WO2001003515A9 - Method of obtaining protein isolates and concentrates from colostrum - Google Patents

Method of obtaining protein isolates and concentrates from colostrum

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Publication number
WO2001003515A9
WO2001003515A9 PCT/NZ2000/000120 NZ0000120W WO0103515A9 WO 2001003515 A9 WO2001003515 A9 WO 2001003515A9 NZ 0000120 W NZ0000120 W NZ 0000120W WO 0103515 A9 WO0103515 A9 WO 0103515A9
Authority
WO
WIPO (PCT)
Prior art keywords
colostrum
serum
skim
deplete
protein
Prior art date
Application number
PCT/NZ2000/000120
Other languages
French (fr)
Other versions
WO2001003515A1 (en
WO2001003515A8 (en
Inventor
Peter Graeme Hobman
Robert Philip Boswell
Joanne Campbell Hutchinson
Georgina Aureen Klarwill
Original Assignee
New Zealand Co Operative Dairy
Peter Graeme Hobman
Robert Philip Boswell
Joanne Campbell Hutchinson
Georgina Aureen Klarwill
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by New Zealand Co Operative Dairy, Peter Graeme Hobman, Robert Philip Boswell, Joanne Campbell Hutchinson, Georgina Aureen Klarwill filed Critical New Zealand Co Operative Dairy
Priority to AU57196/00A priority Critical patent/AU5719600A/en
Publication of WO2001003515A1 publication Critical patent/WO2001003515A1/en
Publication of WO2001003515A9 publication Critical patent/WO2001003515A9/en
Publication of WO2001003515A8 publication Critical patent/WO2001003515A8/en

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Classifications

    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
    • A23J1/00—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites
    • A23J1/20—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from milk, e.g. casein; from whey
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C9/00—Milk preparations; Milk powder or milk powder preparations
    • A23C9/14—Milk preparations; Milk powder or milk powder preparations in which the chemical composition of the milk is modified by non-chemical treatment
    • A23C9/142—Milk preparations; Milk powder or milk powder preparations in which the chemical composition of the milk is modified by non-chemical treatment by dialysis, reverse osmosis or ultrafiltration
    • A23C9/1422—Milk preparations; Milk powder or milk powder preparations in which the chemical composition of the milk is modified by non-chemical treatment by dialysis, reverse osmosis or ultrafiltration by ultrafiltration, microfiltration or diafiltration of milk, e.g. for separating protein and lactose; Treatment of the UF permeate
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C9/00—Milk preparations; Milk powder or milk powder preparations
    • A23C9/20—Dietetic milk products not covered by groups A23C9/12 - A23C9/18
    • A23C9/206—Colostrum; Human milk
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/20—Milk; Whey; Colostrum
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/04—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies from milk
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C2210/00—Physical treatment of dairy products
    • A23C2210/20—Treatment using membranes, including sterile filtration
    • A23C2210/206—Membrane filtration of a permeate obtained by ultrafiltration, nanofiltration or microfiltration
    • 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
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/10—Immunoglobulins specific features characterized by their source of isolation or production
    • C07K2317/12—Immunoglobulins specific features characterized by their source of isolation or production isolated from milk

Definitions

  • This invention relates to a method of producing protein isolates and concentrates by microfiltration of colostrum, and the production of products fortified with immunoglobulins therefrom.
  • WPC whey protein concentrates
  • WPI whey protein isolates
  • WPC and WPI can be used as a source of bioactive materials.
  • Bioactive molecules, and in particular immunoglobulins can be sourced from milk, milk serum, colostrum or colostral serums.
  • Colostral serum is colostrum with a reduced fat and casein content.
  • the immunoglobulin concentrations in colostrum are generally much higher than in milk. This concentration declines with time postpartum, from > 50 mg/ml on day 1 to about 1 5 mg/ml on day 3. This compares with a concentration of ⁇ 1 mg/ml in milk.
  • the main immunoglobulin present is IgGi with much smaller concentrations of IgA and IgM.
  • Immunoglobulins have particular application in the treatment of various diseases. For example, IgA, present in the form of secretory IgA in the epithelial cells of mucous membrane, attacks bacteria and viruses, thus playing an important role in local immunity. Oral administration of secretory IgA can therefore provide immunological protection.
