OILSEED PROTEIN CONCENTRATE AND PROCESS FOR THE PRODUCTION THEREOF

Abstract

A process for producing a protein concentrate from oilseed from a plant of a family selected from the group consisting of Brassicaceae, Cannabaceae, Pedaliaceae, Rosaceae, and Fabaceae, preferably Brassicaceae. The method comprises the successive steps of: a) providing a presscake from at least partially dehulled oilseed; b) washing said presscake by mixing it with a first hydrous-alcohol solvent to obtain a washed presscake, and eventually separating said washed presscake from said first hydrous-alcohol solvent, to obtain a washed, an eventually separated, presscake, wherein said first hydrous-alcohol solvent has an alcohol concentration ranging from 60 to 75% w/w; and c) washing said washed, and eventually separated presscake by mixing it with a last hydrous-alcohol solvent to obtain a protein-containing solid and separating said protein-containing solid from said last hydrous-alcohol solvent to obtain a protein concentrate, wherein the alcohol concentration of the final hydrous-alcohol solvent is about the concentration of the azeotrope.

Claims

1. A process for producing a protein concentrate from oilseed from a plant of a family selected from Brassicaceae, Cannabaceae, Pedaliaceae, Rosaceae, or Fabaceae, said process comprising the successive steps of: a) providing a presscake from at least partially dehulled oilseed; b) washing said presscake by mixing it with a first hydrous-alcohol solvent to obtain a washed presscake, and eventually separating said washed presscake from said first hydrous-alcohol solvent, to obtain a washed, and eventually separated, presscake, wherein said first hydrous-alcohol solvent has an alcohol concentration ranging from 60% to 75% w/w; and c) washing said washed, and eventually separated presscake by mixing it with a last hydrous-alcohol solvent to obtain a protein-containing solid and separating said protein-containing solid from said last hydrous-alcohol solvent to obtain a protein concentrate, wherein the alcohol concentration of the final hydrous-alcohol solvent is about the concentration of the azeotrope.

2. The process according to claim 1, wherein the seed is a seed of plant of the family Brassicaceae.

3. The process according to claim 1, wherein said separated washed presscake of step b) is submitted before step c) to a first additional washing step b1) by mixing it with a second hydrous-alcohol solvent and then separating it from said second hydrous-alcohol solvent.

4. The process according to claim 1, wherein said separated washed presscake of step b1) is submitted before step c) to a second additional washing step b2) by mixing it with a third hydrous-alcohol solvent and then separating it from said third hydrous-alcohol solvent.

5. The process according to claim 1, wherein the presscake of step a) has an oil content ranging from 4% to 25% dry w/w.

6. The process according to claim 1, wherein the oilseed presscake is obtained by double pressing with a preliminary cooking step carried out at a temperature ranging from 70° C. to 110° C., said double pressing being carried out using a first pressing temperature ranging from 50° C. to 110° C., and a second pressing temperature of less than 110° C. but which is higher than the first pressing temperature.

7. The process according to claim 1, wherein said hydrous-alcohol solvent comprises an alcohol which is a C1 to C3 aliphatic alcohol.

8. The process according to claim 1, wherein said first hydrous-alcohol solvent has a concentration in alcohol ranging from 60% w/w to the alcohol azeotrope, and/or wherein said final hydrous-alcohol solvent has a concentration in alcohol ranging from 60% w/w to the alcohol azeotrope.

9. The process according to claim 1, wherein at least one washing step is carried out a temperature ranging from 50° C. to 70° C.

10. The process according to claim 1, wherein said step c) comprises a desolventizing step whereby said last hydrous-alcohol solvent is completely removed from said protein-containing solid.

11. The process according to claim 1, further comprising at least one of the following steps prior to step a): i) cleaning the seed, iii) flaking the seed, and/or iv) cooking the seed.

12. The process according to claim 1, wherein said protein concentrate is subjected to at least one sieving step, optionally preceded by at least one milling step.

13. An oilseed protein concentrate obtained, or obtainable, by the process according to claim 1.

14. An oilseed protein concentrate according to claim 13, said concentrate comprising a protein content from 40% to 69% on dry matter w/w, a mono- and di-saccharide total content of less than 1% w/w on DM, and a total oligosaccharide content of less than 1% dry matter w/w, wherein said oilseed is from a plant from a family selected from Brassicaceae, Cannabaceae, Pedaliaceae, Rosaceae, or Fabaceae.

