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WO2000049899A1 - Agents de revetement reticules a base de caseinate et de lactoserum - Google Patents

Agents de revetement reticules a base de caseinate et de lactoserum Download PDF

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Publication number
WO2000049899A1
WO2000049899A1 PCT/CA2000/000161 CA0000161W WO0049899A1 WO 2000049899 A1 WO2000049899 A1 WO 2000049899A1 CA 0000161 W CA0000161 W CA 0000161W WO 0049899 A1 WO0049899 A1 WO 0049899A1
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WO
WIPO (PCT)
Prior art keywords
films
caseinate
whey
protein
composition
Prior art date
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.)
Ceased
Application number
PCT/CA2000/000161
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English (en)
Inventor
Monique Lacroix
Mircea-Alexandru Mateescu
Geneviève DELMAS-PATTERSON
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Institut National de La Recherche Scientifique INRS
Universite du Quebec a Montreal
Original Assignee
Institut National de La Recherche Scientifique INRS
Universite du Quebec a Montreal
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Priority to AU26546/00A priority Critical patent/AU2654600A/en
Publication of WO2000049899A1 publication Critical patent/WO2000049899A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08L—COMPOSITIONS OF MACROMOLECULAR COMPOUNDS
    • C08L89/00—Compositions of proteins; Compositions of derivatives thereof
    • C08L89/005—Casein
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23B—PRESERVATION OF FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES; CHEMICAL RIPENING OF FRUIT OR VEGETABLES
    • A23B2/00—Preservation of foods or foodstuffs, in general
    • A23B2/70—Preservation of foods or foodstuffs, in general by treatment with chemicals
    • A23B2/704—Preservation of foods or foodstuffs, in general by treatment with chemicals in the form of gases, e.g. fumigation; Compositions or apparatus therefor
    • A23B2/708—Preservation of foods or foodstuffs, in general by treatment with chemicals in the form of gases, e.g. fumigation; Compositions or apparatus therefor in a controlled atmosphere, e.g. partial vacuum, comprising only CO2, N2, O2 or H2O
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23B—PRESERVATION OF FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES; CHEMICAL RIPENING OF FRUIT OR VEGETABLES
    • A23B4/00—Preservation of meat, sausages, fish or fish products
    • A23B4/10—Coating with a protective layer; Compositions or apparatus therefor
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23B—PRESERVATION OF FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES; CHEMICAL RIPENING OF FRUIT OR VEGETABLES
    • A23B4/00—Preservation of meat, sausages, fish or fish products
    • A23B4/14—Preserving with chemicals not covered by groups A23B4/02 or A23B4/12
    • A23B4/18—Preserving with chemicals not covered by groups A23B4/02 or A23B4/12 in the form of liquids or solids
    • A23B4/20—Organic compounds; Microorganisms; Enzymes
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23B—PRESERVATION OF FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES; CHEMICAL RIPENING OF FRUIT OR VEGETABLES
    • A23B5/00—Preservation of eggs or egg products
    • A23B5/06—Coating eggs with a protective layer; Compositions or apparatus therefor
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23B—PRESERVATION OF FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES; CHEMICAL RIPENING OF FRUIT OR VEGETABLES
    • A23B7/00—Preservation of fruit or vegetables; Chemical ripening of fruit or vegetables
    • A23B7/14—Preserving or ripening with chemicals not covered by group A23B7/08 or A23B7/10
    • A23B7/153—Preserving or ripening with chemicals not covered by group A23B7/08 or A23B7/10 in the form of liquids or solids
    • A23B7/154—Organic compounds; Microorganisms; Enzymes
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23B—PRESERVATION OF FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES; CHEMICAL RIPENING OF FRUIT OR VEGETABLES
    • A23B7/00—Preservation of fruit or vegetables; Chemical ripening of fruit or vegetables
    • A23B7/16—Coating with a protective layer; Compositions or apparatus therefor
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23B—PRESERVATION OF FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES; CHEMICAL RIPENING OF FRUIT OR VEGETABLES
    • A23B9/00—Preservation of edible seeds, e.g. cereals
    • A23B9/14—Coating with a protective layer; Compositions or apparatus therefor
    • 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
    • A23J3/00—Working-up of proteins for foodstuffs
    • A23J3/04—Animal proteins
    • A23J3/08—Dairy proteins
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23P—SHAPING OR WORKING OF FOODSTUFFS, NOT FULLY COVERED BY A SINGLE OTHER SUBCLASS
    • A23P20/00—Coating of foodstuffs; Coatings therefor; Making laminated, multi-layered, stuffed or hollow foodstuffs
    • A23P20/10—Coating with edible coatings, e.g. with oils or fats
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23P—SHAPING OR WORKING OF FOODSTUFFS, NOT FULLY COVERED BY A SINGLE OTHER SUBCLASS
    • A23P20/00—Coating of foodstuffs; Coatings therefor; Making laminated, multi-layered, stuffed or hollow foodstuffs
    • A23P20/20—Making of laminated, multi-layered, stuffed or hollow foodstuffs, e.g. by wrapping in preformed edible dough sheets or in edible food containers
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08H—DERIVATIVES OF NATURAL MACROMOLECULAR COMPOUNDS
    • C08H1/00—Macromolecular products derived from proteins
    • C—CHEMISTRY; METALLURGY
    • C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
    • C09D—COATING COMPOSITIONS, e.g. PAINTS, VARNISHES OR LACQUERS; FILLING PASTES; CHEMICAL PAINT OR INK REMOVERS; INKS; CORRECTING FLUIDS; WOODSTAINS; PASTES OR SOLIDS FOR COLOURING OR PRINTING; USE OF MATERIALS THEREFOR
    • C09D189/00—Coating compositions based on proteins; Coating compositions based on derivatives thereof
    • C09D189/005—Casein
    • 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