  • immunisation of an animal for example a cow
  • specific antigens will boost the production of specific antibodies, for example IgG and/or IgA, which may then be "harvested” from the milk or colostrum of that animal.
  • Casein is a potentially valued fraction of milk or colostrum.
  • colostrum refers to the mammary secretion from a cow during the first 4 days postpartum, the term is used to include colostrum in any form including fresh or frozen whole colostrum and preconcentrated colostrum.
  • serum deplete includes complete or partial depletion of whey proteins.
  • shkim colostrum refers to colostrum which has had the fat and cellular and other particulate material removed.
  • impaired ratio of casein:whey protein reflects serum depletion such that proportionately there is a higher casein content.
  • a method of producing a protein concentrate and/or serum isolate from colostrum including:
  • skim colostrum stream taking a skim colostrum stream; and - microfiltration of the skim colostrum stream to produce a permeate rich in colostral serum proteins and a serum deplete retentate including an enhanced ratio of casein:whey protein.
  • the skim colostrum stream may be formed by separation of fat and other particulate and cellular material from colostrum, or by reconstituting skim colostrum powder in water.
  • the permeate may be further processed by ultrafiltration and/or diafiltration to produce a colostrum serum isolate.
  • the retentate may optionally be concentrated before drying to produce a serum deplete colostrum protein concentrate.
  • the method may further include temperature adjustment of the skim colostrum to a temperature lower than that which may cause denaturation of immunoglobulins.
  • the temperature is ⁇ 70°C.
  • the temperature may be adjusted to substantially 50°C or substantially 10°C.
  • the method may further include maintaining the pH of the skim colostrum in the range 4.6 to 8.5.
  • the pH may be maintained in its natural range. Preferably in the range substantially 6 to 7.
  • the microfiltration may involve membranes having a cut- off of 0.05 to 1 .4 ⁇ m.
  • the membrane cut-off may be substantially 0.1 ⁇ m.
  • Microfiltration may preferably involve a ceramic microfiltration membrane.
  • a method of producing a colostrum serum isolate from colostrum including the steps of:
  • concentration of the permeate may involve ultrafiltration and/or diafiltration.
  • the colostrum serum isolate formed in accordance with the method may include at least 70% protein.
  • At least 20% of the protein may be immunoglobulins.
  • a serum deplete colostrum protein concentrate from colostrum including the steps of:
  • skim colostrum stream taking a skim colostrum stream; microfiltration of the skim colostrum stream to produce a serum deplete retentate including an enhanced ratio of casein:whey protein; optionally concentrating the retentate; and drying the optionally concentrated retentate to produce a serum deplete colostrum protein concentrate.
  • a method of producing a colostrum powder deplete in whey proteins from colostrum including the steps of:
  • skim colostrum stream taking a skim colostrum stream; microfiltration of the skim colostrum stream to produce an intermediate permeate rich in colostral serum proteins and a serum deplete retentate having an enhanced ratio of casein:whey protein; - concentration of said intermediate permeate by ultrafiltration and/or diafiltration to produce a further permeate; mixing the further permeate with the serum deplete retentate; optionally concentrating the mixed further permeate/retentate; and - drying the optionally concentrated mixture to produce a colostrum powder deplete in whey proteins.
  • the starting material may comprise colostrum from bovine immunised or hyperimmunised before collection of the colostrum.
  • a colostrum serum isolate including a protein content of at least 70.%.
  • the protein content of the colostrum serum isolate may include at least 20% immunoglobulins.
  • a colostrum protein concentrate which is serum deplete and includes an enhanced ratio of casei whey protein.
  • FIGURE 1 Is a flow diagram showing the main steps of the process of the invention in one preferred form. Detailed Description of the Invention
  • the process of the invention enables the production of colostrum serum isolate (CSI) and serum deplete (SD) colostrum protein concentrate (CPC) by microfiltration, and those products may then be further processed to purify particular bioactive substances.
  • the production of CSI and (SD) CPC from colostrum occurs in parallel as the (SD) CPC is formed from the serum deplete stream resulting from microfiltration.
  • Colostrum powders are typically used in nutritional supplement, functional food and pharmaceutical applications where they confer a wide range of benefits.
