Reduction of Vicine and / or Convicine in Fava Ingredients and Food Products

Abstract

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

Description

[Technical Field]

[0001] The present invention relates generally to the field of faba ingredients or food products prepared with faba ingredients. In particular, the present invention relates to faba ingredients or food products having reduced levels of vicine and / or convicine, and methods / processes for producing such faba ingredients or food products.

[0002] [Background technology] Faba beans are a very good source of protein for food products, especially plant-based food products. One drawback is that the naturally occurring antinutritional factors vicin and convicin make faba (and its derived components, such as protein concentrates and isolates) unsuitable for consumption by faba bean poisoning sufferers. Reducing the levels of vicin and convicin would allow for the incorporation of faba into food products that may be safe for consumption, especially for individuals suffering from faba bean poisoning.

[0003] Various strategies have been explored to reduce the levels of vicine and convicine in faba.

[0004] One strategy involves the use of microorganisms to cleave and reduce vicine and convicine in fava by fermentation. However, fermentation takes a long time to achieve significant reduction of vicine and convicine. Furthermore, fermentation is not a specific reaction and may target compounds other than vicine and convicine in fava. This non-specificity may alter the sensory and nutritional properties of fava. Specifically, fermentation may impart undesirable flavors, including fermented flavors. In addition, fermentation reduces the pH of fava, which may be undesirable from a stability standpoint or for food applications.

[0005] Another strategy involves using enzymes to cleave and reduce the vicine and convicine in faba by hydrolysis. For example, β-glucosidase can be used to cleave the glucose group from the vicine and convicine molecules. The reaction products are the aglycone forms of vicine and convicine and glucose.

[0006] In the art, enzymatic degradation of bicine and convicine is generally performed at acidic pH. In particular, demonstrated enzymatic degradation of bicine and convicine requires several hours (2-4 hours) at acidic pH (pH 4.6 or pH 5) (Arbid and Marquardt 1985; Pulkkinen 2019).

[0007] The use of an acidic pH is not advantageous. In fact, an acidic pH reduces the stability of the proteins in the fava during processing and / or shelf life. In fact, fava bean proteins, which have an isoelectric point of about 4.5, are poorly soluble at acidic pH and prone to uncontrolled aggregation, precipitation and sedimentation, making an acidic pH undesirable during processing and / or shelf life. An acidic pH negatively affects the sensory properties and functionality of ingredients and food products prepared with fava.The effect of pH on the properties and functionality of faba protein has been well documented in the literature; Otegui, I. 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 characterization of concentrated emulsion gels stabilized 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. 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.

[0008] Furthermore, one of skill in the art will recognize that the activity of an enzyme is pH dependent: an enzyme that is active at an acidic pH may not be active, or may have significantly reduced activity, at other pHs, including neutral pH.

[0009] For food applications, it is desirable to carry out enzymatic reactions in food matrices at neutral, near-neutral or slightly alkaline pH, especially at a pH above 6, with short reaction times.

[0010] It is therefore desirable to provide a method for reducing, preferably eliminating, vicine and / or convicin in fava ingredients and in food products prepared with such fava ingredients by enzymatic treatment, wherein the enzymatic solution desirably results in a significant reduction, preferably elimination, of vicine and / or convicin in a short period of time at a neutral, near-neutral, or slightly alkaline pH, particularly a pH above 6.

[0011] Any reference herein to a prior art document should not be taken as an admission that such prior art is well known or forms part of the common general understanding in the art.

[0012] [Summary of the Invention] The present invention aims to improve upon the current state of the art, and in particular to provide methods, food ingredients and food products that overcome the problems of the prior art and address the needs noted above, or at least to provide useful alternatives.

[0013] The inventors have surprisingly found that the object of the invention can be achieved by the subject matter of the independent claims. The dependent claims develop the inventive idea further.

[0014] Thus, a first aspect of the present invention is a method for reducing the vicine and / or convicine content in a fava component, comprising the steps of: a) providing a liquid fava component or suspending the fava component in an aqueous liquid to form a liquid fava component, wherein the fava component comprises vicine and / or convicine; b) adding at least one bicin degrading enzyme and / or convicin degrading enzyme to the fava component, wherein the pH of the fava component is 6.0 or higher; c) incubating the fava component with at least one bicin degrading enzyme and / or convicin degrading enzyme to obtain a treated fava component; d) optionally partially or completely inactivating the bicin- and / or convicin-degrading enzymes in the treated fava component; e) optionally cooling the treated fava component; The present invention proposes a method including:

[0015] A second aspect of the present invention proposes a processed fava component obtainable by the method of the first aspect of the present invention.

[0016] A third aspect of the present invention proposes a fava component, the sum of the vicine and convicine contents of which is less than 600 mg per kg of fava component, preferably less than 450 mg per kg of fava bean protein.

[0017] A fourth aspect of the present invention proposes a food product comprising a processed fava component according to the second aspect of the present invention or a fava component according to the third aspect of the present invention.

[0018] A fifth aspect of the present invention is a method for producing a food product, comprising the steps of: a) providing a food composition, the food composition comprising a fava component and at least one additional food ingredient, the fava component of the food composition comprising vicine and / or convicine; b) adding at least one bicin-degrading enzyme and / or convicin-degrading enzyme to the food composition, wherein the pH of the food composition is 6.0 or higher; c) incubating the food composition with at least one bicin-degrading enzyme and / or convicin-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; The present invention proposes a method including:

[0019] A sixth aspect of the present invention is a method of producing a food product, comprising the steps of: a) providing a food composition, the food composition comprising the treated fava component according to any one of claims 9 or 11-12, or the fava component according to any one of claims 10-12, and also comprising 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 present invention proposes a method including:

[0020] A seventh aspect of the present invention proposes a food product obtainable by a method according to the fifth aspect of the present invention or the sixth aspect of the present invention.

[0021] It has been discovered that in fava components, or food products containing fava components, it is possible to achieve a significant reduction of vicine and / or convicine in a short period of time by enzymatic treatment with bicine-degrading enzymes and / or convicine-degrading enzymes at neutral, near-neutral or slightly alkaline pH, particularly at a pH above 6.

[0022] These and other aspects, features and advantages of the present invention will become more apparent to those skilled in the art from the following detailed description of the embodiments of the invention taken in conjunction with the accompanying drawings. [Brief explanation of the drawings]

[0023] [Figure 1] Figure 1 shows the percentage of bicine reduction / degradation by enzymes E1-E6 at 60°C and pH 6.4 in fava suspensions including fava concentrates containing low levels of vicine / convicine. The initial vicine content (100%) before enzyme treatment in fava concentrates with low levels of vicine / convicine was 661 mg / kg of fava component. [Figure 2]Figure 1 shows the percentage of convicin reduction / degradation by enzymes E1-E6 at 60°C and pH 6.4 in fava suspensions including fava concentrates containing low levels of vicin / convicin. The initial convicin content (100%) before enzyme treatment in fava concentrates with low levels of vicin / convicin was 176 mg / kg of fava component. [Figure 3] Figure 1 shows the percentage of bicine reduction / degradation by enzymes E1-E6 at 60°C and pH 6.4 in a fava suspension containing a fava concentrate containing high levels of vicine / convicine. The initial vicine content (100%) before enzyme treatment in a fava concentrate with high levels of vicine / convicine was 19363 mg / kg of fava component. [Figure 4] Figure 1 shows the percentage of convicin reduction / degradation by enzymes E1-E6 at 60°C and pH 6.4 in a fava suspension containing a fava concentrate containing high levels of vicin / convicin. The initial convicin content (100%) before enzyme treatment in a fava concentrate with high levels of vicin / convicin was 5508 mg / kg of fava component. [Figure 5] Figure 1 shows the percentage of bicine reduction / degradation by enzymes E2 and E3 at 50°C, 55°C, 60°C and 65°C and pH 6.4 in fava suspensions including fava concentrates containing low levels of bicine / convicin. The initial bicine content (100%) before enzyme treatment in fava concentrates with low levels of bicine / convicin was 661 mg / kg of fava component. [Figure 6] Figure 1 shows the percentage of convicin reduction / degradation by enzymes E2 and E3 in fava suspensions including fava concentrates containing low levels of vicin / convicin at 50°C, 55°C, 60°C and 65°C and at pH 6.4. The initial convicin content (100%) before enzyme treatment in fava concentrates with low levels of vicin / convicin was 176 mg / kg of fava component. [Figure 7]Figure 1 shows the percentage of bicine reduction / degradation by enzymes E2 and E3 in a fava suspension including a fava concentrate containing high levels of bicine / convicine at 50°C, 55°C, 60°C and 65°C and at pH 6.4. The initial bicine content (100%) before enzyme treatment in the fava concentrate with high levels of bicine / convicine was 19363 mg / kg fava component. [Figure 8] Figure 1 shows the percentage of convicin reduction / degradation by enzymes E2 and E3 in a fava suspension including a fava concentrate containing high levels of vicin / convicin at 50°C, 55°C, 60°C and 65°C and at pH 6.4. The initial convicin content (100%) before enzyme treatment in a fava concentrate with high levels of vicin / convicin was 5508 mg / kg fava component. [Figure 9] Figure 1 shows the percentage of bicine reduction / degradation by enzymes E2 and E3 at 60°C and pH 6.4 in a food product, specifically a vegetable milk analogue containing a fava concentrate with low levels of vicine and convicine. The initial vicine content (100%) before enzyme treatment in the fava concentrate with low levels of vicine / convicine, respectively, was 661 mg / kg fava component. [Figure 10] Figure 1 shows the percentage of convicin reduction / degradation by enzymes E2 and E3 at 60°C and pH 6.4 in a food product, specifically a vegetable milk analogue containing a fava concentrate with low levels of vicin and convicin. The initial convicin content (100%) before enzyme treatment in the fava concentrate with low levels of vicin / convicin was 176 mg / kg of fava component.