Definitions

  • This invention relates to water-insoluble protein-based covering agents, including coatings and films, methods of preparation and their use in the food industry.
  • Edible films and coatings are capable of offering solutions to these concerns by regulating the mass transfer of water, oxygen, carbon dioxide, lipid, flavor, and aroma movement in food systems (McHugh, T. H.& Krochta, J. M., Food Technology 1994, 48(1):97-103; and Chen, supra!995).
  • Edible films and coatings based on water-soluble proteins are typically water-soluble themselves and exhibit excellent oxygen, lipid and flavor barrier properties; however, they are poor moisture barriers.
  • Edible films and coatings are capable of solving the barrier problems of these and a variety of other food systems. See: Kester, et al, Food Technol., l 1986, 40:47-59; Mezgheni, et al, J. Ag and Food Chem., 1998 46:318-324; and Ressouany, et al, J. Ag and Food Chem., 1998, 46: 1618-1623.
  • Edible agricultural products such as fresh, frozen, whole or cut, fruits and vegetables, meat, fish, eggs, grains, nuts, and inedible agricultural products such as living plants, plant products, and ornamentals are subject to loss of quality over time from moisture loss, enhanced respiration and senescence, and browning and oxidative degradation. Other deterimental effects to agricultural products can result from microbial attack and moisture penetration.
  • the time these products are available in a fresh and attractive form can be extended, if respiration can be slowed down by limiting availability of oxygen or if the carbon dioxide level can be maintained at an optimum level.
  • Many edible products and plant materials have components which are vulnerable to oxidation, with resultant loss in quality, as oxygen diffuses into the tissue of the food or plant material. For example, fresh and frozen fish, frozen fruits and vegetables, nuts, and ornamentals have a limited shelf-life which is due to such oxidation.
  • the time these products are available in a quality form can be extended, if oxidation can be slowed down by limiting diffusion of oxygen into the product.
  • Browning of many food products is a major problem for the food industry. Until recently, both enzymatic and nonenzymatic browning in foods could be inhibited by application of sulfites. However, health concerns have limited their application (Sapers, Food Technology, 47, 75-84, 1993). Other techniques, including modified atmosphere packaging (MAP) and vacuum packaging have been considered. While this approach can delay browning, excessive reduction of oxygen will damage the product by inducing anaerobic metabolism, leading to breakdown and off-flavor formation. Furthermore, the removal of oxygen also entails a risk that conditions in the product might become favorable for the growth of Clostridium botulinum (Sapers, Food Technology, 47, 75-84, 1993).
  • MAP modified atmosphere packaging
  • vacuum packaging vacuum packaging
  • Edible films have been proposed for use on foods to control respiration, reduce oxidation, or limit moisture loss.
  • Coatings for edible products include wax emulsions (U.S. Pat. No. 2,560,820 to Recker and U.S. Pat. No. 2,703,760 to Cunning); coatings of natural materials including milk solids (U.S. Pat. No. 2,282,801 to Musher), lecithin (U.S. Pat. No.
  • Dried foods, low moisture baked products, intermediate moisture foods and high moisture foods all exhibit potential for improvement through the use of edible coatings and films.
  • Dried foods e.g., dried vegetables and dried meats
  • low moisture baked products e.g., crackers, cookies and cereals
  • Low moisture baked foods are also susceptible to moisture uptake from moist fillings and toppings. Such changes can result in loss of sensory acceptability of the food product, as well as a reduced shelf-life.
  • Many dried and baked products are also susceptible to oxidation, lipid migration and volatile flavor loss.
  • Substrates which are high in moisture content and have high moisture at the surface are particularly vulnerable to loss of quality due to moisture loss.
  • Examples are fruits and vegetables and other foods, and plant products which have exposed tissue surfaces created by peeling, cutting, etc. such as peeled and/or sliced apples, sliced tomatoes, peeled eggs, fish filets, and cut-stem flowers. Because their natural skins, peels, and shells, which normally act to retard moisture loss have been removed, these products lose their quality quickly.
  • High moisture food components also typically lose moisture to lower moisture components.
  • Oxidation and flavor loss are also problematic to high moisture food systems. The respiration rates of whole fruits and vegetables often dictate their shelf lives. Minimally processed fruits and vegetables are often subject to unacceptable levels of oxidative browning.
  • Water-insoluble edible films and coatings offer numerous advantages over water-soluble edible films and coatings for many food product applications. Increasing levels of covalent crosslinking in water-insoluble edible films and coatings result in better barriers to water, oxygen, carbon dioxide, lipids, flavors and aromas in food systems. Film mechanical properties are also improved. Many foods, such as fruits and vegetables, are exposed to water during shipping and handling. In these cases, water-insoluble films and coatings remain intact; whereas, water-soluble films and coatings dissolve and lose their barrier and mechanical properties. Edible films in the form of wraps, such as sandwich bags, also require water-insolubility.
  • Edible coatings based on waxes, polysaccharides and proteins have been developed in order to preserve food quality and freshness. Proteins act as a cohesive, structural matrix in multi- component systems to provide films and coatings having good mechanical properties. Plasticizer addition improves film mechanical properties. Such edible films could help to reduce food dehydration.
  • Edible films can be formulated as composite films of heterogeneous nature i.e. formed starting from a mixture of polysaccharides, proteins and/or lipids. This approach allows for the beneficial use of the functional characteristics of each film component.
  • the preparation of composite films imposes an emulsification of the lipidic material in an aqueous phase.
  • the preparation technique of hydrophobic films influences its barrier properties.
  • a film formed starting from a dispersed distribution of the hydrophobic material offers weak barrier properties to steam, compared to films with a continuous layer (Martin-Polo et al, 1992).
  • a dispersed distribution is due to the difference in polarity between the support (example: methyl cellulose) and the hydrophobic material (technique of emulsion).
  • Biodegradation is a process by which bacteria, moulds, yeasts and their enzymes consume a substance as a source of food so that the original form of this substance disappears (Klemchuk, P. 1990, Pol. Degra. Stab., 27, 183-202). Under appropriate conditions, a biodegradation process from two to three years is a reasonable period for the assimilation and the complete disappearance of the product (Klemchuk, supra).
  • Pseudomonas is recognized as being a type of bacteria which can synthesize a very diverse number of enzymes. Being psychrotrophic, it is responsible for the putrefaction of refrigerated foods.
  • the ratio of caseinate-to-whey ranges from 1 :99 to 99: 1, and is adjusted to meet the covering characteristics of the substrate.
  • Additives can be included in the film in order to further tailor the covering agent to its substrate. This covering agent can be applied to agricultural products, and foodstuffs.
  • this invention provides a composition for use as a covering agent, comprising: (i) a caseinate salt; (ii) whey protein; and optionally (iii) a plastisizing agent, and/or (iv) a polysaccharide, wherein said caseinate and whey molecules are cross-linked to form a covering agent.
  • this invention provides a process for the preparation of a covering agent, comprising the steps of: : (i) preparing an aqueous solution comprising a caseinate salt, whey protein and, optionally, a plastisizing agent and/or a polysaccharide; (ii) producing a substantially degassed solution by treating said aqueous solution to remove dissolved air; and (iii) subjecting said degassed aqueous solution to a chemical, or thermal step and/or an irradiation step, wherein said chemical, or heating and/or irradiation step causes crosslinking of caseinate and whey to produce said covering agent.
  • Figure 1 shows elution curves for calcium caseinate (alanate 380): a), native; b), heated at 90°C for 30 minutes; or c), irradiated at 32 kGy.
  • Figure 2 shows elution curves for commercial whey proteins (CWP): a), native; b), heated at 90°C for 30 minutes; or c), irradiated at 32 kGy.
  • CWP commercial whey proteins
  • Figure 3 presents elution curves for whey protein isolate (WPI) and calcium caseinate with ratio of 50- 50: a) control; b), heated at 90°C for 30 minutes; c), irradiated at 32 kGy; or d), combined heat and irradiation treatment.
  • WPI whey protein isolate
  • Figure 4 shows fraction of insoluble matter in function of the irradiation dose. Results are expressed as the percentage in solid yield after soaking the films 24 hours in water.
