WO2023052590A1 - Isolat de protéine de colza - Google Patents

Isolat de protéine de colza Download PDF

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Publication number
WO2023052590A1
WO2023052590A1 PCT/EP2022/077294 EP2022077294W WO2023052590A1 WO 2023052590 A1 WO2023052590 A1 WO 2023052590A1 EP 2022077294 W EP2022077294 W EP 2022077294W WO 2023052590 A1 WO2023052590 A1 WO 2023052590A1
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WIPO (PCT)
Prior art keywords
oil
protein isolate
rapeseed protein
native
native rapeseed
Prior art date
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PCT/EP2022/077294
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English (en)
Inventor
Nicolas Jean-Robert ABELLO
Rudolf Franciscus Wilhelmus Cornelis Van Beckhoven
Marco Alexander Van Den Berg
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Dsm Ip Assets B.V.
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Publication date
Application filed by Dsm Ip Assets B.V. filed Critical Dsm Ip Assets B.V.
Publication of WO2023052590A1 publication Critical patent/WO2023052590A1/fr

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Classifications

    • 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
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/17—Amino acids, peptides or proteins
    • A23L33/185—Vegetable proteins
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
    • A23J1/00—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites
    • A23J1/14—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from leguminous or other vegetable seeds; from press-cake or oil-bearing seeds
    • 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
    • A23J3/14—Vegetable proteins
    • 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
    • A23L11/00—Pulses, i.e. fruits of leguminous plants, for production of food; Products from legumes; Preparation or treatment thereof
    • A23L11/30—Removing undesirable substances, e.g. bitter substances
    • 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
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/40—Complete food formulations for specific consumer groups or specific purposes, e.g. infant formula

