Reduction of vicine and / or convicine in faba ingredients and food products

Abstract

The invention relates to a method of reducing vicine and / or convicine content in a faba ingredient. A liquid faba ingredient is provided and then at least one vicine and / or convicine-degrading enzyme is added to the liquid faba ingredient having a pH of 6.0 or higher. The faba ingredient is then incubated with the at least one vicine and / or convicine-degrading enzyme. The invention also relates to faba ingredient, to food products prepared with faba ingredient and method for producing such food products.

Description

[0001] REDUCTION OF VICINE AND / OR CONVICINE IN FABA INGREDIENTS AND FOOD PRODUCTS

[0002] TECHNICAL FIELD

[0003] The present invention relates generally to the field of faba ingredient or food product prepared with faba ingredient. In particular, it relates to faba ingredient or food product with reduced level of vicine and / or convicine and method / process for producing such a faba ingredient or such a food product.

[0004] BACKGROUND OF THE INVENTION

[0005] Faba bean is a very good source of protein for food products, in particular plant-based food products. One drawback is that naturally present antinutritional factors vicine and convicine make faba (and its derived ingredients, e.g. protein concentrate and isolate) unsuitable for consumption by sufferers of favism. Reducing the levels of vicine and convicine would enable the incorporation of faba into food products that may be safe for consumption, especially for individuals suffering from favism.

[0006] Different strategies are explored to reduce levels of vicine and convicine in faba.

[0007] One strategy consists of the use of microorganism to cleave and to reduce vicine and convicine in faba via fermentation. However, fermentation requires long time to achieve a significant reduction of vicine and convicine. In addition, fermentation is not a specific reaction and may target compounds other than vicine and convicine in faba. This may modify the sensory and nutritional properties of faba. In particular, it may impart undesirable flavours, including fermented flavours. In addition, fermentation reduces the pH of faba which may be undesirable from a stability standpoint or for food application.

[0008] Another strategy consists of the use of enzyme to cleave and to reduce vicine and convicine in faba via hydrolysis. For example, p-glucosidase may be used to cleave the glucose group from vicine and convicine molecules. The reaction products are the aglycone forms of vicine and convicine and glucose.

[0009] Enzymatic degradation of vicine and convicine is generally performed in the art at acidic pH. In particular, documented enzymatic degradation of vicine and convicine take several hours (2h-4h) at acidic pH (pH 4.6 or pH 5) (Arbid and Marquardt 1985; Pulkkinen 2019).

[0010] The use of acidic pH is not advantageous. Indeed, this decreases the stability of proteins in faba over processing and / or the shelf-life. Indeed, under acidic pH, faba bean proteins which have a isoelectric point around 4.5 are more prone to poor solubility, uncontrolled aggregation, precipitation and sedimentation which is undesirable over processing and / or the shelf-life. This adversely affects the sensory properties and functionality of ingredient and food products prepared with faba. The effect of pH on properties and functionality of faba proteins is well reported in the literature: Otegui, I. et et al., (1997). Properties of spray-dried and freeze-dried faba bean protein concentrates, International Journal of Food Science & Technology, 32(6):439-443; Martin Vogelsang-O'Dwyer, M. et al., (2020). Comparison of Faba Bean Protein Ingredients Produced Using Dry Fractionation and Isoelectric Precipitation: Techno-Functional, Nutritional and Environmental Performance. Foods, 9(3), 322; Hu et al., (2023). Formation and characterisation of concentrated emulsion gels stabilised by faba bean protein isolate and its applications for 3D food printing. Colloids and Surfaces A: Physicochemical and Engineering Aspects, 671, 131622; Mohanan, A. et al., (2020), Utilization of pulse protein-xanthan gum complexes for foam stabilization: The effect of protein concentrate and isolate at various pH. Food Chemistry, 316, 126282.

[0011] In addition, those skilled in the art will recognise that the activity of an enzyme is pH dependant. Enzymes that are active at acidic pH, may not be active or have considerably reduced activity, at other pHs, including neutral pH.

[0012] For food applications, it is desired to perform the enzymatic reaction in a food matrix at neutral, nearly neutral or mild alkaline pH, in particular pH above 6 and with short reaction times.

[0013] It would therefore be desirable to provide a method for reducing, preferably eliminating, vicine and / or convicine in faba ingredient and in food products prepared with such faba ingredient via enzymatic treatment. It is desirable that the enzymatic solution leads to a significant reduction, preferably elimination, of vicine and / or convicine at neutral, nearly neutral or mild alkaline pH, in particular pH above 6 within a short time.

[0014] Any reference to prior art documents in this specification is not to be considered an admission that such prior art is widely known or forms part of the common general knowledge in the field.

[0015] SUMMARY OF THE INVENTION

[0016] The object of the present invention is to improve the state of the art, and in particular to provide methods, food ingredients and food products that overcome the problems of the prior art and addresses the needs described above, or at least to provide a useful alternative. The inventors were surprised to see that the object of the present invention could be achieved by the subject matter of the independent claims. The dependent claims further develop the idea of the present invention.

[0017] Accordingly, a first aspect of the invention proposes a method of reducing vicine and / or convicine content in a faba ingredient, said method comprising the steps of: a) Providing a faba ingredient which is liquid or suspending a faba ingredient into an aqueous liquid to form a faba ingredient which is liquid, wherein the faba ingredient comprises vicine and / or convicine, b) Adding at least one vicine and / or convicine-degrading enzyme to the faba ingredient, wherein the faba ingredient has a pH of 6.0 or higher, c) Incubating the faba ingredient with at least one vicine and / or convicine-degrading enzyme to obtain a treated faba ingredient, d) Optionally, partially or fully inactivating the vicine and / or convicine-degrading enzyme in the treated faba ingredient, e) Optionally, cooling the treated faba ingredient.

[0018] A second aspect of the invention proposes a treated faba ingredient which is obtained with the method of the first aspect of the invention.

[0019] A third aspect of the invention proposes a faba ingredient, wherein the sum of vicine and convicine content is less than 600 mg per kg of faba ingredient, preferably less than 450 mg per kg of faba bean protein.

[0020] A fouth aspect of the invention proposes a food product which comprises the treated faba ingredient according to the second aspect of the invention or the faba ingredient according to the third aspect of the invention.

[0021] A fifth aspect of the invention proposes a method for producing a food product which comprises the step of: a) Providing a food composition, wherein the food composition comprises faba ingredient and at least one additional food ingredient, and wherein the faba ingredient of the food composition comprises vicine and / or convicine, b) Adding at least one vicine and / or convicine-degrading enzyme to the food composition, wherein the food composition has a pH of 6.0 or higher, c) Incubating the food composition with the at least one vicine and / or convicine- degrading enzyme to obtained a treated food composition, d) Optionally, homogenizing the treated food composition, e) Heat-treating the treated food composition, f) Cooling the treated food composition to obtain a food product.

[0022] A sixth aspect of the invention proposes a method for producing a food product which comprises the step of: a) Providing a food composition, wherein the food composition comprises the treated faba ingredient according to any one of claims 9 or 11-12 or the faba ingredient according to any one of claims 10 to 12, and also comprises at least one additional food ingredient, b) Optionally, homogenizing the food composition, c) Heat-treating the food composition, d) Cooling the food composition to obtain a food product.

[0023] A seventh aspect of the invention proposes a food product obtained by the method according to the fifth aspect of the invention or the sixth aspect of the invention.

[0024] It has been discovered that it was possible to achieve significant reduction of vicine and / or convicine at neutral, nearly neutral or mild alkaline pH, in particular pH above 6 within a short time in faba ingredient or food product comprising faba ingredient via enzymatic treatment with vicine and / or convicine-degrading enzyme.

[0025] These and other aspects, features and advantages of the invention will become more apparent to those skilled in the art from the detailed description of embodiments of the invention, in connection with the attached drawings.

[0026] BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 shows percentage of vicine reduction / degradation by enzymes E1-E6 at 60°C and at a pH 6.4 in faba suspension comprising faba concentrate containing low levels of vicine / convicine. The initial vicine content (100%) in faba concentrate with low levels of vicine / convicine before enzymatic treatment was 661 mg / kg of faba ingredient.

[0028] Figure 2 shows percentage of convicine reduction / degradation by enzymes E1-E6 at 60°C and at a pH 6.4 in faba suspension comprising faba concentrate containing low levels of vicine / convicine. The initial convicine content (100%) in faba concentrate with low levels of vicine / convicine before enzymatic treatment was 176 mg / kg of faba ingredient.

[0029] Figure 3 shows percentage of vicine reduction / degradation by enzymes E1-E6 at 60°C and at a pH 6.4 in faba suspension comprising faba concentrate containing high levels of vicine / convicine. The initial vicine content (100%) in faba concentrate with high levels of vicine / convicine before enzymatic treatment was 19363 mg / kg of faba ingredient.

[0030] Figure 4 shows percentage of convicine reduction / degradation by enzymes E1-E6 at 60°C and at a pH 6.4 in faba suspension comprising faba concentrate containing high levels of vicine / convicine. The initial convicine content (100%) in faba concentrate with high levels of vicine / convicine before enzymatic treatment was 5508 mg / kg of faba ingredient.

[0031] Figure 5 shows percentage of vicine reduction / degradation by enzymes E2 and E3 at 50°C, 55°C, 60°C and 65°C and at a pH 6.4 in faba suspension comprising faba concentrate containing low levels of vicine / convicine. The initial vicine content (100%) in faba concentrate with low levels of vicine / convicine before enzymatic treatment was 661 mg / kg of faba ingredient.

[0032] Figure 6 shows percentage of convicine reduction / degradation by enzymes E2 and E3 at 50°C, 55°C, 60°C and 65°C and at a pH 6.4 in faba suspension comprising faba concentrate containing low levels of vicine / convicine. The initial convicine content (100%) in faba concentrate with low levels of vicine / convicine before enzymatic treatment was 176 mg / kg of faba ingredient.

[0033] Figure 7 shows percentage of vicine reduction / degradation by enzymes E2 and E3 at 50°C, 55°C, 60°C and 65°C and at a pH 6.4 in faba suspension comprising faba concentrate containing high levels of vicine / convicine. The initial vicine content (100%) in faba concentrate with high levels of vicine / convicine before enzymatic treatment was 19363 mg / kg of faba ingredient.

