EP1890556A1 - Conjugues de pectine - Google Patents

Conjugues de pectine

Info

Publication number
EP1890556A1
EP1890556A1 EP06757795A EP06757795A EP1890556A1 EP 1890556 A1 EP1890556 A1 EP 1890556A1 EP 06757795 A EP06757795 A EP 06757795A EP 06757795 A EP06757795 A EP 06757795A EP 1890556 A1 EP1890556 A1 EP 1890556A1
Authority
EP
European Patent Office
Prior art keywords
pectin
amino acids
oligopeptide
phenolic
composition according
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.)
Withdrawn
Application number
EP06757795A
Other languages
German (de)
English (en)
Inventor
Johannes Wilhelmus Leonardus Boumans
Randolph Peter Happe
Martin Anton Bos
Ricardo Marinus Albertus Nagtegaal
Peter Degn
Merete Faergemand
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Nederlandse Organisatie voor Toegepast Natuurwetenschappelijk Onderzoek TNO
Original Assignee
Nederlandse Organisatie voor Toegepast Natuurwetenschappelijk Onderzoek TNO
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 Nederlandse Organisatie voor Toegepast Natuurwetenschappelijk Onderzoek TNO filed Critical Nederlandse Organisatie voor Toegepast Natuurwetenschappelijk Onderzoek TNO
Priority to EP06757795A priority Critical patent/EP1890556A1/fr
Publication of EP1890556A1 publication Critical patent/EP1890556A1/fr
Withdrawn legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12C—BEER; PREPARATION OF BEER BY FERMENTATION; PREPARATION OF MALT FOR MAKING BEER; PREPARATION OF HOPS FOR MAKING BEER
    • C12C5/00—Other raw materials for the preparation of beer
    • C12C5/02—Additives for beer
    • 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
    • A23J3/10—Casein
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L27/00—Spices; Flavouring agents or condiments; Artificial sweetening agents; Table salts; Dietetic salt substitutes; Preparation or treatment thereof
    • A23L27/60—Salad dressings; Mayonnaise; Ketchup
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
    • A23L29/10—Foods or foodstuffs containing additives; Preparation or treatment thereof containing emulsifiers
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
    • A23L29/20—Foods or foodstuffs containing additives; Preparation or treatment thereof containing gelling or thickening agents
    • A23L29/206—Foods or foodstuffs containing additives; Preparation or treatment thereof containing gelling or thickening agents of vegetable origin
    • A23L29/231—Pectin; Derivatives thereof
    • 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
    • A23P30/00—Shaping or working of foodstuffs characterised by the process or apparatus
    • A23P30/40—Foaming or whipping
    • 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
    • A23C2210/30—Whipping, foaming, frothing or aerating dairy products
    • 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