Definitions

  • the present invention relates to a native rapeseed protein isolate.
  • Rapeseed is one of the most important oilseeds in the world (number 3 after soybean and palm oil). Rapeseed contains high amounts of oil (30 to 45 %) and protein (20 to 30 %). However anti-nutritional compounds such as glucosinolates, polyphenols and phytic acid are also present in rapeseed. Therefore rapeseed has received less attention for human nutrition due to the presence of such anti-nutritional compounds. However newer technologies can now be used to eliminate such compounds.
  • oilseed cake also known as oilseed meal
  • oilseed meal as a by-product from cold-pressing and optionally extracting oil from the rapeseed seeds.
  • the oilseed cake has a high-protein content which can be further extracted to produce rapeseed protein isolate.
  • Rapeseed protein isolate is now being suggested as an alternative to other proteins for human food use due to having a balanced amino acid profile on par with many animal proteins and superior to most vegetable proteins. Furthermore native rapeseed protein isolate has good potential functional properties such as emulsifying, foaming and gelling abilities. All these properties suggest that rapeseed seeds are a valuable source of high-quality protein isolate for utilization in the food processing industry, and can also be used as a good alternative to soybean derivatives and other plant and animal products.
  • WO 2008/094434 discloses the use of wheat protein isolates as an alternative to the use of egg yolk protein in compositions.
  • wheat protein isolates may not be desirable for those with gluten allergies.
  • Rapeseed (Brassica Napus), also known as rape, oilseed rape, rapa, rappi, rapaseed (and in the case of one particular group of cultivars, canola) is a bright yellow flowering member of the family Brassicaceae (mustard or cabbage family), (Wanasundara, 2011).
  • rapeseed plants form an elongated pod with two chambers separated by a membrane with a single row of seeds within each chamber. The pods are contain 15 to 30 small, spherical seeds.
  • the seeds of the Brassica Napus species are brown to black when mature. There are about 115,000 seeds per pound. Seeds are about 0.8 to 2.4 mm in diameter and although seed size may vary with variety and environmental effects the variation is minor when compared to the size and shape of an elongated wheat grain approximately 6 mm in length and 3 to 3.5 mm in width.
  • plant proteins might provide undesired plant protein flavours, such as characteristic odour, characteristic taste, lingering taste or astringency.
  • Rapeseed is unfortunately no exception to that. Particularly, rapeseed protein isolates might have undesired plant protein flavours as well as certain food applications containing the rapeseed protein isolate, which will make these food products less appreciated by the consumer. Hence there is a need in the art for a rapeseed protein isolate that solves these problems.
  • this objective is met by providing a native rapeseed protein isolate comprising an oil content of 1 to 15 wt. % on dry matter of the rapeseed protein isolate, wherein the isolate has a protein content of at least 90 wt.% (calculated as Dumas N x 6.25) on a dry weight basis, and/or wherein the isolate comprises an amount of moisture of less than 10 wt.%.
  • the present inventors found that adding oil to the rapeseed protein isolate reduces the undesired plant protein flavour of the isolate, as well as in applications of the protein isolate.
  • the rapeseed protein isolate of the invention is a native rapeseed protein isolate.
  • native is meant that the protein is not deliberately hydrolysed and that the protein is in its properly folded shape (in its native “conformation” or "structure”).
  • the term isolate means that on a dry basis, 85 wt. % of the total weight of the isolate is protein. This is calculated using the Dumas method with a nitrogen conversion factor of 6.25. Or using nitrogen content determination method such as Dumas combustion.
  • the non-protein content of the protein isolate includes non-protein compounds such as anti- nutritional substances, fibre and other components like the present oil.
  • the present native protein isolate has a protein content of at least 90 wt.% (calculated as Dumas N x 6.25) on a dry weight basis, preferably at least 91 , 92, 93, 94, 95, 96, 97, 98, or at least 99 wt.% on a dry weight basis (calculated as Dumas N x 6.25).
  • the amount of protein and oil do not exceed 100 wt. % of the protein isolate.
  • the amount of protein can be overestimated reaching a percentage above 100%.
  • Nx6.25 is the worldwide standard in commerce, scientists recognise that this overstates the value of plant proteins.
  • the recognized nitrogen conversion factor for soy protein is Nx5.71 , but there is currently no such recognised factor for rapeseed.
  • oil means an oil that is liquid at room temperature.
  • oil is a plant oil.
  • the oil is chosen from the group consisting of rapeseed oil, avocado oil, corn oil, olive oil, soya bean oil, sunflower oil, grapeseed oil, palm oil, peanut oil, walnut oil, coconut oil, line seed oil, camelina oil, groundnut oil, cotton seed oil, safflower oil, sesame oil and rice bran oil.
  • rapeseed oil avocado oil, corn oil, olive oil, soya bean oil, sunflower oil, grapeseed oil, palm oil, peanut oil, walnut oil, coconut oil, line seed oil, camelina oil, groundnut oil, cotton seed oil, safflower oil, sesame oil and rice bran oil.
  • the oil is rapeseed oil.
  • the present native rapeseed protein isolate comprises an oil content of 1.5 to 12 wt. % on dry matter of the rapeseed protein isolate.
  • the present native rapeseed protein isolate comprises an oil content of to 2.5 to 10 wt. % on dry matter of the rapeseed protein isolate.
  • the amount of oil is 1 to 9 wt. %, 2 to 8 wt. %, 2.5 to 7 wt. %, 3 to 6.5 wt. % or 4 to 6 wt. % on dry matter of the rapeseed protein isolate.
  • the amount of oil is 0.5, 1 , 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5 and/or 10 wt. % on dry matter of the rapeseed protein isolate.