[0024] [Mode for Carrying Out the Invention] As used herein, the terms "comprise," "comprising," and the like are to be interpreted in an inclusive sense, i.e., "including, but not limited to," rather than in an exclusive or exhaustive sense.

[0025] As used herein, all numerical ranges should be understood to include every integer within the range.

[0026] As used herein, the singular articles "a," "an," and "the" include plural referents unless the context clearly dictates otherwise.

[0027] As used herein, the term "substantially free" means that no more than about 0.1% by weight, preferably no more than 0.01% by weight, of the excluded material remains. "Completely free" typically means that only trace amounts of the excluded material are present, preferably no detectable amounts.

[0028] Unless otherwise stated, all percentages herein refer to weight percentages, where applicable.

[0029] Unless otherwise defined, all technical and scientific terms have the same meaning and should be given the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0030] The term "faba ingredient" refers to an ingredient that comprises, preferably consists of, a compound derived from faba, preferably from the faba bean, also commonly called broad bean, fava bean, or field bean.

[0031] The term "whole faba bean" refers to faba beans that are intact and have not been subjected to any process of mechanical disruption (e.g., milling, grinding) and / or fractionation to remove fiber, carbohydrates and / or protein. Whole faba beans comprise the germ and endosperm, and optionally the husk. Preferably, whole faba beans comprise the germ, endosperm, and husk. In particular, whole faba beans are not faba bean flour, faba bean protein concentrate, and faba bean protein isolate. For the avoidance of doubt, the term "whole faba bean" does not exclude faba beans that have been subjected to a husk removal process.

[0032] The term "fava bean flour" refers to a composition in powder form having a fava bean protein content of between 5% and 49.9% protein by weight, preferably between 20% and 49.9% protein by weight.

[0033] As used herein, the term "faba bean protein concentrate" refers to a composition having a faba bean protein content of between 50% and 79.9% by weight.

[0034] As used herein, the term "faba bean protein isolate" refers to a composition having a faba bean protein content of between 80.0% and 99.0% by weight.

[0035] As used herein, the term "bicin- and / or convicin-degrading enzyme" refers to an enzyme that degrades vicin and / or convicin into compounds that are no longer vicin and / or convicin, respectively.

[0036] As used herein, the term "β-glucosidase (e.g., EC 3.2.1.21)" refers to a type of glycosyl hydrolase that catalyzes the hydrolysis of terminal non-reducing β-D-glucosyl residues with the release of β-D-glucose.

[0037] As used herein, the term "α-amylase (e.g., EC 3.2.1.1)" refers to an enzyme that catalyzes the endohydrolysis of (1→4)-α-D-glucosidic linkages in polysaccharides having three or more (1→4)-α-linked D-glucose units.

[0038] As used herein, the term "identical" in the context of amino acid sequences means that the amino acid residues in two or more sequences are the same when aligned for maximum correspondence. The length of amino acid sequence identity comparison may be over a contiguous 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 many different algorithms and methods known to those of skill in the art that can be used to measure amino acid sequence identity. By way of example and not limitation, the percentage (%) of sequence identity of two or more amino acid sequences can be calculated using CLUSTAL W (version 1.82, version 2), CLUSTAL omega, BLAST, EMBOSS matcher, or MULTALIN after aligning the sequences to achieve the maximum percent sequence identity and introducing gaps, if necessary, without considering any conservative substitutions as part of the sequence identity.

[0039] As used herein, the term "meat analog product," which may also be referred to as a meat substitute, meat substitute, imitation meat, fake meat, simulated meat, or vegetarian meat, refers herein to a food product that contains plant-derived ingredients, is made exclusively from vegetarian ingredients, preferably vegan ingredients, and has qualities similar to those of a corresponding real meat product in terms of appearance, taste, flavor, and texture. Non-limiting examples of real meat products include products derived from poultry, beef, pork, fish, and mixtures thereof.

[0040] As used herein, the term "dairy analog product" refers herein to a food product that contains plant-derived ingredients, is dairy-free, and has similar qualities in appearance and texture to the corresponding authentic dairy product. Preferably, the dairy analog product is made exclusively from vegan ingredients.

[0041] As used herein, the term "dairy analog" refers herein to a food product that contains plant-derived ingredients, is dairy-free, and has similar qualities in appearance and texture to the corresponding real dairy milk. Preferably, the milk analog is made only from vegan ingredients.

[0042] As used herein, the term "cream analog" refers herein to a food product that contains plant-derived ingredients, is dairy-free, and has qualities similar to the corresponding real dairy cream in terms of appearance and texture. Preferably, the cream analog is made only from vegan ingredients.

[0043] As used herein, the term "egg analog product" refers herein to a food product that contains plant-derived ingredients, is egg-free, and has similar qualities in appearance and texture to the corresponding real egg product. Preferably, the egg analog product is made only from vegan ingredients.

[0044] As used herein, the term "vegetarian" refers to an edible composition that does not contain meat, such as fish.

[0045] As used herein, the term "vegan" refers to an edible composition that does not contain any animal products or animal-derived products.

[0046] 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, etc.), pectin, gums (e.g., gellan gum, locust bean gum, carob gum, guar gum, etc.), hydrocolloids (e.g., alginates, agar, etc.), and fibers.

[0047] As used herein, the term "sweetener" refers to an ingredient that imparts or increases sweetness in a food product or ingredient. Examples of sweeteners include agave syrup, brown sugar, coconut sugar, maple syrup, rice syrup, oat syrup, corn syrup, dextrose, fructose, glucose, honey, invert sugar, maltose, molasses, sucrose, steviol glycosides, and mixtures thereof.

[0048] 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.

[0049] As used herein, the term "flavoring" refers to an ingredient that imparts, enhances, masks, or modifies the taste and / or odor of a food product or food ingredient.

[0050] As used herein, the term "aqueous liquid" refers to a composition that contains at least 70% water by weight. The aqueous liquid may be milk, a milk analog, a cream analog, water, or a combination thereof.

[0051] As used herein, the term "seed" refers to seeds that are distinct from legume seeds, commonly called pulses, and that are distinct from nut seeds, commonly called nuts.

[0052] In a first aspect, the present invention relates to a method for reducing the vicine and / or convicin content in fava components, preferably a method for completely removing vicine and / or convicin from the fava components.

[0053] The method includes the step a) of providing a fava component that is a liquid.

[0054] Alternatively, the method includes step a) of suspending the fava component in an aqueous liquid to form a liquid fava component.

[0055] In one embodiment, the aqueous liquid in step a) has a temperature of at least 45° C., preferably 45° C. to 80° C., more preferably 50° C. to 75° C. For example, the aqueous liquid in step a) has a temperature of 60° C.

[0056] In a preferred embodiment, the aqueous liquid is water.

[0057] In one embodiment, at least 0.5% by weight of the fava component is suspended in the aqueous liquid in step a), preferably at least 1% by weight, more preferably at least 2% by weight, and even more preferably at least 2.5% by weight. In one embodiment, at most 50% by weight of the fava component is suspended in the aqueous liquid in step a), preferably at most 25% by weight, more preferably at most 15% by weight, even more preferably at most 10% by weight, and most preferably at most 8% by weight.

[0058] The fava component comprises vicine and / or convicine. Preferably, the sum of the vicine and convicine contents in the fava component is at least 450 mg / kg of fava component, preferably at least 600 mg / kg of fava component, more preferably 600 mg to 45,000 mg / kg of fava component, more preferably 600 to 35,000 mg / kg of fava component, even more preferably 800 to 35,000 mg / kg of fava component, even more preferably 800 to 30,000 mg / kg of fava component, even more preferably 800 to 20,000 mg / kg of fava component, even more preferably 800 to 10,000 mg / kg of fava component, and most preferably 800 to 2,000 mg / kg of fava component.

[0059] Furthermore, the fava component comprises fava bean protein. In particular, the fava component comprises at least 1% by weight, preferably 1% to 99% by weight, more preferably 10% to 99% by weight, even more preferably 20% to 99% by weight, even more preferably 30% to 99% by weight, even more preferably 50% to 99% by weight of fava bean protein.

[0060] In some other embodiments, the fava component may comprise at least 1% by weight, preferably 1% to 45% by weight, more preferably 10% to 45% by weight, even more preferably 20% to 45% by weight, and even more preferably 30% to 45% by weight of fava bean protein.

[0061] In a preferred embodiment, the fava ingredient is whole fava beans, fava bean flour, fava bean protein concentrate, or fava bean protein isolate. If the fava ingredient is whole fava beans, the whole fava beans are ground before step a). The whole fava beans are ground by wet milling or dry milling, preferably by wet milling. The whole fava beans may be cooked before wet or dry milling.