[0034] Figure 8 shows percentage of convicine reduction / degradation by enzymes E2 and E3 at 50°C, 55°C, 60°C and 65°C and at a pH 6.4 in faba suspension comprising faba concentrate containing high levels of vicine / convicine. The initial convicine content (100%) in faba concentrate with high levels of vicine / convicine before enzymatic treatment was 5508 mg / kg of faba ingredient.

[0035] Figure 9 shows percentage of vicine reduction / degradation by enzymes E2 and E3 at 60°C and at a pH 6.4 in a food product, in particular plant-based milk analogue, containing faba concentrate with low levels of vicine and convicine. The initial vicine content (100%) in faba concentrate with low levels of vicine / convicine before enzymatic treatment was respectively 661 mg / kg of faba ingredient.

[0036] Figure 10 shows percentage of convicine reduction / degradation by enzymes E2 and E3 at 60°C and at a pH 6.4 in a food product, in particular plant-based milk analogue, containing faba concentrate with low levels of vicine and convicine. The initial convicine content (100%) in faba concentrate with low levels of vicine / convicine before enzymatic treatment was 176 mg / kg of faba ingredient.

[0037] DETAILED DESCRIPTION OF THE INVENTION

[0038] As used in the specification, the words "comprise", "comprising" and the like are to be construed in an inclusive sense, that is to say, in the sense of "including, but not limited to", as opposed to an exclusive or exhaustive sense.

[0039] As used in the specification, all numerical ranges should be understood to include each whole integer within the range.

[0040] As used in the specification, the singular forms "a", "an", and "the" include plural referents unless the context clearly dictates otherwise.

[0041] As used in the specification, the term "substantially free" means that no more than about 0.1, preferably 0.01 weight percent of the excluded material remains. "Entirely free" typically means that at most only trace amount of the excluded material is present, and preferably, no detectable amount is present.

[0042] Unless noted otherwise, all percentages in the specification refer to weight percent, where applicable.

[0043] Unless defined otherwise, all technical and scientific terms have and should be given the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0044] The term "faba ingredient" is an ingredient comprising, preferably consisting of compounds originated from faba, preferably faba bean. Faba bean is also commonly designated as broad bean, fava bean, or field bean. The term "whole faba bean" refers to faba bean which is intact and has not undergone any step of mechanical disruption (e.g. milling, grinding) and / or fractionation to remove fibers, carbohydrates and / or proteins. A whole faba beam comprises the germ and the endosperm, optionally and the hull. Preferably, a whole faba bean comprises the germ, the endosperm, and the hull. In particular, a whole faba bean is not faba bean flour, faba bean protein concentrate and faba bean protein isolate. For avoidance of doubt, the term "whole faba bean" does not exclude faba bean which has undergone a step of removal of hull.

[0045] The term "faba bean flour" refers to a composition, in powder form, comprising a faba bean protein content from 5wt% to 49.9wt% proteins, preferably 20wt.% to 49.9wt.% proteins.

[0046] As used herein, the term "faba bean protein concentrate" refers to a composition comprising a faba bean protein content from 50wt% to 79.9wt%.

[0047] As used herein, the term "faba bean protein isolate" refers to a composition comprising a faba bean protein content from 80.0wt% to 99.0wt%.

[0048] As used herein, the term "vicine and / or convicine-degrading enzyme" refers to enzyme which degrades vicine and / or convicine into a compound which is respectively no longer vicine and / or convicine.

[0049] As used herein, the term "beta-glucosidase (e.g., EC 3.2.1.21)" refers to a type of glycosyl hydrolases that catalyses the hydrolysis of terminal, non-reducing beta-D-glucosyl residues with the release of beta-D-glucose.

[0050] As used herein, the term "alpha-amylase (e.g. EC 3.2.1.1)" refers to an enzyme that catalyses the endohydrolysis of the (l->4)-alpha-D-glucosidic linkage in polysaccharides possessing three or more (1— >4)-a Ipha-linked D-glucose units.

[0051] As used herein, the term "identical" in the context of amino acid sequences refers to the amino acid residues in the two or multiple sequences that are the same when aligned for maximum correspondence. The length of amino acid sequence identity comparison may be over a stretch of at least 9 amino acid residues, at least 20 amino acid residues, at least 24 amino acid residues, at least 28 amino acid residues, at least 32 amino acid residues, or at least 36 or more amino acid residues. There are a number of different algorithms and methods well known to those skilled in the art that can be used to measure amino acid sequence identity. By way of example and without being limitative, the percentage (%) sequence identity of two or multiple amino acid sequences may be calculated using CLUSTALW (version 1.82, version 2), CLUSTAL omega, BLAST, EMBOSS matcher or MULTALIN, after aligning the sequences and introducing gaps if necessary, to accomplish the maximum percent sequence identity, without considering any conservative substitutions as part of the sequence identity.

[0052] As used herein, the term "meat analogue product", sometimes also called meat alternative, meat substitute, mock meat, faux meat, imitation meat or vegetarian meat, refers herein to a food product which comprises ingredients of plant origin, which is exclusively made from vegetarian ingredients, preferably vegan ingredients, and which has qualities as to appearance, taste, flavor, and texture as the corresponding real meat product. Non-limitating examples of real meat product include products derived poultry, beef, pork, fish and mixtures thereof.

[0053] As used herein, the term "dairy analogue product" refers herein to a food product which comprises ingredients of plant origin, which is free from dairy and which has qualities as to appearance, and texture as the corresponding real dairy product. Preferably, the dairy analogue product is exclusively made from vegan ingredients.

[0054] As used herein, the term "milk analogue" refers herein to a food product which comprises ingredients of plant origin, which is free from dairy and which has qualities as to appearance, and texture as the corresponding real dairy milk. Preferably, the milk analogue is exclusively made from vegan ingredients.

[0055] As used herein, the term "cream analogue" refers herein to a food product which comprises ingredients of plant origin, which is free from dairy and which has qualities as to appearance, and texture as the corresponding real dairy cream. Preferably, the cream analogue is exclusively made from vegan ingredients.

[0056] As used herein, the term "egg analogue product" refers herein to a food product which comprises ingredients of plant origin, which is free from egg and which has qualities as to appearance, and texture as the corresponding real egg product. Preferably, the egg analogue product is exclusively made from vegan ingredients.

[0057] As used herein, the term "vegetarian" refers to an edible composition which is devoid of meat, including fish.

[0058] As used herein, the term "vegan" refers to an edible composition which is entirely devoid of animal products, or animal derived products.

[0059] As used herein, the term "texturizing agent" refers to ingredients other than plant proteins that contribute to viscosity. Examples include starches (e.g., tapioca starch, corn starch, rice starch, potato starch, cassava starch, corn flour, oat flour, and the like), pectins, gums (e.g., gellan gum, locust bean gum, carob bean gum, guar gum, and the like), hydrocolloids (e.g., alginate, agar, and the like), fibers.

[0060] As used herein, the term "sweetening agent" refers to an ingredient that impart or increase the sweet taste in a food product or food ingredient. Examples of sweetening agent include agave syrup, brown sugar, coconut sugar, maple syrup, rice syrup, oat syrup, corn syrup, dextrose, fructose, glucose, honey, invert sugar, maltose, molasse, sucrose, steviol glycosides and a mixture thereof.

[0061] As used herein, the term "bulking agent" refers to an ingredient that increases the bulk-volume of a food product or ingredient without affecting its taste.

[0062] As used herein, the term "flavouring agent" refers to an ingredient that imparts, increases, masks or modifies taste and / or odor of a food product or food ingredient.

[0063] As used herein, the term "aqueous liquid" refers to an ingredient which comprises at least 70wt% of water. It may be milk, milk analogue, cream analogue, water or a combination thereof.

[0064] As used herein, the term "seed" refers to seeds different from legume seeds, which are generally called pulse and different from nut seeds, which are generally called nuts.

[0065] In a first aspect, the invention relates to a method of reducing vicine and / or convicine content in a faba ingredient. Preferably, the method is a method for completely removing vicine and / or convicine in a faba ingredient.

[0066] The method comprises a step a) of providing a faba ingredient which is liquid.

[0067] Alternatively, the method comprises a step a) of suspending a faba ingredient into an aqueous liquid to form a faba ingredient which is liquid.

[0068] In an embodiment, the aqueous liquid has a temperature of at least 45°C, preferably 45°C to 80°C, more preferably 50°C to 75°C in step a). For example, the aqueous liquid has a temperature of 60°C in step a).

[0069] In a preferred embodiment, the aqueous liquid is water.

[0070] In an embodiment, at least 0.5wt%, preferably at least lwt%, more preferably at least 2wt% faba ingredient, even more preferably at least 2.5wt% faba ingredient, is suspended into the aqueous liquid in step a). In an embodiment, at most 50wt%, preferably at most 25wt%, more preferably at most 15wt%, even more preferably at most 10wt%, most preferably at most 8wt% faba ingredient is suspended into the aqueous liquid in step a). The faba ingredient comprises vicine and / or convicine. Preferably, the sum of vicine and convicine content in the faba ingredient is of at least 450 mg per kg of faba ingredient, preferably at least 600mg per kg of faba ingredient, more preferably 600 mg to 45000 mg per kg of faba ingredient, more preferably 600 to 35000 mg per kg of faba ingredient, even more preferably 800 to 35000 mg per kg of faba ingredient, even more preferably 800 to 30000 mg per kg of faba ingredient, even more preferably 800 to 20000 mg per kg of faba ingredient , even more preferably 800 to 10000 mg per kg of faba ingredient, most preferably 800 to 2000 mg per kg of faba ingredient.

[0071] In addition, the faba ingredient comprises faba bean protein. In particular, the faba ingredient comprises at least lwt%, preferably lwt% to 99wt%, more preferably 10wt% to 99wt%, even more preferably 20 to 99wt.%, even more preferably 30 to 99wt%, even more preferably 50 to 99wt% faba bean protein.

[0072] In some other embodiment, the faba ingredient may comprise at least lwt%, preferably lwt% to 45wt%, more preferably 10wt% to 45wt%, even more preferably 20 to 45wt.%, even more preferably 30 to 45wt% faba bean protein.

[0073] In a preferred embodiment, the faba ingredient is whole faba bean, faba bean flour, faba bean protein concentrate or faba bean protein isolate. Where the faba ingredient is whole faba bean, the whole faba beans are ground before step a). The whole faba bean are ground by wet milling or dry milling, preferably wet milling. The whole faba bean may be cooked before wet or dry milling.