  • a powder formed from the (SD) CPC derived from the present invention is likely to have an elevated content of complex lipids. It has an elevated immunoglobulin content, reduced levels of serum protein and consequently an enhanced ratio of casein:whey protein. It has the potential to be used in functional food applications.
  • the immunoglobulin content of colostrum decreases progressively postpartum.
  • the starting material for the method of the invention is, or is derived from, colostrum obtained during the first 2 days postpartum.
  • CSI derived from the method of the present invention has application in, for example, cultured foods, sports applications, medical and dietetic applications, dairy desserts, dry mix beverages, ready to drink beverages and prepared foods.
  • CSI offers an advantage in terms of the higher concentration of immunoglobulins and other small bioactive molecules. It also has excellent clarity when in solution as a result of the low fat content.
  • the starting material for this process is a skim colostrum stream from colostrum. It will be appreciated that this may be fresh, dried or frozen. The colostrum undergoes a series of process conditions as seen in Figure 1 .
  • Colostrum is passed through a separator to remove the fat and other particulate and cellular material, resulting in a skim colostrum stream and a cream stream.
  • the skim stream (or whole colostrum stream) may be pasteurised for microbial hygiene prior to microfiltration (MF).
  • the MF process is carried out preferably using ceramic MF membranes with 0.05- 1 .4/vm cut-off, but preferably 0.1/vm.
  • the temperature is maintained between 4 and 70°C, and preferably about 50°C.
  • the pH of the MF feed is maintained between pH 4.6 and 8.5, and preferably at a natural pH in the range 6 to 7.
  • Additives e.g. filter aids
  • the retentate from MF may optionally be concentrated (by ultrafiltration/diafiltration) prior to, optionally, evaporating, and then drying, producing (SD) CPC.
  • the permeate from MF may be further concentrated by processes such as ultrafiltration, nanofiltration, reverse osmosis or evaporation.
  • Ultrafiltration is the preferred method and may be performed between 5 °C and 70 °C, preferably about 10 °C, using membranes with a molecular weight cut-off of between 1 kD and 100kD, but preferably about 10kD to increase the solids.
  • Diafiltration (DF) may also be used.
  • the concentrated MF permeate may then be evaporated (optional), and dried, resulting in CSI, which has a high concentration of immunoglobulins.
  • the permeate from microfiltration may be mixed back with the serum deplete retentate.
  • the mixture may then optionally be concentrated prior to evaporation and drying.
  • Spray dried skim colostrum (IMMULAC t ) wee reconstituted with demineralised Water to 12% total BolidSi
  • the pH of the solution was neutral, at about 6.7-6.8.
  • the solution was then heated to 50°C prior to microfiltration.
  • the pilot plant used was Tetra Pak Filtration System MFS1 9, fitted with an SCT (Societe des Ceramiques Techniques) membrane (0.1 ⁇ m ceramic membrane with 4mm channel spacers).
  • the crossflow rate was 6-7 m/s.
  • a Uniform Transmembrane Pressure (UTP) system was used to reduce fouling and to achieve optimal conditions over the entire memb ane area.
  • the temperature was maintained at 50°C throughout the process.
  • the permeate operating pressure difference was 1 .6 bar.
  • the retentate operating pressure difference was 1 .9 bar.
  • the retentate from the membranes was collected and spray dried. This fraction of the colostrum forms the (SD) CPC.
  • the permeate from microfiltration was then ultrafiltered at 50°C using 5kD spiral wound membranes to total solids of 9.5%. 1 00% diafiltration water was added during the ultrafiltration.
  • the ultrafiltered permeate was then spray dried, resulting in the CSI, which is high in immunoglobulins.
  • compositional results for the (SD) CPC and CSI are shown in Table I, in comparison with skim colostrum powder, UF skim colostrum powder and milk protein concentrate (MPC) which comes from the ultrafiltration of skim milk.
  • Table I shows that the lactose content in CSI is much lower than that in skim milk colostrum and UF skim colostrum, and the lactose content of (SD) CPC is lower than skim colostrum, but higher than UF colostrum.
  • the (SD) CPC is comparable to skim colostrum powder in composition. However the protein content is higher and would be enriched in casein. It is anticipated that the protein content will also include an elevated level of immunoglobulins in comparison with standard skim colostrum powder.