  • Figure 5 demonstrates the puncture strength of unirradiated and irradiated (32 kGy) whey protein isolate (WPI)- calcium caseinate films. Ratios express the proportion in WPI or calcium caseinate for a formulation based on 5% w/w total protein solution. For instance, the formulation 25-75 represents 1.25g WPI protein and 3.75g calcium caseinate protein per lOOg protein solution.
  • Fiugre 6 shows the puncture strength of unirradiated and irradiated (32 kGy) commercial whey protein-calcium caseinate films. Ratios express the proportion in CWP or calcium caseinate for a formulation based on 5% w/w total protein.
  • Figure 7 shows the viscoelasticity coefficient for unirradiated and irradiated (32 kGy) CWP- calcium caseinate films.
  • Figure 11 shows mold contamination (%) of coated/non-coated strawberries. Coating based on 5% w/w mixed proteins (whey and calcium caseinate) and 2.5% w/w glycerol.
  • Figure 16 shows the results of viscoelasticity (coefficient de viscoelasticite) on various caseinate-whey films, ranging from 1 :99 to 99:1 caseinate:whey and crosslinked using either heat or irradiation.
  • Figure 17 shows the results of puncture strength studies (force de rupture) on various caseinate-whey films, ranging from 1:99 to 99:1 caseinate:whey and crosslinked using either heat or irradiation.
  • Figure 18 shows the results of results of viscoelasticity (coefficient de viscoelasticite) on various caseinate-whey films, ranging from 1:99 to 99:1 caseinate:whey and crosslinked using heat
  • Figure 19 shows the results of studies (deformation a la rupture (mm) ) on various caseinate- whey films, ranging from 1 :99 to 99: 1 caseinate: whey and crosslinked using heat.
  • Figure 20 the results of puncture strength studies (force de rupture) on various caseinate-whey films, ranging from 1:99 to 99: 1 caseinate:whey and crosslinked using heat.
  • Figure 21 shows the results of a comparative study of various properties of films based on milk proteins.
  • Figure 22 presents results of a study demonstrating influence of storage time on the antioxidative properties of calcium caseinate films containing essential oils from rosemary and thyme.
  • Figure 23 presents results of a study demonstrating effects of incorporation of lecithin on the antioxidative properties of calcium caseinate films containing essential oils from rosemary and thyme.
  • Figure 24 presents results of a study demonstrating antioxidative activities of calcium caseinate films made with water and water/ethanol extracts of dry spices from rosemary and thyme.
  • Figure 25 presents results of a study demonstrating influence of physical cross-linking on the water vapor permeability of films made with calcium caseinate and whey protein isolate.
  • Figure 26 presents results of a study demonstrating the influence of physical cross-linking on the water vapor permeability of films made with calcium caseinate and whey protein concentrate.
  • This invention provides a caseinate-whey covering agent, wherein the ratio of caseinate:whey can be varied from 1:99 to 99:1 in order to optimize the characteristics of the covering agent and the final product to the requirements of the product to be covered.
  • the total concentration of protein in solution can be varied in order to meet the requirements of the substrate to be covered.
  • Other additives can be included to bestow further properties to the covering agent.
  • WPI whey protein isolate
  • WPC whey protein concentrate
  • WVP water vapor permeability
  • PG propylene glycol
  • TEG triethylene glycol
  • CMC carboxy methyl cellulose
  • BSA bovine serum albumin
  • coating refers to a thin film which surrounds the coated object. Coatings will not typically have the mechanical strength to exist as stand-alone films and are formed by applying a diluted component mixture to an object and evaporating excess solvent.
  • film refers to a stand-alone thin layer of material which is flexible and which can be used as a wrapping.
  • Films of the present invention are preferably formed from an emulsified mixture of two proteins, optionally in combination with a lipid and/or a plasticizer.
  • dissolved gases refers to any gases, including oxygen, nitrogen, and air which become entrapped in the emulsified fluid mixture prior to crosslinking.
  • disulfide formation refers to the formation of new — S--S-- bonds which can occur either intermolecularly or intramolecularly. These bonds can be formed in the proteins used in preparation of the films and coatings of the present invention by several routes. Disulfide formation can take place via thiol oxidation reactions wherein the free sulfhydryl groups of cysteine residues become oxidized and form disulfide bonds. Additionally, thiol-disulfide exchange reactions can take place wherein existing intramolecular disulfide bonds are broken by heat, chemical or enzymic means and allowed to form new disulfide bonds which are a mixture of the intermolecular and intramolecular variety.
  • lipid component refers to all oils, waxes, fatty acids, fatty alcohols, monoglycerides and triglycerides having long carbon chains of from 10 to 20 or more carbon atoms, which are either saturated or unsaturated.
  • lipid components are beeswax, paraffin, carnuba wax, stearic acid, palmitic acid and hexadecanol.
  • plasticizer refers to compounds which increase the flexibility of films and which have been approved for use in foods.
  • Preferred plasticizers are polyalcohols such as glycerol, sorbitol and polyethylene glycol.
  • protein refers to isolated proteins having either cysteine and/or cystine residues which are capable of undergoing thiol-disulfide interchange reactions and/or thiol oxidation reactions, or proteins having tyrosine residues which are capable of undergoing covalent crosslinkage to form bityrosine moieties.
  • Preferred proteins are those which are isolated from milk, the most preferred proteins being casein and whey.
  • Bovine casein is an abundant, economic and easily accessible protein. Casein alone roughly accounts for 80% of the total proteins in cow's milk (Schmidt and Morris, 1984). It can be isolated from skimmed milk either by acidification with mineral acid, or by acidification with mixed bacterial cultures (Nuillemard and Al, 1989). It is a phosphoprotein with amphiphilic characteristics which binds strongly to the Ca 2+ and Zn 2+ ions (Schmidt and Morris, 1984; Nuillemard et al., 1989). Due to their absorbent character, casein films do not produce an effective barrier to moisture. On the other hand, it can act as an emulsifying agent and create a stable casein-lipid emulsion (Avena-Bustillos and Krochta, 1993).
  • casein-based films can be improved by the polymerization of the protein with calcium (Ca +2 ) but also by adjusting the pH of the medium at the isoelectric point of casein.
  • the adjustment at the isoelectric point optimizes the protein-protein interactions, modifies the molecular configuration and would influence the mass transfer properties (Krochta, 1991 and Avena-Bustillos and Krochta, 1993).
  • Bovine casein is composed of four major proteinic complexes, named ⁇ sl an d2, ⁇ - and ⁇ -caseins.
  • a casein molecule consists of a primarily hydrophobic core of ⁇ and of ⁇ - casein and surrounded by ⁇ -casein on the surface (Schmidt and Morris, 1984).
  • the stability of micelles is ensured by the ⁇ -caseins and the calcium colloidal phosphates found on the periphery (Schmidt and Morris, 1984).
  • Casein contains many uniformly distributed proline residues. That gives it an open structure thus limiting the formation of alpha helixes and beta layers (Modler, 1985).
  • Caseinates are obtained either by the acidification with mineral acid (HC1 or H 2 S0 4 ), or by the acidification by mixed cultures made up o ⁇ Streptococcus subspecies lactis and/or cremoris, at the isoelectric point of casein (pH of 4,6).
  • the neutralization of the insoluble precipitates of casein or lactic acids by alkalis allows for the dissolution in salts of sodium of calcium, potassium, magnesium, or ammonium (Schmidt and Morris, 1984; Nuillemard et al, 1989; McHugh and Krochta, 1994).
  • the solubilized caseinates are dehydrated thereafter. Salts of caseins thus obtained are soluble above pH 5.5.
  • Whey mainly ⁇ -lactalbumine and ⁇ -lactoglobuline
  • Whey small-milk proteins
  • Whey mainly ⁇ -lactalbumine and ⁇ -lactoglobuline
  • Whey small-milk proteins
  • thermoirreversible gel which is pH-dependent and heat sensitive
  • heating of whey proteins at temperatures between 70 and 85° C and to a concentration higher than 5% forms a thermoirreversible gel.
  • This gel develops by the formation of new intermolecular disulphide bonds (Nuillemard and Al, 1989).
  • the gelling process of whey proteins is strongly influenced by the pH of the medium during heating since a pH > 6.5 decreases the intermolecular interactions (Schmidt and Morris, 1984; Xiong, 1992). High ionic forces seem to increase proteinic stability probably through an increase of the proteins' capacity of hydration (solubility) (Xiong, 1992).
  • WPI whey protein isolate
  • WPC whey protein concentrate
  • This covering agent differs from those known in the art, primarily because it is comprised of two proteins, caseinate and whey, whose ratio is determined to generate a covering agent with characteristics that are optimal for the product to protect.
  • the ratio of caseinate-to-whey is choosen to optimise the mechanical characteristics of the covering agent in accordance with the requirements for the food product it is intended to protect.