[0062] The fava component may be mixed with at least one additional food ingredient before step b). For example, the at least one additional food ingredient may be selected from the list consisting of sweeteners, tempering agents, buffer salts, bulking agents, fiber, fat, minerals, vitamins, antioxidants, colorants, flavorings, aqueous liquids, proteins other than fava bean protein, or combinations thereof. Preferably, the protein other than fava bean protein may be a plant protein other than fava bean protein, a dairy protein, or a mixture thereof. More preferably, the protein other than fava bean protein may be selected from the list consisting of a cereal protein, a seed protein, a nut protein, a pulse protein, a dairy protein, or a mixture thereof. The pulse protein is different from the fava bean protein. Even more preferably, the protein other than fava bean protein may be selected from the list consisting of a cereal protein, a pulse protein, a dairy protein, or a mixture thereof. Examples of pulse proteins include chickpea protein, pea protein, lentil protein, or a mixture thereof. Examples of grain proteins include oat protein, rice protein, wheat protein, barley protein, rye protein, buckwheat protein, quinoa protein, millet protein, corn protein, or a mixture thereof. Examples of seed proteins include hemp seed protein, canola seed protein, flaxseed protein, pumpkin seed protein, sunflower seed protein, cottonseed protein, safflower seed protein, squash seed protein, chia seed protein, or a mixture thereof. Examples of nut proteins 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 proteins include casein, whey protein, or a mixture thereof.

[0063] In one embodiment, the fat may be dairy fat or vegetable fat, examples of which include cocoa butter, coconut oil, rapeseed oil, canola oil, walnut oil, grapeseed oil, peanut oil, olive oil, sunflower seed oil, or mixtures thereof.

[0064] In one 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 protein of the fava component consists of fava bean protein. Most preferably, the protein of the fava component consists exclusively of fava bean protein.

[0065] The method further comprises step b) of adding at least one bicin- and / or convicin-degrading enzyme to the fava component.

[0066] The at least one bicin- and / or convicin-degrading enzyme is selected from the list consisting of β-glucosidase, an enzyme with β-glucosidase side activity, or a mixture thereof. It has been observed that β-glucosidase, as well as enzymes with β-glucosidase side activity, allow for an effective reduction of vicine and / or convicin in the fava component at a pH above 6.

[0067] β-glucosidase includes any enzyme that has the function of a β-glucosidase, ie, the hydrolysis of β-glycosidic bonds to terminal non-reducing residues in glucosides and oligosaccharides with the release of glucose.

[0068] In a preferred embodiment, the enzyme with β-glucosidase side activity is selected from the list consisting of glucosidase, xylanase, cellulase, hemicellulase, amylase, glucanase, pectinase, mannanase, or mixtures thereof. Preferably, the enzyme with β-glucosidase side activity is selected from the list consisting of xylanase, cellulase, or mixtures thereof.

[0069] In one embodiment, the at least one bicin-degrading enzyme and / or convicin-degrading enzyme comprises, and preferably consists of, a vicin-degrading enzyme and / or convicin-degrading enzyme derived from almond, particularly Prunus dulcis, and / or a vicin-degrading enzyme and / or convicin-degrading enzyme derived from Thermotoga maritima. More preferably, the at least one bicin-degrading enzyme and / or convicin-degrading enzyme comprises, and preferably consists of, a β-glucosidase derived from almond, particularly Prunus dulcis, and / or a β-glucosidase derived from Thermotoga maritima. Even more preferably, the at least one bicin-degrading enzyme and / or convicin-degrading enzyme comprises, and preferably consists of, a vicin-degrading enzyme and / or convicin-degrading enzyme derived from Thermotoga maritima. Most preferably, the at least one bicin- and / or convicin-degrading enzyme comprises, and preferably consists of, a β-glucosidase derived from Thermotoga maritima.

[0070] The presence of vicine and / or convicine degrading enzymes, particularly β-glucosidases derived from almonds, especially Prunus dulcis, and / or Thermotoga maritima, offers significant advantages. These enzymes have been observed to effectively reduce the levels of vicine and / or convicine in favas, achieving high reduction rates in the range of more than 80% at pH levels above 6. Remarkably, β-glucosidase from Thermotoga maritima shows exceptional effectiveness in nearly eliminating vicine and convicine in favas at pH levels above 6.

[0071] In an even more preferred embodiment, the at least one bicin-degrading enzyme and / or convicin-degrading enzyme is one or more β-glucosidases having an amino acid sequence that is 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 sequences 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. In a most preferred embodiment, the at least one bicin-degrading enzyme and / or convicin-degrading enzyme is one or more β-glucosidases having an amino acid sequence that is 100% identical to one or more amino acid sequences 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.

[0072] The presence of bicin and / or convicin degrading enzymes having the above sequences provides significant advantages. The above enzymes have been observed to effectively reduce the levels of vicin and / or convicin in fava, achieving high reduction rates in the range of more than 80% at pH levels above 6. Notably, the enzymes of SEQ ID NO: 1 and SEQ ID NO: 3 show exceptional effectiveness in nearly removing vicin and convicin in fava at pH levels above 6. In one embodiment, at least one enzyme different from the bicin and / or convicin degrading enzymes may also be further added to the fava component in step b).

[0073] The pH of the fava component is 6.0 or higher, preferably 6.0 to 9.0, more preferably 6.0 to 7.5. The fava component generally already has this pH in step a), i.e., the original pH of the fava component is 6.0 or higher, preferably 6.0 to 9.0, more preferably 6.0 to 7.5. If the pH of the fava component is not 6.0 or higher, the pH of the fava component may be adjusted between steps a) and b) to reach a pH of 6.0 or higher, preferably 6.0 to 9.0, more preferably 6.0 to 7.5.

[0074] It has been discovered that a short enzymatic treatment at a pH above 6 can significantly reduce the amount of vicine and / or convicine.

[0075] The dosage of bicin and / or convicin degrading enzymes added to the fava component will vary depending, inter alia, on the temperature during incubation, the volume or amount of fava component to be processed, and the activity of the enzymes. Generally, this dosage should be at least 0.1% (w / v) based on the volume of the liquid fava component. Preferably, this dosage of bicin and / or convicin degrading enzymes may be between 0.1% and 1% (w / v) based on the volume of the liquid fava component.

[0076] The method comprises a step c) of incubating the fava component with at least one bicin and / or convicin degrading enzyme to obtain a treated fava component. This incubation step c) is carried out at a pH above 6.0, preferably between 6.0 and 9.0, preferably between 6.0 and 7.5. The incubation step c) may be carried out at a temperature of at least 50°C, and optionally between 50 and 65°C.

[0077] The present invention allows for a significant reduction of vicine and / or convicine in a short reaction time.

[0078] The incubation step is carried out for less than 2 hours, preferably less than 90 minutes, more preferably 15 to 90 minutes.

[0079] It has been observed that even at a pH above 6, such enzymes were effective in significantly reducing the amount of vicine and / or convicine in the fava component.

[0080] In particular, the content of vicine and / or convicin in the fava component is reduced by the at least one bicine- and / or convicin-degrading enzyme in step c) 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%, and most preferably 100%. In other words, the treated fava component has a vicine and / or convicin content that 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%, and most preferably 100% less than the vicine and / or convicin content in the fava component.

[0081] Because the process is carried out at a pH above 6, the proteins, particularly the fava bean proteins, remain intact and retain good stability and good functionality during shelf life and / or processing, making the fava ingredient suitable for use in food applications.

[0082] Thus, in one embodiment, the proteins of the processed fava component, particularly the fava bean proteins, are intact, "intact" referring to proteins that do not aggregate in an uncontrolled manner, particularly proteins that do not precipitate or sediment.

[0083] The process (i.e., method) includes an optional step d) of partially or completely, preferably completely, inactivating at least one bicin- and / or convicin-degrading enzyme in the treated fava component. In one embodiment, step d) is not optional. In a preferred embodiment, the inactivation step d) is carried out by heat treating the treated fava component. The heat treatment is carried out at a temperature of at least 70°C, preferably between 70°C and 140°C, more preferably between 80°C and 110°C. Preferably, the heat treatment is carried out for a period of between 3 seconds and 20 minutes.

[0084] The process (i.e., method) includes an optional step d) of cooling the treated fiber component, for example, to a storage temperature for the treated fiber component, such as room temperature.

[0085] The process (i.e., method) may include evaporating the treated fiber component after step c), or after step d), or after step e).

[0086] The process (i.e., method) may include, after step c), or after step d), or after step e), drying the treated fava ingredient to obtain a powdered treated fava ingredient. The drying step may be carried out by drum drying, roller drying, spray drying, or freeze drying. The drying step may also be carried out by any other food drying technique known in the art. Converting the ingredient to a powder may be advantageous, as powders generally have good physicochemical stability and a long shelf life.

[0087] The evaporation and / or drying step may also make it possible to inactivate at least one bicine- and / or convicine-degrading enzyme, in which case the inactivation step d) is optional.

[0088] The present process (i.e., method) allows for a significant reduction of vicine and / or convicine in fava components in a limited time at a pH above 6. The resulting treated fava components retain good stability and functionality and can be used in food applications.