  • the CSI has a total protein composition more similar to that of a WPI, but with a low lactose and fat content. Moreover, the nature or profile of the protein content is different from that for a milk-derived WPI.
  • a serum deplete protein concentrate and a protein isolate can be produced from colostrum by microfiltration without forming a whey intermediate.
  • Purification of commercially useful quantities of bioactive materials, such as immunoglobulins, from the (SD) CPC and CSI is also possible.

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Abstract

This invention relates to a method of producing protein isolates and concentrates by microfiltration of colostrum, and the production of products fortified with immunoglobulins. The method involves taking a skim colostrum stream, derived from colostrum by the separation of fat and other particulate and cellular material, or by reconstituting skim colostrum powder and water, and microfiltration of that skim colostrum stream to produce a permeate rich in colostral serum proteins and a serum deplete retentate including an enhanced ratio of casein:whey protein. The invention also relates to a method of producing a colostrum powder deplete in whey proteins from colostrum in which an intermediate permeate from the microfiltration, rich in colostral serum proteins, is concentrated and then mixed back with the serum deplete retentate, the mixture optionally being concentrated before drying to produce a colostrum powder deplete in whey proteins. The invention also includes a colostral serum isolate including a protein content of at least 70 %, a colostrum protein concentrate which is serum deplete and includes an enhanced ratio of casein:whey protein and a colostrum powder deplete in whey proteins.

Description

METHOD OF OBTAINING PROTEIN ISOLATES AND CONCENTRATES
FROM COLOSTRUM
Technical Field
This invention relates to a method of producing protein isolates and concentrates by microfiltration of colostrum, and the production of products fortified with immunoglobulins therefrom.
Background to the Invention
There are two major types of whey products: whey protein concentrates (WPC) which generally have a protein content > 35% and whey protein isolates (WPI) which generally have a protein content > 80%. WPC is manufactured from whey by ultrafiltration whilst WPI is manufactured from whey by microfiltration or ion exchange. Alternatively WPI can be manufactured by microfiltration of skim milk.
As well as being important from a nutritional point of view, WPC and WPI can be used as a source of bioactive materials.
Bioactive molecules, and in particular immunoglobulins can be sourced from milk, milk serum, colostrum or colostral serums. Colostral serum is colostrum with a reduced fat and casein content. The immunoglobulin concentrations in colostrum are generally much higher than in milk. This concentration declines with time postpartum, from > 50 mg/ml on day 1 to about 1 5 mg/ml on day 3. This compares with a concentration of < 1 mg/ml in milk. The main immunoglobulin present is IgGi with much smaller concentrations of
Figure imgf000003_0001
IgA and IgM. Immunoglobulins have particular application in the treatment of various diseases. For example, IgA, present in the form of secretory IgA in the epithelial cells of mucous membrane, attacks bacteria and viruses, thus playing an important role in local immunity. Oral administration of secretory IgA can therefore provide immunological protection.
The separation and purification of bioactive materials from milk and colostrum has significant commercial advantages over the sourcing of such active materials from human blood. Blood serum antibodies are difficult to collect and it is therefore difficult to produce commercial quantities for use in medications. Furthermore, secretory IgA is more stable against proteases and acidic conditions than serum IgA.
It is recognised that immunisation of an animal, for example a cow, with specific antigens will boost the production of specific antibodies, for example IgG and/or IgA, which may then be "harvested" from the milk or colostrum of that animal.
However, known methods of separating and purifying immunoglobulins from milk (via the formation of WPC and WPI) or colostrum have not proved entirely satisfactory in that the antibodies isolated may not be sufficiently stable to undergo the kind of processing required to produce a pharmaceutical composition, the yield of antibodies may be poor, and problems of lactose intolerance may arise in some individuals treated with antibodies derived from milk products as a result of residual lactose associated with the separated antibodies.
In recent years there has been a growth in demand for colostrum powders, and there is a growing awareness of the potential of colostrum as a functional food. As a result there is a need for improvements in the methods of processing colostrum in particular to produce discrete fractions suitable for a variety of applications.