[0074] The faba ingredient may be mixed with at least one additional food ingredient before step b). For example, the at least one additional food ingredient is selected from the list consisting of sweetening agent, texturizing agent, buffering salt, bulking agent, fiber, fat, mineral, vitamin, antioxidant, colour, flavouring agent, aqueous liquid, protein different from faba bean protein, or a combination thereof. Preferably, the protein different from faba bean protein may be plant protein different from faba bean protein, dairy protein or a mixture thereof. More preferably, the protein different from faba bean protein may be selected from the list consisting of cereal protein, seed protein, nut protein, pulse protein, dairy protein or a mixture thereof. The pulse protein is different from faba bean protein. Even more preferably, the protein different from faba bean protein may be selected from the list consisting of cereal protein, pulse protein, dairy protein or a mixture thereof. Examples of pulse protein include chickpea protein, pea protein, lentil protein or a mixture thereof. Examples of cereal protein include oat protein, rice protein, wheat protein, barley protein, rye protein, buckwheat protein, quinoa protein, millet protein, maize protein or a mixture thereof. Examples of seed protein include hemp seed protein, canola seed protein, flaxseed protein, pumpkin seed protein, sunflower seed protein, cotton seed protein, safflower seed protein, squash seed protein, chia seed protein or a mixture thereof. Examples of nut protein include walnut protein, almond protein, pine nut protein, peanut protein, hazelnut protein, cashew nut protein, pistachio protein, tiger nut protein, pecan nut protein, macadamia nut protein or a mixture thereof. Examples of dairy protein include caseins, whey proteins or a mixture thereof.

[0075] In an embodiment, the fat may be milk fat or vegetable fat. Examples of vegetable fat include cocoa butter, coconut oil, rapeseed oil, canola oil, walnut oil, grapeseed oil, peanut oil, olive oil, sunflower seed oil or a mixture thereof.

[0076] In an embodiment, at least 10%, preferably at least 30%, more preferably at least 50%, more preferably at least 80%, more preferably at least 90%, even more preferably at least 95% of the proteins of the faba ingredient consist of faba bean protein. Most preferably, the proteins of the faba ingredient consist only of faba bean protein.

[0077] The method further comprises a step b) of adding at least one vicine and / or convicine- degrading enzyme to the faba ingredient.

[0078] The at least one vicine and / or convicine-degrading enzyme is selected from the list consisting of beta-glucosidase, enzyme having beta-glucosidase side activity or a mixture thereof. It has been observed that not only beta-glucosidase but also enzyme with betaglucosidase side activity allows effective reduction of vicine and / or convicine in faba ingredient, even at pH above 6.

[0079] Beta-glucosidase encompasses any enzyme having the function of a beta-glucosidase, i.e. that of hydrolysing beta-glycosidic bonds to terminal non-reducing residues in glucosides and oligosaccharides, with release of glucose.

[0080] In a preferred embodiment, the enzyme having beta-glucosidase side activity is selected from the list consisting of glucosidase, xylanase, cellulase, hemicellulase, amylase, glucanase, pectinase, mannanase or a mixture thereof. Preferably, the enzyme having betaglucosidase side activity is selected from the list consisting of xylanase, cellulase, or a mixture thereof.

[0081] In an embodiment, the at least one vicine and / or convicine-degrading enzyme comprises, preferably consists of vicine and / or convicine-degrading enzyme coming from almond, in particular Prunus dulcis and / or vicine and / or convicine-degrading enzyme coming from Thermotoga maritima. More preferably, the at least one vicine and / or convicine- degrading enzyme comprises, preferably consists of beta-glucosidase coming from almond, in particular Prunus dulcis and / or beta-glucosidase coming from Thermotoga maritima. Even more preferably, the at least one vicine and / or convicine-degrading enzyme comprises, preferably consists of vicine and / or convicine-degrading enzyme coming from Thermotoga maritima. Most preferably, the at least one vicine and / or convicine-degrading enzyme comprises, preferably consists of beta-glucosidase coming from Thermotoga maritima.

[0082] The presence of vicine and / or convicine-degrading enzymes, in particular betaglucosidases derived from almond, in particular Prunus dulcis and / or Thermotoga maritima, offer significant advantages. These enzymes have been observed to effectively reduce vicine and / or convicine levels in faba, achieving high reduction rates ranging above 80% at pH levels above 6. Notably, the beta-glucosidases from Thermotoga maritima demonstrate exceptional efficacy in nearly eliminating vicine and convicine in faba at pH levels above 6.

[0083] In an even more preferred embodiment, the at least one vicine and / or convicine- degrading enzyme is one or more beta-glucosidase(s) having amino acid sequence(s) that is / are at least 70%, at least 80%, at least 90%, at least 95%, at least 98%, at least 99% identical to one or more amino acid sequence(s) selected from the group consisting of SEQ ID NO: 1, SEQ NO: 2, SEQ NO: 3, SEQ NO: 4, and SEQ ID NO: 5, preferably selected from SEQ ID NO: 1, SEQ ID NO:2 and SEQ ID NO: 3, even more preferably selected from SEQ ID NO: 1 and SEQ ID NO: 3 or even more preferably selected from SEQ ID NO: 1 and SEQ ID NO:2. In a most preferred embodiment, the at least one vicine and / or convicine-degrading enzyme is one or more beta-glucosidase(s) having amino acid sequence(s) that is / are 100% identical to one or more amino acid sequence(s) selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, and SEQ ID NO: 5, preferably selected from SEQ ID NO: 1, SEQ ID NO: 2 and SEQ ID NO: 3, even more preferably selected from SEQ ID NO: 1 and SEQ ID NO: 3 or even more preferably selected from SEQ ID NO: 1 and SEQ ID NO:2.

[0084] The presence of vicine and / or convicine-degrading enzymes having the above sequences offer significant advantages. These enzymes have been observed to effectively reduce vicine and / or convicine levels in faba, achieving high reduction rates ranging above 80% at pH levels above 6. Notably, the enzymes with SEQ ID NO: 1 and SEQ ID NO: 3 demonstrate exceptional efficacy in nearly eliminating vicine and convicine in faba at pH levels above 6. In an embodiment, at least one enzyme different from vicine and / or convicine- degrading enzyme may also be further added to the faba ingredient in step b). The faba ingredient has a pH of 6.0 or higher, preferably a pH of 6.0 to 9.0, more preferably a pH of 6.0 to 7.5. The faba ingredient has generally already this pH in step a), i.e. the native pH of the faba ingredient is of 6.0 or higher, preferably of 6. O to 9.0, more preferably of 6.0 to 7.5. If the pH of the faba ingredient is different than 6.0 or higher, the pH of the faba ingredient may be adjusted to reach a pH of of 6.0 or higher, preferably a pH of 6.0 to 9.0, preferably a pH of 6.0 to 7.5 between step a) and step b).

[0085] It has been discovered that it is possible to decrease significantly the amount of vicine and / or convicine via enzymatic treatment at pH above 6 in a short time.

[0086] The dosage of vicine and / or convicine-degrading enzyme that is added to the faba ingredient, is dependent, inter alia, on the temperature during the incubation, the volume or amount of faba ingredient to be processed, and the activity of the enzyme. Generally, this dosage should be at least 0.1% (w / v) based on the volume of liquid faba ingredient. Preferably, this dosage of vicine and / or convicine-degrading enzyme may be of 0.1% to 1% (w / v) based on the volume of liquid faba ingredient.

[0087] The method comprises a step c) of incubating the faba ingredient with the at least one vicine and / or convicine-degrading enzyme to obtain a treated faba ingredient. This incubation step c) is performed at a pH of 6.0 or higher, preferably a pH of 6.0 to 9.0, preferably a pH of 6.0to 7.5. The incubation step c) may be performed at a temperature of at least 50°C, between 50-65°C.

[0088] The invention allows a significant decrease in vicine and / or convicine with a short reaction time.

[0089] The incubation step is performed for less than 2 hours, preferably less than 90 minutes, more preferably 15 minutes to 90 minutes.

[0090] Even at pH above 6, it has been observed that enzymes were effective at significantly reducing the amount of vicine and / or convicine in faba ingredients.

[0091] In particular, the content of vicine and / or convicine in the faba ingredient is reduced by at least 30%, preferably at least 50%, more preferably at least 80%, more preferably at least 85%, more preferably at least 90%, more preferably at least 95%, even more preferably at least 98%, most preferably 100% by the at least one vicine and / or convicine-degrading enzyme in step c). In other words, the treated faba ingredient has vicine and / or convicine content which is at least 30%, preferably at least 50%, more preferably at least 80%, more preferably at least 85%, more preferably at least 90%, more preferably at least 95%, even more preferably at least 98%, most preferably 100% less than the vicine and / or convicine content in the faba ingredient.

[0092] As the process is performed at pH above 6, the proteins, in particular faba bean protein, are intact such that they preserve good stability and good functionality over shelf-life and / or processing. This good stability and functionality make the faba ingredient suitable for use in food applications.

[0093] Hence, in an embodiment, the proteins, in particular faba bean proteins, of the treated faba ingredient are intact. By "intact", it refers to proteins which are not aggregated in an uncontrolled manner, in particular which are not precipitated or sedimented.

[0094] The process (i.e. method) comprises an optional step d) of partially or fully, preferably fully, inactivating the at least one vicine and / or convicine-degrading enzyme in the treated faba ingrdient. In an embodiment, the step d) is not optional. In a preferred embodiment, the inactivation step d) is performed by heat-treating the treated faba ingredient. The heattreatment is performed at a temperature of at least 70°C, preferably 70°C to 140°C, more preferably 80°C to 110°C. Preferably, the heat-treatment is performed for 3 seconds to 20 minutes.

[0095] The process (i.e. method) comprises an optional step d) of cooling the treated faba ingredient. For example, the treated faba ingredient is cooled down to the storage temperature of the treated faba ingredient, for example to room temperature.

[0096] The process (i.e. method) may comprise a step of evaporating the treated faba ingredient after step c) or after step d) or after step e).

[0097] The process (i.e. method) may comprise a step of drying the treated faba ingredient after step c) or after step d) or after step e) to obtain a treated faba ingredient which is a powder. The drying step may be performed by drum drying, roller drying, spray drying or freeze drying. The drying step may also be performed by any other food drying technologies well known in the art. The transformation of the ingredient into powder may be advantageous as powders have generally good physico-chemical stability and extended shelf-life.