[0089] In one embodiment, the process (i.e., method) does not include the addition of microorganisms, particularly lactic acid bacteria. The process (i.e., method) does not include a fermentation step, particularly before step c) or step d). A significant reduction in vicine and / or convicine is obtained without fermentation. Fermentation is undesirable for the reasons described in the background of the present invention.

[0090] The processed fava component may be liquid, semi-liquid or powder.

[0091] The processed fava component may have the same characteristics as those provided herein for the fava component, except that the amount of vicine and / or convicine in the processed fava component is significantly lower.

[0092] In one embodiment, the method does not include the step of adding an organic solvent. In particular, the treated fiber component may be free of organic solvents. 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-dimethoxyethane (glyme, DME), dimethylformamide (DMF), dimethyl sulfoxide (DMSO), 1,4 dioxane, 1,2-dichloroethane, ethanol, ethyl acetate, ethylene glycol, glycerin, heptane, hexamethylphosphoramide (HMPA), hexane, methanol, methylene chloride, N-methyl-2-pyrrolidinone (NMP), nitromethane, naphthalene, pentane, 1-propanol, 2-propanol, pyridine, toluene, triethylamine, tricine, tris, tetrahydrofuran, o-xylene, m-xylene, p-xylene, and combinations thereof.

[0093] A significant reduction of vicine / convicine in fava is achieved without the use of organic solvent extraction, which allows freedom from organic solvents that can adversely affect the properties of the fava components.

[0094] In some embodiments, the method does not include the step of adding amylase before step c), hi some embodiments, the method does not include any step of adding amylase.

[0095] In some embodiments, the method does not include rinsing the fava component with an aqueous liquid, preferably water, prior to step b) or step c). In further embodiments, the method does not include rinsing the fava component with an aqueous liquid, preferably water, at a temperature above 50°C, preferably between 50°C and 90°C, prior to step b) or step c).

[0096] The method of the present invention achieves a significant reduction in vicine / convicine without the need for any rinsing step. The absence of a rinsing step prior to the enzyme treatment is advantageous. Indeed, the absence of such a rinsing step simplifies the process by limiting the number of steps, saves energy and resources, and is therefore more sustainable. Furthermore, the absence of such a rinsing step avoids altering the quality of the fava components. Indeed, rinsing can alter the quality of the fava components, especially at high temperatures.

[0097] In a second aspect, the present invention relates to a processed fiber component obtainable by the method of the first aspect of the invention. The characteristics of the fiber component of the third aspect of the invention apply to the processed fiber component of the second aspect of the invention.

[0098] The processed fava component may be liquid, semi-liquid or powder.

[0099] In a third aspect, the present invention relates to a fava component.

[0100] The fava component contains limited amounts of vicine and / or convicine. The sum of the vicine and convicine contents in the fava component is less than 600 mg / kg of fava component, preferably less than 450 mg / kg of fava component, more preferably less than 300 mg / kg of fava component, even more preferably less than 125 mg / kg of fava component, even more preferably less than 100 mg / kg of fava component, and even more preferably less than 50 mg / kg of fava component. In a preferred embodiment, the fava component is substantially free of vicine and / or convicine, preferably completely free of vicine and / or convicine.

[0101] In one embodiment, the fava component comprises a substantial amount of fava bean protein. The fava component comprises fava bean protein. In particular, the fava component comprises at least 1% by weight of fava bean protein, preferably 1% to 99% by weight, more preferably 10% to 99% by weight, even more preferably 30% to 99% by weight, and most preferably 50% to 99% by weight.

[0102] The fava component may contain, in addition to fava bean protein, a protein other than the fava bean protein. Preferably, the protein other than the fava bean protein may be a plant protein other than the fava bean protein, a milk protein, or a mixture thereof. More preferably, the protein other than the fava bean protein may be selected from the list consisting of a cereal protein, a seed protein, a nut protein, a pulse protein, a milk protein, or a mixture thereof. The pulse protein is different from the fava bean protein. Even more preferably, the protein other than the fava bean protein may be selected from the list consisting of a cereal protein, a pulse protein, a milk protein, or a mixture thereof. Examples of pulse proteins include chickpea protein, pea protein, lentil protein, or a mixture thereof. Examples of cereal proteins include oat protein, rice protein, wheat protein, barley protein, rye protein, buckwheat protein, quinoa protein, millet protein, corn protein, or a mixture thereof. Examples of seed proteins include hemp seed protein, canola seed protein, flaxseed protein, pumpkin seed protein, sunflower seed protein, cottonseed protein, safflower seed protein, squash seed protein, chia seed protein, or a mixture thereof. Examples of nut proteins 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 milk proteins include casein, whey protein, or a mixture thereof.

[0103] In one 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 protein of the fava component consists of fava bean protein. Most preferably, the protein of the fava component consists exclusively of fava bean protein.

[0104] In one embodiment, the pH of the fava component is 6.0 or higher, preferably pH 6.0-9.0, preferably pH 6.0-7.5.

[0105] Thus, in one embodiment, the proteins of the fava component, particularly the fava bean proteins, are intact, "intact" referring to proteins that do not aggregate in an uncontrolled manner, particularly proteins that do not precipitate or sediment.

[0106] In a preferred embodiment, the fava component is not fermented.

[0107] In one embodiment, the fava component may include sweeteners, texturizing agents, buffer salts, bulking agents, fiber, fat, minerals, vitamins, antioxidants, colorants, flavoring agents, aqueous liquids, or combinations thereof. The fat may be as provided above.

[0108] In one embodiment, the fava component is a liquid, semi-liquid, or powder.

[0109] The fava component contains small amounts of vicine and / or convicine. The fava component retains good stability and functionality and can be used in food applications.

[0110] In one embodiment, the fava component is an enzyme-treated fava component, i.e., a fava component that has been treated with at least one bicin- and / or convicin-degrading enzyme, which may be a bicin- and / or convicin-degrading enzyme as disclosed in the first aspect of the present invention.

[0111] The features of the fiber component of the third aspect of the invention apply to the processed fiber components of the first and second aspects of the invention, and vice versa.

[0112] In some embodiments, the Faba component may be free of organic solvents. 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-dimethoxyethane (glyme, DME), dimethylformamide (DMF), dimethyl sulfoxide (DMSO), 1,4 dioxane, 1,2-dichloroethane, ethanol, ethyl acetate, ethylene glycol, glycerin, heptane, hexamethylphosphoramide (HMPA), hexane, methanol, methylene chloride, N-methyl-2-pyrrolidinone (NMP), nitromethane, naphthalene, pentane, 1-propanol, 2-propanol, pyridine, toluene, triethylamine, tricine, tris, tetrahydrofuran, o-xylene, m-xylene, p-xylene, and combinations thereof.

[0113] In a fourth aspect, the present invention relates to a food product comprising the processed fava component according to the second aspect of the invention or the fava component according to the third aspect.

[0114] The food product may be selected from the list consisting of dairy products, milk analog products, meat analog products, egg analog products, nutritional products, confectionery products, pet food, vegetable purees, soups, beverages, baby food, medical nutritional products, dietary supplement products, or combinations thereof. The dairy product may be selected from the list consisting of yogurt, milk, cream, ice cream, dairy desserts, cheese, dairy drinks, milk powder, condensed milk, powdered milk drinks, infant formula, or combinations thereof. The dairy analog product may be selected from the list consisting of yogurt analog, milk analog, cream analog, ice cream analog, cheese analog, dairy drink analog, dairy dessert analog, milk powder analog, condensed milk analog, powdered milk drink analog, infant formula analog, or combinations thereof.

[0115] In some embodiments, the food product may be free of organic solvents. 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-dimethoxyethane (glyme, DME), dimethylformamide (DMF), dimethyl sulfoxide (DMSO), 1,4 dioxane, 1,2-dichloroethane, ethanol, ethyl acetate, ethylene glycol, glycerin, heptane, hexamethylphosphoramide (HMPA), hexane, methanol, methylene chloride, N-methyl-2-pyrrolidinone (NMP), nitromethane, naphthalene, pentane, 1-propanol, 2-propanol, pyridine, toluene, triethylamine, tricine, tris, tetrahydrofuran, o-xylene, m-xylene, p-xylene, and combinations thereof.

[0116] In a fifth aspect, the present invention relates to a method for producing a food product, which may be the food product provided in the fourth aspect of the invention.

[0117] The method comprises the step a) of providing a food composition.

[0118] The food composition comprises a fava component. The fava component of the food composition comprises vicine and / or convicine. Preferably, the sum of the vicine and convicine content in the fava component is at least 450 mg / kg of fava component, preferably at least 600 mg / kg of fava component, more preferably 600 mg-45,000 mg / kg of fava component, more preferably 600-35,000 mg / kg of fava component, even more preferably 800-10,000 mg / kg of fava component, and most preferably 800-2,000 mg / kg of fava component.

[0119] The fava component may be the fava component provided in the first aspect of the invention.

[0120] In one embodiment, the food composition may comprise at least 0.5% by weight of the fava component, preferably 0.5% to 50% by weight, more preferably 1% to 50% by weight, even more preferably 1% to 25% by weight, even more preferably 1% to 15% by weight, even more preferably 1% to 10% by weight, and most preferably 1% to 8% by weight.

