase GSH concentration. Glutathione reductase maintains 5s GSH in its predominant reduced form This serves both to maintain this functional state and also to control 1 cellular concent: tion since reduced glutathione (GSH) cannot cross the membrane, whereas the oxidized form (GSSG) can and does efflux, resulting in decreased total glutathione. Besides these enzymes, gamma glutamyltranspeptidase (GGT) is important in GSH metabolism. GGT serves as a salvage pathway for glutamyl moieties at the cell membrane level, passing them back into the cytosol to be used in GSH synthesis. Increased activity of this enzyme has been associated with elevated GSH concentration in a number of cell lines and malignant tissues.
The effects of a small increment in cellular GSH may be greater than expected. For example, there are many reports of human and murine tumor cell lines selected in vitro for resistance to a variety of chemotherapeutic agents. In a number of these cell lines cellular GSH is increased consistently by 2-fold compared to the drug sensitive parental cell line, despite the fact 0O that the level of drug resistance is often much greater, o e.g. as much as 30-fold. In these cell lines, depletion of cellular GSH by selective inhibition of synthesis restores drug sensitivity to the resistent cells. This S" is effective only if the GSH depletion is maintained 25 throughout the drug-treatment period.
Given the fact that cellular GSH is very tightly regulated, that a 2-fold increase may be maximal, and that the effect of small increments in GSH may be amplified by a variety of GSH-utilizing enzymes glutathione Speroxidase, glutathione-S-transferase), the reproducible sees,: change in GSH concentration observed in animals fed S the whey-rich diet is likely to have biological importance.
The chronic nature of this augmentation may contribute 3 significantly to this effect.
13 1 Relevant to the present invention are recent data indicating specifically that a lack of the GSH precursor, cysteine, rather than a decrease in biosynthetic enzyme activities is responsible for the deficiency of GSH noted in aging animals. Similarly, the fall in cytosolic GSH in the liver of chronic ethanol fed rats does not appear to be caused by a limitation in the capacity of -glutamylcysteine synthetase activity.
Summary and Signification of the Invention a. The glutathione promoting activity of dietary whey protein concentrate is dependant on the glutamylcysteine groups contained in the beta-lactoglobulin and serum albumin fractions and possibly in the IgG fraction.
b. The preservation of the disulfide bond (which involves the cysteine) may be crucial to the release, upon digestion, of intact glutamylcysteine peptide for 2)b absorption by the intestinal mucosa. Denaturation, on the other hand, by unfolding the protein molecule, exposes the glutamylcysteine sequence to the digestive enzymes with subsequent release of either of the single aminoacids or other peptide combinations.
This would be consistent with the observed absence of glutathione promoting activity in the denatured form of whey protein concentrate.
The present invention provides a method to remove from 9O whey protein the alpha-lactalbumin (table 1).
o• Oral administration of the whey protein fraction or fractions found to release most glutamylcysteine groups upon digestion, with the purpose of increasing host 35 cellular glutathione. The total amount required would be significantly less than what is needed to obtain 14 1 similar objectives using undenatured whey protein concentrate. Hence the product could be administered either in capsule or as a powder to be easily (undenatured) dissolved in water or other liquids.
.0:6
Claims (18)
1. Method of producing a biologically active whey protein concentrate composition comprising a suitable concentration of undenatured whey protein concentrate comprising the following steps: immediately after milking cooling the milk to a temperature in the range of 20C to 10 0 C, after *notler cleaning of the milk, precipitation of the curd by reducing the pH to about 4.6 with lactic acid initially at 20 0 C, addition of rennet and raising the temperature to about 30 0 C for 20 minutes to promote expulsion of whey from the p.rd, allowing the agitation in a vat to resolve at low speed, thermal treatment of the pasteurization type of the 0 remaining product in the vat and agitating at high speed, irradiation -nd separation of the whey and ultrafiltration of the whey using a membrane having a molecular weight cut off of substantially 17,000 or less to retain B-lactoglobulin and serum albumin, said method being characterized in that the fraction of whey protein concentrate is not heated and the material from which it is derived is slowly agitated to minimize protein denaturation and that said ultrafiltration is carried out in a production line comprising up to 20 frame-type modules holding a large number of said membranes achieving a final undenatured protein concentrate in dry matter, wherein said ultrafiltration is carried out at a temperature in the range of 4 0 C to 200C.
2. Method according to claim 1, characterized in that the retentate from the ultrafiltration is subjected to further 3u 16 ultrafiltration using a membrane having a molecular weight cut off of substantially 100,000 to form a concentrate which is further dried to form a powder.
3. Method according to either claim 1 or 2 characterized in that the retentate from the ultrafiltration is subjected to further ultrafiltration using a membrane having a molecular weight cut off of substantially 500,000 to form a concentrate which is further dried to form a powder.