  • the invention relates to conjugates of phenolic polymeres, methods for preparing the same and use of conjugates of phenolic polymeres in among others foodstuffs.
  • it relates to surface active compositions comprising phenolic polymeres that are covalently cross-linked to a proteinaceous substance.
  • Phenolic polymeres are polymeres comprising one or more phenolic substituents.
  • Preferred phenolic substituents are ferulic acid, caffeic acid, coumaric acid, vanillic acid and cinnamic acid.
  • the polymer is preferably a polysaccharide comprising phenolic substituents.
  • Naturally occuring phenolic polymeres comprise polymeres of monosaccharides, such as pectins, heteroxylans, arabinoxylans as well as hemicellulosic material. Also polymers to which phenolic groups have been added chemically or enzymatically are within the scope of the invention.
  • Pectin (E440) is a heterogeneous grouping of acidic structural polysaccharides. It is a natural substance and, as a component of the middle lamella and the primary cell wall, an important structural element of all plants. It acts as a bonding substance with supporting and stabilizing functions and controls, due to its great swelling ability and hydrocolloid nature, the plant's water system. Pectin mainly consists of D-galacturonic acid molecules which are linked to each other by ⁇ -(l,4)-glycosidic bonds to become polygalacturonic acid. The carboxyl groups are partially esterified with methanol. In some cases, the secondary alcohol groups of the pectins can also carry acetyl groups.
  • pectins also ferolyl groups can be found.
  • neutral sugars such as galactose, arabinose or xylose which are linked as side chains to the pectin macromolecule and the rupture of the main chain caused by rhamnose make pectin to a branched zig-zag-shaped heteropolysaccharide.
  • Pectins are mainly used as gelling agents, but can also act as thickener, water binder and stabilizer.
  • Low methoxyl pectins ( ⁇ 50% esterified) form thermoreversible gels in the presence of calcium ions and at low pH (3 - 4.5) whereas high methoxyl pectins rapidly form thermally irreversible gels in the presence of sufficient (e.g. 65% by weight) sugars such as sucrose and at low pH ( ⁇ 3.5); the lower the methoxyl content, the slower the set.
  • the degree of esterification can be (incompletely) reduced using commercial pectin methylesterase, leading to a higher viscosity and firmer gelling in the presence of Ca 2+ ions.
  • the polysaccharide pectin has been used as a food additive, a thickening agent, and a gelling agent (Rolin C.
  • pectin In: Whistler RL, Bemiller JN, eds. Industrial Gums: Polysaccharides and Their Derivatives. New York, NY: Academic Press; 1993:257-293).
  • pectin can reduce interfacial tension between an oil phase and a water phase and may be used for the preparation of emulsion (Leroux J, Langendorff V, Schick G, Vaishnav V, Mazoyer J. Emulsion stabilizing properties of pectin. Food Hydrocolloids. 2003;17:455-462).
  • a specific class of pectin is represented by beet pectin, in particular sugar beet pectin.
  • Sugar beet pectin is one of the few polysaccharides which contain ferulic acid. It is attached to the 0-2 position of (l,5)-linked arabinose • residues in the arabinan side-chains and it can also be found to be attached to the 0-6 position of galactose residues in (l,4)-linked galactans. It is well known that several oxidizing agents, including, oxidative enzymes, are capable of initiating oxidative cross-linking of beet pectins, via the formation of ferulic acid dehydrodimers.
  • pectin in the food industry, for example use as a fat replacer or as an emulsifier.
  • WO96/03440 discloses pectin derived from sugar beet, which is modified using a laccase enzyme. The phenolic groups of the sugar beet pectins are modified such that a cross-linked gel is provided. It is taught in WO96/03440 that the gel may be used in foodstuffs as thickening, stabilising, setting or viscosity regulating agent or that the gel may be used in medicinal or agricultural applications.
  • EP0656176 discloses a calcium-sensitive pectin. Calcium is added to the pectin to form ionic bonds between the pectin.
  • WO00/40098 discloses a fat replacer comprising a pectin composition wherein at least a population of the pectin molecules is covalently cross-linked to one another. Boeriu, CG. et al. (J. Agricult. Food Chem. 2004, 52:21) describe a method to efficiently produce conjugates between arabinoxylans and beta- casein. It does not report on any functional properties of the conjugates formed, but suggests that they may have improved or novel functional properties.
  • the EA is lower than that of the unmodified protein! It is true that conjugation increases the EA compared to unmodified mixtures of WPI and pectin. It is well known that proteins, in particular milk proteins, have good emulsifying properties. Oligopeptides, as described in the present invention, do not have good surface active properties (see examples 3, 4 and 5), nor do phenolic polymers like sugar beet pectin. The surface active properties of the conjugates of the present invention therefore are much better than that of the separate components and mixtures thereof, which is in contrast to those described in Einhorn-Stoll et ⁇ l.
  • US2004/0009285 describes a process for the preparation of an emulsion, by adding pectin and an oxidative enzyme, resulting in in situ oxidation (and cross-linking) of pectin during or after formation of the oil- water emulsion.