One method of processing milk or colostral whey, involving microfiltration, is described in United States patent specification no. US 5,707,678. The process described in that specification is designed to avoid denaturation of immunoglobulins. The method involves careful monitoring and control of temperature and pH levels. It uses fresh, frozen or dried milk, milk serum, colostrum or colostral serum as the starting ingredient in the production of protein concentrates. The process also involves initial separation of cream or fat. The defatted milk, milk serum, colostrum or colostral serum is then acidified to precipitate casein, at a pH in the range 4.5 to 5.0. The casein is separated off leaving a whey which is then subjected to microfiltration, optionally after ultrafiltration.
That method involves denaturation and removal of casein, and the formation of a whey intermediate. Casein is a potentially valued fraction of milk or colostrum.
It is an object of the present invention to provide a method of obtaining protein concentrates and/or isolates, and fortified products containing immunoglobulins or other bioactive molecules, from colostrum or preconcentrated colostrum which reduces or overcomes the above- mentioned problems or which at least provides the public with a useful alternative.
Other objects of the invention may become apparent from the following description which is given by way of example only.
In this specification "colostrum" refers to the mammary secretion from a cow during the first 4 days postpartum, the term is used to include colostrum in any form including fresh or frozen whole colostrum and preconcentrated colostrum. The term "serum deplete" includes complete or partial depletion of whey proteins. The term "skim colostrum" refers to colostrum which has had the fat and cellular and other particulate material removed. The phrase "enhanced ratio of casein:whey protein" reflects serum depletion such that proportionately there is a higher casein content.
Summary of the Invention
According to one aspect of the present invention there is provided a method of producing a protein concentrate and/or serum isolate from colostrum, the method including:
taking a skim colostrum stream; and - microfiltration of the skim colostrum stream to produce a permeate rich in colostral serum proteins and a serum deplete retentate including an enhanced ratio of casein:whey protein.
The skim colostrum stream may be formed by separation of fat and other particulate and cellular material from colostrum, or by reconstituting skim colostrum powder in water.
In one preferred form of the method the permeate may be further processed by ultrafiltration and/or diafiltration to produce a colostrum serum isolate.
In a further preferred method the retentate may optionally be concentrated before drying to produce a serum deplete colostrum protein concentrate. Preferably the method may further include temperature adjustment of the skim colostrum to a temperature lower than that which may cause denaturation of immunoglobulins. Preferably, the temperature is < 70°C.
Preferably, the temperature may be adjusted to substantially 50°C or substantially 10°C.
Preferably the method may further include maintaining the pH of the skim colostrum in the range 4.6 to 8.5.
Preferably, the pH may be maintained in its natural range. Preferably in the range substantially 6 to 7.
Preferably, the microfiltration may involve membranes having a cut- off of 0.05 to 1 .4 μm.
Preferably, the membrane cut-off may be substantially 0.1μm.
Microfiltration may preferably involve a ceramic microfiltration membrane.
According to a further aspect of the present invention there is provided a method of producing a colostrum serum isolate from colostrum, the method including the steps of:
taking a skim colostrum stream; microfiltration of the skim colostrum stream to separate out casein and lipids, and produce a permeate which is rich in colostral serum proteins; and - concentration of the permeate to produce a colostral serum isolate. Preferably, concentration of the permeate may involve ultrafiltration and/or diafiltration.
Preferably, the colostrum serum isolate formed in accordance with the method may include at least 70% protein.
Preferably at least 20% of the protein may be immunoglobulins.
According to a further aspect of the invention there is provided a method of producing a serum deplete colostrum protein concentrate from colostrum, the method including the steps of:
taking a skim colostrum stream; microfiltration of the skim colostrum stream to produce a serum deplete retentate including an enhanced ratio of casein:whey protein; optionally concentrating the retentate; and drying the optionally concentrated retentate to produce a serum deplete colostrum protein concentrate.
According to a further aspect of the invention there is provided a method of producing a colostrum powder deplete in whey proteins from colostrum, the method including the steps of:
- taking a skim colostrum stream; microfiltration of the skim colostrum stream to produce an intermediate permeate rich in colostral serum proteins and a serum deplete retentate having an enhanced ratio of casein:whey protein; - concentration of said intermediate permeate by ultrafiltration and/or diafiltration to produce a further permeate; mixing the further permeate with the serum deplete retentate; optionally concentrating the mixed further permeate/retentate; and - drying the optionally concentrated mixture to produce a colostrum powder deplete in whey proteins.