  • the present native rapeseed protein isolate comprises saturated fatty acids, unsaturated fatty acids, omega fatty acids and/or trans fatty acids.
  • the present oil comprises saturated fatty acids, unsaturated fatty acids, omega fatty acids and/or trans fatty acids.
  • the saturated fatty acids comprise caproic acid, caprylic acid, capric acid, lauric acid, myristic acid, palmitic acid, stearic acid and/or arachidic acid.
  • the unsaturated fatty acids comprise monounsaturated (MLIFA) and/or polyunsaturated fatty acids (PLIFA).
  • the monounsaturated fatty acids comprise palmitoleic acid, oleic acid and/or erucic acid.
  • the omega fatty acids comprise omega-9 fatty acids, omega-6 fatty acids and/or omega-3 fatty acids.
  • the omega-3 fatty acids comprise eicosapentaenoic acid, docosahexaenoic acid, alphalinolenic acid, hexadecatrienoic acid, stearidonic acid, eicosatrienoic acid, eicosatetraenoic acid, heneicosapentaenoic acid, docosapentaenoic acid, tetracosapentaenoic and/or tetracosahexaenoic acid.
  • the omega-6 fatty acids comprise linoleic acid, gamma-linoleic acid, arachidonic acid, docosatetetraenoic acid, tetracosatetraenoic acid, tetracosapentaenoic acid and/or docosapentaenoic acid.
  • the present oil content is determined by FAME analysis, calculated as sum of individual fatty acids expressed as triglyceride equivalents, preferably according to AOAC method 996.06 (Association of Official Analytical Chemists).
  • the predominant storage proteins found in rapeseed seeds are cruciferins and napins.
  • Cruciferins are globulins and are the major storage protein in the seed.
  • Cruciferin is composed of 6 subunits and has a total molecular weight of approximately 300 kDa.
  • Napins are albumins and are a low molecular weight storage protein with a molecular weight of approximately 14 kDa.
  • Napins are more easily solubilized and in for example EP 1715752 a process is disclosed to separate out the more soluble napin fraction, preferably to at least 85 wt.%. Napins are primarily proposed for use in applications where solubility is key.
  • Rapeseed proteins can be also divided into various fractions according to the corresponding sedimentation coefficient in Svedberg units (S). This coefficient indicates the speed of sedimentation of a macromolecule in a centrifugal field.
  • S Svedberg units
  • the main reported fractions are: 12S, 7S and 2S.
  • Napin is a 2S albumin
  • cruciferin is a 12S globulin.
  • the present native rapeseed protein isolate comprises 40 to 65 wt. % cruciferins and 35 to 60 wt. % napins.
  • the present native rapeseed protein isolate comprises 40 to 55 wt. % cruciferins and 45 to 60 wt. % napins.
  • the present native rapeseed protein isolate comprises 60 to 95 wt. % cruciferins and 5 to 40 wt. % napins, such as 60 to 80 wt. % cruciferins and 20 to 40 wt. % napins.
  • the present native rapeseed protein isolate comprises 65 to 75 wt. % cruciferins and 25 to 35 wt. % napins.
  • the present native rapeseed protein isolate comprises 85 to 95 wt. % cruciferins and 5 to 15 wt. % napins.
  • the present native rapeseed protein isolate comprises 0 to 20 wt. % cruciferins and 80 to 100 wt. % napins, such as 0 to 15 wt. % cruciferins and 85 to 100 wt. % cruciferins, such as 0 to 10 wt. % cruciferins and 90 to 100 wt. % napins.
  • the present native rapeseed protein isolate comprises 1 to 5 wt. % cruciferins and 95 to 100 wt. % napins.
  • the amounts of cruciferins and napins are calculated based on the total amount of protein in the present isolate. Or alternatively, the amounts of cruciferins and napins are calcuated based on the sum of cruciferins and napins. Preferably, the amounts of cruciderins and napins are determined by size exclusion chromatography (SEC).
  • SEC size exclusion chromatography
  • the amounts of cruciderins and napins are determined by size exclusion chromatography (SEC) using the following test: samples of protein isolate are dissolved in a 500 mM NaCI saline solution and analyzed by High Performance SEC using the same solution as the mobile phase, followed by detection using UV absorbance at 220 nm, wherein the relative contribution of cruciferin and napin (wt. %) was calculated as the ratio of the peak area of each protein with respect to the sum of both peak areas, and absolute quantitation of cruciferin and napin was calculated using the response factor determined with a bovine serum albumin protein solution as an external calibration standard.
  • SEC size exclusion chromatography
  • the present native rapeseed protein isolate comprises 40 to 65 wt. % 12S and 35 to 60 wt. % 2S.
  • the present native rapeseed protein isolate comprises 40 to 55 wt. % 12S and 45 to 60 wt. % 2S.
  • the present native rapeseed protein isolate comprises 60 to 80 wt. % 12S and 20 to 40 wt. % 2S.
  • the present native rapeseed protein isolate comprises 65 to 75 wt. % 12S and 25 to 35 wt. % 2S.
  • the present native rapeseed protein isolate comprises 0 to 20 wt. % 12S and 80 to 100 wt. % 2S, such as 0 to 10 wt. % 12S and 90 to 100 wt. % 2S.
  • the present native rapeseed protein isolate comprises 1 to 5 wt. % 12S and 95 to 100 wt. % 2S.
  • the present native rapeseed protein isolate comprises 0.5 to 5 wt. % 12S and 80 to 90 wt. % 2S.
  • the amounts of 12S and 2S is determined by sedimentation velocity analytical ultracentrifugation (SV-ALIC) analysis.
  • the amounts of 12S and 2S is determined by sedimentation velocity analytical ultracentrifugation (SV-ALIC) analysis using the following test: samples of protein isolate are dissolved in a 3.0% (or 500 mM) NaCI saline solution and amounts determined using interference optics, as described in example 19 of patent US 8,623,445.
  • the present native rapeseed protein isolate comprises a conductivity in a 2 wt.% aqueous solution of less than 9000 pS/cm over a pH range of 2 to 12. More preferably the conductivity of the native rapeseed protein isolate in a 2 wt. % aqueous solution is less than 4000 pS/cm over a pH range of 2.5 to 11.5. For comparison the conductivity of a 5 g/l NaCI aqueous solution is around 9400 pS/cm. Preferably conductivity is measured with a conductivity meter, for example Hach senslON+ EC71.
