Plant-derived or fungus-derived particles loaded with protein

Abstract

The present invention relates to a method for loading proteins into plant-derived or fungal-derived particles, preferably plant-derived particles from vegetables, fruits and fruiting bodies of edible fungi, said method comprising: (a) providing plant-derived or fungal-derived particles, preferably plant-derived particles; (b) providing a soluble protein capable of being immobilized in a plant-derived or fungal-derived particle, preferably a plant-derived particle; (c) adding the soluble protein provided in step (b) to the plant-derived or fungal-derived particles, preferably plant-derived particles, provided in step (a), preferably followed by mixing; (d) contacting the plant-derived or fungal-derived particles, preferably plant-derived particles, with the protein in the mixture provided in step (c) under conditions in which the protein remains soluble to load the plant-derived or fungal-derived particles, preferably plant-derived particles, with the protein; (e) immobilizing at least a portion of the protein in a plant-derived or fungal-derived particle, preferably a plant-derived particle, The protein-loaded plant-derived or fungal derived particles, preferably plant-derived particles, obtained in step (e) comprise at least 35% by weight of protein, based on the dry weight of the protein-loaded plant-derived or fungal derived particles, preferably plant-derived particles, obtained in step (e).The present invention further relates to protein-loaded plant-derived or fungal derived particles, preferably plant-derived particles from vegetables, fruits and fruiting bodies of edible fungi.

Description

Technical Field

[0001] Field of the Invention The present invention relates to a method for loading proteins onto plant-derived or fungus-derived particles, preferably plant-derived particles, plant-derived or fungus-derived particles loaded with proteins, preferably plant-derived particles, plant-derived or fungus-derived particles, preferably plant-derived particles loaded with proteins obtainable via the method, a food comprising or consisting of the plant-derived or fungus-derived particles, preferably plant-derived particles loaded with proteins, and the use of the plant-derived or fungus-derived particles, preferably plant-derived particles loaded with proteins as a food ingredient.

Background Art

[0002] Background of the Invention In the human diet, few nutrients are as important as proteins. Proteins are the main building blocks of the human body. They are used to create muscles, tendons, organs, and skin, as well as enzymes, hormones, neurotransmitters, and other molecules that perform many important functions. Proteins are composed of amino acids as building blocks. The human body produces some of these amino acids, but other amino acids known as essential amino acids must be obtained through the diet. Generally, animal proteins provide all essential amino acids in appropriate ratios. Thus, meat and fish are important sources of protein for human consumption.

[0003] However, the increasing global consumption of meat and fish is closely related to long-term sustainability issues, because it is known to have an adverse impact on the environment and to put additional pressure on scarce resources. For this, see, for example, H. Dagevos and J. Voordouw, Sustainability and meat consumption: is reduction realistic?, Sustainability: Science, Practice and Policy, 9(2), Summer 2013, pp 60-69.

[0004] Proteins derived from plant materials may potentially form a major protein source for food applications. Thus, much research effort has been devoted to the development of technologies for isolating plant-derived proteins and to the development of new products based on plant-derived proteins, such as products that are sensorially similar to meat or fish. Such products preferably also have a protein content similar to that of meat or fish. Meat typically has a protein content of 50-85 wt% based on dry matter. Fish typically has a protein content of 50-85 wt% based on dry matter.

[0005] A well-known example of a product positioned as a meat substitute or used as an ingredient in meat substitutes is textured or texturized vegetable protein (TVP), also known as textured soy protein (TSP) or soy meat. TVP is a defatted soy flour product, i.e., a by-product of soy oil extraction. It is often used as a meat substitute or extender. It has a protein content comparable to that of certain meats. The process for manufacturing TVP requires extrusion, which causes changes in the structure of soy protein resulting in a fibrous, sponge-like matrix similar to the texture of meat. TVP can be used to create vegetarian or vegan versions of traditional meat-based dishes.