  • this on first sight may seem to include hetero- coupling, it is known that the reaction rate between two feruloyl groups is much higher than between two tyrosines or between a feruloyl group and a tyrosine (see figure 1 in J. Agric. Food Chem. 49, 2503-10).
  • tyrosines in intact proteins are relatively poor substrates for this reaction because of accessibility.
  • US2002/004089 describes polymers suitable for use as edible films, comprising casein and pectin in a cross-linked 3-dimensional network. There is no report on surface active properties of the products formed and only intact proteins (casein) are included. The products formed in my view are entirely different from the conjugates we claim. Products formed by D7 will be water- insoluble and will have very poor surface active properties, if any. Thus, the products formed in this prior art document are entirely different from the conjugates of the present invention.
  • the present invention provides a novel surface active composition comprising phenolic polymeres that are cross-linked to oligopeptides.
  • a surface active composition lowers the surface tension of the medium in which it is dissolved, and/or the interfacial tension with other phases, and, accordingly, is positively adsorbed at the liquid/vapour and/or at other interfaces.
  • surfactant is also applied correctly to sparingly soluble substances, which lower the surface tension of a liquid by spreading spontaneously over its surface.
  • oligopeptide refers to a peptide of about 3 to about 30 amino acids, preferably of about 5 to about 20 amino acids, more preferably of about 7 to about 15 amino acids, most preferably of about 10 amino acids.
  • a composition of the invention has surprisingly good surface active properties and is advantageously used, e.g. as emulsifier or foam stabilizer, in a foodstuff.
  • the phenolic polymere is a pectin comprising phenolic substituents. It is thought that attachment of oligopeptides provide a hydrophobic moiety to the predominantly hydrophilic pectin structure. The presence of the peptide moiety thus enhances the hydrophobic nature of the molecule, giving it a surface active character with potential as an interfacial agent in oil/water emulsions and air/water foams. Accordingly, composition wherein the oligopeptide has a high hydrophobicity (i.e. low hydrophilicity) are preferred.
  • the average hydrophobicity of a given peptide can be calculated by several methods known in the art. For example, the software available free of charge at http://www.innovagen.se/custom-peptide-synthesis/peptide-property- calculator/peptide-property-calculator.asp can be used to deduce the average hydrophilicity value of a peptide.
  • the average hydrophilicity value of a peptide according to this method is given by a number between +3 and -3.
  • a negative value implies that the average amino acid has a hydrophobic nature, whereas a positive value implies that the average amino acid has a hydrophilic nature.
  • the average hydrophilicity of the oligopeptide is lower than zero, preferably lower than -0.3, preferably lower than -0.5, more preferably lower than -0.7.
  • phenolic polymers such as pectins comprising phenolic substituents can be cross-linked to any type of oligopeptide. It may be a plurality of the same oligopeptide (homo- oligopeptides), or to a mixture of different oligopeptides (hetero-oligopep tides).
  • Synthetic peptides may be used which are synthesized according to standard procedures, for example solid phase peptide synthesis.
  • Oligopeptides may contain one or more amino acid analogues, such as a D-amino acid analogue.
  • An advantage of synthetic peptides is that they can be custom-made with respect to peptide length and amino acid sequence.
  • oligopeptides obtained from a natural source may be preferred.
  • a protein hydrolysate can be used as a source of oligopeptides. Since a composition of the invention is particularly useful in foodstuff, edible proteins are the preferred sources of oligopeptides.
  • the composition of the invention comprises pectin that is cross-linked to oligopeptide derived from an edible protein.
  • pectin that is cross-linked to oligopeptide derived from an edible protein.
  • milk derived proteins such as whey or casein are preferred because of their taste contribution.
  • Oligopeptides derived from casein are particularly preferred.
  • Casein hydrolysates are widely available and represent a relatively inexpensive source of oligopeptides.
  • casein hydrolysates are a good source for hydrophobic peptides. Casein represents approximately 80% of total milk protein.
  • casein proteins are alpha(sl) and alpha(s2) -caseins, ⁇ -casein, and kappa-casein.
  • the distinguishing property of all caseins is their low solubility at pH 4.6.
  • the high number of proline residues in caseins causes particular bending of the protein chain and inhibits the formation of close-packed, ordered secondary structures.
  • Casein hydrolysates can be prepared by methods known in the art (Enzymic hydrolysis of food proteins, J. Adler-Nissen (1986) Elsevier Applied Science Publishers LTD). This typically involves either enzyme-mediated hydrolysis or chemical (acid) cleavage of peptidic bonds.
  • proteolytic enzymes proteolytic enzymes (proteases) such as trypsin or trypsin like proteases can be used to obtain oligopeptides.