15. A method for preparing a food product, a feed product or an aquafeed product, comprising adding and/or mixing the oilseed protein concentrate of claim 13 to other ingredients to prepare a food product, a feed product or an aquafeed product.

16. A food ingredient or a foodstuff comprising the oilseed protein concentrate according to claim 13.

17. The process according to claim 1, wherein the seed is a seed of the genus Brassica.

18. The process according to claim 1, wherein the oilseed presscake is obtained by double pressing with a preliminary cooking step carried out at a temperature ranging from 80° C. to 86° C., said double pressing being carried out using a first pressing temperature ranging from 67° C. to 72° C., and a second pressing temperature of 80° C. to 90° C.

19. The process according to claim 1, wherein said hydrous-alcohol solvent comprises ethanol, isopropanol, methanol, or a mixture thereof.

20. The process according to claim 1, wherein all of the washing steps are carried out a temperature ranging from 50° C. to 70° C.

Description

BRIEF DESCRIPTION OF THE FIGURES

[0103] FIG. 1 shows the minimum Gelling concentration of the rapeseed protein concentrate of Example 1 (top) and example 2 (bottom).

EXAMPLE 1—PRODUCTION OF RAPESEED PROTEIN CONCENTRATE (RPC)

[0104] The starting material was French Rapeseed (Brassica napus) and its composition is presented in Table 1

TABLE-US-00001 TABLE 1 Analysis of the rapeseed sample. Analysis French Rapeseed Moisture (%).sup.a 6.8% Crude Protein (% DM).sup.b 21.0% Oil Content (% DM).sup.c 46.0% .sup.adetermined by the moisture balance method NF EN ISO 6498 (2012) .sup.bDry Weight, or Dry Matter (DM) protein content, was determined by the Dumas method (N × 6.25) using a DUMATHERM apparatus (Gerhardt GmbH & CO KG (Germany)) (Norme AFNOR) NF EN ISO 16634-1. .sup.cdetermined by the Soxhlet method (ISO734: 2016).

Dehulling

[0105] The rapeseed was dehulled using an impact huller. The mixture (almond, hulls) obtained was separated using a Denis D50 separator (from the French company DENIS). The following settings were used: [0106] Upper screen: wide open; [0107] Lower screen: 1 mm round; [0108] Front suction: 1; and [0109] Rear suction: 0

[0110] After hulls separation, the kernels obtained had a purity of 84%. To measure the purity, an internal image analysis method was used. This method consists in placing a sample of dehulled seeds in a scanner and capturing the image. The obtained image is processed with the image processing software Image J. The difference in colour or the proportion of yellow surface when compared to reference images corresponds to the kernel purity of the obtained mixture.

[0111] The mass balance in the dehulling process in given in table 2.

TABLE-US-00002 TABLE 2 Dehulling step Flow Weight (kg) Rapeseed In 1007 Dehulled seed Out 713 Recycled seed Out 102 Hulls 174 Losses 18

Pressing

[0112] The dehulled kernels were crushed to extract the oil by using a double pressing process in semi-continuous mode using a screw press MBU75 sold by the French Company OLEXA (Arras, FR) and for the second stage a small press from the same company: MBU 20.

[0113] The rapeseed were first preheated at 80-86° C. to feed the MBU75 press. For the first pressing step, the temperature within the press was stabilised at 70° C. and the rotation speed was stabilised at 23 Hz. The outflow of press cake and oil was around 60-70 kg/h and 39-44 kg/h, respectively. During the second pressing step where the smaller press (MBU 20) was used, the rotating frequency was set at 26 Hz and the temperature stabilised around 86° C. In these conditions, the rapeseed cake flow rate was around 54-59 Kg/h and the oil flow rate around 8.5-10 Kg/h. The composition of the final rapeseed press cake (pressed meal) thus obtained is presented in Table 3.

TABLE-US-00003 TABLE 3 Dry Protein Glucosinolate matter Fat content Solubility (μmol/g of (wt %) (%/DM) (%/DM) (%) dry solid) 92.50 11.47 38.42 57.9 35.9

[0114] Hydrous-Alcohol Solvent Extraction/Protein Purification

[0115] 4 different washing steps, each using a solvent having different alcohol contents, were successively carried out on the rapeseed press cake in order to wash, or purify, the proteins. These solvents were a mixture of water and ethanol having the following ethanol proportions 65%, 70%, 80% and 96% by weight. These solvent/washing steps and the key parameters for each step are presented in Table 4. 91 Kg of the Rapeseed press cake were used for extraction and protein concentration.