  • a 99: 1 caseinate-whey covering agent can be either a coating or a film, depending upon how the protein is crosslinked.
  • the source of the whey has a large impact on the characteristics of the covering agent: commercially obtained whey tends to be more denatured, versus whey produced in a laboratory by microfiltration which tends to form coverings with greater puncture strength.
  • Example IN presents studies demonstrating the effect of varying the ratio of caseinate to whey.
  • a food grade plasticizer is added to the denatured protein solution.
  • the food grade plasticizer serves to increase both the mechanical strength of the film and its flexibility.
  • the plasticizer is preferably a polyalcohol, for example, sorbitol, glycerol, triethylene glycol or polyethylene glycol.
  • the amount of food grade plasticizer which is added will typically be about 1 to 15 % by weight in solution, preferably about 2 to 10 % by weight in solution. In other embodiments, it may be desirable to include agents such as emulsifiers, lubricants, binders, or de-foaming agents to influence the spreading characteristics of the coating agent.
  • the viscoelasticity and puncture strength of films and coatings can be measured to determine the mechanical properties, which can also be correlated with transmission electron microscopy observations.
  • the mechanical strength of protein solutions can be increased by the formation of cross-links which confer elastomeric properties to the material as well as improve the water resistance of such protein films (Brault et al., J. Agric. Food Chem., 24(8), 2964-2969, 1997). Size-exclusion chromatography can be performed on cross-linked solutions to determine the molecular weight distribution of the cross-linked proteins.
  • Film thickness can be measured using commercially available instrumentation such as a Mitutoyo Digimatic Indicator (Tokyo, Japan) by measuring random positions around a film. For example, measuring six random positions around a sample film, should provide a film with a thickness in the range of 45 - 60 ⁇ m.
  • Molecular weight determination of the cross-linked proteins can be determined using size- exclusion chromatography.
  • size-exclusion chromatography is performed on a soluble protein fraction using a Varian Nista 5500 HPLC coupled with a Narian Auto Sampler model 9090, with detection of the protein solution performed using a standard UN detector set at 280 nm.
  • a Supelco Progel TSK GMPW column followed by two Waters Hydrogel columns (2000 and 500) is used for the molecular weight determination of the cross-linked proteins, wherein the total molecular weight exclusion limit is 25 x 10 6 daltons based on linear polyethylene glycol (PEG).
  • the eluant (80% v/v aqueous and 20% v/v acetonitrile) is flushed through the columns at a flow rate of 0.8 mL per minute.
  • the molecular weight calibration curve is established using a series of protein molecular weight markers (Sigma, MW-GF-1000, USA) ranging from 2 x 10 6 daltons to 29 000 daltons. All soluble protein solutions ( 0.5 % w/v ) are filtered on 0.45 ⁇ m prior to injection.
  • Insolubility measurements can be performed as in the following example, wherein the average dry weight of the films is determined on seven films by drying them in an oven at 45 °C until constant weight was achieved (6 or 7 days). Seven more films are dropped in 100 mL of boiling water for 30 minutes. The flasks are removed from the heat and the films remain in the water for another 24 hours. After 24 hours, the solid films are removed and dried in the oven as previously described. Results are calculated using the following formula: [ Dry Weight (solid residues)/ Dry Weight (untreated film)] x 100
  • the puncture strength of a film can be determined by measuring the 'breaking load' and 'strain at failure' which are calculated simultaneously for the samples, by recording the application of pressure to a film, which is then converted into units of force (N). Puncture tests can be carried out using a Stevens LFRA Texture Analyzer Model TA/1000 (NY, USA), as described previously by Gontard et al. ( J. Food Sci., 57 (1), 190-195, 1992). In this example, films are equilibrated for 48 hours in a dessicator containing a saturated NaBr solution ensuring 56% relative humidity.
  • a cylindrical probe ( 0.2 cm diameter ) is moved perpendicularly at the film surface at a constant speed (1 mm/sec) until it passes through the film.
  • Strength and deformation values at the puncture point are used to determine hardness and deformation capacity of the film. In order to avoid any thickness variation, the puncture strength values are divided by the thickness of the film. The force-deformation curves are recorded.
  • the viscoelasticity of a film can be measured by the relaxation curve obtained following the application of a force to the film.
  • An important characteristic sought in film products is elasticity, hence a film having a low relaxation coefficient is preferable.
  • Niscoelastic properties can be evaluated using relaxation curves. The same procedure as the used for the puncture test can be used, but the probe is stopped and maintained at 3 mm deformation.
  • the parameter 7(1 min) (F°-F')/F° where F° and F 1 were forces record initially and after 1 min of relaxation, respectively [Peleg, M., J. Food Sci. 1979, 44 (1), 277].
  • a low relaxation coefficient (Y ⁇ O) indicates high film elasticity whereas a high coefficient (7-> 1) indicates high film viscosity.
  • Heats of Wetting can be determined by obtaining isothermic measures using disposable glass ampules in a calorimetre SetaramTM C80.
  • a known amount of the sample which is dessicated for a minimum of 24 hours, is placed into a vacuum sealed ampule and then placed into a water filled cell equipped with TeflonTM joints, to prevent water evaporation.
  • the ampule with the cell is placed into the calorimetre and when the thermic equilibrium is obtained, the ampule is broken. Water from the cell enters the ampule due to the negative pressure and reacts with the sample.
  • the registered measurement is then converted into joules per gram giving the heat of wetting of the samples measured.
  • TEM Transmission electron microscopy
  • dry films are first immersed in a solution of 2.5% glutaraldehyde in cadodylate buffer, washed and postfixed in 1.3% osmium tetroxide in collidine buffer. Samples are then dehydrated in acetone (25, 50, 75, 95 and 100%) before embedding in a SPURRTM resin. Polymerization of the resin proceeds at 60 °C for 24 hours. Sections are made with an ultramicrotome (LKB 2128 UltrotomeTM) using a diamond knife and transferred on Formvar- carbon coated grids. Sections are stained 20 minutes with uranyl acetate (5% in 50% ethanol) and 5 minutes with lead citrate. Grids are observed with an HitachiTM 7100 transmission electron microscope operated at an accelerating voltage of 75 keN.
  • Water vapor permeability can be determined in a manner similar to U.S. Patent 5543164. Briefly, test cups were made out of Plexiglas such that the bottom of the cup had an outside diameter of 8.2 cm., the area of the cup mouth was 78.5 cm 2 , and the well inside the cup had a depth of 1.2 cm. Silicon sealant (High Vacuum Grease, Dow Corning, Midland, Mich., U.S.A.) and four screws, symmetrically located around the cup circumference, were used to seal films into test cups. Desiccator cabinets were purchased from Fisher Scientific, Inc. (Fair Lawn, N.J., U.S.A.) and variable speed motors with attached fans were installed. These cabinets were placed in a 24° C controlled temperature room.
  • Air speeds were measured using a Solomat anemometer (Stamford, Conn., U.S.A.). Fan speeds were set to achieve air speeds of 500 ft/min in the cabinets.
  • Each cabinet contained an Airguide hygrometer (Chicago, 111., U.S.A.) to monitor the relative humidity conditions within the cabinets. Prior to each experiment, cabinets were equilibrated to 0% relative humidity (RH) using calcium sulfate Drierite desiccant (Fisher Scientific, Inc., Fair Lawn, N.J., U.S.A.). Six milliliters of distilled water or equivalent amounts of saturated salt solutions were placed in the bottoms of the test cups to expose the film to a high percentage relative humidity inside the test cups.
  • Oxygen permeability can be determined for caseinate-whey coverings on a commercial unit such as a MOCON OXTRAN 2-20 (Minneapolis, Minn., U.S.A.). This system provides the flexibility of testing films under a variety of relative humidity and temperature conditions.
  • Tests to determine the antioxidant properties of the covering agent can be performed to optimize this criteria depending on the requirements of the foodstuff.
  • Evaluation of the antioxidative properties of a film or coating can be measured using a model allowing the release of oxidative species by electrolysis of saline buffer. In this method, measurements can be performed following a modified procedure of the DPD (N,N-diethyl-/7-phenylenediamine) colorimetric method reported by Dumoulin et al, (Arzneim-Forsch/Drug Res., 46, 855-861, 1996). Films can be cut in pieces of equal thickness all measuring 0.8 x 2.5 cm.
  • Tests to determine whether the coating delays enzymatic browning can be performed to optimize this criteria for certain foodsuffs.