  • the present native rapeseed protein isolate comprises a solubility of at least 88 % when measured over a pH range from 3 to 10 at a temperature of 23 +1-2 °C.
  • This is also known as the soluble solids index (SSI).
  • solubility is calculated by:
  • Protein solubility (%) (concentration of protein in supernatant (in g/l) I concentration of protein in total dispersion (in g/l)) x 100.
  • the solubility is measured using the following test: -sufficient protein isolate to supply 0.8 g of protein is weighed into a beaker; -a small amount of demineralized water is added to the powder and the mixture is stirred until a smooth paste is formed;
  • the dispersion is slowly stirred for at least 30 min using a magnetic stirrer
  • the pH is determined and adjusted to the desired level (2, 3, 4, etc.) with NaOH or HOI;
  • the pH of the dispersion is measured and corrected periodically during 60 minutes stirring; -after 60 minutes of stirring, an aliquot of the protein dispersion is reserved for protein concentration determination (Kjeldahl or Dumas analysis; N x 6.25), another portion of the sample is centrifuged at 20,000 G for 2 min;
  • Protein solubility (%) (concentration of protein in supernatant (in g/l) I concentration of protein in total dispersion (in g/l)) x 100.
  • the present native rapeseed protein isolate has a phytate level less than 5 wt.%, preferably less than, 4, 3, 2, 1 , 0.5, 0.4, 0.3, 0.2. 0.1 or less than 0.01 wt. %.
  • the present native rapeseed protein isolate has a phytate level of 0.01 to 4, 0.05 to 3, 0.1 to 1 wt.%.
  • the phytate level is measured using Eurofins method QD495, based on Ellis et al, Analytical Biochemistry Vol. 77:536-539 (1977), or 31 P-NMR determination.
  • the present native rapeseed protein isolate has a phenolic content of less than 1 wt.% on dry matter expressed as sinapic acid equivalents. Preferably less than 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1 , 0.05 or less than 0.01 wt.% on dry matter expressed as sinapic acid equivalents.
  • the present native rapeseed protein isolate comprises ⁇ 10 ppm gliadin.
  • the native rapeseed protein isolate comprises less than 5 ppm gliadin and most preferably no gliadin can be detected.
  • gliadin content is determined using sandwich ELISA from R-Biopharm (cat no R7001 , lot 14434) used according to the manufacturer’s instructions to determine the gliadin ppm in extracts.
  • the present native rapeseed protein isolate comprises an amount of moisture of less than 9%, such as less than 8, 7, 6, 5, 4, 3, 2, or 1%.
  • the native rapeseed protein isolate of the invention can be made by for example blending the present rapeseed protein isolate with the present oil, up to the desired amounts.
  • the present invention relates to a food product comprising a native rapeseed protein isolate as defined herein.
  • Rapeseed protein isolate was prepared from cold-pressed rapeseed oil seed meal as described in WO 2018/007492; the protein content was 90% (w/w).
  • the resultant rapeseed protein isolate comprised in the range of from 40 to 65% (w/w) cruciferins and 35 to 60% (w/w) napins, contained less than 0.26% (w/w) phytate and had a solubility of at least 88% when measured over a pH range from 3 to 10 at a temperature of 23 ⁇ 2°C.
  • rapeseed protein isolate was dispersed in 50ml solutions with various concentrations of rapeseed oil (Brassica Oil, Deventer, The Netherlands). The details of the oil concentrations are shown in table 1 below.
  • Odour intensity dissolved rapeseed protein isolate has a particular odour, the strength of a 2% solution in tap-water was set at 10
  • rapeseed protein isolate was added to 49ml rapeseed oil-tap water dispersions according to the concentrations in table 1.
  • the dispersions were slowly stirred for at least 30 min using a magnetic stirrer. After 30 mins the odour and taste of the solutions were evaluated and scored relatively to the 2% rapeseed protein isolate solution in tap water (i.e. the 0% oil in table 1).
  • the odour intenstity and lingering aftertaste decreased, whereas the watery taste (more bland, neutral taste) increased. This improved further with increasing oil concentrations.
  • the addition of rapeseed oil reduces undesired plant protein flavours and increases a more bland, neutral flavour.
  • Odour intensity dissolved rapeseed protein isolate has a particular odour, the strength of a 2% solution in tap-water was set at 10
  • Odour intensity dissolved rapeseed protein isolate has a particular odour, the strength of a 2% solution in tap-water was set at 10
  • rapeseed protein isolate was added to 49ml rice bran oil-tap water dispersions according to the concentrations in table 3. The dispersions were slowly stirred for at least 30 min using a magnetic stirrer. After 30 mins the odour and taste of the solutions were evaluated and scored relatively to the 2% rapeseed protein isolate solution in tap water (i.e. the 0% oil in table 2). Surprisingly, already after addition of 2% rice bran oil, both odour intenstity and the taste intensity were reduced, which further reduced with increasing oil concentrations. Hence, the addition of rice bran oil reduces undesired plant protein flavours.
  • Odour intensity dissolved rapeseed protein isolate has a particular odour, the strength of a 2% solution in tap-water was set at 10
  • the % dry matter was determined, the DUMAS method was used to determine the wt% of total nitrogen on dry matter, the wt% total protein on dry matter was calculated as Dumas N x 6.25, and the wt% of total fat on dry matter was determined with the FAME method (see table 5).
  • Dissolved rapeseed protein isolate (the reference sample) has a characteristic taste, which has a certain intensity and astringency (i.e., drying-out, roughening and puckery sensation felt in the mouth).
  • the intensity of a 2% solution in tap-water was set at 10. Both aspects are gradually reduced with increasing amounts of oil present.