TABLE-US-00004 TABLE 4 Parameters Washing1 Washing2 Washing3 Washing4 Ethanol content in 65% 70% 80% 96% solvent in wt % Solvent: rapeseed press 8.2:1 8.1:1 8.1:1 7.5:1 cake ratio (weight) Mass of solvent (Kg) 748 733 735 681 Extraction temperature 65° C.   65° C.   65° C.   65° C.   (° C.) Extraction time (minutes) 72 min. 60 min. 60 min. 60 min.

[0116] The washing steps which are separately described below were used to remove the oil from the cake, as well as small soluble components such as polyphenols, sugars, salts etc. The washing, or extracting, steps were carried out in an agitated filter tank (Guedu—capacity 400 litres—supplier De Dietrich). This device is equipped with a vertically mobile stirrer, a jacket for hot steam, or water, circulation in order to heat the mix and a cotton cloth (De Dietrich) having a 10 μm mesh allowing for liquid—solid separation. During each washing step, or extraction, step, the pressed meal was immersed in the solvent at a specific temperature, according to a determined time, and under an agitation of 21-24 rpm.

[0117] Step 1: Ethanol 65%

[0118] The first solvent used was 65% ethanol in water. The temperature set was ranging from 55-65° C. The rapeseed press cake:liquid solvent ratio was 1:8 (w/w) and the total processing time included solvent heating was about 72 minutes. A solid/liquid separation step was then carried out by filtration (percolation).

[0119] Step 2: Ethanol 70%

[0120] The solid from the extraction step 1 obtained was then mixed with a 70% ethanol solution according to a rapeseed press cake:liquid solvent ratio of 1:8. The temperature of this washing step was 65° C. The total processing time included solvent heating was about 90 minutes. A solid/liquid separation step was then carried out by filtration (percolation).

[0121] Step 3: Ethanol 80%

[0122] The solid from extraction step 2 was mixed with an 80% ethanol solution according to a solid:liquid weight ratio of 1:8. The extraction temperature was around 65° C. and the extraction time, including solvent heating, was 90 minutes. A solid/liquid separation step was then carried out by filtration (percolation).

[0123] Step 4: Ethanol 96%

[0124] The solid from extraction step 3 obtained was mixed with a 96% ethanol solution according to a solid:liquid weight ratio of 1:7.5. The extraction temperature was around 65° C. and the extraction time, including solvent heating, was 90 minutes. A solid/liquid separation step was then carried out by filtration (percolation).

[0125] Desolventizing

[0126] The solid fraction obtained by filtration (percolation) from step 4 was desolventised (i.e. the solvent is removed) in the filter tank used to carry out the extraction steps (Guedu; capacity 400 litres—De Dietrich Process Systems). Upon agitation, a vacuum is applied (0.8 bar) at 53° C. for more than 10 hours. A final rapeseed concentrate is thus obtained. The dry matter percentage of the obtained concentrate was 88.4%.

[0127] The mass balance of each extraction step is shown in Table 5 below:

TABLE-US-00005 TABLE 5 Flux Extraction step 1 2 3 4 in Press cake weight  91 Kg 1 1 1 Solvent weight 748 Kg 733 Kg 735 Kg 681 Kg Out Miscella weight 660 Kg 660 Kg 742 Kg 683 Kg Miscella dry matter 1.94% 0.85% 0.31% 0.57%

EXAMPLE 2—DRY FRACTIONATION (SIEVING) OF THE RAPESEED PROTEIN CONCENTRATE OBTAINED IN EXAMPLE 1

[0128] The concentrate obtained by the process of Example 1 was or deagglomerated and then sieved using a 180 microns sieve in order to obtained a protein-rich or protein containing fraction. The fiber-rich fraction is retained by the sieve and the protein-rich fraction pass through the sieve.

[0129] The protein-rich fraction obtained after sieving was analysed. The protein content of this fraction was determined as described in Example 1. It has a protein content of 55.5% on dry matter (to compare to the protein content of 51.3% before sieving; see Example 3 below).