[0121] 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 sweeteners, tempering agents, buffer salts, bulking agents, fiber, fat, minerals, vitamins, antioxidants, colorants, flavorings, aqueous liquids, proteins other than fiber bean protein, or combinations thereof. Preferably, the proteins other than fiber bean protein may be plant proteins other than fiber bean protein, dairy proteins, or mixtures thereof. More preferably, the proteins other than fiber bean protein may be selected from the list consisting of cereal proteins, seed proteins, nut proteins, pulse proteins, dairy proteins, or mixtures thereof. The pulse proteins are different from fiber bean protein. Even more preferably, the proteins other than fiber bean protein may be selected from the list consisting of cereal proteins, pulse proteins, dairy proteins, or mixtures thereof. Examples of pulse proteins include chickpea protein, pea protein, lentil protein, or mixtures thereof. Examples of grain proteins include oat protein, rice protein, wheat protein, barley protein, rye protein, buckwheat protein, quinoa protein, millet protein, corn protein, or a mixture thereof. Examples of seed proteins include hemp seed protein, canola seed protein, flaxseed protein, pumpkin seed protein, sunflower seed protein, cottonseed protein, safflower seed protein, squash seed protein, chia seed protein, or a mixture thereof. Examples of nut proteins 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 milk proteins include casein, whey protein, or a mixture thereof.

[0122] In one embodiment, the fat may be the fat provided in the first aspect of the invention.

[0123] 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 sweeteners, tempering agents, buffer salts, bulking agents, fiber, fat, minerals, vitamins, antioxidants, colorants, flavorings, aqueous liquids, proteins other than fiber bean protein, or combinations thereof. Preferably, the protein other than fiber bean protein may be a plant protein other than fiber bean protein, a dairy protein, or a mixture thereof. More preferably, the protein other than fiber bean protein may be selected from the list consisting of a cereal protein, a seed protein, a nut protein, a pulse protein, a dairy protein, or a mixture thereof. The pulse protein is different from fiber bean protein. Even more preferably, the protein other than fiber bean protein may be selected from the list consisting of a cereal protein, a pulse protein, a dairy protein, or a mixture thereof. Examples of pulse proteins include chickpea protein, pea protein, lentil protein, or a mixture thereof. Examples of grain proteins include oat protein, rice protein, wheat protein, barley protein, rye protein, buckwheat protein, quinoa protein, millet protein, corn protein, or a mixture thereof. Examples of seed proteins include hemp seed protein, canola seed protein, flaxseed protein, pumpkin seed protein, sunflower seed protein, cottonseed protein, safflower seed protein, squash seed protein, chia seed protein, or a mixture thereof. Examples of nut proteins 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 milk proteins include casein, whey protein, or a mixture thereof.

[0124] In one embodiment, the fat may be the fat provided in the first aspect of the invention.

[0125] In one embodiment, the food composition may comprise at least 0.5% by weight, preferably at least 2% by weight, more preferably between 2% and 25% by weight of protein, preferably fava bean protein.

[0126] In one embodiment, the protein of the food composition comprises 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% fava bean protein. Most preferably, the protein of the food composition consists exclusively of fava bean protein.

[0127] In one embodiment, the food composition may be liquid, semi-liquid, or pasty. For example, to prepare a beverage, the food composition may be liquid, whereas to prepare a meat analog product, the food composition may be semi-liquid or pasty. Preferably, the food composition is liquid.

[0128] The method further comprises step b) of adding at least one bicin- and / or convicin-degrading enzyme to the food composition.

[0129] The at least one bicin and / or convicin degrading enzyme may be a bicin and / or convicin degrading enzyme as disclosed in the first aspect of the present invention.

[0130] In one embodiment, in step b) at least one enzyme different from the bicin-degrading enzyme and / or the convicin-degrading enzyme may also be added to the food composition.

[0131] The pH of the food composition is 6.0 or higher, preferably 6.0 to 9.0, preferably 6.0 to 7.5. The food composition generally already has the above pH in step a), i.e., the original pH of the food composition is 6.0 or higher, preferably 6.0 to 9.0, preferably 6.0 to 7.5. If the pH of the food composition is not 6.0 or higher, the pH of the food composition may be adjusted between steps a) and b) to reach a pH of 6.0 or higher, preferably 6.0 or higher, preferably 6.0 to 9.0, preferably 6.0 to 7.5.

[0132] The dosage of bicin- and / or convicin-degrading enzymes added to a food composition will vary depending, inter alia, on the temperature during incubation, the amount or volume of the food composition being processed, the amount of fava component in the food composition, and the activity of the enzymes. Generally, this dosage should be at least 0.1% by weight based on the weight of the fava component in the food composition. Preferably, this dosage of bicin- and / or convicin-degrading enzymes may be between 0.1% and 5% by weight based on the weight of the fava component in the food composition.

[0133] The method further comprises step c) of incubating the food composition with at least one bicin-degrading enzyme and / or convicin-degrading enzyme to obtain a treated food composition. This incubation step c) is carried out at a pH of 6.0 or higher, preferably at a pH of 6.0 to 9.0, preferably at a pH of 6.0 to 7.5. Incubation step c) may be carried out at a temperature of at least 50°C, and optionally at a temperature of 50 to 65°C.

[0134] The present invention allows for a significant reduction of vicine and / or convicine in a short reaction time.

[0135] The incubation step is carried out for less than 2 hours, preferably less than 90 minutes, more preferably 15 to 90 minutes.

[0136] Even at a pH above 6, the enzyme has been observed to be effective in significantly reducing the amount of vicine and / or convicine in food compositions containing fava ingredients.

[0137] In particular, the content of vicine and / or convicin in the food composition is reduced by the at least one bicine-degrading enzyme and / or convicin-degrading enzyme in step c) 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%, and most preferably 100%. In other words, the treated food composition has a vicine and / or convicin content that 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%, and most preferably 100% less than the vicine and / or convicin content in the food composition.

[0138] Because the enzymatic treatment is carried out at a pH above 6, the proteins, especially the faba bean proteins, are intact and have good stability and good functionality for further processing into food products and during the shelf life of the food products.

[0139] Thus, in one embodiment, the proteins, in particular the faba bean proteins, of the treated food composition obtained after step c) are intact, "intact" referring to proteins that do not aggregate in an uncontrolled manner, in particular proteins that do not precipitate or sediment.

[0140] The method further comprises an optional step d) of homogenizing the treated food composition. In a preferred embodiment, the homogenizing step d) is not optional. The homogenizing step may be carried out at a pressure greater than 50 bar. Preferably, the homogenizing step may be carried out at a pressure between 50 bar and 700 bar. Even more preferably, the homogenizing step may be carried out at a pressure between 50 bar and 500 bar. More preferably, the homogenizing step may be carried out at a pressure between 50 bar and 300 bar, between 100 bar and 300 bar, or between 150 bar and 300 bar.

[0141] In a preferred embodiment, the homogenization step may be carried out at a temperature of 50°C to 80°C, preferably 50°C to 70°C. More preferably, the homogenization step may be carried out at a temperature of 55°C to 65°C.

[0142] The method further comprises step e) of heat-treating the treated food composition. Preferably, the heat treatment is carried out 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 carried out at a temperature of 80°C to 145°C, preferably 80°C to 125°C, for 3 seconds to 20 minutes, preferably 3 seconds to 90 seconds.

[0143] This heat treatment step allows for extending the shelf life of the food product while partially or completely inactivating at least one bicin-degrading enzyme and / or convicin-degrading enzyme in the treated food composition.

[0144] The homogenization step d) can be upstream or downstream of the heat treatment step e). In one embodiment, the method can include two homogenization steps: a first homogenization step d) upstream of the heat treatment and a second homogenization step downstream of the heat treatment. The pressure and temperature conditions given for the homogenization step d) also apply to the second homogenization step.

[0145] The method further comprises step e) of cooling the treated food composition to obtain a food product. For example, the treated food composition is cooled to a storage temperature for the food product, for example to room temperature, or under refrigerated conditions, i.e., between 2°C and 14°C.

[0146] The method may further comprise the step of evaporating the treated food composition after step d) or step e) or step f).

[0147] The method may further comprise, after step d) or step e) or step f), drying the treated food composition to form a food product in powder form. The drying step may be carried out by drum drying, roller drying, spray drying, or freeze drying. The drying step may also be carried out by any other food drying technique well known in the art. Converting a food product into a powder may be advantageous, as powders generally have good physicochemical stability and a long shelf life.

[0148] In a preferred embodiment, the drying step is preceded by an evaporation step as provided herein.

[0149] The treated food composition may have the same characteristics as those provided herein for the food composition, except that the amount of vicine and / or convicine in the treated food composition is significantly lower.

[0150] In one embodiment, the method does not include the step of adding an organic solvent. In particular, the treated food compositions and food products may be free of organic solvents. 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-dimethoxyethane (glyme, DME), dimethylformamide (DMF), dimethyl sulfoxide (DMSO), 1,4 dioxane, 1,2-dichloroethane, ethanol, ethyl acetate, ethylene glycol, glycerin, heptane, hexamethylphosphoramide (HMPA), hexane, methanol, methylene chloride, N-methyl-2-pyrrolidinone (NMP), nitromethane, naphthalene, pentane, 1-propanol, 2-propanol, pyridine, toluene, triethylamine, tricine, tris, tetrahydrofuran, o-xylene, m-xylene, p-xylene, and combinations thereof.