[0006] US Patent Publication No. 2020 / 0288733A1 relates to dried fruits and vegetables with increased protein and a jerky-like texture. A process is disclosed in which whole or sliced vegetables are optionally blanched in hot water and cooled, where the resulting vegetables are contacted with a protein-containing marin typically having a low molecular weight and high solubility to obtain protein-injected vegetables. The resulting product is dried to a low AW value. Examples are based on dried products, including mushrooms injected with algal protein and sugar resulting in a 23% sugar content and a maximum protein content of 16.7%, mushrooms injected with whey protein and sugar resulting in a 23% sugar content and a maximum protein content of 25.1%, eggplants injected with algal protein and sugar resulting in a 23% sugar content and a maximum protein content of 11.8%, eggplants injected with whey protein and sugar resulting in a 23% sugar content and a maximum protein content of 18.9%, mushrooms injected with pea protein and sugar resulting in a 49% sugar content and a maximum protein content of 7.1%, and mushrooms injected with canola protein and sugar resulting in a 49% sugar content and a maximum protein content of 15.0%. In US Patent Publication No. 2020 / 0288733A1, the amount of injection varies based on the protein source, but it was concluded that blanching improves the amount of protein injection. A hypothesis was put forward that this improvement was achieved because blanching changes or expands the cell structure, thereby making it easier for the protein to access by injection.

[0007] It is an object of the present invention to provide a novel plant-derived food rich in protein, preferably rich in plant-derived protein.

[0008] It is a further object of the present invention to provide a plant-derived food that is not manufactured using extrusion and is fortified with protein, preferably fortified with plant-derived protein.

[0009] It is yet another object of the present invention to provide a novel plant-derived food rich in protein, preferably plant-derived protein, which can be used as or in place of meat and / or fish substitutes or alternatives. Summary of the Invention Means for Solving the Problems

[0010] Summary of the Invention The inventors have unexpectedly found that one or more of these objects can be met by loading soluble proteins into plant-derived or fungus-derived particles, preferably plant-derived particles from vegetables, fruits and fruiting bodies of edible fungi, and then immobilizing at least a portion of the protein in the plant-derived or fungus-derived particles, preferably plant-derived particles, using heat treatment, acid treatment, isoelectric precipitation or combinations thereof.

[0011] The immobilization step results in a product with very limited protein leakage when exposed to a liquid, aqueous or wet environment.

[0012] Accordingly, in a first aspect, the present invention provides a method for loading proteins into plant-derived or fungus-derived particles, preferably plant-derived particles from vegetables, fruits and fruiting bodies of edible fungi, the method comprising: (a) providing plant-derived or fungus-derived particles, preferably plant-derived particles; (b) providing a soluble protein that can be immobilized in the plant-derived or fungus-derived particles, preferably plant-derived particles; (c) adding the soluble protein provided in step (b) to the plant-derived or fungus-derived particles provided in step (a), preferably followed by mixing; (d) contacting the plant-derived or fungus-derived particles, preferably plant-derived particles, with the protein in the mixture provided in step (c) under conditions such that the protein remains soluble, to load the protein into the plant-derived or fungus-derived particles, preferably plant-derived particles. (e) immobilizing at least a part of the protein in particles derived from plants or fungi, preferably particles derived from plants; and The particles derived from plants or fungi, preferably particles derived from plants, loaded with the protein obtained in step (e) contain at least 35% by weight of the protein based on the dry weight of the particles derived from plants or fungi, preferably particles derived from plants, loaded with the protein obtained in step (e).

[0013] In a second aspect, the present invention provides particles derived from plants or fungi, preferably particles derived from plants, loaded with a protein, which can be obtained via a process as defined herein.

[0014] In a third aspect, the present invention provides particles derived from plants or fungi, preferably particles derived from plants, loaded with a protein, comprising a continuous matrix of a plant-derived or fungal-derived material containing the protein distributed throughout the matrix, preferably plant-derived particles from vegetables, fruits and fruiting bodies of edible fungi, and the particles derived from plants or fungi, preferably particles derived from plants, loaded with the protein provided (i) 3 to 97% by weight of water and 97 to 3% by weight of dry matter (the amount of water and the amount of dry matter together constitute 100% by weight of the particles derived from plants or fungi, preferably particles derived from plants, loaded with the protein) based on the total weight of the particles derived from plants or fungi, preferably particles derived from plants, loaded with the protein; and (ii) 20 to 65% by weight of the plant-derived or fungal-derived material based on the dry weight of the particles derived from plants or fungi, preferably particles derived from plants, loaded with the protein; and (iii) at least 35% by weight of the protein based on the dry weight of the particles derived from plants or fungi, preferably particles derived from plants, loaded with the protein; and (iv) 0 to 20% by weight of additional components based on the dry weight of the particles derived from plants or fungi, preferably particles derived from plants, loaded with the protein, The amount of plant - derived or fungus - derived material, protein, and further components together constitutes 100% by weight of the dry matter of the protein - loaded plant - derived or fungus - derived particles, preferably plant - derived particles. At least a part of the protein is immobilized in a continuous matrix of the plant - derived or fungus - derived material.