In one preferred form of method of the invention the starting material may comprise colostrum from bovine immunised or hyperimmunised before collection of the colostrum.
According to a further aspect of the present invention there is provided a colostrum serum isolate including a protein content of at least 70.%.
Preferably the protein content of the colostrum serum isolate may include at least 20% immunoglobulins.
According to a further aspect of the invention there is provided a colostrum protein concentrate which is serum deplete and includes an enhanced ratio of casei whey protein.
Other aspects of the invention may become apparent from the following description which is given by way of example only and with reference to the accompanying examples.
Brief Description of the Figures
FIGURE 1 : Is a flow diagram showing the main steps of the process of the invention in one preferred form. Detailed Description of the Invention
In broad terms, the process of the invention enables the production of colostrum serum isolate (CSI) and serum deplete (SD) colostrum protein concentrate (CPC) by microfiltration, and those products may then be further processed to purify particular bioactive substances. The production of CSI and (SD) CPC from colostrum occurs in parallel as the (SD) CPC is formed from the serum deplete stream resulting from microfiltration. Colostrum powders are typically used in nutritional supplement, functional food and pharmaceutical applications where they confer a wide range of benefits. A powder formed from the (SD) CPC derived from the present invention is likely to have an elevated content of complex lipids. It has an elevated immunoglobulin content, reduced levels of serum protein and consequently an enhanced ratio of casein:whey protein. It has the potential to be used in functional food applications.
It will be appreciated by those skilled in the art that the immunoglobulin content of colostrum decreases progressively postpartum. To produce end products with a preferred higher immunoglobulin content, the starting material for the method of the invention is, or is derived from, colostrum obtained during the first 2 days postpartum.
CSI derived from the method of the present invention has application in, for example, cultured foods, sports applications, medical and dietetic applications, dairy desserts, dry mix beverages, ready to drink beverages and prepared foods. CSI offers an advantage in terms of the higher concentration of immunoglobulins and other small bioactive molecules. It also has excellent clarity when in solution as a result of the low fat content. The starting material for this process is a skim colostrum stream from colostrum. It will be appreciated that this may be fresh, dried or frozen. The colostrum undergoes a series of process conditions as seen in Figure 1 .
Colostrum is passed through a separator to remove the fat and other particulate and cellular material, resulting in a skim colostrum stream and a cream stream. It will be appreciated that the process may alternatively start with a spray dried skim colostrum powder which is then reconstituted to form the skim colostrum stream. The skim stream (or whole colostrum stream) may be pasteurised for microbial hygiene prior to microfiltration (MF). The MF process is carried out preferably using ceramic MF membranes with 0.05- 1 .4/vm cut-off, but preferably 0.1/vm. The temperature is maintained between 4 and 70°C, and preferably about 50°C. The pH of the MF feed is maintained between pH 4.6 and 8.5, and preferably at a natural pH in the range 6 to 7. Additives (e.g. filter aids) may or may not be added.
The retentate from MF may optionally be concentrated (by ultrafiltration/diafiltration) prior to, optionally, evaporating, and then drying, producing (SD) CPC. The permeate from MF may be further concentrated by processes such as ultrafiltration, nanofiltration, reverse osmosis or evaporation. Ultrafiltration is the preferred method and may be performed between 5 °C and 70 °C, preferably about 10 °C, using membranes with a molecular weight cut-off of between 1 kD and 100kD, but preferably about 10kD to increase the solids. Diafiltration (DF) may also be used. The concentrated MF permeate may then be evaporated (optional), and dried, resulting in CSI, which has a high concentration of immunoglobulins.
In order to produce a colostrum powder with reduced whey proteins the permeate from microfiltration (preferably after concentration by ultrafiltration/diafiltration), or a portion of it, may be mixed back with the serum deplete retentate. The mixture may then optionally be concentrated prior to evaporation and drying.
The method of the Invention will nbw be described by reference to a specific example.