[0098] The evaporation and / or drying step may also allow to inactivate the at least-one vicine and / or convicine-degrading enzyme. In this event, the inactivation step d) is optional.

[0099] The process (i.e. method) of the invention allows to reduce significantly the vicine and / or convicine of faba ingredient at pH above 6 in a limited time. The obtained treated faba ingredient keeps good stability and functionality and may be used for food application. In an embodiment, the process (i.e. method) does not comprise any addition of microorganism, in particular lactic acid bacteria. The process (i.e. method) does not comprise any step of fermentation, in particular before step c) or d). A significant reduction of vicine and / or convicine is obtained without fermentation. Fermentation is undesirable for the reasons provided in the background of the invention.

[0100] The treated faba ingredient may be liquid, semi-liquid or powder.

[0101] The treated faba ingredient may have the same features as provided herein for the faba ingredient, except the amount of vicine and / convicine which are significantly lower in the treated faba ingredient.

[0102] In an embodiment, the method does not comprise any step of addition of organic solvent. In particular, the treated faba ingredient may be free from any organic solvent. For example, the organic solvent may be selected from the list consisting of acetic acid, acetone, acetonitrile, benzene, 1-butanol, 2-butanol, 3-butanone, t-butyl alcohol, carbon tetrachloride, chlorobenzene, chloroform, cyclohexane, diethylene glycol, diethyl ether, diethylene glycol dimethyl ether, 1,2-dimethoxy-ethane (glyme, DIVIE), dimethyl-formamide (DMF), dimethyl sulfoxide (DMSO), 1,4 dioxane, 1,2-dicholoroethane, ethanol, ethyl acetate, ethylene glycol, glycerin, heptane, hexamethylphosphoramide (HMPA), hexane, methanol, methylene chloride, / V-methyl-2-pyrrolidinone (NMP), nitromethane, naphthalene, pentane, 1-propanol, 2-propanol, pyridine, toluene, triethyl amine, tricine, tris, tetrahydrofuran, o-xylene, m- xylene, p-xylene and combination thereof.

[0103] A significant reduction of vicine / convicine in faba is achieved without using any extraction with organic solvent. The method allows to be emancipated from organic solvents which may negatively affect the properties of the faba ingredient.

[0104] In some embodiments, the method does not comprise any step of addition of amylase before step c). In some embodiments, the method does not comprise any step of addition of amylase.

[0105] In some embodiments, the method does not comprise any step of rinsing the faba ingredient before step b) or step c) with an aqueous liquid, preferably water. In a further embodiment, the method does not comprise any step of rinsing the faba ingredient before step b) or step c) with an aqueous liquid, preferably water, at a temperature above 50°C, preferably between 50°C and 90°C.

[0106] The method of the invention achieves a significant reduction of vicine / convicine without requiring any rinsing step. The absence of rinsing step before enzymatic treatment is advantageous. Indeed, this simplifies the process by limiting the number of steps, saves energy and resources, and is therefore more sustainable. Moreover, this avoids altering the quality of the faba ingredient. Indeed, rinsing, in particular at elevated temperature, may alter the quality of the faba ingredient.

[0107] In a second aspect, the invention relates to a treated faba ingredient which is obtained with the method of the first aspect of the invention. The features of the faba ingredient of the third aspect of the invention applies to the treated faba ingredient of the present second aspect of the invention.

[0108] The treated faba ingredient may be liquid, semi-liquid or powder.

[0109] In a third aspect, the invention relates to a faba ingredient.

[0110] The faba ingredient comprises a limited amount of vicine and / or convicine. The sum of vicine and convicine content in the faba ingredient is less than 600 mg per kg of faba ingredient, preferably less than 450 mg per kg of faba ingredient, more preferably less than 300 mg per kg of faba ingredient, even more preferably less than 125 mg per kg of faba ingredient, even more preferably less than lOOmg per kg of faba ingredient, even more preferably less than 50 mg per kg of faba ingredient. In a preferred embodiment, the faba ingredient is substantially free, preferably entirely free from vicine and / or convicine.

[0111] In an embodiment, the faba ingredient comprises substantial amount of faba bean protein. The faba ingredient comprises faba bean protein. In particular, the faba ingredient comprises at least lwt%, preferably lwt% to 99wt%, more preferably 10wt% to 99wt%, even more preferably 30wt% to 99wt%, most preferably 50wt% to 99wt% faba bean protein.

[0112] In addition to faba bean protein, the faba ingredient may also comprise protein different from faba bean protein. Preferably, the protein different from faba bean protein may be plant protein different from faba bean, dairy protein or a mixture thereof. More preferably, the protein different from faba bean protein may be selected from the list consisting of cereal protein, seed protein, nut protein, pulse protein, dairy protein or a mixture thereof. The pulse protein is different from faba bean protein. Even more preferably, the protein different from faba bean protein may be selected from the list consisting of cereal protein, pulse protein, dairy protein or a mixture thereof. Examples of pulse protein include chickpea protein, pea protein, lentil protein or a mixture thereof. Examples of cereal protein include oat protein, rice protein, wheat protein, barley protein, rye protein, buckwheat protein, quinoa protein, millet protein, maize protein or a mixture thereof. Examples of seed protein include hemp seed protein, canola seed protein, flaxseed protein, pumpkin seed protein, sunflower seed protein, cotton seed protein, safflower seed protein, squash seed protein, chia seed protein or a mixture thereof. Examples of nut protein include walnut protein, almond protein, pine nut protein, peanut protein, hazelnut protein, cashew nut protein, pistachio protein, tiger nut protein, pecan nut protein, macadamia nut protein or a mixture thereof. Examples of dairy protein include caseins, whey proteins or a mixture thereof.

[0113] In an embodiment, at least 10%, preferably at least 30%, more preferably at least 50%, more preferably at least 80%, more preferably at least 90%, even more preferably at least 95% of the proteins of the faba ingredient consist of faba bean protein. Most preferably, the proteins of the faba ingredient consist only of faba bean protein.

[0114] In an embodiment, the faba ingredient has a pH of of 6.0 or higher, preferably a pH of 6.0 to 9.0, preferably a pH of 6.0 to 7.5.

[0115] Hence, in an embodiment, the protein, in particular faba bean protein, of the faba ingredient are intact. By "intact", it refers to proteins which are not aggregated in an uncontrolled manner, in particular which are not precipitated or sedimented.

[0116] In a preferred embodiment, the faba ingredient is not fermented.

[0117] In an embodiment, the faba ingredient may comprise sweetening agent, texturizing agent, buffering salt, bulking agent, fiber, fat, mineral, vitamin, antioxidant, colour, flavouring agent, aqueous liquid or a combination thereof. The fat may be as provided hereinabove.

[0118] In an embodiment, the faba ingredient is liquid, semi-liquid or powder.

[0119] The faba ingredient comprises low amount of vicine and / or convicine. It has good stability and functionality and may be used for food application.

[0120] In an embodiment, the faba ingredient is an enzymatically-treated faba ingredient, i.e. a faba ingredient treated with at least one vicine and / or convicine degrading enzyme. The at least one vicine and / or convicine degrading enzyme may be vicine and / or convicine degrading enzyme as disclosed in the first aspect of the invention.

[0121] The features of faba ingredient of the present third aspect of the invention apply to the treated faba ingredient of the first and second aspect of the invention, and vice versa.

[0122] In some embodiments, the faba ingredient may be free from any organic solvent. For example, the organic solvent may be selected from the list consisting of acetic acid, acetone, acetonitrile, benzene, 1-butanol, 2-butanol, 3-butanone, t-butyl alcohol, carbon tetrachloride, chlorobenzene, chloroform, cyclohexane, diethylene glycol, diethyl ether, diethylene glycol dimethyl ether, 1,2-dimethoxy-ethane (glyme, DIVIE), dimethyl-formamide (DMF), dimethyl sulfoxide (DMSO), 1,4 dioxane, 1,2-dicholoroethane, ethanol, ethyl acetate, ethylene glycol, glycerin, heptane, hexamethylphosphoramide (HMPA), hexane, methanol, methylene chloride, / V-methyl-2-pyrrolidinone (NMP), nitromethane, naphthalene, pentane, 1-propanol, 2-propanol, pyridine, toluene, triethyl amine, tricine, tris, tetrahydrofuran, o-xylene, m- xylene, p-xylene and combination thereof.

[0123] In a fourth aspect, the invention relates to a food product which comprises the treated faba ingredient according to the second or the faba ingredient according to the third aspect of the invention.

[0124] The food product may be selected from the list consisting of dairy product, dairy analogue product, meat analogue product, egg analogue product, nutritional product, confectionery product, pet food, vegetable puree, soup, beverage, baby food, medical nutrition product, food supplement, or a combination thereof. The dairy product may be selected from the list consisting of yogurt, milk, cream, ice cream, dairy dessert, cheese, dairy beverage, milk powder, condensed milk, powdered dairy beverage, infant formula or a combination thereof. The dairy analogue product may be selected from the list consisting of yogurt analogue, milk analogue, cream analogue, ice cream analogue, cheese analogue, dairy beverage analogue, dairy dessert analogue, milk powder analogue, condensed milk analogue, powdered dairy beverage analogue, infant formula analogue or a combination thereof.

[0125] In some embodiments, the food product may be free from any organic solvent. For example, the organic solvent may be selected from the list consisting of acetic acid, acetone, acetonitrile, benzene, 1-butanol, 2-butanol, 3-butanone, t-butyl alcohol, carbon tetrachloride, chlorobenzene, chloroform, cyclohexane, diethylene glycol, diethyl ether, diethylene glycol dimethyl ether, 1,2-dimethoxy-ethane (glyme, DIVIE), dimethyl-formamide (DMF), dimethyl sulfoxide (DMSO), 1,4 dioxane, 1,2-dicholoroethane, ethanol, ethyl acetate, ethylene glycol, glycerin, heptane, hexamethylphosphoramide (HMPA), hexane, methanol, methylene chloride, / V-methyl-2-pyrrolidinone (NMP), nitromethane, naphthalene, pentane, 1-propanol, 2-propanol, pyridine, toluene, triethyl amine, tricine, tris, tetrahydrofuran, o-xylene, m- xylene, p-xylene and combination thereof. In a fifth aspect, the invention relates to a method for producing a food product. The food product may be a food product as provided in the fourth aspect of the invention.

[0126] The method comprises a step a) of providing a food composition.