[0015] In a fourth aspect, the invention relates to a food product comprising, or consisting of, protein - loaded plant - derived or fungus - derived particles, preferably plant - derived particles, which can be obtained by a previously defined or previously defined method.

[0016] In a fifth aspect, the invention relates to the use of protein - loaded plant - derived or fungus - derived particles, preferably plant - derived particles, which can be obtained by a method defined herein or defined herein, as a food ingredient.

[0017] Definition The term "protein - rich" as used in relation to the present invention means that the protein - loaded plant - derived or fungus - derived particles, preferably plant - derived particles, have a higher weight percentage of protein after loading than before loading. The weight percentage of protein after loading is based on the protein already present in the plant - derived or fungus - derived particles, preferably plant - derived particles, before loading and specific to these, and also on the protein loaded into the plant - derived or fungus - derived particles, preferably plant - derived particles. Thus, for the purposes of the present invention, the expressions "protein - loaded" and "protein - rich" are considered interchangeable.

[0018] As used herein, the term "soluble protein" refers to a protein that has (still) a high level of water solubility. The term "soluble protein" as used herein is considered to be the same as the terms "techno-functional protein", "(substantially) native protein" and "(substantially) non-denatured protein" as used herein. For the purposes of the present invention, a protein is considered soluble if it has a water solubility of at least 20%, preferably at least 50%, more preferably at least 70% at pH = 7.0 and T = 20 °C when measured according to the analytical protocol defined in the experimental section. Some proteins have a very limited solubility at pH = 7.0 and T = 20 °C, but have a significant solubility at pH = 3.0 and T = 20 °C. For the purposes of the present invention, a protein is also considered soluble if it has a water solubility of at least 20%, preferably at least 50%, more preferably at least 70% at pH = 3.0 and T = 20 °C when measured according to the analytical protocol defined in the experimental section.

[0019] As used herein, the term "immobilization" refers to the process by which a soluble, i.e., mobile, protein is treated, for example, by heat, pH change, change in pH relative to the isoelectric point, or combinations thereof, to cause precipitation, denaturation and / or coagulation of the protein.

[0020] The term "denaturation" refers to the loss of the native conformation and biological activity of a protein. Denatured proteins have a reduced water solubility and can thus precipitate from an aqueous solution. Denaturation can be induced using physical methods such as heating or repeated freezing and thawing, or using chemical agents such as strong acids or strong bases, i.e., under extreme pH conditions. Depending on the conditions, the denaturation and the corresponding decrease in water solubility can be reversible or irreversible.

[0021] Denaturation is the first step of coagulation. The term "coagulation" refers to the solidification change of proteins, that is, the formation of insoluble aggregates caused by physical and / or chemical factors that bring about denaturation and precipitation. Also, depending on the conditions, coagulation may be reversible or irreversible.

[0022] As used herein, the term "precipitation" relates to the process by which proteins are separated from an aqueous solution. Apart from precipitation due to denaturation as described above, proteins can also typically precipitate by removal of electrostatic repulsion forces and hydration shells. An example is the precipitation of proteins at the isoelectric point, also called "flocculation", by adjusting the pH. Flocculation is typically a reversible process. However, flocculated proteins may subsequently be treated with heat, irreversibly degrading the protein's techno-functional properties such as its water solubility. Precipitation by dehydration can be induced, for example, using alcohol. Furthermore, precipitation of proteins can be induced using certain salts.

[0023] The terms "comprise" and "include" and variations such as "comprises", "comprising", "includes" and "including" used throughout this specification and the appended claims are to be construed in an inclusive sense. These words are intended to convey that, unless otherwise specified, they can include other elements or integers not specifically recited that the context permits.

[0024] As used herein, the articles "a" and "an" refer to the grammatical object of one or more (i.e., one or at least one) articles. For example, "an element" can mean one element or more than one element, unless otherwise specified. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] BRIEF DESCRIPTION OF THE DRAWINGS