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  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Food Science & Technology (AREA)
  • Polymers & Plastics (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
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  • Mycology (AREA)
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  • Agronomy & Crop Science (AREA)
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Abstract

La présente invention concerne un isolat de protéine de colza native comprenant une teneur en huile de 1 à 15 % en poids sur la matière sèche de l'isolat de protéine de colza, l'isolat ayant une teneur en protéines d'au moins 90 % en poids (calculée en tant que Dumas N x 6,25) sur une base de poids sec, et l'isolat comprenant une teneur en humidité inférieure à 10 % en poids.
PCT/EP2022/077294 2021-09-30 2022-09-30 Isolat de protéine de colza WO2023052590A1 (fr)

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EP21200364.4 2021-09-30

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EP1715752A1 (fr) 2004-01-20 2006-11-02 Burcon Nutrascience (MB) Corp. Nouvel isolat de la proteine du colza canola
WO2008094434A2 (fr) 2007-01-26 2008-08-07 Archer-Daniels-Midland Company Compositions comprenant de l'isolat de protéine de froment et procédés s'y rapportant
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US20150073127A1 (en) * 2011-07-28 2015-03-12 Piotr Wnukowski Protein isolation from oil seeds
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US20210137134A1 (en) * 2017-07-10 2021-05-13 Napiferyn Biotech Sp. Z O.O Method for isolation of protein from plant material
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