[0127] The food composition comprises faba ingredient. The faba ingredient of the food composition comprises vicine and / or convicine. Preferably, the sum of vicine and convicine content in the faba ingredient is of at least 450 mg per kg of faba ingredient, preferably at least 600 mg per kg of faba ingredient, more preferably 600 mg to 45000 mg per kg of faba ingredient, more preferably 600 to 35000 mg per kg of faba ingredient, even more preferably 800 to 10000 mg per kg of faba ingredient, most preferably 800 to 2000 mg per kg of faba ingredient.

[0128] The faba ingredient may be a faba ingredient as provided in the first aspect of the invention.

[0129] In an embodiment, the food composition may comprise at least 0.5wt%, preferably from 0.5wt% to 50wt.%, more preferably from lwt% to 50wt%, even more preferably from lwt% to 25wt%, even more preferably lwt% to 15wt%, even more preferably lwt% to 10wt%, most preferably lwt% to 8wt% faba ingredient.

[0130] The food composition comprises at least one additional food ingredients. For example, the at least one additional food ingredient is selected from the list consisting of sweetening agent, texturizing agent, buffering salt, bulking agent, fiber, fat, mineral, vitamin, antioxidant, colour, flavouring agent, aqueous liquid, protein different from faba bean protein, or a combination thereof. Preferably, the protein different from faba bean protein may be plant protein different from faba bean, dairy protein or a mixture thereof. More preferably, the protein different from faba bean protein may be selected from the list consisting of cereal protein, seed protein, nut protein, pulse protein, dairy protein or a mixture thereof. The pulse protein is different from faba bean protein. Even more preferably, the protein different from faba bean protein may be selected from the list consisting of cereal protein, pulse protein, dairy protein or a mixture thereof. Examples of pulse protein include chickpea protein, pea protein, lentil protein or a mixture thereof. Examples of cereal protein include oat protein, rice protein, wheat protein, barley protein, rye protein, buckwheat protein, quinoa protein, millet protein, maize protein or a mixture thereof. Examples of seed protein include hemp seed protein, canola seed protein, flaxseed protein, pumpkin seed protein, sunflower seed protein, cotton seed protein, safflower seed protein, squash seed protein, chia seed protein or a mixture thereof. Examples of nut protein include walnut protein, almond protein, pine nut protein, peanut protein, hazelnut protein, cashew nut protein, pistachio protein, tiger nut protein, pecan nut protein, macadamia nut protein or a mixture thereof. Examples of dairy protein include caseins, whey proteins or a mixture thereof.

[0131] In an embodiment, the fat may be a fat as provided in the first aspect of the invention.

[0132] In a further embodiment, at least one additional food ingredient may be added to the food composition, preferably before step b). For example, the at least one additional food ingredient may be selected from the list consisting of sweetening agent, texturizing agent, buffering salt, bulking agent, fiber, fat, mineral, vitamin, antioxidant, colour, flavouring agent, aqueous liquid, protein different from faba bean protein or a combination thereof. Preferably, the protein different from faba bean protein may be plant protein different from faba bean, dairy protein or a mixture thereof. More preferably, the protein different from faba bean protein may be selected from the list consisting of cereal protein, seed protein, nut protein, pulse protein, dairy protein or a mixture thereof. The pulse protein is different from faba bean protein. Even more preferably, the protein different from faba bean protein may be selected from the list consisting of cereal protein, pulse protein, dairy protein or a mixture thereof. Examples of pulse protein include chickpea protein, pea protein, lentil protein or a mixture thereof. Examples of cereal protein include oat protein, rice protein, wheat protein, barley protein, rye protein, buckwheat protein, quinoa protein, millet protein, maize protein or a mixture thereof. Examples of seed protein include hemp seed protein, canola seed protein, flaxseed protein, pumpkin seed protein, sunflower seed protein, cotton seed protein, safflower seed protein, squash seed protein, chia seed protein or a mixture thereof. Examples of nut protein include walnut protein, almond protein, pine nut protein, peanut protein, hazelnut protein, cashew nut protein, pistachio protein, tiger nut protein, pecan nut protein, macadamia nut protein or a mixture thereof. Examples of dairy protein include caseins, whey proteins or a mixture thereof.

[0133] In an embodiment, the fat may be a fat as provided in the first aspect of the invention.

[0134] In an embodiment, the food composition may comprise at least 0.5wt%, preferably at least 2wt%, more preferably 2wt% to 25wt% protein, preferably faba bean protein.

[0135] In an embodiment, the proteins of the food composition comprise at least 10%, preferably at least 30%, more preferably at least 50%, more preferably at least 80%, more preferably at least 90%, even more preferably at least 95% of faba bean protein. Most preferably, the proteins of the food composition consist only of faba bean protein. In an embodiment, the food composition may be liquid or semi-liquid or pasty. For example, for preparing a beverage, the food composition may be liquid whereas for preparing a meat analogue product, the food composition may be semi-liquid or pasty. Preferably, the food composition is liquid.

[0136] The method further comprises a step b) of adding at least one vicine and / or convicine- degrading enzyme to the food composition.

[0137] The at least one vicine and / or convicine-degrading enzyme may be vicine and / or convicine-degrading enzyme as disclosed in the first aspect of the invention.

[0138] In an embodiment, at least one enzyme different from vicine and / or convicine- degrading enzyme may also be added to the food composition in step b).

[0139] The food composition has a pH of 6.0 or higher, preferably a pH of 6.0 to 9.0, preferably a pH of 6.0 to 7.5. The food composition has generally already this pH in step a), i.e. that the native pH of the food composition is of 6.0 or higher, preferably a pH of 6.0 to 9.0, preferably a pH of 6.0 to 7.5. If the pH of the food composition is different than 6.0 or higher, the pH of the food composition may be adjusted to reach a pH of 6.0 or higher, preferably a pH of 6.0 or higher, preferably a pH of 6.0 to 9.0, preferably a pH of 6.0 to 7.5 between step a) and step b).

[0140] The dosage of vicine and / or convicine-degrading enzyme that is added to the food composition, is dependent, inter alia, on the temperature during the incubation, the amount or volume of food composition to be processed, the amount of faba ingredient in the food composition, and the activity of the enzyme. Generally, this dosage should be at least 0.1wt% based on the weight of faba ingredient in the food composition. Preferably, this dosage of vicine and / or convicine-degrading enzyme may be of 0.1wt% to 5wt% based on the weight of faba ingredient in the food composition.

[0141] The method further comprises a step c) of incubating the food composition with the at least one vicine and / or convicine-degrading enzyme to obtain a treated food composition. This incubation step c) is performed at a pH of 6.0 or higher, preferably a pH of 6.0 to 9.0, preferably a pH of 6.0 to 7.5. The incubation step c) may be performed at a temperature of at least 50°C, between 50-65°C.

[0142] The invention allows a significant decrease in vicine and / or convicine with a short reaction time.

[0143] The incubation step is performed for less than 2 hours, preferably less than 90 minutes, more preferably 15 minutes to 90 minutes. Even at pH above 6, it has been observed that enzymes were effective at significantly reducing the amount of vicine and / or convicine in the food composition comprising faba ingredient.

[0144] In particular, the content of vicine and / or convicine in the food composition is reduced by at least 30%, preferably at least 50%, more preferably at least 80%, more preferably at least 85%, more preferably at least 90%, more preferably at least 95%, even more preferably at least 98%, most preferably 100% by the at least one vicine and / or convicine-degrading enzyme in step c). In other words, the treated food composition has vicine and / or convicine content which is at least 30%, preferably at least 50%, more preferably at least 80%, more preferably at least 85%, more preferably at least 90%, more preferably at least 95%, even more preferably at least 98%, most preferably 100% less than the vicine and / or convicine content in the food composition.

[0145] As the enzymatic treatment is performed at pH above 6, the proteins, in particular faba bean protein, are intact such that they have good stability and good functionality for further processing into a food product and during the shelf-life of the food product.

[0146] Hence, in an embodiment, the protein, in particular faba bean protein, of the treated food composition obtained after step c) are intact. By "intact", it refers to proteins which are not aggregated in an uncontrolled manner, in particular which are not precipitated or sedimented.

[0147] The method further comprises an optional step d) of homogenizing the treated food composition. In a preferred embodiment, the step d) of homogenization is not optional. The homogenization step may be performed at a pressure above 50 bar. Preferably, the homogenization step may be performed at a pressure of 50 bar to 700 bar. Further preferably, the homogenization step may be performed at a pressure of 50 bar to 500 bar. More preferably, the homogenization step may be performed at a pressure of 50 to 300 bar, from 100 to 300 bar or from 150 to 300 bar.

[0148] In a preferred embodiment, the homogenization step may be performed at a temperature from 50°C to 80°C, preferably from 50°C to 70°C. More preferably, the homogenization step may be performed at a temperature from 55°C to 65°C.

[0149] The method further comprises a step e) of heat-treating the treated food composition. Preferably, the heat treatment is performed at a temperature of 70°C to 150°C, preferably 75°C to 145°C for 3 seconds to 20 minutes, preferably 3 seconds to 90 seconds. More preferably, the heat treatment is performed at a temperature from 80°C to 145°C, preferably 80°C to 125°C, for 3 seconds to 20 minutes, preferably 3 seconds to 90 seconds.

[0150] This heat treatment step allows to extend the shelf-life of the food product while partially or fully inactivating the at least one vicine and / or convicine-degrading enzyme in the food treated food composition.

[0151] The homogenization step d) may be upstream or downstream the heat treatment step e). In an embodiment, the process may comprise two homogenization step, a first homogenization step d) which is upstream the heat-treatment and a second homogenization step which is downstream the heat-treatment. The pressure and temperature conditions provided for the homogenization step d) also apply to the second homogenization step.

[0152] The method further comprises a step e) of cooling the treated food composition to obtain a food product. For example, the treated food composition is cooled down to the storage temperature of the food product, for example to room temperature or under chilled conditions, i.e., between 2°C to 14°C.

[0153] The method may further comprise a step of evaporating the treated food composition after step d) or e) or f).

[0154] The method may further comprise a step of drying the treated food composition after step d) or e) or f) to form a food product in powder form. The drying step may be performed by drum drying, roller drying, spray drying or freeze drying. The drying step may also be performed by any other food drying technologies well known in the art. The transformation of the food product into powder may be advantageous as powders have generally good physico-chemical stability and extended shelf-life.

[0155] In a preferred embodiment, the drying step is preceded by an evaporation step as provided herein.