Example 1
Spray dried skim colostrum (IMMULACt ) wee reconstituted with demineralised Water to 12% total BolidSi The pH of the solution was neutral, at about 6.7-6.8. The solution was then heated to 50°C prior to microfiltration. The pilot plant used was Tetra Pak Filtration System MFS1 9, fitted with an SCT (Societe des Ceramiques Techniques) membrane (0.1 μm ceramic membrane with 4mm channel spacers). The crossflow rate was 6-7 m/s. A Uniform Transmembrane Pressure (UTP) system was used to reduce fouling and to achieve optimal conditions over the entire memb ane area. This Involved the use of a high velocity permeate circulation cαhcϋi ehtly With the retentate inside the module, but outside the element. This gave uniform TMP over the entire area due to the space between elements on the permeate side being filled with plastic grains. The high velocity circulation of the permeate causes a pressure drop inside the channels. The pressure drop on the permeate side was regulated by the permeate pump and was constant during the operation of the plant.
The temperature was maintained at 50°C throughout the process. The permeate operating pressure difference was 1 .6 bar. The retentate operating pressure difference was 1 .9 bar. The retentate from the membranes was collected and spray dried. This fraction of the colostrum forms the (SD) CPC. The permeate from microfiltration was then ultrafiltered at 50°C using 5kD spiral wound membranes to total solids of 9.5%. 1 00% diafiltration water was added during the ultrafiltration. The ultrafiltered permeate was then spray dried, resulting in the CSI, which is high in immunoglobulins.
The compositional results for the (SD) CPC and CSI are shown in Table I, in comparison with skim colostrum powder, UF skim colostrum powder and milk protein concentrate (MPC) which comes from the ultrafiltration of skim milk.
TABLE I
Figure imgf000013_0001
[* calculated.]
Table I shows that the lactose content in CSI is much lower than that in skim milk colostrum and UF skim colostrum, and the lactose content of (SD) CPC is lower than skim colostrum, but higher than UF colostrum. The (SD) CPC is comparable to skim colostrum powder in composition. However the protein content is higher and would be enriched in casein. It is anticipated that the protein content will also include an elevated level of immunoglobulins in comparison with standard skim colostrum powder.
The CSI has a total protein composition more similar to that of a WPI, but with a low lactose and fat content. Moreover, the nature or profile of the protein content is different from that for a milk-derived WPI.
Analysis of the IgG content was performed using HPLC. This showed the IgG/ total protein content of the liquid CSI prior to UF to be 48.9% .
Reverse phase chromatography analysis of the CSI revealed the proportions of various protein components, shown in Table II (IgA content was not measured).
TABLE II
Figure imgf000014_0001
Higher levels of immunoglobulin present in the CSI and lower levels of lactose should help to reduce the problems previously associated with having to administer large quantities of pharmaceutical composition in order to achieve the desired levels of antibody administration. The protein yields achieved with the method of the invention, in this particular example, were:
CSI - 0.057 kg protein/kg protein in IMMULAC™ powder, (SD) CPC - 0.935 kg protein/kg protein in IMMULAC™ powder.
Thus, by the method of the invention a serum deplete protein concentrate and a protein isolate can be produced from colostrum by microfiltration without forming a whey intermediate. Purification of commercially useful quantities of bioactive materials, such as immunoglobulins, from the (SD) CPC and CSI is also possible.
Where in the foregoing description, reference has been made to specific components or integers of the invention having known equivalents then such equivalents are herein incorporated as if individually set forth.
Although this invention has been described by way of example and with reference to possible embodiments thereof, it is to be understood that modifications or improvements may be made thereto without departing from the scope or spirit of the invention.

Claims

A method of producing a protein concentrate and/or serum isolate from colostrum, the method including:
taking a skim colostrum stream; and microfiltration of the skim colostrum stream to produce a permeate rich in colostral serum proteins and a serum deplete retentate including an enhanced casein:whey protein ratio.
A method according to claim 1 further including preparing the skim colostrum stream by separation of fat and other particulate and cellular material from colostrum.
A method according to claim 1 further including preparing the skim colostrum stream by reconstituting skim colostrum powder in water.
A method according to any one of claims 1 to 3 further including ultrafiltration and/or diafiltration of the permeate to produce a colostrum serum isolate.
A method according to claim 4 further including drying the retentate to produce a serum deplete colostrum protein concentrate.
A method according to claim 5 wherein the retentate is concentrated by ultrafiltration and/or diafiltration prior to drying.