  • Color measurements can be taken to demonstrate whether a coating efficiently delays enzymatic browning by acting as oxygen barrier. In one example, can be taken every thirty minutes for a total experimental period of five hours. The color can be read using a ColormetTM sperctrocolorimeter (Instrumar Engineering Ltd., St. John's, NF., Canada) using the standard (1976) CIELABTM color system. Lightness is reported as L* and the HUE angle value is given by tan "1 (b*/a*). As the HUE angle decreases, red pigmentation increases. The a* axis (red) corresponds to a HUE angle of 0°. Color measurements can be taken once on each slice of fruit or vegetable, for example, for between 8 and 12 readings per data point.
  • Additives including chelating agents, such as ascorbic acid and calcium disodium EDTA, antibacterial agents, flavorings, vitamins and minerals, etc can be included in the coating agent to optimize the characteristics of the covering.
  • a lipid or edible oil component can be incorporated into the covering agent
  • the lipid component can be a fatty acid, a fatty alcohol, a wax, a triglyceride, a monoglyceride or any combination thereof.
  • fatty acids which are useful in the present invention are stearic acid, palmitic acid, myristic acid and lauric acid.
  • fatty alcohols which can be used in the present invention are stearyl alcohol and hexadecanol.
  • Waxes which are useful in the present invention include beeswax, carnuba wax, microcrystalline wax and paraffin wax.
  • the lipid component will typically be present in an amount of from 1 to 30% by weight in solution, preferably about 2 to 15% by weight in solution.
  • the method of removal will typically involve subjecting the solution to reduced pressures by means of a vacuum pump or water aspirator.
  • the first step is the formation of an aqueous denatured protein solution.
  • the protein Prior to denaturation, the protein will typically be solubilized in an aqueous solution in a concentration range of from ? to ?% by weight, preferably about 5% by weight.
  • crosslinking of a proteinic solution can be achieved by a variety of methods including irradiation, heat, chemical and enzymic.
  • the preferred crosslinking treatments of the present invention being irradiation and heating, resulting in inter- and intra-molecular linkages including bityrosine residues and thiol-disulfide bridges.
  • hydroxyl radicals Upon radiolysis of an aqueous protein solution, hydroxyl radicals are generated. Aromatic amino acids react readily with these hydroxyl radicals. For example, tyrosine amino acids react with hydroxyl radicals to produce tyrosyl radicals. These may then react with other tyrosyl radicals or with tyrosine molecules to form stable biphenolic compounds, in which the phenolic moieties are linked through a covalent bond. The 2',2-biphenol bityrosine moiety exhibits a characteristic fluorescence, which provides a means of monitoring the formation of such crosslinks. The formation of bityrosine is one mechanism for causing protein aggregation, although other crosslinks can be formed.
  • the gamma irradiation treatment presents a number of conveniences, including the production of sterile goods.
  • the aqueous protein solution is heated to a temperature above the denaturation temperature of the particular protein for a period of time sufficient to initiate crosslinkage reactions, which are predominantly disulfide bridges.
  • These thiol-disulfide interchange and thiol oxidation reactions can be either intramolecular or intermolecular.
  • the precise temperature and length of time for a given protein can be determined empirically, but will typically involve temperatures of from about 70 to 95°c, preferably from about 75 to 85°c and a length of time of up to 3 hours, preferably from about 15 to 45 minutes.
  • the result of this reaction is a solution of a denatured protein having a mixture of intermolecular and intramolecular disulfide crosslinks.
  • the denatured protein solution may applied to a food item and water is evaporated to form a coating for the food item.
  • the method of application is not critical and will depend upon the particular food item. Suitable application methods include dipping, brushing and spraying. Similarly, the method of evaporation is not critical. Water can be removed by standing in air at ambient temperature. Alternatively, water can also be removed by gently warming the coated food item and exposing it to a stream of air or other suitable gas such as nitrogen.
  • dissolved gases are removed from the aqueous protein solution prior to denaturing the protein.
  • the removal of dissolved gases prevents formation of air bubbles in the films and increases both the mechanical strength of the film and the ability of the film to control mass transfer in foods.
  • the method selected for removal of dissolved gases is not critical, however, a preferred method involves subjecting the solution to reduced pressures by means of a vacuum pump or water aspirator.
  • the present invention also provides foodstuffs and packagings coated with the coating agents of the instant invention.
  • the following examples are provided by way of illustration and not by way of limitation.
  • the following examples describe the effect of combined physical treatments (heat and irradiation) on the mechanical and structural properties of milk protein-based covering agents.
  • the effects of gamma-irradiation and thermal treatment of caseinate and whey proteins solutions has been studied using size-exclusion chromatography.
  • the puncture strength and the viscoelastic properties of film formulations containing different protein ratios has been correlated with transmission electron microscopy observations.
  • This example demonstrates the mechanical properties of cross-linked edible films based on calcium caseinate and two type of whey proteins (commercial and isolate).
  • the present study focuses on the effect of combined physical treatments (heat and irradiation) on the mechanical and structural properties of milk protein-based edible films).
  • Cross-linking of the proteins was carried out using thermal and radiative treatments.
  • the effects of gamma-irradiation and thermal treatment of calcium caseinate and whey protein solutions was studied using size- exclusion chromatography.
  • the puncture strength and the viscoelastic properties of film formulations containing different protein ratios was correlated with transmission electron microscopy observations.
  • Whey protein isolate (WPI, 90.57% w/w protein) was obtained from the Food Research Center of Agriculture and Agri-food Canada and the commercial whey protein concentrate (Sapro-75, 76.27% w/w protein) was purchased from Saputo cheeses Ltd (Montreal, Quebec, Canada). Whey protein isolate was produced from permeate obtained by tangential membrane microfiltration. Fresh skim milk was microfiltered three-fold at 50 °C using an MF pilot cross-flow unit as described previously by St-Gelais et al. (1995).
  • the proteins contained in the permeate were concentrated twenty-five-fold at 50 °C by ultrafiltration using a UF pilot unit equipped with a Romicon membrane (PM 10, total surface area 1.3 m 2 ).
  • the concentrate was diafiltered five-fold by constant addition of water and freeze-dried before use in order to obtain WPI.
  • Carboxymethyl cellulose sodium salt (CMC, low viscosity) was obtained from Sigma Chemicals (St.Louis, MO, USA).
  • Glycerol 99.5%, reagent grade
  • Acetronitrile (99.95%) was obtained from Anachemia Chemicals (Montreal, Quebec, Canada). All products were used as received without further purification.
  • Film thickness was measured using a Mitutoyo Digimatic Indicator (Tokyo, Japan) at six random positions around the film. Depending on the formulation and irradiation dose, the average film thickness was in the range of (45-60) ⁇ 2 ⁇ m.
  • Size-exclusion chromatography was performed on the soluble protein fraction using a Varian Vista 5500 HPLC coupled with a Varian Auto Sampler model 9090. Proteins were determined using a standard UV detector set at 280 nm. Two Supelco Progel TSK PWH and GMPW columns followed by two Waters Hydrogel columns (2000 and 500) were used for the molecular weight determination of the cross-linked proteins. The total molecular weight exclusion limit was 25 x 10 6 daltons based on linear polyethylene glycol (PEG). The eluant (80%) v/v aqueous and 20% v/v acetonitrile) was flushed through the columns at a flow rate of 0.8 mL per minute.
  • PEG linear polyethylene glycol
  • the molecular weight calibration curve was established using a set of protein molecular weight markers MW-GF-1000 (Sigma) ranging from 2 x 10° daltons to 29 000 daltons. All soluble protein solutions (0.5 % w/v ) were filtered on 0.45 ⁇ m nylon membrane filters (VWR, Nalge, Mississauga, Ontario, Canada) prior to injection.
  • Puncture tests were carried out using a Stevens LFRA Texture Analyzer Model TA/1000 (NY, USA), as described previously by Gontard et al. (1992). Films were equilibrated for 48 hours in a dessicator containing a saturated NaBr solution ensuring 56% relative humidity. A cylindrical probe ( 0.2 cm diameter) was moved perpendicularly to the film surface at a constant speed (1 mm/sec) until it passed through the film. Strength and deformation values at the puncture point were used to determine hardness and deformation capacity of the film. In order to avoid any thickness variation, the puncture strength values were divided by the thickness of the film. The force-deformation curves were recorded. Viscoelastic properties were evaluated using relaxation curves. The same procedure was used, but the probe was stopped and maintained at 3 mm deformation. The parameter Y was calculated using the equation:
  • Dry films were first immersed in a solution of 2.5% glutaraldehyde in cacodylate buffer, washed and postfixed in 1.3% osmium tetroxide in collidine buffer. Samples were then dehydrated in acetone (25, 50, 75, 95 and 100%) before embedding in a SPURR resin. Polymerization of the resin proceeded at 60 °C for 24 hours. Sections were made with an ultramicrotome (LKB 2128 Ultratome) using a diamond knife and transferred on Formvar- carbon coated grids. Sections were stained 20 minutes with uranyl acetate (5% in 50% ethanol) and 5 minutes with lead citrate. Grids were observed with an Hitachi 7100 transmission electron microscope operated at an accelerating voltage of 75 keV.
  • LLB 2128 Ultratome ultramicrotome
  • Figure 1 shows the elution curves obtained for native, heated or irradiated calcium caseinate. Heating calcium caseinate at 90°C for 30 minutes increased the molecular weight 3 to 4-fold ( Figure 1, b). However, when the protein was submitted to gamma-irradiation at a dose of 32 kGy, cross-linking occurred and the molecular weight distribution peak shifted to higher molecular weights. Based on the protein calibration curve, the molecular weight distribution of the cross-linked soluble calcium caseinate fraction was > 2 x 10 6 daltons, an increase greater than 60-fold (Figure 1, c). Previous studies demonstrated that gamma-irradiation induced the formation of bityrosine (Davies, J. A., I. General aspects. J.
  • Figure 3 shows the molecular mass changes in the case of a 50%-50% mixture of whey protein isolate and caseinate before ( Figure 3, a) or after heating ( Figure 3, b), or irradiated ( Figure 3, c), or heating at first then treated with irradiation ( Figure 3, d). About 40% of the protein was cross-linked ( > 10 x 10 daltons) in the combined heating and irradiation treatment ( Figure 3, d).
  • Figure 4 shows the results obtained for calcium caseinate films irradiated at different doses.
  • the proportion of the insoluble fraction increases with the irradiation dose up to 32 kGy ⁇ when 70% of the film remained insoluble after 24 hours.
  • These results are supported by the size exclusion chromatography results ( Figure 1, 2 and 3) which suggest that a maximum cross-linking density was obtained at about 32 kGy.
  • the size-exclusion chromatography results combined with the solubility measurements indicate that the irradiation of calcium caseinate led to the formation of an insoluble fraction of high molecular weight which accounts for 70%) of the dry matter and a soluble protein fraction of molecular weight > 2 x 10 6 . Ressouany et al.
  • the present research shows that gamma-irradiation, which induces the cross-linking of tyrosine residues in a manner similar to peroxidase (Matheis and Whitaker, J. Food Biochem. 1987, 11, 309-327), is a method specific enough for the development of edible films, and particularly cost-efficient when used on a large-scale basis.
  • protein cross-linking by ⁇ -irradiation increased water-resistance, and it has been demonstrated that tyrosine-tyrosine cross-links improved the mechanical resistance of these films (Mezgheni et al., 1998; Ressouany et al., 1998).
  • a dose of 32 kGy was chosen in order to evaluate the effect of ⁇ -irradiation on the mechanical properties of edible films based on calcium caseinate and whey proteins.
  • Figure 5 shows the puncture strength variations of films cast from solutions containing different whey protein isolate-calcium caseinate ratios (5% w/w total protein solution). For instance, a protein ratio of 50-50 corresponds to 2.5% WPI protein and 2.5% calcium caseinate protein. Addition of WPI in the formulations did not significantly affect the puncture strength of the films up to a WPI-calcium caseinate ratio of 50-50. At higher WPI concentrations, the puncture strength of the films was significantly reduced (p ⁇ 0.05) and reached a minimal value of 0.04 N/ ⁇ m for the films based on WPI only. Gamma-irradiation significantly increased (p ⁇ 0.05) the mechanical properties of the films by inducing cross-links between protein chains.
  • Figure 7 shows the viscoelasticity coefficient of films irradiated or unirradiated.
  • a low viscoelasticity coefficient means that the material is highly elastic while a high coefficient indicates that the material is more viscous and easily distorted.
  • ⁇ -irradiation decreases the viscoelasticity coefficient of caseinate films resulting in a more elastic material.
  • An addition of whey proteins (CWP) by 25% of total total protein did not change the viscoelasticity coefficient (p ⁇ 0.05). No statistical differences (p > 0.05) were found between films unirradiated or irradiated. However, the decrease from the 0-100 to the 50-50 formulations was found to be statistically significant (p ⁇ 0.05).
  • FIG. 8 shows the micrographs that were obtained for cross-sections of films made from calcium caseinate. The micrographs show that the structure of these films is highly porous. Similar observations were made by Frinault et al. (J. Food Sci. 1997, 62 (4), 744-747) on casein films prepared by a modified wet spinning process. However, the microstructure of the films that were cast from irradiated solutions ( Figure 8, b) is clearly more dense than the films cast from unirradiated solutions ( Figure 8, a). Cross-links, which are present in the irradiated films, increase the molecular proximity of the protein chains.
  • the films made of CWP only (100-0) have a granular structure and contain numerous dense masses that may be attributed to impurities such as fat, lactose and mineral salts. Addition of calcium caseinate to the formulations rendered their microstructure smoother and slicker.
  • major differences are seen between the micrographs of films 50-50 ( Figure 9, a) and 75-25 ( Figure 9, b) in terms of pore size.
  • the pores are obviously much larger in the case of the films cast from a solution containing a protein ratio of 75-25.
  • the variations in pore size distribution of these films might be correlated in part, with the variations in puncture strength.
  • the internal structure might be indicative of the protein phase behavior.
  • This example compares the effectiveness of edible whey: caseinate coverings (based on 5 % w/w protein and 2.5% w/w glycerol) to ⁇ -irradiation treatment to reduce water loss and mold growth in fruit
  • the results demonstrate that both treatments are effective in reducing water loss and mold growth, and that whey: caseinate coatings are more effective than those based on calcium caseinate alone.
  • ⁇ -irradiation was used in combination with edible coatings for possible synergistic effects between the two treatments.
  • this example also demonstrates that the addition of calcium chloride or polysaccharides to the protein formulations increases their effectiveness by further delaying mold growth.
  • Strawberries were choosen as an exemplary fruit because strawberry decay resulting from mold growth is a common problem during fruit storage. Rot caused by Rhizopus sp. and Aspergillus sp. are mainly accountable for fruit loss. Because strawberries are especially sensitive to mold growth, its shelf life is of 2 days when stored at 15 °C. In order to control fruit decay and losses, many studies have been done in order to develop new preservation methods. Among those tested, gamma-irradiation has proven effective in reducing bacterial and mold contamination as well as delaying the ripening of climacteric fruits (Kader, A. A., Food Technology, 6, 117-121).
  • Baccaunaud and Chapon (INFOS-Centre technique interprof essionnel des fruits et legumes, 9, 43-54, 1985) have shown that modified atmosphere packaging (MAP) followed by ⁇ -irradiation at 2 kGy extended the shelf life of strawberries to over a month when stored at 4 °C, as compared to 14 days for heat-treated fruits (40 °C for 10 minutes) combined with irradiation (2 kGy).
  • MAP modified atmosphere packaging
  • ⁇ -irradiation at 2 kGy extended the shelf life of strawberries to over a month when stored at 4 °C, as compared to 14 days for heat-treated fruits (40 °C for 10 minutes) combined with irradiation (2 kGy).
  • Coating formulations were based on 5 % w/w protein and 2.5% w/w glycerol.
  • Calcium caseinate New Zealand Milk Products, Santa Rosa, CA, USA
  • CaCl 2 (0.125% w/w) (BDH Chemicals Ltd., Montreal, Quebec, Canada) or a mixture of polysaccharides (0.1% agar and 0.1% pure pectin) were also added to the coating formulations.
  • the agar was purchased from Sigma Chemicals (St. Louis, MO, USA) and commercial liquid pectin (Certo brand) was obtained from Kraft Canada inc. (Cobourg, Ontario, Canada).
  • Weight loss determination was calculated on each strawberry case. Each case was weighed and the ratio of the final weight on the initial weight was determined. The number of fruits rejected due to mold growth was determined each day of analysis.
  • Figure 11 shows the results obtained for irradiated coating formulations based on a mixture of calcium caseinate and whey proteins. It can be seen that 90% of fruit contamination was obtained on day 20 for the mixed-proteins coated fruits while a similar number was reached on day 17 for the pure calcium caseinate formulation (control + film 32 kGy) ( Figure 2). The addition of whey proteins in the formulation delayed mold apparition by another 3 days. Similarly, CaCl 2 or polysaccharides (agar and pectin) were added to the mixed-proteins formulation. It can be seen that the addition of salt or polyssacharides improved the coating formulations' efficiency by further reducing mold growth on strawberries.