  • the hydrolysate of casein consists of peptides with an average chain length of about 3 to about 30 amino acids, preferably of about 5 to about 20 amino acids, more preferably of about 7 to about 15 amino acids, most preferably of about 10 amino acids.
  • pectin includes fractions of pectin, one or more compounds from the class of compounds known as pectins, and derivatives thereof. All plant raw materials with high pectin content is suitable for the production of pectins.
  • the extraction of soluble pectin on industrial scale is generally carried out by means of acid hydrolysis resulting in liquid pectin.
  • This liquid pectin can be dried to obtain pectin extracts or the pectin can be isolated from the aqueous solution by alcohol precipitation. Afterwards it is dried, grinded and sieved to a defined particle size.
  • the prime sources are apple pomace, citrus peels and sugar beet chips.
  • the various raw materials yield different amounts of extractable pectin.
  • Pectins extracted from various raw materials differ in molecular structure (i.e. molecular weight, degree of esterification, acetyl content, neutral sugar content, distribution of the methoxylated carboxyl groups) and therefore possess different functional properties.
  • molecular structure i.e. molecular weight, degree of esterification, acetyl content, neutral sugar content, distribution of the methoxylated carboxyl groups
  • any type of pectin is suitably used as surface active substance according to the invention.
  • the pectin material and/or the pectin composition contain at least a population of pectin which have phenolic substituents.
  • the pectin comprising phenolic substituents is obtainable from or is obtained from members of the plant family Chenopodiaceae (which includes beets, spinach and mange wurzels). More preferably, the pectin material and/or the pectin composition is obtainable from or is obtained from beets, yet more preferably, sugar beets.
  • the phenolic substituents are phenolic acid substituents.
  • a composition according to the invention comprises pectin cross-linked to a oligopeptide, wherein the pectin has a molecular weight ranging from about 10,000 kDa to about 500,000 kDa, preferably from about 50.000 kDa to about 250.000 kDa, more preferably from about 75.000 kDa to about 150.000 kDa, most preferably about 100.000 kDa.
  • Pectin may be extracted from plants, e.g. using the isolation method described in WO00/40098.
  • Pectin is commercially available from various suppliers, for instance from Danisco A/S or CP Kelco
  • the relative presence of oligopeptide moieties and phenolic polymers, such as pectins pectin molecules in a cross-linked composition can vary.
  • the weight ratio of phenolic polymers, such as pectin to oligopeptide in the composition ranges from about 100:1 to about 6:1.
  • oligopeptides can be cross-linked to phenolic polymers, such as pectin via any type of intermolecular linkage.
  • the cross-links are provided between a phenolic moiety of the polymer and a tyrosine, lysine or cysteines moiety of the peptide.
  • Cross links of this type can be achieved enzymatically.
  • the teaching of WO00/40098 can be used which discloses the use of various cross-linking agents, including enzymes and chemical cross-linking agents such as oxidants such as peroxides, persulfates or perchlorates: aldehydes such as formalin or glutaric aldehyde and epoxides.
  • cross-linking agent is an enzyme.
  • the enzyme is selected oxidoreductases (i.e. enzymes employing molecular oxygen as acceptor) such as laccases (EC 1.10.3.2) capable of catalysing oxidation of phenolic groups, phenol-oxidising oxidoreductases (EC 1.10.3.1), glucoseoxidases, hexoseoxidases, peroxidases (EC 1.11.1.7), mixtures and derivatives thereof.
  • laccases EC 1.10.3.2
  • glucoseoxidases hexoseoxidases
  • peroxidases EC 1.11.1.7
  • mixtures and derivatives thereof mixtures and derivatives thereof.
  • Commercial laccase preparation may be obtained from Novozymes, Biocatalysts or J ⁇ lich Fine Chemicals.
  • the enzyme is laccase.
  • Laccases can be obtained from a variety of microbial sources, notably bacteria and fungi including filamentous fungi and yeasts.
  • reaction mixture containing the polymers, the peptides and laccase will be contacted to atmospheric air. This will normally suffice to ensure an adequate supply of oxygen for oxidation, although forcible aeration of a reaction mixture with air, or possibly even substantially pure oxygen, may be advantageous under certain conditions.
  • the oxidizing agent is hydrogen peroxide, methylperoxide, ethylperoxide or sodium perborate.
  • hydrogen peroxide can be generated in situ by an enzyme system comprising an oxidoreductase such as glucose oxidase, sulfhydryl oxidase, vanillyl alcohol oxidase etc. and an appropriate substrate.
  • Plant peroxidases are the preferred peroxidases, of which horseradish peroxidase is the preferred peroxidase.
  • an enzymatic cross-linking procedure is employed.
  • cross-linking of oligopeptides to pectin in a batch wise procedure resulted in the desired heterologous pectin- oligopeptide cross-linked composition.
  • a high degree of cross-linking between pectin and oligopeptides can be obtained by mixing all necessary substrates and reagents together without running the risk of unwanted formation of pectin-pectin and peptide-peptide cross-linked compositions. This is unlike the situation in EP 1169922, which discloses a method for enzymatically cross-linking proteins and phenolic polymers.