[0151] A significant reduction of vicine / convicine in fava is achieved without the use of organic solvent extraction, which allows freedom from organic solvents that can adversely affect the properties of fava components, food compositions and food products.

[0152] In some embodiments, the method does not include the step of adding an amylase prior to step c), hi some embodiments, the method does not include the step of adding an amylase.

[0153] In some embodiments, the method does not include rinsing the fava component and / or food composition with an aqueous liquid, preferably water, prior to step b) or step c).In further embodiments, the method does not include rinsing the fava component and / or food composition with an aqueous liquid, preferably water, at a temperature above 50°C, preferably between 50°C and 90°C, prior to step b) or step c).

[0154] The method of the present invention achieves a significant reduction of vicine / convicine in fava without the need for any rinsing step. The absence of a rinsing step prior to enzymatic treatment is advantageous. Indeed, the absence of such a rinsing step simplifies the process by limiting the number of steps, saves energy and resources, and is therefore more sustainable. Furthermore, the absence of such a rinsing step avoids altering the quality of the fava ingredients, food compositions, and even food products. Indeed, rinsing can alter the quality of the fava ingredients, food compositions, and even food products, especially at high temperatures.

[0155] In a sixth aspect, the present invention relates to a method for producing a food product, which may be a food product provided in the fourth or fifth aspect of the present invention.

[0156] The method includes the step of providing a food composition.

[0157] The food composition comprises a treated fava component according to the second aspect of the invention or a fava component according to the third aspect of the invention.

[0158] In one embodiment, the food composition may comprise at least 0.5% by weight of the fava component according to the third aspect of the invention or the processed fava component according to the second aspect, preferably 0.5% to 50% by weight, more preferably 1% to 50% by weight, even more preferably 1% to 25% by weight, even more preferably 1% to 15% by weight, even more preferably 1% to 10% by weight, and most preferably 1% to 8% by weight.

[0159] In one 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 sweeteners, tempering agents, buffer salts, bulking agents, fiber, fat, minerals, vitamins, antioxidants, colorants, flavorings, aqueous liquids, proteins other than fiber bean protein, or combinations thereof. Preferably, the proteins other than fiber bean protein may be plant proteins other than fiber bean protein, dairy proteins, or mixtures thereof. More preferably, the proteins other than fiber bean protein may be selected from the list consisting of cereal proteins, seed proteins, nut proteins, pulse proteins, dairy proteins, or mixtures thereof. The pulse proteins are different from fiber bean protein. Even more preferably, the proteins other than fiber bean protein may be selected from the list consisting of cereal proteins, pulse proteins, dairy proteins, or mixtures thereof. Examples of pulse proteins include chickpea protein, pea protein, lentil protein, or mixtures thereof. Examples of grain proteins include oat protein, rice protein, wheat protein, barley protein, rye protein, buckwheat protein, quinoa protein, millet protein, corn protein, or a mixture thereof. Examples of seed proteins include hemp seed protein, canola seed protein, flaxseed protein, pumpkin seed protein, sunflower seed protein, cottonseed protein, safflower seed protein, squash seed protein, chia seed protein, or a mixture thereof. Examples of nut proteins 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 milk proteins include casein, whey protein, or a mixture thereof.

[0160] In one embodiment, the fat may be the fat provided in the first aspect of the invention.

[0161] 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 fiber bean protein, sweetener, conditioner, buffer salt, bulking agent, fiber, fat, mineral, vitamin, antioxidant, colorant, flavoring, aqueous liquid, or a combination thereof. Preferably, the protein different from fiber bean protein, preferably the protein different from fiber bean protein, may be a plant protein different from fiber bean protein, a dairy protein, or a mixture thereof. More preferably, the protein different from fiber bean protein may be selected from the list consisting of a cereal protein, a seed protein, a nut protein, a pulse protein, a dairy protein, or a mixture thereof. The pulse protein is different from fiber bean protein. Even more preferably, the protein different from fiber bean protein may be selected from the list consisting of a cereal protein, a pulse protein, a dairy protein, or a mixture thereof. Examples of pulse proteins include chickpea protein, pea protein, lentil protein, or a mixture thereof. Examples of grain proteins include oat protein, rice protein, wheat protein, barley protein, rye protein, buckwheat protein, quinoa protein, millet protein, corn protein, or a mixture thereof. Examples of seed proteins include hemp seed protein, canola seed protein, flaxseed protein, pumpkin seed protein, sunflower seed protein, cottonseed protein, safflower seed protein, squash seed protein, chia seed protein, or a mixture thereof. Examples of nut proteins 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 proteins include casein, whey protein, or a mixture thereof.

[0162] In one embodiment, the fat may be the fat provided in the first aspect of the invention.

[0163] In one embodiment the food composition comprises at least 0.5% by weight, preferably at least 2% by weight, more preferably between 2% and 25% by weight of protein, preferably fava bean protein.

[0164] In one embodiment, the protein of the food composition comprises 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% fava bean protein. Most preferably, the protein of the food composition consists exclusively of fava bean protein.

[0165] In one embodiment, the pH of the food composition may be 6.0 or higher, preferably pH 6.0 to 9.0, more preferably pH 6.0 to 7.5.

[0166] In one embodiment, the food composition may be liquid, semi-liquid, or pasty. For example, to prepare a beverage, the food composition may be liquid, whereas to prepare a meat analog product, the food composition may be semi-liquid or pasty. Preferably, the food composition is liquid.

[0167] The method further comprises an optional step b) of homogenizing the food composition. In a preferred embodiment, the homogenizing step b) is not optional. The homogenizing step may be carried out at a pressure greater than 50 bar. Preferably, the homogenizing step may be carried out at a pressure between 50 bar and 700 bar. Even more preferably, the homogenizing step may be carried out at a pressure between 50 bar and 500 bar. More preferably, the homogenizing step may be carried out at a pressure between 50 bar and 300 bar, between 100 bar and 300 bar, or between 150 bar and 300 bar.

[0168] In a preferred embodiment, the homogenization step may be carried out at a temperature of 50°C to 80°C, preferably 50°C to 70°C. More preferably, the homogenization step may be carried out at a temperature of 55°C to 65°C.

[0169] The method further comprises step c) of heat-treating the food composition. Preferably, the heat treatment is carried out 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 carried out at a temperature of 80°C to 145°C, preferably 80°C to 125°C, for 3 seconds to 20 minutes, preferably 3 seconds to 90 seconds.

[0170] The homogenization step b) can be upstream or downstream of the heat treatment step c). In one embodiment, the method can include two homogenization steps: a first homogenization step b) that is upstream of the heat treatment step c) and a second homogenization step that is downstream of the heat treatment step c). The pressure and temperature conditions given for the homogenization step b) also apply to the second homogenization step.

[0171] The method further comprises a step d) of cooling the food composition to obtain a food product, for example, the food composition is cooled to a storage temperature for the food product, for example to room temperature, or under refrigerated conditions, i.e., between 2°C and 14°C.

[0172] The method may further comprise the step of evaporating the food composition after step b) or step c) or step d).

[0173] The method may further comprise, after step b) or step c) or step d), drying the food composition to form a food product in powder form. The drying step may be carried out by drum drying, roller drying, spray drying, or freeze drying. The drying step may also be carried out by any other food drying technique well known in the art. Converting a food product into a powder may be advantageous, as powders generally have good physicochemical stability and a long shelf life.

[0174] In a preferred embodiment, the drying step is preceded by an evaporation step as provided herein.

[0175] In one embodiment, the method does not include the step of adding an organic solvent. In particular, the food composition may be free of organic solvents. 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-dimethoxyethane (glyme, DME), dimethylformamide (DMF), dimethyl sulfoxide (DMSO), 1,4 dioxane, 1,2-dichloroethane, ethanol, ethyl acetate, ethylene glycol, glycerin, heptane, hexamethylphosphoramide (HMPA), hexane, methanol, methylene chloride, N-methyl-2-pyrrolidinone (NMP), nitromethane, naphthalene, pentane, 1-propanol, 2-propanol, pyridine, toluene, triethylamine, tricine, tris, tetrahydrofuran, o-xylene, m-xylene, p-xylene, and combinations thereof.

[0176] A significant reduction of vicine / convicine in fava is achieved without the use of organic solvent extraction. The method allows freedom from organic solvents that can adversely affect the properties of fava components, food compositions and food products.

[0177] In some embodiments, the method does not include the step of adding an amylase prior to step c), hi some embodiments, the method does not include the step of adding an amylase.

[0178] In some embodiments, the method does not include rinsing the fava component and / or food composition with an aqueous liquid, preferably water, prior to step b) or step c).In further embodiments, the method does not include rinsing the fava component and / or food composition with an aqueous liquid, preferably water, at a temperature above 50°C, preferably between 50°C and 90°C, prior to step b) or step c).

[0179] The method of the present invention achieves a significant reduction of vicine / convicine in fava without the need for any rinsing step. The absence of a rinsing step prior to enzymatic treatment is advantageous. Indeed, the absence of such a rinsing step simplifies the process by limiting the number of steps, saves energy and resources, and is therefore more sustainable. Furthermore, the absence of such a rinsing step avoids altering the quality of the fava ingredients, food compositions, and even food products. Indeed, rinsing can alter the quality of the fava ingredients, food compositions, and even food products, especially at high temperatures.