[0156] The treated food composition may have the same features as provided herein for the food composition, except the amount of vicine and / convicine which are significantly lower in the treated food composition.

[0157] In an embodiment, the method does not comprise any step of addition of organic solvent. In particular, the treated food composition and the food product may be free from organic solvent. For example, the organic solvent may be selected from the list consisting of acetic acid, acetone, acetonitrile, benzene, 1-butanol, 2-butanol, 3-butanone, t-butyl alcohol, carbon tetrachloride, chlorobenzene, chloroform, cyclohexane, diethylene glycol, diethyl ether, diethylene glycol dimethyl ether, 1,2-dimethoxy-ethane (glyme, DIVIE), dimethyl- formamide (DMF), dimethyl sulfoxide (DMSO), 1,4 dioxane, 1,2-dicholoroethane, ethanol, ethyl acetate, ethylene glycol, glycerin, heptane, hexamethylphosphoramide (HMPA), hexane, methanol, methylene chloride, / V-methyl-2-pyrrolidinone (NMP), nitromethane, naphthalene, pentane, 1-propanol, 2-propanol, pyridine, toluene, triethyl amine, tricine, tris, tetrahydrofuran, o-xylene, m-xylene, p-xylene and combination thereof.

[0158] A significant reduction of vicine / convicine in faba is achieved without using any extraction with organic solvent. The method allows to be emancipated from organic solvents which may negatively affect the properties of the faba ingredient, the food composition and the food product.

[0159] In some embodiments, the method does not comprise any step of addition of amylase before step c). In some embodiments, the method does not comprise any step of addition of amylase.

[0160] In some embodiments, the method does not comprise any step of rinsing the faba ingredient and / or the food composition before step b) or step c) with an aqueous liquid, preferably water. In a further embodiment, the method does not comprise any step of rinsing the faba ingredient and / or the food composition before step b) or step c) with an aqueous liquid, preferably water, at a temperature above 50°C, preferably between 50°C and 90°C.

[0161] The method of the invention achieves a significant reduction of vicine / convicine in faba without requiring any rinsing step. The absence of rinsing step before enzymatic treatment is advantageous. Indeed, this simplifies the process by limiting the number of steps, saves energy and resources, and is therefore more sustainable. Moreover, this avoids altering the quality of the faba ingredient, the food composition and even the food product. Indeed, rinsing, in particular at elevated temperature, may alter the quality of the faba ingredient, the food composition and even the food product.

[0162] In a sixth aspect, the invention relates to a method for producing a food product. The food product may be a food product as provided in the fourth or fifth aspect of the invention.

[0163] The method comprises the step of providing a food composition.

[0164] The food composition comprises the treated faba ingredient according to the second aspect or the faba ingredient according to the third aspect of the invention.

[0165] In an embodiment, the food composition may comprise at least 0.5wt%, preferably from 0.5wt% to 50wt%, more preferably from lwt% to 50wt%, even more preferably from lwt% to 25wt%, even more preferably lwt% to 15wt%, even more preferably lwt% to 10wt%, most preferably lwt% to 8wt% faba ingredient according to the third aspect of the invention or treated faba ingredient according to the second aspect.

[0166] In an embodiment, the food composition comprises at least one additional food ingredient. For example, the at least one additional food ingredient is selected from the list consisting of sweetening agent, texturizing agent, buffering salt, bulking agent, fiber, fat, mineral, vitamin, antioxidant, colour, flavouring agent, aqueous liquid, protein different from faba bean protein, or a combination thereof. Preferably, the protein different from faba bean protein may be plant protein different from faba bean, dairy protein or a mixture thereof. More preferably, the protein different from faba bean protein may be selected from the list consisting of cereal protein, seed protein, nut protein, pulse protein, dairy protein or a mixture thereof. The pulse protein is different from faba bean protein. Even more preferably, the protein different from faba bean protein may be selected from the list consisting of cereal, pulse, dairy protein or a mixture thereof. Examples of pulse protein include chickpea protein, pea protein, lentil protein or a mixture thereof. Examples of cereal protein include oat protein, rice protein, wheat protein, barley protein, rye protein, buckwheat protein, quinoa protein, millet protein, maize protein or a mixture thereof. Examples of seed protein include hemp seed protein, canola seed protein, flaxseed protein, pumpkin seed protein, sunflower seed protein, cotton seed protein, safflower seed protein, squash seed protein, chia seed protein or a mixture thereof. Examples of nut protein include walnut protein, almond protein, pine nut protein, peanut protein, hazelnut protein, cashew nut protein, pistachio, tiger nut protein, pecan nut protein, macadamia nut protein or a mixture thereof. Examples of dairy protein include caseins, whey proteins or a mixture thereof.

[0167] In an embodiment, the fat may be a fat as provided in the first aspect of the invention.

[0168] In a further embodiment, at least one additional food ingredient may be added to the food composition, preferably before step b). For example, the at least one additional food ingredient may be selected from the list consisting of faba bean protein, sweetening agent, texturizing agent, buffering salt, bulking agent, fiber, fat, mineral, vitamin, antioxidant, colour, flavouring agent, aqueous liquid, or a combination thereof. Preferably, the protein different from faba bean protein Preferably, the protein different from faba bean protein may be plant protein different from faba bean, dairy protein or a mixture thereof. More preferably, the protein different from faba bean protein may be selected from the list consisting of cereal protein, seed protein, nut protein, pulse protein, dairy protein or a mixture thereof. The pulse protein is different from faba bean protein. Even more preferably, the protein different from faba bean protein may be selected from the list consisting of cereal protein, pulse protein, dairy protein or a mixture thereof. Examples of pulse protein include chickpea protein, pea protein, lentil protein or a mixture thereof. Examples of cereal protein include oat protein, rice protein, wheat protein, barley protein, rye protein, buckwheat protein, quinoa protein, millet protein, maize protein or a mixture thereof. Examples of seed protein include hemp seed protein, canola seed protein, flaxseed protein, pumpkin seed protein, sunflower seed protein, cotton seed protein, safflower seed protein, squash seed protein, chia seed protein or a mixture thereof. Examples of nut protein include walnut protein, almond protein, pine nut protein, peanut protein, hazelnut protein, cashew nut protein, pistachio protein, tiger nut protein, pecan nut protein, macadamia nut protein or a mixture thereof. Examples of dairy protein include caseins, whey proteins or a mixture thereof.

[0169] In an embodiment, the fat may be a fat as provided in the first aspect of the invention.

[0170] In an embodiment, the food composition comprises at least 0.5wt%, preferably at least 2wt%, more preferably 2wt% to 25wt% protein, preferably faba bean protein.

[0171] In an embodiment, the proteins of the food composition comprise at least 10%, preferably at least 30%, more preferably at least 50%, more preferably at least 80%, more preferably at least 90%, even more preferably at least 95% of faba bean protein. Most preferably, the proteins of the food composition consist only of faba bean protein.

[0172] In an embodiment, the food composition may have a pH of 6.0 or higher, preferably a pH of 6.0 to 9.0, more preferably a pH of 6.0 to 7.5.

[0173] In an embodiment, the food composition may be liquid or semi-liquid or pasty. For example, for preparing a beverage, the food composition may be liquid whereas for preparing a meat analogue product, the food composition may be semi-liquid or pasty. Preferably, the food composition is liquid.

[0174] The method further comprises an optional step b) of homogenizing the food composition. In a preferred embodiment, the step b) of homogenization is not optional. The homogenization step may be performed at a pressure above 50 bar. Preferably, the homogenization step may be performed at a pressure of 50 bar to 700 bar. Further preferably, the homogenization step may be performed at a pressure of 50 bar to 500 bar. More preferably, the homogenization step may be performed at a pressure of 50 to 300 bar, from 100 to 300 bar or from 150 to 300 bar. In a preferred embodiment, the homogenization step may be performed at a temperature from 50°C to 80°C, preferably from 50°C to 70°C. More preferably, the homogenization step may be performed at a temperature from 55°C to 65°C.

[0175] The method further comprises a step c) of heat-treating the food composition. Preferably, the heat treatment is performed at a temperature of 70°C to 150°C, preferably 75°C to 145°C for 3 seconds to 20 minutes, preferably 3 seconds to 90 seconds. More preferably, the heat treatment is performed at a temperature from 80°C to 145°C, preferably 80°C to 125°C, for 3 seconds to 20 minutes, preferably 3 seconds to 90 seconds.

[0176] The homogenization step b) may be upstream or downstream the heat treatment step c). In an embodiment, the process may comprise two homogenization steps, a first homogenization step b) which is upstream the heat treatment step c) and a second homogenization step which is downstream the heat treatment step c). The pressure and temperature conditions provided for the homogenization step b) also apply to the second homogenization step.

[0177] The method further comprises a step d) of cooling the food composition to obtain a food product. For example, the food composition is cooled down to the storage temperature of the food product, for example to room temperature or under chilled conditions, i.e. between 2°C to 14°C.

[0178] The method may further comprise a step of evaporating the food composition after step b) or c) or d).

[0179] The method may further comprise a step of drying the food composition after step b) or c) or d) to form a food product in powder form. The drying step may be performed by drum drying, roller drying, spray drying or freeze drying. The drying step may also be performed by any other food drying technologies well known in the art. The transformation of the food product into powder may be advantageous as powders have generally good physico-chemical stability and extended shelf-life.

[0180] In a preferred embodiment, the drying step is preceded by an evaporation step as provided herein.

[0181] In an embodiment, the method does not comprise any step of addition of organic solvent. In particular, the food composition may be free from organic solvent. For example, the organic solvent may be selected from the list consisting of acetic acid, acetone, acetonitrile, benzene, 1-butanol, 2-butanol, 3-butanone, t-butyl alcohol, carbon tetrachloride, chlorobenzene, chloroform, cyclohexane, diethylene glycol, diethyl ether, diethylene glycol dimethyl ether, 1,2-dimethoxy-ethane (glyme, DIVIE), dimethyl-formamide (DMF), dimethyl sulfoxide (DMSO), 1,4 dioxane, 1,2-dicholoroethane, ethanol, ethyl acetate, ethylene glycol, glycerin, heptane, hexamethylphosphoramide (HMPA), hexane, methanol, methylene chloride, / V-methyl-2-pyrrolidinone (NMP), nitromethane, naphthalene, pentane, 1-propanol, 2-propanol, pyridine, toluene, triethyl amine, tricine, tris, tetrahydrofuran, o-xylene, m- xylene, p-xylene and combination thereof.