  • the procedure needs to be controlled such that the formation of hetero-cross-linked products (i.e. protein-phenolic polymer) is favoured, while minimizing the formation of peptide-peptide and/or phenolic polymer-phenolic polymer cross-links.
  • the protein must be present initially in a high ratio as compared to the phenolic polymer.
  • a small quantities of the substrate By subsequent stepwise or continuous addition a small quantities of the substrate, desired cross-linking products could be achieved.
  • a cross-linked pectin composition of the present invention may be prepared in a similar fashion, it surprisingly appeared that the use of oligopeptides makes the rather complex controlled procedure of EPl 169922 redundant.
  • the invention also provides a method of batch- wise cross- linking a oligopeptide and a phenolic polymer comprising mixing said oligopeptide and said polymer with an enzyme and oxidizing agent suitable for said enzyme in a solvent, wherein said peptide has a chain length of about 3 to about 30 amino acids, preferably of about 5 to about 20 amino acids, more preferably of about 7 to about 15 amino acids, most preferably of about 10 amino acids.
  • the oligopeptide can be derived from a hydrolysate of casein.
  • a method of the invention is not limited to cross-linking peptides to pectin. Rather, any type of polymeric materials having phenolic substituents can be used as a starting material.
  • the invention furthermore provides a use of the cross-linked polymer/pep tide composition as emulsifier, and/or foam stabiliser.
  • the invention also provides a foodstuff comprising a composition according to the invention.
  • the term "foodstuff may include food for human and/or animal consumption.
  • the foodstuff is for example selected from products made from or containing milk (such as cheese, fresh cheese, yoghurt ,sour cream or ice cream), low fat spread, mayonnaise, sauces, meat and meat products, poultry products, fish products, fruit spreads, bakery products including bakery fillings, margarine, reconstituted fruits, jams, fruit preparations, fruit fillings, ripples, fruit sauces, stewed fruit, coffee whitener, instant fruit dessert, confectionery (such as marsh mallow), and fine foods (such as dressings including salad dressings; ketchup, vinaigrette dressings and soups).
  • the foodstuff may be a beverage.
  • the beverage may be a drinking yoghurt, a fruit juice, a beverage concentrate or a fruit based beverage.
  • a composition of the invention is advantageously used as foam stabiliser in liquid or solid foodstuff, for example in aerated (frozen) confection products such as ice cream and mousse-type desserts.
  • a composition is used as a foam stabiliser in beer.
  • Foam occurs on dispensing the beer as a result of the formation of CO2 bubbles released by the reduction in pressure.
  • the CO2 bubbles collect surface active materials as they rise. These surface active substances have a low surface tension. This means that, within limits, they can increase their surface area and also, after the bubbles have risen, they form an elastic skin around the gas bubble. The greater the amount of dissolved CO2, the more foam is formed. But foam formation is not the same as foam stability. Foam is only stable in the presence of these surface active substances, such as a composition according to the present invention.
  • the present invention may be used in the preparation of a starting reagent or an intermediate in the preparation of a foodstuff. Alternatively, present invention may be used in the preparation of a foodstuff itself.
  • the pectin composition may be incorporated in materials other than foodstuffs. For example, the pectin composition may be incorporated in cosmetics (such as cream) and pharmaceuticals.
  • Laccase activity as defined herein is determined on the basis of spectrophotometric measurements of the oxidation of ABTS under aerobic conditions. The intensity of colour produced in the oxidation reaction is measured at 410 nm.
  • Dilution buffer 100 mM sodium acetate buffer, pH 5.0, containing BSA (100 mg/1) and 0.01% Tween 20. This buffer is used to dilute laccase samples.
  • Reagent buffer 100 mg ABTS in 20 ml of 100 mM sodium acetate buffer pH 5.0 (prepare fresh daily).
  • aerate the reagent buffer with oxygen 900 ⁇ liter of reagent buffer with 100 ⁇ liter (diluted) sample and measure the change in extinction at 410 nm and at 25 °C over time. Calculate the activity using a molar extinction coefficient of 36.800.
  • laccase unit is the amount of enzyme that catalyses the conversion of 1 ⁇ M of ABTS per minute at 25 0 C.
  • Example 1 Preparation of sugar beet pectin Teknisk
  • Fresh sugar beet pulp was obtained from a sugar factory (Danisco Sugar and Sweetener in Assens, Denmark). Extraction of sugar beet pectin from sugar beet pulp was performed as described in Buchholt et al. (2004) Carbohydrate Pol. 58, 149-161.
  • a solution was prepared containing 0.67% (w/v) pectin Teknisk and 0.33% (w/v) casein hydrolysate CE90GBT (DMV International, Veghel, The Netherlands).
  • the solution was heated to 40 0 C, kept at constant pH of 6.0 using a pH stat, 570 Units/litre laccase (Trametes versicolor; Wacker Chemie) was added and the solution was incubated for 16 hours. The solution was then freeze-dried.