Fig. 1a

Fig. 1b

Fig. 2a

Fig. 2b

Fig. 3a

Fig. 3b

BEST MODE FOR CARRYING OUT THE INVENTION

[0026] Detailed Description Method for loading proteins into plant-derived or fungus-derived particles, preferably plant-derived particles In a first aspect, the present invention relates to a method for loading a protein onto a plant-derived or fungus-derived particle, preferably a plant-derived particle from a vegetable, fruit, and edible fungus fruiting body, the method comprising: (a) providing a plant-derived or fungus-derived particle, preferably a plant-derived particle; (b) providing a soluble protein that can be immobilized in a plant-derived or fungus-derived particle, preferably a plant-derived particle; (c) adding the soluble protein provided in step (b) to the plant-derived or fungus-derived particle provided in step (a), preferably a plant-derived particle, and preferably then mixing; (d) contacting the plant-derived or fungus-derived particle, preferably a plant-derived particle, with the protein in the mixture provided in step (c) under conditions such that the protein remains soluble, to load the protein onto the plant-derived or fungus-derived particle, preferably a plant-derived particle; (e) immobilizing at least a part of the protein in the plant-derived or fungus-derived particle, preferably a plant-derived particle, and the plant-derived or fungus-derived particle loaded with the protein obtained in step (e), preferably a plant-derived particle, contains at least 35% by weight of protein based on the dry weight of the plant-derived or fungus-derived particle loaded with the protein obtained in step (e). The expression "contains at least xx% by weight of protein based on the dry weight of the plant-derived or fungus-derived particle loaded with the protein obtained in step (e)" refers to the weight of the protein determined using the Kjeldahl method with a conversion factor of 6.25 divided by the dry weight of the plant-derived or fungus-derived particle loaded with the protein obtained in step (e).

[0027]

[0028] ​In a preferred embodiment, the plant-derived or fungus-derived particles loaded with the protein obtained in step (e), preferably the plant-derived particles, contain at least 38% by weight, more preferably at least 40% by weight, even more preferably at least 42% by weight, for example, at least 44% by weight, at least 46% by weight, at least 48% by weight, at least 50% by weight, at least 52% by weight, at least 54% by weight, at least 56% by weight, at least 58% by weight, at least 60% by weight, or at least 62% by weight of the protein, based on the dry weight of the plant-derived or fungus-derived particles loaded with the protein obtained in step (e), preferably the plant-derived particles.

[0029] In another preferred embodiment, the plant-derived or fungus-derived particles loaded with the protein obtained in step (e), preferably the plant-derived particles, contain 35 - 80% by weight, more preferably 38 - 80% by weight, even more preferably 40 - 80% by weight, for example, 42 - 80% by weight, 44 - 80% by weight, or 46 - 80% by weight of the protein, based on the dry weight of the plant-derived or fungus-derived particles loaded with the protein obtained in step (e), preferably the plant-derived particles.

[0030] In yet another preferred embodiment, the plant-derived or fungus-derived particles loaded with the protein obtained in step (e), preferably the plant-derived particles, contain 35 - 75% by weight, for example, 35 - 65% by weight, 35 - 60% by weight, 35 - 56% by weight, 35 - 50% by weight, or 35 - 46% by weight of the protein, based on the dry weight of the plant-derived or fungus-derived particles loaded with the protein obtained in step (e), preferably the plant-derived particles.

[0031] Step (a): Providing plant-derived or fungus-derived particles, preferably plant-derived particles In step (a) of the process as defined herein, plant-derived or fungus-derived particles, preferably plant-derived particles from vegetables, fruits, and the fruiting bodies of edible fungi, are provided.

[0032] In a preferred embodiment, the plant-derived or fungus-derived particles provided in step (a), preferably the plant-derived particles, are rehydratable, meaning that they can absorb and retain an amount of water that is at least 5 times their own dry weight, more preferably at least 10 times their own dry weight, even more preferably at least 15 times their own dry weight, for example, at least 20 times their own dry weight, at least 25 times their own dry weight, or at least 30 times their own dry weight.

[0033] In another preferred embodiment, the plant-derived or fungus-derived particles provided in step (a), preferably the plant-derived particles, are rehydratable, meaning that they can absorb and retain an amount of water that is 2 to 33 times their own dry weight, more preferably 5 to 30 times their own dry weight, even more preferably 8 to 25 times their own dry weight, for example, 10 to 20 times their own dry weight.

[0034] In one embodiment, the plant-derived or fungus-derived particles, preferably the plant-derived particles, include the whole vegetable, the whole fruit, or the whole fruiting body of an edible fungus, excluding any of the stem, leaves, and skin.