EXAMPLE 3—CHARACTERIZATION OF THE RAPESEED PROTEIN CONCENTRATES OF THE INVENTION

3.1 Composition of the Rapeseed Protein Concentrates (RPC) Obtained in Example 1 and 2 (Unsieved and Sieved RPC)

[0130]

TABLE-US-00006 TABLE 6 Composition of non-sieved and sieved rapeseed protein concentrate Non-sieved Sieved RPC RPC Analysis and method used (example 1) (example 2) Dry Matter (%) - Dessic 4 h/103° C. NF ISO 6496 (10/2011) 91.8 91.9 Protein (% DM) - NF EN ISO 16634 (2008) 51.3 55.5 Lipids (% DM) - CEE98/64 (light petroleum) 1998 2.4 3.2 Ash (% DM) - at 550° C. NF v 18-101 1977 9.0 9.0 Phosphorus (% DM) - ICP microwave NF EN ISO 15621 (2012) 1.6 1.7 Cellulose (% DM) - Wendee Gravimetric NF V 03-040 7.8 8.1 Neutral Detergent Fibre- NDF (% DM) - Acid hydrolysis NF 26.9 18.0 V 18-122 (2013) Energy (kJ/g DM) - bomb calorimeter test 15.1 15.3 Acid Detergent Fibre - ADF (% DM) - Acid Hydrolysis NF V 18.0 10.9 18-122 (2013) Acid detergent Lignin - ADL (% DM) - Meth. Acid hydrolysis 8.7 3.4 NF V18-122 (2013) Glucosinolates (μmol/g DM) - dil. methanol - Internal 3.8 ND Enzymatic method ISO 9167 - HPLC (2013) Polyphenols (% DM) 0.1 0.1 Phytic acid (% DM) - Brooks et al., 2001 5.3 5.8 Arginine - HPLC - NF EN ISO 13903 (2005) 2.9 3.3 Histidine - HPLC - NF EN ISO 13903 (2005) 1.3 1.5 Isoleucine - HPLC - NF EN ISO 13903 (2005) 1.9 2.1 Leucine - HPLC - NF EN ISO 13903 (2005) 3.5 3.9 Lysine - - HPLC - NF EN ISO 13903 (2005) -react 3.0 3.3 ninhydrin/detect. 570 nm. Methionine - Method XP V 18-113 NF EN ISO 13903 (2005) 1.0 1.2 Phenylalanine - NF EN ISO 13903 (2005) 1.9 2.1 Threonine - NF EN ISO 13903 (2005) 2.2 2.4 Valine - HPLC - NF EN ISO 13903 (2005) 2.4 2.7 Sugar profiles - Luff Schoorl Glucose (%/DM) <0.2 <0.2 method as described in UE Fructose (%/DM) <0.2 <0.2 Regulation 152/2009 Sucrose (%/DM) 0.22 0.22 Lactose (%/DM) <0.2 <0.2 Maltose (%/DM) <0.2 <0.2

[0131] Total polyphenols content: measured by the following colorimetric protocol:

Reference

[0132] 0.15 g of gallic acid (from Riedel Haen, ref. 27645) mixed with 100 ml of water is further diluted in 900 mL of water (ratio 1/10). Then 50 mL vials containing 0.0 mL, 0.5 mL, 1 mL and 2 mL of this solution are further diluted with 30 mL of water. A 1 mL aliquot from each vial is poured in a 50 ml vial and 30 ml of water are added. Then, 2.5 ml of Folin-Ciocalteu reagent (2N Sigma ref. F9252) is added to the vial and the vial is agitated. 7.5 ml of a solution of sodium carbonate (20% (w:w) of anhydrous sodium carbonate) are then added. After 30 seconds, the mixture is agitated and then let to rest for 1 H30 mn at room temperature. The total volume is then adjusted to 100 mL by adding more water to the vials. Absorbance of these solutions is measured at 765 nm using UV/VIS spectrophotometer against distilled water and a standard or calibration curve is obtained.

[0133] Samples:

[0134] The samples are prepared by grinding each of the solid samples with a ball mill. The powder used has a maximum size of the particles of 1 mm. 1 g of this powder is weighted in a 100 mL vial. Close to 100 mL of a mixture of MeOH/water/acetone/HC(1N), having the relative proportion 40/38/20/2, (v:v), is added to the sample. The vial's content is agitated for 1 h at 60° C. in a bain-marie. The total volume is then adjusted to 100 mL by adding more solvent.