  • the product is named pectin conjugate in all examples described below.
  • several other cross-linked products were made as controls.
  • 0.67% pectin Teknisk was incubated with laccase under identical conditions, giving PxP.
  • 0.33% casein hydrolysate CE90GBT was incubated with laccase under identical conditions, giving HxH.
  • Foam was prepared according to the whipping method described by Caessens (1999, Enzymatic hydrolysis of ⁇ -casein and ⁇ -lactoglobulin, PhD Thesis, WAU). A solution was whipped for 70 s at 3500 rpm. The resulting foam was poured in the cuvet of the Foam Analyser. The size of the cuvet is 45 x 57 x 134 mm. Perspex and/or teflon cubes were placed in the cuvet to elevate the foam to the height of the camera.
  • Foam was analyzed in transmission and reflection mode.
  • the transmission mode light passed straight through the cuvet to a camera at the opposite side of the light source.
  • a prism of 90° is placed on the 57 mm side of the cuvet.
  • Light source and camera are placed in such a way that an image of the foam at the glass side can be taken in reflection.
  • the camera is a Mega Pixel Progressive Scan Camera (1300 x 1030 pixels). After placing the cuvet in the Foam Analyser 50 images were taken at an interval of 60 s. From images taken in transmission foam height was determined.
  • Foams were prepared with 0.1% pectin conjugate in water. As controls, a mixture of pectin (0.067%) and casein hydrolysate (0.033%) coded P+H, cross- linked pectin coded PxP and cross-linked casein hydrolysate coded HxH were included. Results of foam analysis are shown in figure 1.
  • Foam volume as a function in time of pectin conjugate in beer Beer with poor foaming properties was obtained from Diageo, Dublin, Ireland. Pectin conjugate in several concentrations was added to the beer. As a reference, PGA (commercial name Biofoam, Quest International) was included. Results of foam volume in time are shown in figure 2.
  • All emulsions prepared contained 30% sunflower oil and an aqueous phase containing polysaccharide in 20 mM phosphate pH 6.0. Each emulsion was 15 ml in volume. Emulsions were prepared by mixing oil into polysaccharide solution using an Ultra Turrax (Polytron, Switzerland) for 60 seconds at
  • the mixture was homogenized by 10 passes through a Delta lab-scale homogenizer operating at a pressure of 150 bar.
  • Emulsions were prepared using an aqueous phase containing 0.25% pectin conjugate. As controls, emulsions were prepared using an aqueous phase containing either 0.25% pectin, or a mixture of pectin and protein hydrolysate CE90GBT in a ratio 2:1 and a total dry weight content of 0.25%.
  • Emulsions were characterized after one week of storage at room temperature by determining particle size distribution (see figure 3) and turbidity (see figure 4).
  • Results of particle size distribution after one week of storage can also be depicted by the following numbers.
  • Emulsion stability was characterized by determining particle size distribution after 1 week of storage at room temperature.
  • Emulsions prepared with guar gum were highly unstable, making it not possible to obtain particle size distribution after 1 week of storage.
  • pectin conjugates were prepared using several hydrolysates. All protein hydrolysates were from DMV International (Veghel, The Netherlands).
  • Salad dressing was produced according to the following method and with addition of PGA or pectin conjugate. Contents in g
  • Phosphoric acid 8.5% 200,0 200,0 200,0
  • Pectin conjugate was prepared as described in Example 2, however, now using defined peptides instead of casein hydrolysate CE90GBT. Two defined peptides were used, strongly varying in hydrophobicity, but similar molecular weight and each containing one tyrosine residue.
  • a pectin conjugate was produced with each of the two peptides and the functionality in emulsion stability was examined according to the methods described in example 5.
  • Beverage emulsions were prepared by making a premix of Gum Arabic solution (standard) or various alternatives (see table) and citrus oil using an ultraturrax. Then, sodium benzoate and citric acid was mixed in and the volume was made up with water.
  • a small scale high pressure homogenizer (Delta Instruments, type HU- 3.0) was used in order to make small amounts of the emulsion. After homogenization the emulsions were checked by microscopy and the average particle size was about 1 micron.
  • Beverage emulsions were prepared using various combinations between gum Arabic, Xanthan gum, Locus bean gum and Pectin conjugate (see table).
  • Pectin conjugate was prepared as described in Example 2.
  • the viscosity of the emulsion was determined and emulsion stability was examined as described in Example 5 and after 2 weeks of storage at room temperature.
  • Samples 1, 7 and 13 are identical and are the positive controls containing only gum Arabic as stabilizer.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Food Science & Technology (AREA)
  • Polymers & Plastics (AREA)
  • Health & Medical Sciences (AREA)
  • Nutrition Science (AREA)
  • Zoology (AREA)
  • Biochemistry (AREA)
  • Organic Chemistry (AREA)
  • Dispersion Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Wood Science & Technology (AREA)
  • Jellies, Jams, And Syrups (AREA)
  • General Preparation And Processing Of Foods (AREA)