  • Calcium caseinate (alanate 380) was provided by New Zealand Milk Products (Santa Rosa, CN USA). Concentrated whey protein powder was obtained from Les Fromages Saputo Ltee. (St-Hyacinthe, Quebec, Canada). Glycerol (99,5%, reagent grade) was purchased from American Chemicals ltd (Montreal, Quebec, Canada), carboxymethyl cellulose sodium salt (CMC, low viscosity), and calcium chloride (CaCl 2 , laboratory reagent) was obtained from BDH Chemicals (Montreal, Quebec, Canada). All products were used as received without further purification.
  • a dipping solution formation was prepared to generate 5 % (w/w) protein (calcium caseinate or whey protein powder in a 50:50 ratio), 2,5% (w/w) glycerol, 0,25% (w/w) CMC and 0,125% (w/w) CaCl 2 were diluted in water and mixed to obtain homogeneous solutions.
  • Mclntosh apples Quebec, Canada
  • washed potatoes Canada #1 product from Prince Edward Island, prepared by Emballages D.L. Inc, Laval, Qc, Canada
  • Five slices were cut from three potatoes and apples, dipped one minute in the protein solutions and laid in petri dishes. Control potatoes and apples were cut and laid without dipping in the dishes an exposed to atmospheric air. The experiment was repeated three times.
  • Color measurements were taken every five minutes for a total experimental period of 130 minutes. The color was read using a Colormet spectrocolorimeter (Instrumar Engineering Ltd, St. John's, ⁇ F, Canada) using the standard (1976) CIELAB color system. Lightness is reported as L* and the HUE angle value is given by tan "1 (b*/a*). As the HUE angle decreases, red pigmentation increases. The a* axis (red) corresponds the a HUE angle of 0°. Color measurements were taken once on each slice (potato or apple) for a total of 15 readings per data.
  • Figure 12 shows the variation of the lightness parameter (L*) in function of time for coated potato slices.
  • L* the lightness parameter
  • For the uncoated control slices, an increase in lightness is noted for the first fifteen minutes. This feature is probably due to the exudation of natural juices that contribute to increase the surface's luminosity. Then, as enzymatic browning occurs, the brightness of the uncoated potato slices starts to progressively decrease with time for the remaining experimental period. The lightness value for perfect white is 100 while L* 0 corresponds to black. The loss of whiteness associated with enzymatic browning can be estimated for the entire experimental period. Contrary to the uncoated potato slices, the coated potato slices did not show any evidence of darkening. A slight increase in lightness was even noticed for both types of coated potato slices.
  • Figure 13 shows the HUE angle variation for uncoated and coated potato slices.
  • red pigmentation becomes more pronounced.
  • the control (uncoated) slices undergo rapid enzymatic browning as seen by the sharp decrease of the HUE. The sharpest decrease was noted within the first 45 minutes. Following that decrease, the HUE stabilized for the remainder of the experimental period.
  • the coated slices only a slight variation of the HUE was noted for the entire experimental time period although those small color changes are not coupled with a darkening of the potato slices ( Figure 7).
  • Figures 14 and 15 show the lightness (L*) and HUE angle results obtained for apple slices. Similarly to what was observed for potato slices, L* rapidly decreased with time for the uncoated apple slices.. For the coated apple slices, the lightness parameter remained rather constant showing that the protein coatings effectively protected the fruit from oxygen. As for the HUE (Fgure 15), results show that for all types of apple slices, the angle decreased slightly with time. That effect seems to be somewhat less noticeable in the case of the whey coating. As the HUE decreases, red pigmentation develops. However, previous results ( Figure 14) show that those small color fluctuations are not associated with darkening (lower L*).
  • cysteine a sulfhydryl-containing amino acid was used as a polyphenol oxidase inhibitor by acting as a coupling agent with quinones forming stable, colorless compounds (Dudley and Hotchkiss, Journal of Food Biochemistry, 13, 65-75, 1989).
  • Calcium caseinate (Alanate 380, 91.8% w/w protein) was provided by New Zealand Milk Product Inc. (Santa Rosa, CN USA).
  • Commercial whey protein concentrate (Sapro-75, 76.27% w/w protein) was purchased from Saputo Cheeses Ltd (Montreal, Quebec, Canada).
  • Whey protein isolate (WPI, 92.52%) w/w protein) was obtained from the Food Research Center of Agriculture Canada, wherein it was produced from permeate obtained by tangential membrane microfiltration.
  • Fresh skim-milk was microfiltered three-fold at 50 °C using a MF pilot cross-flow unit as described previously by St-Gelais et al., (MilchBib 1995, 50 (11), 614-619).
  • the proteins contained in the permeate were concentrated twenty-five-fold at 50 °C by ultrafiltration using an UF pilot unit equipped with a Romicon membrane (PM 10, total surface area 1.3 m 2 ).
  • the concentrate was diafiltered five-fold by constant addition of water and freeze-dried before use in order to obtain WPI.
  • the components of the films are solubilized in distilled water, under stirring, and the solutions are heated at 90°C for 30 minutes. They are then degassed under vacuum to remove dissolved air and flushed under gas according to Brault et al. (J. Agric. Food Chem. , 45 (8), 2964- 2969,1997). Irradiation of the solutions at a total dose of 32 kGy is performed in a 0 Co underwater calibrator unit (UC-15b; 17.33 kGy/hour) (MDS Nordion, Kanata, Ontario, Canada) at the Canadian Irradiation Center. Films are cast by pipetting 5 mL of the solution onto smooth rimmed 8.5 cm internal diameter Petri dishes sitting on a leveled surface. Solutions are spread evenly and allowed to dry overnight at room temperature (20 ⁇ 2°C) in a climatic chamber (45-50% RH). Dried films are peeled intact from the casting surface.
  • the antioxidant capacities of the films were determined by the DPD (N, N-diethyl-p- phenylenediamine) colorimetric method described by Dumoulin et al., 1996.
  • Small rectangular pieces of films (1 x 3 cm) were introduced in electrolysis cells containing 3 ml of Krebs- Henseleit (KH) buffer and electrolyzed at 10 mA DC for lmn using a model 1000/500 power supply (Biorad, Richmond, CA, USA) to generated oxygen free radicals (OFR).
  • Scavenging capacity (%) 100-[ (ODsample / ODcontrol).100], where ODsample and ODcontrol are optical densities of electrolyzed samples and positive control, respectively.
  • EXAMPLE VI Cross-linking effect on water vapor permeability of whey protein isolate, whey protein concentrate, and calcium caseinate films.
  • Calcium caseinate (Alanate 380; 91.8% protein on weight basis) was obtained from New Zealand Milk Product Inc. (Santa Rosa, CA, USA). Whey protein concentrate (Sapro-75, 76.27% protein on whey basis) and whey protein isolate (90.57%) protein on weight basis) were obtained from Saputo Cheeses Ltd (Montreal, Quebec, Canada) and the Food Research and Development Centre (St-Hyacinthe, Quebec, Canada), respectively.
  • Calcium caseinate was solubilized in distilled water in presence of low viscosity carboxymethyl cellulose (2.5% w/v; Sigma Chemicals, St-Louis, MO, USA) and reagent grade glycerol (2.5% w/v; American Chemicals Ltd, Montreal, Quebec). Whey protein concentrate and whey protein isolate added to the solution to obtain various casein/whey protein ratios (100/0, 75/25, 50/50, 25/75, and 0/100) with a total protein concentration of 5% (w/v) in the film forming solutions.
  • the film forming solutions were cast directly onto smooth petri plates (8.5 cm, I.D.) and allowed to dry overnight at 20 ⁇ 1°C in a climatic chamber (45-50%, R Humidity).
  • the irradiated films were obtained in the same manner, but the film forming solutions were first irradiated at a total dose of 32 kGy in a 60Co underwater calibrator unit (UC-15b) (MSD, Nordion, Laval, Quebec) with a mean rate dose of 17.33 kGy/h before casting. Both the unirradiated and the irradiated film were used in the permeability measurements.
  • the film thickness was determined using a Digimatic indicator micrometer (Mitutoyo, Tokyo, Japan). Measurement were taken at five locations and the means values were used for permeability calculations. The thichness off the films averaged 66-95 ⁇ 3.6 (m depending on the formulation.
  • WVP Water Vapor Permeability of the films was determined gravimetrically at 23 °C using a modified ASTM (1983) procedure.
  • the test films were sealed to glass cups contained phosphorus pentoxide cristals (Sigma Chemicals, St-Louis, MO, USA) with exposed film area of 13.40 cm 2 .
  • the cups were placed in dessicators with werenot at 23 °C under 100% RH (21.59 mmHg water vapor pressure) with distilled water of 56% RH (9.82 mmHg water vapor pressure) with saturated sodium bromide solution (Sigma Chemicals, St-Louis, MO).
  • the water vapor transferred through the film and absorbed by the desiccant was determined by the weight gain of the phosporus pentoxide.