[0135] A 1 mL aliquot from each vial is poured in a 50 ml vial and 30 ml of water are added. Then, 2.5 ml of Folin-Ciocalteu reagent (2N Sigma ref. F9252) is added to the vial and the vial is agitated. 7.5 ml of a solution of sodium carbonate (20% (w:w) of anhydrous sodium carbonate) are then added. After 30 seconds, the mixture is agitated and then let to rest for 1H30 mn at room temperature. The total volume is then adjusted to 100 mL by adding more water to the vials. Absorbance of these solutions is measured at 765 nm using UV/VIS spectrophotometer against distilled water. The total phenolic content was read against the calibration curve and the results are expressed as micrograms per milliliter of gallic acid equivalents (GAE) (μg/mL of GAE) or converted on percentage. The measures are duplicated.

3.2 Functional Characteristics: Measurement Methods

[0136] a) Emulsifying Capacity

[0137] The emulsifying capacity represents the maximum amount of oil that can be emulsified as an oil-in-water emulsion by a protein sample. A 1% (w/w) protein equivalent (PE) was prepared in water. After 1 h of solubilisation by stirring in a Rheax (orbital shaker), the protein solution was centrifuged at 6000 rpm with an IKA shaker. Sunflower oil was continuously added in the solution at a flow of 50 mL/min to form an oil-in-water emulsion. The amount of oil required by trigger phase inversion (formation of a water-in-oil emulsion) was assessed visually. Sodium casein is the reference for this test.

[0138] b) Protein Solubility

[0139] The protein solubility was tested on the supernatant obtained after centrifugation 15000 g (10 min.) of a 2% (w/w) protein equivalent solution obtained as mentioned in point a above. The solubility of the supernatant was tested at pH 3, 4, 5, 6, 7 and 8. The amount of nitrogen was measured on the supernatant obtained after centrifugation and the proportion of protein's solubilised estimated by Kjeldahl method (i.e. [protein]=6.25 [nitrogen]).

[0140] c) Apparent Viscosity

[0141] Apparent viscosity is defined as the viscosity of a protein solution measured at a shear rate of 100 s.sup.−1. A 10% or 15% (w/w) protein solution of the concentrate was prepared in water and solubilised in a Rheax (orbital shaker). Viscosity of the solution thus obtained was measured at 25° C. at a shear rate of 100 s.sup.−1 for 600 seconds using a Vane cup geometry on a DHR-2 rheometer (TA).

[0142] d) Differential Scanning Calorimetry (DSC)—Thermal Stability

[0143] DSC is used to measure the thermal stability of a sample. A 10% (w/w) protein aqueous solution was prepared and solubilised during 1 hour in a Rheax (orbital shaker). It was then placed in a DSC cell, heated from 30° C. to 140° C. at a gradient of 0.4° C./min, and cooled from 140° C. to 30° C. at a gradient of 2.5° C./min. When an endothermic peak is observed, the following parameters are measured: [0144] Denaturation temperature (Td); [0145] Specific heat (ΔH), which is the enthalpy associated to the denaturation of protein; [0146] Onset temperature of denaturation (T onset); [0147] End temperature of denaturation (T offset)

[0148] e) Water Holding Capacity

[0149] Water Holding Capacity (WHC) is the amount of water that can be bound by protein. A 2% (w/w) of ingredient is dissolved in water and the solution is and solubilised at 500 rpm during 1 h. The solution was then centrifuged at 5000 g during 30 min. The amount of water in the supernatant was measured. The amount of water in pellet was deduced by difference. WHC is calculated as the mass of water held in pellet per mass of sample (g/g).

[0150] f) Minimum Gelling Concentration of a Rapeseed Protein Concentrates

[0151] Minimum gelling concentration was measured by preparing solutions of the concentrate of example 1 and 2 from 2% (w/w) protein content to 20% protein content, in increment of 2%, in test tubes (PR-18009) at pH 6. After solubilisation, the solutions were heated 1 h in a water-bath at 85° C. and then cooled 2 h at 4° C. A solution was considered to have formed a gel if it behaved like a liquid before heating (i.e., free-flowing) and did not flow when test-tube was put upside-down after heating.