Abstract

L'invention concerne des conjugués de polymères phénoliques, des procédés de préparation de ceux-ci et une utilisation des conjugués de polymères phénoliques dans, par exemple, des produits alimentaires. L'invention concerne une composition à surface active comprenant un polymère phénolique, de préférence, une pectine, réticulé à un oligopeptide. Celui-ci peut être dérivé d'un hydrolysate de caséine renfermant des peptides d'une longueur de chaîne moyenne d'environ 3 à environ 30 acides aminés, de préférence d'environ 5 à environ 20 acides aminés, idéalement d'environ 7 à environ 15 acides aminés, plus idéalement encore d'environ 10 acides aminés. L'invention concerne, en outre, un procédé de réticulation par lots d'un oligopeptide et d'un polymère phénolique consistant à mélanger le peptide et le polymère avec une enzyme et un agent oxydant conçu pour l'enzyme dans un solvant, le peptide présentant une longueur de chaîne d'environ 3 à environ 30 acides aminés, de préférence, d'environ 5 à environ 20 acides aminés, idéalement d'environ 7 à environ 15 acides aminés et encore plus idéalement d'environ 10 acides aminés.
EP06757795A 2005-06-10 2006-06-12 Conjugues de pectine Withdrawn EP1890556A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
EP06757795A EP1890556A1 (fr) 2005-06-10 2006-06-12 Conjugues de pectine

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
EP05076366A EP1731043A1 (fr) 2005-06-10 2005-06-10 Conjugués de pectines
PCT/NL2006/000288 WO2006132529A1 (fr) 2005-06-10 2006-06-12 Conjugues de pectine
EP06757795A EP1890556A1 (fr) 2005-06-10 2006-06-12 Conjugues de pectine

Publications (1)

Publication Number Publication Date
EP1890556A1 true EP1890556A1 (fr) 2008-02-27

Family

ID=35240978

Family Applications (2)

Application Number Title Priority Date Filing Date
EP05076366A Withdrawn EP1731043A1 (fr) 2005-06-10 2005-06-10 Conjugués de pectines
EP06757795A Withdrawn EP1890556A1 (fr) 2005-06-10 2006-06-12 Conjugues de pectine

Family Applications Before (1)

Application Number Title Priority Date Filing Date
EP05076366A Withdrawn EP1731043A1 (fr) 2005-06-10 2005-06-10 Conjugués de pectines

Country Status (2)

Country Link
EP (2) EP1731043A1 (fr)
WO (1) WO2006132529A1 (fr)