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Abstract

L'invention concerne des agents de revêtement solubles à l'eau, à base de caséinate et de lactosérum. L'invention concerne également des procédés de formation d'une gamme de revêtements et de pellicules dans lesquelles la proportion de caséinate, par rapport au lactosérum, est choisie de manière à optimiser les caractéristiques recherchées de l'agent de revêtement et, éventuellement, l'ajout d'autres composés. Les agents de revêtement de l'invention, qui sont comestibles, sont utiles dans l'industrie alimentaire et permettent d'allonger la durée de conservation de différents fruits et légumes, d'éliminer la contamination bactérienne de la viande, et de réduire la quantité de matériaux d'emballage nécessaires.
PCT/CA2000/000161 1999-02-22 2000-02-22 Agents de revetement reticules a base de caseinate et de lactoserum Ceased WO2000049899A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU26546/00A AU2654600A (en) 1999-02-22 2000-02-22 Caseinate-whey crosslinked covering agent

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CA2262310 1999-02-22
CA2,262,310 1999-02-22

Publications (1)

Publication Number Publication Date
WO2000049899A1 true WO2000049899A1 (fr) 2000-08-31

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Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CA2000/000161 Ceased WO2000049899A1 (fr) 1999-02-22 2000-02-22 Agents de revetement reticules a base de caseinate et de lactoserum

Country Status (2)

Country Link
AU (1) AU2654600A (fr)
WO (1) WO2000049899A1 (fr)

Cited By (17)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2001037683A3 (fr) * 1999-11-24 2001-10-18 Mateescu M Alexandru Films biologiques a base de proteines et de polysaccharides
NL1019890C2 (nl) * 2002-02-01 2003-08-07 Tno Vochtwerende barrière.
WO2010120356A1 (fr) * 2009-04-13 2010-10-21 The University Of Vermont And State Agricultural College Adhésifs pour bois écologiques à base de protéine de lactosérum et leurs procédés de production et d'utilisation
US8551544B2 (en) 2005-07-13 2013-10-08 Archer Daniels Midland Company Protein isolate compositions and uses thereof
CN103609670A (zh) * 2013-11-08 2014-03-05 渤海大学 一种营养可食的果蔬复合涂膜保鲜剂的制备方法
CN104920583A (zh) * 2014-03-20 2015-09-23 柳州长远食品配料科技有限公司 一种用于肉制品和水产品的无磷保水剂及其制备方法
WO2016005481A1 (fr) * 2014-07-11 2016-01-14 Knauf Insulation Sprl Liant contenant une protéine de lactosérum
WO2016016373A1 (fr) * 2014-08-01 2016-02-04 Instituto Tecnológico Del Embalaje, Transporte Y Logística (Itene) Emballage pour champignons, fruits et légumes frais
DE102017202887A1 (de) 2017-02-22 2018-08-23 Uwe Vorreiter Beschichtungslösung zur Beschichtung eines Verpackungsmittels zur Erzeugung einer Barriereschicht, Verpackungsmittel sowie ein Verfahren zu dessen Herstellung
CN111868174A (zh) * 2017-12-19 2020-10-30 拉茨蒂普斯公司 基于酪蛋白和/或酪蛋白酸盐的可生物降解的热塑性塑料
EP3895538A1 (fr) * 2016-11-07 2021-10-20 Polynatural Holding SpA Compositions d'enrobage et leurs procédés d'utilisation
WO2021252403A1 (fr) * 2020-06-07 2021-12-16 Comestaag Llc Compositions de revêtement protecteur pour denrées périssables et procédés, systèmes, kits et articles revêtus associés
CN114158612A (zh) * 2021-12-21 2022-03-11 广东广益科技实业有限公司 一种槟榔上光剂、其制备方法及应用
WO2022144483A1 (fr) 2020-12-30 2022-07-07 Universitat De València (60%) Souche de lactobacillus plantarum, utilisation en tant que probiotique et produit bioactif dérivé de celle-ci
US11582979B2 (en) 2020-06-07 2023-02-21 Comestaag Llc Selectively treating plant items
WO2025051976A1 (fr) * 2023-09-08 2025-03-13 Lactips Film hydrosoluble biodégradable à base de caséine et/ou de caséinate pour la production d'emballages ou d'étiquettes et son procédé d'obtention
WO2025055959A1 (fr) * 2023-09-12 2025-03-20 Bing Biotech Limited Encapsulation de composition à deux enzymes pour prévenir, traiter et/ou atténuer la veisalgie et les symptômes qui y sont associés

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EP0477832A1 (fr) * 1990-09-24 1992-04-01 Societe Des Produits Nestle S.A. Composition de brunissement pour la cuisson aux micro-ondes
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MCHUGH T H ET AL: "MILK-PROTEIN-BASED EDIBLE FILMS AND COATINGS", FOOD TECHNOLOGY,US,INSTITUTE OF FOOD TECHNOLOGISTS. CHICAGO, vol. 48, no. 1, 1 January 1994 (1994-01-01), pages 97 - 103, XP000423512, ISSN: 0015-6639 *

Cited By (23)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2001037683A3 (fr) * 1999-11-24 2001-10-18 Mateescu M Alexandru Films biologiques a base de proteines et de polysaccharides
NL1019890C2 (nl) * 2002-02-01 2003-08-07 Tno Vochtwerende barrière.
WO2003063620A3 (fr) * 2002-02-01 2003-11-20 Tno Couche de separation resistante a l'humidite
US8551544B2 (en) 2005-07-13 2013-10-08 Archer Daniels Midland Company Protein isolate compositions and uses thereof
WO2010120356A1 (fr) * 2009-04-13 2010-10-21 The University Of Vermont And State Agricultural College Adhésifs pour bois écologiques à base de protéine de lactosérum et leurs procédés de production et d'utilisation
CN103609670A (zh) * 2013-11-08 2014-03-05 渤海大学 一种营养可食的果蔬复合涂膜保鲜剂的制备方法
CN103609670B (zh) * 2013-11-08 2015-02-04 渤海大学 一种营养可食的果蔬复合涂膜保鲜剂的制备方法
CN104920583A (zh) * 2014-03-20 2015-09-23 柳州长远食品配料科技有限公司 一种用于肉制品和水产品的无磷保水剂及其制备方法
WO2016005481A1 (fr) * 2014-07-11 2016-01-14 Knauf Insulation Sprl Liant contenant une protéine de lactosérum
WO2016016373A1 (fr) * 2014-08-01 2016-02-04 Instituto Tecnológico Del Embalaje, Transporte Y Logística (Itene) Emballage pour champignons, fruits et légumes frais
EP3895538A1 (fr) * 2016-11-07 2021-10-20 Polynatural Holding SpA Compositions d'enrobage et leurs procédés d'utilisation
DE102017202887A1 (de) 2017-02-22 2018-08-23 Uwe Vorreiter Beschichtungslösung zur Beschichtung eines Verpackungsmittels zur Erzeugung einer Barriereschicht, Verpackungsmittel sowie ein Verfahren zu dessen Herstellung
DE102017202887B4 (de) 2017-02-22 2024-05-29 Uwe Vorreiter Beschichtungslösung zur Beschichtung eines Verpackungsmittels zur Erzeugung einer Barriereschicht, ein Verfahren zu deren Herstellung sowie deren Verwendung
CN111868174A (zh) * 2017-12-19 2020-10-30 拉茨蒂普斯公司 基于酪蛋白和/或酪蛋白酸盐的可生物降解的热塑性塑料
US12286536B2 (en) 2017-12-19 2025-04-29 Lactips Biodegradable thermoplastic material made from casein and/or caseinate
WO2021252403A1 (fr) * 2020-06-07 2021-12-16 Comestaag Llc Compositions de revêtement protecteur pour denrées périssables et procédés, systèmes, kits et articles revêtus associés
US11582979B2 (en) 2020-06-07 2023-02-21 Comestaag Llc Selectively treating plant items
WO2022144483A1 (fr) 2020-12-30 2022-07-07 Universitat De València (60%) Souche de lactobacillus plantarum, utilisation en tant que probiotique et produit bioactif dérivé de celle-ci
CN114158612A (zh) * 2021-12-21 2022-03-11 广东广益科技实业有限公司 一种槟榔上光剂、其制备方法及应用
CN114158612B (zh) * 2021-12-21 2024-01-05 广东广益科技实业有限公司 一种槟榔上光剂、其制备方法及应用
WO2025051976A1 (fr) * 2023-09-08 2025-03-13 Lactips Film hydrosoluble biodégradable à base de caséine et/ou de caséinate pour la production d'emballages ou d'étiquettes et son procédé d'obtention
FR3152809A1 (fr) * 2023-09-08 2025-03-14 Lactips Film hydrosoluble biodégradable à base de caséine et/ou de caséinate pour la production d’emballages ou d’étiquettes et son procédé d’obtention
WO2025055959A1 (fr) * 2023-09-12 2025-03-20 Bing Biotech Limited Encapsulation de composition à deux enzymes pour prévenir, traiter et/ou atténuer la veisalgie et les symptômes qui y sont associés

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