4. Method of producing an undenatured whey protein concentrate according to claim 1, characterized in that the: whey protein concentrate is after ultrafiltration free of lactose, salts and water. Method according to any one of the preceding claims, wherein the temperature of ultrafiltration in step and/or the temperature of cooling in step is 4°C.
6. Biologically active whey protein composition obtained by the method as claimed in any one of the preceding claims comprising a whey protein concentrate containing a whey protein isolated mixture having protein and/or peptide components containing glutamylcysteine groups wherein said components are present in -ubstantially undenatured state and wherein the biological activity of the whey protein concentrate is restricted to its undenatured conformation.
7. Whey protein composition according to claim 6, wherein said whey protein isolate mixture contains beta-Lactoglobulin and/or serum albumin and/or immunoglobulin.
8. Whey protein composition according to either of claims S6 or 7, wherein the alpha-Lactalbumin fraction is removed.
9. Whey protein composition according to any one of claims 6 to 8, wherein the whey protein isolate mixture is further concentrated and refined to medicinal purity. 17 Whey protein composition according to any one of claims 6 to 9, wherein the whey protein concentrate has immunoenhancing proteins being heat-labile and insensitive to digestion and wherein the immunoenhancing properties depend upon the undenatured state of the peptides and proteins contained in said whey protein mixture.
11. Whey protein composition according to claim 6, wherein the whey protein concentrate is one selected from the group consisting of bovine and/or goat and/or sheep and/or human whey protein concentrate.
12. Whey protein composition according to any one claims 6 to 11, wherein amount of whey protein concentrate is 18 to 28g whey protein/l00g diet.
13. Whey protein composition according to any one of claims 6 to 12, wherein amount of whey protein concentrate is 20g whey protein/100g diet. :2g C
14. Whey protein composition according to any one of claims 6 to 13 comprising in combination said whey protein concentrate together with vitamins B 1 and B 2 in excess of minimum daily requirements. Whey protein composition according to claim 14 wherein the vitamin amounts are 1.5 to 2.0mg B 1 and 1.5 to B 2 /100g diet.
16. Milk or milk protein formulation comprising a whey protein composition according to any one of claims 6 to 15 in a concentration of 18 to 28g whey protein/l00g diet.
17. Method of treatment of humans and/or animals for the condition of lactose malabsorption; to increase the rate of replenishment, and/or concentration levels of glutathione; or to improve the host resistance comprising administration of therapeutically or prophylactically effective amount of whey protein composition according to any one of claims 6 to i 410 6pn^'~ 18
18. Method of treatment of colon cancer comprising administration of a therapeutically or prophylactically effective amount of whey protein composition according to any one of claims 6 to
19. Food or food supplement comprising whey protein composition according to any one of claims 6 to 15 adapted for specific nutritional requirements and a neutral vehicle.
20. Food or food supplement according to claim 19 further comprising one or more additional food materials.
21. Pharmaceutical composition comprising whey protein composition according to any one of claims 6 to 15 and a neutral vehicle suitable for oral feeding. DATED: 21 April 1993 PHILLIPS ORMONDE FITZPATRICK Patent Attorneys For: n1.C4 <A. PROF. GUSTAVO BOUNOUS and PROF. PHIL GOLD 2* 0000 o o *s S *S
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US288971 | 1988-12-23 | ||
| US07/288,971 US4974018A (en) | 1984-06-11 | 1988-12-23 | Exposure method and exposure apparatus |
Publications (2)
| Publication Number | Publication Date |
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| AU4715689A AU4715689A (en) | 1990-06-28 |
| AU638441B2 true AU638441B2 (en) | 1993-07-01 |
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| AU47156/89A Ceased AU638441B2 (en) | 1988-12-23 | 1989-12-21 | Biologically active whey protein composition, a method for producing it and use of the composition |
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB1495940A (en) * | 1975-02-18 | 1977-12-21 | Grindstedvaerket As | Anti-cancer active whey fraction |
| EP0320152A2 (en) * | 1987-12-11 | 1989-06-14 | Express Foods Group (International) Limited | Isolation of an immunoglobulin rich fraction from whey |
| AU596632B2 (en) * | 1986-01-13 | 1990-05-10 | Lanier Industries, Inc. | Immunologically active whey fraction and recovery process |
-
1989
- 1989-12-21 AU AU47156/89A patent/AU638441B2/en not_active Ceased
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB1495940A (en) * | 1975-02-18 | 1977-12-21 | Grindstedvaerket As | Anti-cancer active whey fraction |
| AU596632B2 (en) * | 1986-01-13 | 1990-05-10 | Lanier Industries, Inc. | Immunologically active whey fraction and recovery process |
| EP0320152A2 (en) * | 1987-12-11 | 1989-06-14 | Express Foods Group (International) Limited | Isolation of an immunoglobulin rich fraction from whey |
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| AU4715689A (en) | 1990-06-28 |
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