Families Citing this family (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR3011841B1 (fr) 2013-10-14 2016-09-16 Agronomique Inst Nat Rech Polymere phenolique a liaisons biaryles 5-5, procede pour sa preparation et utilisations
CN107846939B (zh) * 2015-07-31 2022-08-12 不二制油集团控股株式会社 多糖类-蛋白质复合体的制造方法
US20200305445A1 (en) * 2017-10-26 2020-10-01 Basf Se Protein hydrolysates as emulsifier for baked goods
WO2019114967A1 (fr) * 2017-12-14 2019-06-20 Symrise Ag Conjugués de protéine et de polyuronide et leur utilisation en tant qu'émulsifiants
CN108013475B (zh) * 2017-12-26 2021-06-08 华中农业大学 一种多肽-多糖复合乳液及其制备方法
US20240049704A1 (en) * 2021-03-23 2024-02-15 Animal Health Concepts B.V. Emulsion

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1998009537A1 (fr) * 1996-09-09 1998-03-12 Kiwitech Limited Polymere caseine acide ou sel soluble non toxique de cette derniere, et pectine a haute teneur de methoxyle
EP1169922A1 (fr) * 2000-07-03 2002-01-09 Instituut Voor Agrotechnologisch Onderzoek (Ato-Dlo) Méthode pour réticuler par action enzymatique des protéines et des polymères phénoliques
FR2817870B1 (fr) * 2000-12-08 2006-07-28 Skw Nature Products Holding Fr Polymeres conjugues a base de pectines et leur application comme emulsifiants
US7297359B2 (en) * 2001-02-26 2007-11-20 Conopco, Inc. Process for the preparation of a foamed product and products obtainable by this process
CA2490312C (fr) * 2002-06-24 2011-07-26 Unilever Plc Emulsion de type aqueux stable

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of WO2006132529A1 *

Also Published As

Publication number Publication date
EP1731043A1 (fr) 2006-12-13
WO2006132529A1 (fr) 2006-12-14

Similar Documents

Publication Publication Date Title
CN101267744B (zh) 低pH充气产品
US8465911B2 (en) Native potato protein isolates
US7879382B2 (en) Stabilizers and compositions and products comprising same
US20060204454A1 (en) Method for improving the functional properties of a globular protein, protein thus prepared, use thereof and products containing the protein
AU2017220193A1 (en) Functional mung bean-derived compositions
EP1890556A1 (fr) Conjugues de pectine
WO2008026940A1 (fr) concentrés de protéines de lait appauvris en calcium stabilisateurs d'aliments
WO2006016809A1 (fr) Méthode de réticulation enzymatique d’une protéine, protéine réticulée ainsi obtenue et son utilisation.
AU2015345302B2 (en) Composition for preparing a frozen confection
US5274079A (en) Protein partial degradation products that are useful as surface active agents and dispersing agents
US8425970B2 (en) Edible aerated oil-and-water emulsion
WO2000040098A1 (fr) Composition de pectine utilisee comme substitut de matiere grasse et emulsifiant
EP1600207B1 (fr) Emulsifiant, procede de production dudit emulsifiant et composition emulsifiee utilisant ledit emulsifiant
JPH1014494A (ja) 酸性飲食品、酸性飲食品用酸性クリームおよび粉末
US5273773A (en) Protein partial degradation products
CN103491799B (zh) 充气食品
JP7355488B2 (ja) ゲル状食品の製造方法
EP3890514B1 (fr) Assaissonement
JPH01281043A (ja) クリーム状乳加工品の品質改良剤
WO2024058666A1 (fr) Mousse sans gélatine
Farahmand Utilization of inulin as fat replacer in cake and mayonnaise
WO2002082922A1 (fr) Substance pectique modifiee
MXPA97001517A (en) Low emulsified alliers in gr

Legal Events

Date Code Title Description
PUAI Public reference made under article 153(3) epc to a published international application that has entered the european phase

Free format text: ORIGINAL CODE: 0009012

17P Request for examination filed

Effective date: 20071210

AK Designated contracting states

Kind code of ref document: A1

Designated state(s): AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HU IE IS IT LI LT LU LV MC NL PL PT RO SE SI SK TR

DAX Request for extension of the european patent (deleted)
17Q First examination report despatched

Effective date: 20080314

DAX Request for extension of the european patent (deleted)
STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: THE APPLICATION IS DEEMED TO BE WITHDRAWN

18D Application deemed to be withdrawn

Effective date: 20080725