[0182] A significant reduction of vicine / convicine in faba is achieved without using any extraction with organic solvent. The method allows to emancipate from organic solvents which may negatively affect the properties of the faba ingredient, the food composition and the food product.

[0183] In some embodiments, the method does not comprise any step of addition of amylase before step c). In some embodiments, the method does not comprise any step of addition of amylase.

[0184] In some embodiments, the method does not comprise any step of rinsing the faba ingredient and / or the food composition before step b) or step c) with an aqueous liquid, preferably water. In a further embodiment, the method does not comprise any step of rinsing the faba ingredient and / or the food composition before step b) or step c) with an aqueous liquid, preferably water, at a temperature above 50°C, preferably between 50°C and 90°C.

[0185] The method of the invention achieves a significant reduction of vicine / convicine in faba without requiring any rinsing step. The absence of rinsing step before enzymatic treatment is advantageous. Indeed, this simplifies the process by limiting the number of steps, saves energy and resources, and is therefore more sustainable. Moreover, this avoids altering the quality of the faba ingredient, the food composition and even the food product. Indeed, rinsing, in particular at elevated temperature, may alter the quality of the faba ingredient, the food composition and even the food product.

[0186] In a seventh aspect, the invention relates to a food product obtained by the method according to the fifth aspect of the invention or the sixth aspect of the invention.

[0187] The food product may be food product as disclosed in the fourth, fifth and sixth aspect of the invention.

[0188] Protein sequences

[0189] • SEQ ID NO: 1 (length: 444) MKKFPEGFLWGVATASYQIEGSPLADGAGMSIWHTFSHTPGNVKNGDTGDVACDHYNRWKEDIEIIEK

[0190] LGVKAYRFSISWPRILPEGTGRVNQKGLDFYNRIIDTLLEKGITPFVTIYHWDLPFALQLKGGWANREIAD

[0191] WFAEYSRVLFENFGDRVKNWITLNEPWVVAIVGHLYGVHAPGMRDIYVAFRAVHNLLRAHARAVKVFR

[0192] ETVKDGKIGIVFNNGYFEPASEKEEDIRAVRFMHQFNNYPLFLNPIYRGDYPELVLEFAREYLPENYKDDM

[0193] SEIQEKIDFVGLNYYSGHLVKFDPDAPAKVSFVERDLPKTAMGWEIVPEGIYWILKKVKEEYNPPEVYITEN

[0194] GAAFDDVVSEDGRVHDQNRIDYLKAHIGQAWKAIQEGVPLKGYFVWSLLDNFEWAEGYSKRFGIVYVD

[0195] YSTQKRIVKDSGYWYSNVVKNNGLED

[0196] • SEQ ID NO: 2 (length: 446)

[0197] MNVKKFPEGFLWGVATASYQIEGSPLADGAGMSIWHTFSHTPGNVKNGDTGDVACDHYNRWKEDIEII

[0198] EKLGVKAYRFSISWPRILPEGTGRVNQKGLDFYNRIIDTLLEKGITPFVTIYHWDLPFALQLKGGWANREIA

[0199] DWFAEYSRVLFENFGDRVKNWITLNEPWVVAIVGHLYGVHAPGMRDIYVAFRAVHNLLRAHARAVKVF

[0200] RETVKDGKIGIVFNNGYFEPASEKEEDIRAVRFMHQFNNYPLFLNPIYRGDYPELVLEFAREYLPENYKDD

[0201] MSEIQEKIDFVGLNYYSGHLVKFDPDAPAKVSFVERDLPKTAMGWEIVPEGIYWILKKVKEEYNPPEVYIT

[0202] ENGAAFDDVVSEDGRVHDQNRIDYLKAHIGQAWKAIQEGVPLKGYFVWSLLDNFEWAEGYSKRFGIVY

[0203] VDYSTQKRIVKDSGYWYSNVVKNNGLED

[0204] • SEQ ID NO: 3 (length: 542)

[0205] MAMQLRSLLLCVLLLLLGFALADTNAAARIHPPVVCANLSRANFDTLVPGFVFGAATASYQVEGAANLD

[0206] GRGPSIWDTFTHKHPEKIADGSNGDVAIDQYHRYKEDVAIMKDMGLESYRFSISWSRVLPNGTLSGGINK

[0207] KGIEYYNNLINELLHNGIEPLVTLFHWDVPQTLEDEYGGFLSNRIVNDFEEYAELCFKKFGDRVKHWTTLN

[0208] EPYTFSSHGYAKGTHAPGRCSAWYNQTCFGGDSATEPYLVTHNLLLAHAAAVKLYKTKYQAYQKGVIGIT

[0209] VVTPWFEPASEAKEDIDAVFRALDFIYGWFMDPLTRGDYPQSMRSLVGERLPNFTKKESKSLSGSFDYIGI

[0210] NYYSARYASASKNYSGHPSYLNDVNVDVKTELNGVPIGPQAASSWLYFYPKGLYDLLCYTKEKYNDPIIYIT

[0211] ENGVDEFNQPNPKLSLCQLLDDSNRIYYYYHHLCYLQAAIKEGVKVKGYFAWSLLDNFEWDNGYTVRFGI

[0212] NYVDYDNGLKRHSKHSTHWFKSFLKKSSRNTKKIRRCGNNNTSATKFVF

[0213] • SEQ ID NO: 4 (length 544)

[0214] MALQFRSLLLCMVLLLLGFALANTNAARTDPPIVCATLNRTHFDTLFPGFTFGAATAAYQLEGAANIDGR

[0215] GPSVWDNFTHEHPEKITDGSNGDVAIDQYHRYKEDVAIMKDMGLDAYRFSISWSRLLPNGKLSGGINKK

[0216] GIEYYNNLTNELLRNGIEPLVTLFHWDVPQALVDEYGGLLSPRIVDDFKAYADLCYKEFGDRVKHWTTLNE

[0217] PYTISNHGYTIGIHAPGRCSDWYNPKCLGGDSGIEPYLVTHYLLLAHAAAVKLYREKYQAYQNGVIGITVVS

[0218] HWFEPASESQQDKDAAFQALDFMYGWFMDPLTRGDYPQIMRSILGARLPNFTEEQSKSLSGSYDYIGV NYYSARYASAYPKDYSVTTPPSYLTDVHVNVTTDLNGVPIGPRAASDWLYVYPKGLYDLVLYTKEKYNDPI MYITENGMDEFNNPKLSLEQALNDGNRIDYYYRHLCYLQAAMKEGANVQGYFAWSLLDNFEWSEGYT VRFGINYIDYDNGLERHSKLSTHWFKSFLKRSSISKKKIRRCGNNNGRATKFVYQI

[0219] • SEQ ID NO: 5 (length 661)

[0220] MRKRQPKTLHLLVMAMQLRSLLLCVLLLLLGFALADTNAAARIHPPVVCANLSRANFDILVPGFVFGAAT ASYQVEGAANLDGRGPSIWDTFTHKHPEKIADGSNGDVAIDQYHRYKEDVAIMKDMGLESYRFSISWSR VLPNGTLSGGINKKGIEYYNNLINELLHNGIEPLVTLFHWDVPQTLEDEYGGFLSNRIVNDFEEYAELCFKKF GDRVKHWTTLNEPYTFSSHGYAKGTHAPGRCSAWYNQTCFGGDSATEPYLVTHNLLLAHAAAVKLYKTK YQAYQKGVIGITVVTPWFEPASEAKEDIDAVFRALDFIYGWFMDPLTRGDYPQSMRSLVGERLPNFTKKE SKSLSGSFDYIGINYYSARYASASKNYSGHPSYLNDVNVDVKSEYKLKTSTELNGVPIGPQAASSWLYFYPK GLYDLLRYTKEKYNDPIIYITENGVDEFNQPNPKLSLCQLLDDSNRIYYYYHHLCYLQAAIKEGVKVKGYFA WSLLDNFEWDNGYTVRFGINYVDYDNGLKRHSKHSTHWFKSFLKKSSRNTKKIRRCGNNNTSATKGALT LFGCGWRDTLCLNIVDPPPKYLLLILRHLGTNEMLVGLDQMWRPIKYDYEVPKQALIFALCQTRLQFGTW EGYDTGLPTPLLHQALSNIALSHCNTT

[0221] Those skilled in the art will understand that they can freely combine all features of the present invention disclosed herein. In particular, features described for the product of the present invention may be combined with the method of the present invention and vice versa. Further, features described for different embodiments of the present invention may be combined.

[0222] Furthermore, where known equivalents exist to specific features, such equivalents are incorporated as if specifically referred in this specification. Further advantages and features of the present invention are apparent from the figures and non-limiting examples.

[0223] EXAMPLES

[0224] Example 1: Degradation of vicine / convicine in a faba suspension comprising faba concentrate via treatment with enzyme preparations possessing beta-glucosidase activity or side-activity at pH of 6.4 and at 60°C

[0225] The following experiments were conducted at 60°C, as a first attempt to test enzyme activities in a faba / water model system. These experiments were first carried out with faba concentrate containing low levels of vicine / convicine. Afterward, faba concentrate with high levels of vicine / convicine was used to test the robustness of this enzymatic approach.

[0226] A faba concentrate with low levels of vicine / convicine was provided. In particular, the initial vicine and convicine content in the faba concentrate containing low levels of vicine / convicine were respectively 661 and 176 mg / kg-faba ingredient.

[0227] 5% W / V of said faba concentrate (60% protein content) with low levels of vicine / convicine was slowly added into a thermomixer containing 60°C Milli-Q water. The water and the faba concentrate were mixed in the thermomixer for 30-60 min to form a faba suspension having a faba protein content of 3wt.%. The pH of the resulting faba suspension was pH 6.4 and no further pH adjustment was done. 0.25% W / V of enzyme preparation E1-E6 that possess betaglucosidase activity or side-activity was added into the faba suspension.

[0228] In particular, enzymes E1-E6 were purchased. Enzyme preparations E1-E4 possessed betaglucosidase activity as their main activity (i.e. E1-E4 are beta-glucosidases); enzyme preparation E5 was a xylanase possessing beta-glucosidase side activity; enzyme preparation E6 was a cellulase possessing beta-glucosidase side activity. The origin of the different enzymes El to E6 is provided in table 1 below. The amino acid sequences for enzymes E2 and E3 are also provided in table 1 below.