[0180] In a seventh aspect, the present invention relates to a food product obtainable by a method according to the fifth aspect of the invention or the sixth aspect of the invention.

[0181] The food product may be a food product as disclosed in the fourth, fifth and sixth aspects of the present invention.

[0182] Protein sequence Sequence number 1 (length: 444) MKKFPEGFLWGVATASYQIEGSPLADGAGMSIWHTFSHTPGNVKNGDTGDVACDHYNRWKEDIEIIEKLGVKAYRFSISWPRILPEGTGRVNQKGLDFYNRIIDTLLEKGITPFVTIYHWDLPFALQLKGGWANREIADWFAEYSRVLFENFGDRVKNWITLNEPWVVAIVGHLYGVHAPGMRDIYVAFRAVHNLLRAHARAVKVFRETVKDGKIGIVFNNG YFEPESEKEEDIRAVRFMHQFNNYPLFLNPIYRGDYPELVLEFAREYLPENYKDDMSEIQEKIDFVGLNYYSGHLVKFDPDAPAKVSFVERDLPKTAMGWEIVPEGIYWILKKVKEEYNPPEVYITENGAAFDDVVSEDGRVHDQNRIDYLKAHIGQAWKAIQEGVPLKGYFVWSLLDNFEWAEGYSKRFGIVYVDYSTQKRIVKDSGYWYSNVVKNNLED

[0183] · sequence number 2(length:446) MNVKKFPEGFLWGVATASYQIEGSPLADGAGMSIWHTFSHTPGNVKNGDTGDVACDHYNRWKEDIEIIEKLGVKAYRFSISWPRILPEGTGRVNQKGLDFYNRIIDTLLEKGITPFVTIYHWDLPFALQLKGGWANREIADWFAEYSRVLFENFGDRVKNWITLNEPWVVAIVGHLYGVHAPGMRDIYVAFRAVHNLLRAHARAVKVFRETVKDGKIGIVFNN GYFEPESEKEEDIRAVRFMHQFNNYPLFLNPIYRGDYPELVLEFAREYLPENYKDDMSEIQEKIDFVGLNYYSGHLVKFDPDAPAKVSFVERDLPKTAMGWEIVPEGIYWILKKVKEEYNPPEVYITENGAAFDDVVSEDGRVHDQNRIDYLKAHIGQAWKAIQEGVPLKGYFVWSLLDNFEWAEGYSKRFGIVYVDYSTQKRIVKDSGYWYSNVVKNNLED

[0184] Sequence number 3 (length: 542) MAMQLRSLLLCVLLLLLGFALADTNAAARIHPPVVCANLSRANFDTLVPGFVFGAATASYQVEGAANLDGRGPSIWDTFTHKHPEKIADGSNGDVAIDQYHRYKEDVAIMKDMGLESYRFSISWSRVLPNGTLSG GINKKGIEYYNNLINELLHNGIEPLVTLFHWDVPQTLEDEYGGFLSNRIVNDFEEYAELCFKKFGDRVKHWTTLNEPYTFSSHGYAKGTHAPGRCSAWYNQTCFGGDSATEPYLVTHNLLLAHAAAVKLYKTKYQA YQKGVIGITVVTPWFEPASEAKEDIDAVFRALDFIYGWFMDPLTRGDYPQSMRSLVGERLPNFTKKESKSLSGSFDYIGINYYSARYASASKNYSGHPSYLNDVNVDVKTELNGVPIGPQAASSWLYFYPKGLYD LLCYTKEKYNDPIIYITENGVDEFNQPNPKLSLCQLLDDSNRIYYYYHHLCYLQAAIKEGVKVKGYFAWSLLDNFEWDNGYTVRFGINYVDYDNGLKRHSKHSTHWFKSFLKKSSRNTKKIRRCGNNNTSATKFVF

[0185] Sequence number 4 (length 544) MALQFRSLLLCMVLLLLGFALANTNAARTDPPIVCATLNRTHFDTLFPGFTFGAATAAYQLEGAANIDGRGPSVWDNFTHEHPEKITDGSNGDVAIDQYHRYKEDVAIMKDMGLDAYRFSISWSRLLPNGKLSGGI NKKGIEYYNNLTNELLRNGIEPLVTLFHWDVPQALVDEYGGLLSPRIVDDFKAYADLCYKEFGDRVKHWTTLNEPYTISNHGYTIGIHAPGRCSDWYNPKCLGGDSGIEPYLVTHYLLLAHAAAVKLYREKYQAYQ NGVIGITVVSHWFEPASESQQDKDAAFQALDFMYGWFMDPLTRGDYPQIMRSILGARLPNFTEEQSKSLSGSYDYIGVNYYSARYASAYPKDYSVTTPPSYLTDVHVNVTTDLNGVPIGPRAASDWLYVYPKGLYD LVLYTKEKYNDPIMYITENGMDEFNNPKLSLEQALNDGNRIDYYYRHLCYLQAAMKEGANVQGYFAWSLLDNFEWSEGYTVRFGINYIDYDNGLERHSKLSTHWFKSFLKRSSISKKKIRRCGNNNGRATKFVYQI

[0186] Sequence number 5 (length 661) MRKRQPKTLHLLVMAMQLRSLLLCVLLLLLGFALADTNAAARIHPPVVCANLSRANFDILVPGFVFGAATASYQVEGAANLDGRGPSIWDTFTHKHPEKIADGSNGDVAIDQYHRYKEDVAIMKDMGLESYRFSISWSRVLPNGTLSGGINKKGIEYYNNLINEL LHNGIEPLVTLFHWDVPQTLEDEYGGFLSNRIVNDFEEYAELCFKKFGDRVKHWTTLNEPYTFSSHGYAKGTHAPGRCSAWYNQTCFGGDSATEPYLVTHNLLLAHAAAVKLYKTKYQAYQKGVIGITVVTPWFEPASEAKEDIDAVFRALDFIYGWFMDPLTRG DYPQSMRSLVGERLPNFTKKESKSLSGSFDYIGINYYSARYASASKNYSGHPSYLNDVNVDVKSEYKLKTSTELNGVPIGPQAASSWLYFYPKGLYDLLRYTKEKYNDPIIYITENGVDEFNQPNPKLSLCQLLDDSNRIYYYYHHLCYLQAAIKEGVKVKGYFA WSLLDNFEWDNGYTVRFGINYVDYDNGLKRHSKHSTHWFKSFLKKSSRNTKKIRRCGNNNTSATKGALTLFGCGWRDTLCLNIVDPPPKYLLLILRHLGTNEMLVGLDQMWRPIKYDYEVPKQALIFALCQTRLQFGTWEGYDTGLPTPLLHQALSNIALSHCNTT

[0187] Those skilled in the art will understand that all features of the invention disclosed herein may be freely combined. In particular, features described for the product of the invention may be combined with the method of the invention, and vice versa. Furthermore, features described for different embodiments of the invention may be combined.

[0188] Furthermore, where known equivalents exist for specific features, such equivalents are incorporated as if specifically referred to herein. Further advantages and features of the present invention will be apparent from the drawings and non-limiting examples.

[0189] [Example] Example 1: Degradation of vicine / convicine in fava suspensions containing fava concentrates by treatment with enzyme preparations having β-glucosidase activity or side activity at pH 6.4 and 60°C

[0190] The following experiments were performed at 60°C as a first attempt to test enzyme activity in a faba / water model system. These experiments were first performed with a faba concentrate containing low levels of bicine / convicine. A faba concentrate with high levels of bicine / convicine was then used to test the robustness of this enzymatic approach.

[0191] A fava concentrate containing low levels of vicine / convicine was prepared, specifically, the initial vicine and convicine contents in the fava concentrate containing low levels of vicine / convicine were 661 mg and 176 mg per kg of fava component, respectively.

[0192] 5% w / v of the above fava concentrate (60% protein content) with low levels of vicine / convicine was slowly added to a thermomixer containing Milli-Q water at 60°C. The water and fava concentrate were mixed in the thermomixer for 30-60 minutes to form a fava suspension with a fava protein content of 3% by weight. The pH of the resulting fava suspension was 6.4, and no further pH adjustment was performed. 0.25% w / v of enzyme preparations E1-E6 with β-glucosidase activity or side activity was added to the fava suspension.

[0193] Specifically, enzymes E1-E6 were purchased. Enzyme preparations E1-E4 had β-glucosidase activity as their primary activity (i.e., E1-E4 are β-glucosidases); enzyme preparation E5 was a xylanase with β-glucosidase side activity; enzyme preparation E6 was a cellulase with β-glucosidase side activity. The origins of the different enzymes E1-E6 are shown in Table 1 below. The amino acid sequences of enzymes E2 and E3 are also shown in Table 1 below.

[0194] [Table 1]

[0195] After adding the enzyme preparation, the suspension was incubated in a thermomixer at 60°C and 900 rpm for 60 minutes to obtain an enzyme-treated fiber composition, which was then heated at 95°C for 15 minutes to inactivate the enzyme.