3.2 Functional Analyses

[0152] a) Emulsifying Capacity

[0153] Emulsifying capacity (oil g) for RPCs was 60 mL for the sieved concentrate of Example 2 and 81 mL for non-sieved concentrate of Example 1. This is respectively 12 and 9 times lower than the emulsifying capacity of sodium caseinate (774 mL).

[0154] b) Nitrogen Solubility Profile

[0155] Both concentrates (example 1 and 2), with and without a sieving step, were insoluble (<20% solubility) at all pH tested.

[0156] c) Viscosity

[0157] The apparent viscosity of a solution at 10% and 15% (w/w) solid in water at 25° C. of the concentrate of Example 1 at a shear rate of 100 s.sup.−1, was 436 cP and 226 cP respectively.

[0158] The apparent viscosity of a solution at 10% and 15% (w/w) solid in water at 25° C. of Example 2 at a shear rate of 100 s.sup.−1, was 14 cP and 141 cP.

[0159] For reference, at this shear rate, the apparent viscosity of water is 1 cP, sunflower oil 30 cP and ketchup 750 cP.

[0160] d) DSC

[0161] No endothermic phenomenon was observed on DSC thermogram for the concentrates of Examples 1 and 2. The concentrates were therefore thermally stable. This suggests that proteins were already denatured.

[0162] e) Water Holding Capacity

[0163] The concentrate of example 1 had a WHC (g/g) of 3.7 g/g, which means that 1 g of RPC can retain 3.7 g of water.

[0164] The concentrate of example 2 had a WHC (g/g) of 4.1 g/g.

[0165] f) Minimum Gelling Concentration of a Rapeseed Protein Concentrates

[0166] The minimum gelling concentration is difficult to distinguish for both RPC as the materials are heterogeneous. At 12% total matter (6% protein content) a solid mass can be observed at the top of the tube and no liquid is observable at the bottom of the tube. No significant difference is clear between the two concentrates except their appearance (see FIG. 1 wherein reference C4633 is the RPC obtained from Example 1 and C4633-1 is the sieved RPC obtained from Example 2).

3.3 Conclusion

[0167] The rapeseed protein concentrate obtained in Examples 1 and 2 are thermally stable, probably denatured, insoluble in water at all pH and have little emulsifying capacity. They have a very high water absorption capacity and form a highly viscous suspension. The viscosity of the RPC obtained from example 1 is higher than that of the sieved RPC obtained in example 2. Due to their functional properties, they can be used as an ingredient for making food and feed products.

EXAMPLE 4—MANUFACTURING OF FISH DIETS

[0168] Ingredients: fishmeal (37.5% w/w), rapeseed protein concentrate (RPC from example 1 or sieved RPC from example 2) (25% w/w), wheat meal (15% w/w), wheat gluten (7.5% w/w), soybean meal (7.5% w/w), fish hydrolysates (for example CPSP® G from Sopropeche (France)) (3.5% w/w), vitamin & mineral premix (3.5% w/w).

[0169] Diets are usually manufactured by extrusion. All powdered ingredients can be mixed in relative proportions accordingly to the target formulation in a double-helix mixer and ground (below a particle size 400 μm) in a micropulverizer hammer mill. Pelletized compounds (pellet size: 2.5 mm) are manufactured with a twin-screw extruder (model BC45, Clextral, France) with a screw diameter of 55.5 mm. Extrusion conditions: feeder rate (80-85 kg/h), screw speed (247-266 rpm), water addition in barrel 1 (345 ml/min), temperature barrel 1 (32-34° C.), temperature barrel 3 (111-117° C.). Extruded pellets are dried in a ventilated drying oven (35° C.) before bagging.

[0170] These fish diets are satisfactory in term of protein content and nutrient digestibility in trout (Oncorhynchus mykiss). The rapeseed protein concentrates according to the invention used at around 25% w/w in a fish diet are a reliable alternative to soy protein concentrate, in particular for trout aquafeed. This also indicates that RPCs of the invention are very interesting raw materials for salmonid feedstuffs.

[0171] The rapeseed protein concentrate (RPC) described in this Example can be replaced in the fish diet, at least in part, by a protein concentrate obtained from a seed of a plant of the family Brassicaceae, in particular of the genus Brassica (e.g., Brassica juncea or Brassica carinata) according to the process of the invention.

REFERENCES

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