[0229] Table 1

[0230] After addition of the enzymatic preparation, the suspension was incubated at 60°C at 900 rpm in a thermomixer for 60 min to obtain an enzyme treated faba composition. Subsequently, this enzyme treated faba composition was heated at 95°C for 15 min to inactivate enzymes. Remaining vicine and / or convicine in the enzyme treated faba composition were extracted with 70% acetonitrile in Milli-Q water and were subjected to measurement by reversed phase high performance chromatography (RP-HPLC) with a diode array detector (DAD) at 273 nm. In particular, separation of vicine and convicine was achieved by an Acquity UPLC® BEH HILIC column (2.1 x 100 mm) with particle size of 1.7pm (Waters, Milford, USA). Elution of vicine and convicine was accomplished by a gradient run for 10 min at a flow rate of 0.3 mL / min: 1) starting from 0% of Eluent A (10% acetonitrile in Milli-Q water, containing 10 mM ammonium formate and 0.1% formic acid) and 100% of Eluent B (90% acetonitrile in Milli-Q water, containing 10 mM ammonium formate and 0.1% formic acid) to 20% of Eluent A and 80% of Eluent B in the first 5 min; 2) the gradient of Eluent B was decreased from 80% to 50% in 1.5 min and held for 1 min; 3) finally the gradient of Eluent B was increased from 50% to 100% in 0.5 min and held for 2 min.

[0231] After elution, the vicine / convicine content was measured by UV at 273nm by a diode array detector. Vicine / convicine content in the resulting enzyme treated faba composition was compared against a control that was treated equally but without the enzyme addition.

[0232] The above experiment was repeated using faba concentrate (60% protein content) containing high levels of vicine / convicine instead of the faba concentrate containing low levels of vicine / convicine to test the robustness of this enzyme method. The faba concentrate with high levels of vicine / convicine (i.e. before incubation) contains 19363 mg vicine per kg-faba ingredient, and 5508 mg convicine per kg-faba ingredient.

[0233] The results are shown in figures 1 to 4.

[0234] Among the chosen enzyme preparations, El was not effective at significantly degrading vicine and convicine levels in faba compositions at pH of 6.4. The reduction of vicine / convicine was lower than 30% for faba composition prepared with faba concentrate comprising low and high levels of vicine / convicine (Figures 1-4).

[0235] E2-E6 achieved a significant reduction of vicine and / or convicine of more than 30% at pH of 6.4 at 60°C. This is the case for faba composition prepared with faba concentrate comprising low and high levels of vicine / convicine. For faba concentrate containing low levels of vicine / convicine, E2, E3, E4 and the cellulase E6 containing beta-glucosidase side activity managed to degrade vicine and / or convicine at a level of more than 50% (Figures 1 and 2). The highest reduction in vicine and convicine was achieved with E2 and E3 (more than 80% reduction).

[0236] For faba concentrate containing high levels of vicine / convicine, E2 and E3 were able to degrade vicine / convicine to more than 80% at native pH of 6.4 (Figures 3 and 4).

[0237] Example 2: Degradation of vicine / convicine in a faba suspension comprising faba concentrate via treatment with enzyme preparations possessing beta-glucosidase activity or side-activity at pH of 6.4 and at a temperature from 50°C to 65°C

[0238] The degradation of vicine / convicine in faba concentrate suspensions was performed as described in Example 1 but at different incubation temperatures 50°C, 55°C, 60°C or 65°C with E2 or E3 to test the efficiency of E2 and E3 at different incubation temperature.

[0239] For these trials, the faba concentrate containing low levels of vicine / convicine of example 1 (figures 5 and 6) and the faba concentrate containing high levels of vicine / convicine of example 1 (figures 7 and 8) were also used. In particular, the initial vicine and convicine content in the faba concentrate containing low levels of vicine / convicine were respectively 661 and 176 mg / kg-faba ingredient; and the initial vicine and convicine content in the faba concentrate containing high levels of vicine / convicine were respectively 19363 and 5508 mg / kg-faba ingredient.

[0240] The results are shown in figures 5 to 8.

[0241] For compositions prepared with faba concentrate containing low and high levels of vicine / convicine, both beta-glucosidases E2 and E3 could reach at least 55% degradation of vicine and convicine at all temperatures. E2 shows outstanding degradation of more than 99% of vicine / convicine at all temperatures. Example 3: Degradation of vicine / convicine in a model recipe containing faba ingredients to produce a food product via enzymatic treatment with enzyme preparations possessing betaglucosidase activity or side-activity at a pH of 6.4 and at a temperature of 60°C

[0242] 2.5% W / V oat flour, 0.2% W / V gellan gum dissolved in 2.5% W / V canola oil, 2.5% W / V sugar, 3.5% W / V faba concentrate with low level of vicine / convicine of example 1 were added into a thermomixer containing 60°C Milli-Q. water. The different ingredients were homogenized together into the thermomixer for 30-60 minutes to form a food composition. The pH of the food composition was pH 6.4 and no further adjustment was made. 0.18% W / V of enzyme preparations E2 and E3 of example 1 were added into the food composition. The food composition was incubated at 60°C at 900 rpm in a thermomixer for 60 min. Subsequently, the food compositions was heated at 95°C for 15 min to inactivate enzymes and to obtain a food product, in particular a plant-based milk analogue.

[0243] Remaining vicine and / or convicine in the food product was extracted with 70% acetonitrile in Milli-Q water and was subjected to measurement by RP-HPLC coupled with DAD at 273 nm as described in example 1. Vicine / convicine content in the resulting food product was compared against a control that was treated equally but without the enzyme addition.

[0244] The results are shown in figures 9 and 10.

[0245] The results show that E2 and E3 were also able to degrade significantly vicine and convicine to more than 80% even in a complex food matrix comprising different ingredients.

[0246] Although the invention has been described by way of example, it should be appreciated that variations and modifications may be made without departing from the scope of the invention as defined in the claims.

Claims

CLAIMS A method of reducing vicine and / or convicine content in a faba ingredient, said method comprising the steps of: a) Providing a faba ingredient which is liquid or suspending a faba ingredient into an aqueous liquid to form a faba ingredient which is liquid, wherein the faba ingredient comprises vicine and / or convicine, b) Adding at least one vicine and / or convicine-degrading enzyme to the faba ingredient, wherein the faba ingredient has a pH of 6.0 or higher, c) Incubating the faba ingredient with the at least one vicine and / or convicine- degrading enzyme to obtain a treated faba ingredient, d) Optionally, partially or fully inactivating the vicine and / or convicine-degrading enzyme in the treated faba ingredient, e) Optionally, cooling the treated faba ingredient. The method according to claim 1, wherein the faba ingredient has a pH between pH 6.0 and pH 9.

0. The method according to any one of the preceding claims, wherein the faba ingredient is whole faba bean, faba bean flour, faba bean protein concentrate or faba bean protein isolate. The method according to any one of the preceding claims, wherein the at least one vicine and / or convicine-degrading enzyme is selected from the list consisting of betaglucosidase, enzyme having beta-glucosidase side activity or a mixture thereof. The method according to any one of the preceding claims, wherein the at least one vicine and / or convicine-degrading enzyme is one or more beta-glucosidases having amino acid sequences that are at least 70% identical to one or more amino acid sequences selected from the group consisting of SEQ ID NO: 1, SEQ NO: 2, SEQ NO: 3, SEQ NO: 4, and SEQ ID NO: 5.

6. The method according to any one of the preceding claims, wherein the at least one vicine and / or convicine-degrading enzyme comprises vicine and / or convicine- degrading enzyme coming from almond.

7. The method according to any one of the preceding claims, one vicine and / or convicine- degrading enzyme comprises vicine and / or convicine-degrading enzyme coming from Thermotoga maritima.

8. The method according to any one of the preceding claims, wherein the content of vicine and / or convicine in the faba ingredient is reduced by at least 30%, preferably at least 50% by the at least one vicine and / or convicine-degrading enzyme in step c).

9. The method according to any one of the preceding claims, which comprises a step of drying the treated faba ingredient after step e) to obtain a faba ingredient which is a powder.

10. The method according to any one of the preceding claims, wherein at least lwt% faba ingredient is suspended into the aqueous liquid in step a).

11. The method according to any one of the preceding claims, wherein the step c) of incubation is performed for less than 2 hours.

12. A treated faba ingredient which is obtained with the method of any one of claims 1 to 11.

13. A faba ingredient, wherein the sum of vicine and convicine content is less than 600 mg per kg of faba ingredient, preferably less than 450 mg per kg of faba bean protein.

14. The treated faba ingredient according to claim 12 or the faba ingredient according to claim 13, wherein the faba ingredient or the treated faba ingredient is liquid, semiliquid or powder.The treated faba ingredient according to claim 12 or 14 or the faba ingredient according to claim 13 or 14, wherein proteins are intact. A food product which comprises the treated faba ingredient according to any one of claims 12 or 14-15 or the faba ingredient according to any one of claims 13 to 15. A method for producing a food product which comprises the step of: a) providing a food composition, wherein the food composition comprises faba ingredient and at least one additional food ingredient, and wherein the faba ingredient of the food composition comprises vicine and / or convicine, b) Adding at least one vicine and / or convicine-degrading enzyme to the food composition, wherein the food composition has a pH of 6.0 or higher, c) Incubating the food composition with the at least one vicine and / or convicine- degrading enzyme to obtain a treated food composition, d) optionally, homogenizing the treated food composition, e) Heat-treating the treated food composition, f) Cooling the treated food composition to obtain a food product. A method for producing a food product which comprises the step of: a) providing a food composition, wherein the food composition comprises the treated faba ingredient according to any one of claims 12 or 14-15 or the faba ingredient according to any one of claims 13 to 15, and also comprises at least one additional food ingredient, b) optionally, homogenizing the food composition, c) Heat-treating the food composition, d) Cooling the food composition to obtain a food product. The method according to claim 17 or 18, wherein the at least one additional food ingredient is selected from the list consisting of protein different from faba bean protein, sweetening agent, texturizing agent, buffering salt, bulking agent, fiber, fat, mineral, vitamin, antioxidant, colour, flavouring agent, aqueous liquid, or a combination thereof.The method according to claim 19, wherein the at least one additional food ingredient comprises protein different from faba bean protein and wherein the protein is a cereal, pulse, or dairy protein, and wherein the pulse protein is different from faba bean protein. A food product obtained by the method of claims 17-20.

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