[0196] The remaining bicine and / or convicine in the enzyme-treated faba composition was extracted with 70% acetonitrile in Milli-Q water and measured by reversed-phase high-performance liquid chromatography (RP-HPLC) using a diode array detector (DAD) at 273 nm. Specifically, the separation of bicine and convicine was carried out using an Acquity UPLC® BEH HILIC column (2.1 × 100 mm) with a 1.7 μm particle size (Waters, Milford, USA). Elution of bicine and convicine was performed with a 10-minute gradient run at a flow rate of 0.3 mL / min: 1) starting from 0% eluent A (10% acetonitrile in Milli-Q water containing 10 mM ammonium formate and 0.1% formic acid) and 100% eluent B (90% acetonitrile in Milli-Q water containing 10 mM ammonium formate and 0.1% formic acid), followed by 20% eluent A and 80% eluent B in the first 5 minutes; 2) the gradient of eluent B was decreased from 80% to 50% in 1.5 minutes and held for 1 minute; 3) finally, the gradient of eluent B was increased from 50% to 100% in 0.5 minutes and held for 2 minutes.

[0197] After elution, the bicine / convicine content was measured by UV at 273 nm using a diode array detector. The bicine / convicine content in the resulting enzyme-treated faba composition was compared with a control treated in the same manner except that no enzyme was added.

[0198] To test the robustness of this enzymatic method, the above experiment was repeated using a fava concentrate containing a high level of vicin / convicin (protein content 60%) instead of the fava concentrate containing a low level of vicin / convicin. The fava concentrate with high levels of vicin / convicin (i.e., before incubation) contained 19,363 mg of vicin per kg of fava component and 5,508 mg of convicin per kg of fava component.

[0199] The results are shown in Figures 1 to 4.

[0200] Of the selected enzyme preparations, E1 was not effective in significantly reducing vicine and convicine levels in fava compositions at pH 6.4. Vicine / convicine reduction was less than 30% for fava compositions prepared with fava concentrates containing low and high levels of vicine / convicine (Figures 1-4).

[0201] E2-E6 achieved a significant reduction of more than 30% of vicine and / or convicine at pH 6.4 and 60° C. This was true for fava compositions prepared with fava concentrates containing low and high levels of vicine / convicine.

[0202] For fava concentrates containing low levels of vicine / convicine, E2, E3, E4, and cellulase E6 with β-glucosidase side activity degraded vicine and / or convicine at levels barely above 50% (Figures 1 and 2). The greatest reduction in vicine and convicine was achieved with E2 and E3 (over 80% reduction).

[0203] For faba concentrates containing high levels of bicine / convicine, E2 and E3 were able to degrade bicine / convicine by more than 80% at the original pH of 6.4 (FIGS. 3 and 4).

[0204] Example 2: Degradation of vicine / convicine in faba suspensions containing faba concentrates by treatment with enzyme preparations having β-glucosidase activity or side activity at pH 6.4 and temperatures between 50°C and 65°C To test the efficiency of E2 and E3 at different incubation temperatures, degradation of vicine / convicine in faba concentrate suspension was carried out using E2 or E3 as described in Example 1, except that the incubation temperatures were 50°C, 55°C, 60°C, or 65°C.

[0205] For the above tests, the fava concentrate containing low levels of vicin / convicin from Example 1 (Figures 5 and 6) and the fava concentrate containing high levels of vicin / convicin from Example 1 (Figures 7 and 8) were also used. Specifically, the initial vicin and convicin contents in the fava concentrate containing low levels of vicin / convicin were 661 mg and 176 mg per kg of fava component, respectively; the initial vicin and convicin contents in the fava concentrate containing high levels of vicin / convicin were 19363 mg and 5508 mg per kg of fava component, respectively.

[0206] The results are shown in Figures 5 to 8.

[0207] For compositions prepared with fava concentrates containing low and high levels of vicine / convicine, both β-glucosidases E2 and E3 were able to achieve at least 55% degradation of vicine and convicine at all temperatures, with E2 showing significant degradation of over 99% vicine / convicine at all temperatures.

[0208] Example 3: Degradation of vicine / convicine by enzymatic treatment with an enzyme preparation having β-glucosidase activity or side activity at pH 6.4 and a temperature of 60°C in a model recipe containing fava ingredients for producing a food product.

[0209] 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, and 3.5% w / v faba concentrate containing low levels of vicine / convicine from Example 1 were added to a thermomixer containing Milli-Q water at 60°C. The different ingredients were combined in the thermomixer and homogenized for 30-60 minutes to form a food composition. The pH of the food composition was pH 6.4 without further adjustment. 0.18% w / v of enzyme preparations E2 and E3 from Example 1 were added to the food composition. The food composition was incubated in the thermomixer at 60°C and 900 rpm for 60 minutes. The food composition was then heated at 95°C for 15 minutes to inactivate the enzymes, resulting in a food product, specifically a plant-based milk analog.

[0210] Residual bicine and / or convicin in the food products was extracted with 70% acetonitrile in Milli-Q water and measured at 273 nm by RP-HPLC coupled with DAD as described in Example 1. The bicine / convicin content in the resulting food compositions was compared with a control treated identically, except that no enzyme was added. The results are shown in Figures 9 and 10.

[0211] The results show that E2 and E3 were able to significantly degrade vicine and convicine by more than 80%, even in complex food matrices containing different ingredients.

[0212] Although the invention has been described by example, it should be understood that variations and modifications can be made without departing from the scope of the invention as defined in the claims.

Claims

1. 1. A method for reducing the vicine and / or convicine content in a fava component, comprising: a) providing a liquid fava component or suspending a fava component in an aqueous liquid to form a liquid fava component, said fava component comprising vicine and / or convicine; b) adding at least one bicin degrading enzyme and / or convicin degrading enzyme to the fava component, wherein the pH of the fava component is 6.0 or higher; c) incubating said fava component with said at least one bicin and / or convicin degrading enzyme to obtain a treated fava component; d) optionally partially or completely inactivating the bicine and / or convicine degrading enzymes in the treated fava component; e) optionally cooling the treated fava component; A method comprising:

2. 2. The method of claim 1, wherein the pH of the fava component is between pH 6.0 and pH 9.

0.

3. 3. The method of claim 1 or 2, wherein the fava ingredient is whole fava beans, fava bean flour, fava bean protein concentrate, or fava bean protein isolate.

4. 4. The method according to claim 1, wherein the at least one bicin- and / or convicin-degrading enzyme is selected from the list consisting of β-glucosidase, an enzyme with β-glucosidase side activity, or a mixture thereof.

5. 5. The method of claim 1, wherein the at least one bicin-degrading enzyme and / or convicin-degrading enzyme is one or more β-glucosidases having an amino acid sequence that is at least 70% identical to one or more amino acid sequences 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.

6. 6. The method of claim 1, wherein the at least one bicin-degrading enzyme and / or convicin-degrading enzyme comprises a vicin-degrading enzyme and / or a convicin-degrading enzyme derived from almonds.

7. The method according to any one of claims 1 to 6, wherein the one type of bicin-degrading enzyme and / or convicin-degrading enzyme comprises a bicin-degrading enzyme and / or a convicin-degrading enzyme derived from Thermotoga maritima.

8. 8. The method according to any one of claims 1 to 7, wherein the content of vicine and / or convicin in the fava component is reduced by at least 30%, preferably at least 50%, in step c) by the at least one bicine- and / or convicin-degrading enzyme.

9. 9. The method according to any one of claims 1 to 8, comprising, after step e), drying the treated fava component to obtain a powdered fava component.

10. 10. The method according to any one of claims 1 to 9, wherein in step a) at least 1% by weight of fava components is suspended in said aqueous liquid.

11. The method according to any one of claims 1 to 10, wherein the incubation step c) is carried out for less than 2 hours.

12. A processed fava component obtained by the method according to any one of claims 1 to 11.

13. A fava component having a sum of vicine and convicine content of less than 600 mg per kg of fava component, preferably less than 450 mg per kg of fava bean protein.

14. 14. The processed fava component of claim 12 or the processed fava component of claim 13, wherein the fava component or the processed fava component is liquid, semi-liquid, or powder.

15. 15. The processed fava component of claim 12 or 14, or the fava component of claim 13 or 14, wherein the protein is intact.

16. A food product comprising the processed fava component of any one of claims 12 or 14-15, or the fava component of any one of claims 13-15.

17. 1. A method of producing a food product, comprising: a) providing a food composition, said food composition comprising a fava component and at least one additional food ingredient, said fava component of said food composition comprising vicine and / or convicine; b) adding at least one bicin-degrading enzyme and / or convicin-degrading enzyme to the food composition, wherein the pH of the food composition is 6.0 or higher; c) incubating the food composition with the at least one bicin-degrading enzyme and / or convicin-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 comprising:

18. 1. A method of producing a food product, comprising: a) providing a food composition, said food composition comprising the treated fava component according to any one of claims 12 or 14-15, or the fava component according to any one of claims 13-15, and also comprising 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; A method comprising:

19. 19. The method of claim 17 or 18, wherein the at least one additional food ingredient is selected from the list consisting of a protein other than a faba bean protein, a sweetener, a texturizing agent, a buffer salt, a bulking agent, fiber, fat, a mineral, a vitamin, an antioxidant, a colorant, a flavoring agent, an aqueous liquid, or a combination thereof.

20. 20. The method of claim 19, wherein the at least one additional food ingredient comprises a protein other than a faba bean protein, the protein being a cereal protein, a pulse protein, or a dairy protein, and the pulse protein is other than a faba bean protein.

21. A food product obtainable by the method according to claims 17 to 20.

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