7. A method according to any one of claims 1 to 6 wherein the temperature of the skim colostrum is kept below the temperature which may cause denaturation of immunoglobulins.
8. A method according to claim 7 wherein the temperature of the skim colostrum is retained below 70°C.
9. A method according to claim 8 wherein the temperature is adjusted to substantially 50°C.
10. A method according to claim 8 wherein the temperature is adjusted to substantially 10°C.
1 1 . A method according to any one of claims 1 to 10 wherein the pH of the skim colostrum is in the range 4.6 to 8.5.
1 2. A method according to claim 1 1 wherein the pH of the skim colostrum is in the range substantially 6 to 7.
1 3. A method according to any one of claims 1 to 1 2 wherein the microfiltration involves membranes having a cut-off of 0.05 to 1 .4μm.
14. A method according to claim 1 3 wherein the membrane cut-off is substantially 0.1 μm.
1 5. A method according to any one of claims 1 to 1 4 wherein microfiltration involves a ceramic microfiltration membrane.
1 6. A method of producing a colostrum serum isolate from colostrum, the method including the steps of: taking a skim colostrum stream; microfiltration of the skim colostrum stream to separate out casein and lipids, and produce a permeate which is rich in colostral serum proteins; and concentration of the permeate to produce a colostral serum isolate.
1 7. A method according to claim 1 6 wherein concentration of the permeate involves ultrafiltration and/or diafiltration.
1 8. A method according to claim 1 7 wherein the colostral serum isolate includes at least 70% proteins.
1 9. A method according to claim 1 8 wherein at least 20% of the protein content is immunoglobulins.
20. A method of producing a serum deplete colostrum protein concentrate from colostrum, the method including the steps of:
taking a skim colostrum stream; microfiltration of the skim colostrum stream to produce a serum deplete retentate including an enhanced casein:whey protein ratio; optionally concentrating the retentate; and drying the optionally concentrated retentate to produce a serum deplete colostrum protein concentrate.
21 . A method according to claim 20 wherein the retentate is concentrated by ultrafiltration and/or diafiltration.
22. A method of producing a colostrum powder deplete in whey proteins from colostrum, the method including the steps of: taking a skim colostrum stream; microfiltration of the skim colostrum stream to produce an intermediate permeate rich in colostral serum proteins and a serum deplete retentate having an enhanced ratio of casein:whey protein; concentration of said intermediate permeate by ultrafiltration and/or diafiltration to produce a further permeate; mixing the further permeate with the serum deplete retentate; optionally concentrating the mixed further permeate/retentate; and drying the optionally concentrated mixture to produce a colostrum powder deplete in whey proteins.
23. A method according to any one of the preceding claims wherein the colostrum is derived from bovine immunised or hyperimmunised animals.
24. A colostrum serum isolate including a protein content of at least 70% .
25. A colostrum serum isolate according to claim 24 wherein at least 20% of the protein content is immunoglobulins.
26. A colostrum serum isolate produced by a method according to any one of claims 1 to 1 9.
27. A colostrum protein concentrate which is serum deplete and includes an enhanced ratio of casein:whey protein.
28. A bblostrϋm pi-bteln concentrate prbdlibed by a method according to any one of clai s 1 to 1 B br 2D oi- 21.
29. A colostrum pαwasr deplete in wney proteins produced doDcrdiHg to thy mbthϋd ϋf aHy bf claim 22.
30. A rtiethb ϋf prbdUαlH ά prøtølH bbrlcbhtrate h /ϋ serum Isoldte substantially ύβ hHtølH dysbH fed and with rbfererice to the dcbϋrh-bøHylHd. tiyure UH M axarHfjle,
31. A metnoα ot proαucmg a colostrum powαer αepiete in whey protein substantially as herein described and with reference to the accompanying figure;
32. A colostrum ββrum Isolate substantially as herein described and With reference iό the accompanying figure and/or example.
3d. A cdlofctrϋm protein concentra e SGtøitflHtliilrø es herein described aHd ith reference to the accompanying figure a;hd/dr example.
PCT/NZ2000/000120 1999-07-07 2000-07-07 Method of obtaining protein isolates and concentrates from colostrum WO2001003515A1 (en)

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