[0035] The average size or average maximum dimension of the plant-derived or fungus-derived particles, preferably plant-derived particles, provided in step (a) is not particularly limited. However, as will be understood by those skilled in the art, the higher the specific surface area (surface area divided by volume) of the plant-derived or fungus-derived particles, preferably plant-derived particles, the faster the loading rate. Thus, in certain preferred embodiments, the plant-derived or fungus-derived particles, preferably plant-derived particles, provided in step (a) have a particle size or maximum dimension of from 0.5 mm to 5 cm, preferably from 1 mm to 3 cm, more preferably from 1 mm to 1 cm. In some embodiments, the plant-derived or fungus-derived particles, preferably plant-derived particles, provided in step (a) have a particle size or maximum dimension of at least 5 mm. This particle size is particularly advantageous when the plant-derived or fungus-derived particles, preferably plant-derived particles, are used in a burger. In some embodiments, the plant-derived or fungus-derived particles, preferably plant-derived particles, provided in step (a) have a particle size or maximum dimension of up to 4 mm. This particle size is particularly advantageous when the plant-derived or fungus-derived particles, preferably plant-derived particles, are used in a confectionery or pastry product such as a cake.

[0036] As will be understood by those skilled in the art, the plant-derived or fungus-derived particles, preferably plant-derived particles, provided in step (a), i.e., before loading the protein, are not necessarily the preferred average size or average maximum dimension of the plant-derived or fungus-derived particles, preferably plant-derived particles, loaded with the protein intended to be applied in or as a food product. In other words, the plant-derived or fungus-derived particles, preferably plant-derived particles, loaded with the protein obtained via the method as previously defined can be subjected to cutting, slicing or grinding in a further step.

[0037] In certain embodiments, the plant-derived or fungus-derived particles are plant-derived particles from vegetables such as sugar beet, carrot, chicory, potato and combinations thereof.

[0038] In another embodiment, the plant-derived or fungus-derived particles are plant-derived particles from fruits such as apples, pineapples, citrus fruits, cranberries, grapes, and combinations thereof.

[0039] In yet another embodiment, the plant-derived or fungus-derived particles are fungus-derived particles from the fruiting bodies of edible fungi such as mushrooms.

[0040] In a preferred embodiment, the plant-derived or fungus-derived particles, preferably plant-derived particles, are particles from vegetables, fruits, and the fruiting bodies of edible fungi selected from the group consisting of sugar beet, carrot, chicory, potato, apple, pineapple, citrus fruits, cranberries, grapes, mushrooms, and combinations thereof.

[0041] In one embodiment, the plant-derived or fungus-derived particles provided in step (a), preferably plant-derived particles, exclude seeds and beans that are already rich in protein, especially seeds and beans such as lupin seeds and soybean beans.

[0042] Preferably, the plant-derived or fungus-derived particles provided in step (a), preferably plant-derived particles, have a moisture content of 30 to 97% by weight, more preferably less than 96% by weight, still more preferably less than 95% by weight, yet still more preferably less than 94% by weight, and most preferably less than 93% by weight, based on the weight of the plant-derived or fungus-derived particles, preferably plant-derived particles.

[0043] In another preferred embodiment, the plant-derived or fungus-derived particles provided in step (a), preferably plant-derived particles, have a moisture content of 40 to 97% by weight, for example, 50 to 97% by weight, 60 to 97% by weight, 70 to 97% by weight, 80 to 97% by weight, or 85 to 97% by weight, based on the weight of the plant-derived or fungus-derived particles, preferably plant-derived particles.

[0044] In yet another preferred embodiment, the plant-derived or fungus-derived particles provided in step (a), preferably the plant-derived particles, have a moisture content of 85 to 96% by weight, for example, 86 to 95% by weight, 87 to 94% by weight, or 88 to 93% by weight, based on the weight of the plant-derived or fungus-derived particles, preferably the plant-derived particles.

[0045] In one embodiment, the plant-derived particles provided in step (a) of the process as defined herein contain at least 70% by weight of soft tissue cell wall material and less than 6% by weight of lignin, based on the dry matter of the plant-derived particles.

[0046] The term "soft tissue cell wall material" is well known in the art. The soft tissue cell wall, also referred to as the "primary cell wall", refers to soft tissue or succulent tissue, which is the most abundant type of cell wall in edible plants. For example, in sugar beet, the soft tissue cells are the most abundant tissue surrounding the secondary vascular tissue (xylem and phloem). In this regard, see P.W. van der Poel et al., Sugar Technology, Verlag Dr Albert Bartens KG, Berlin 1998, page 211. Soft tissue cells contain a relatively thin cell wall compared to the secondary cell wall and are interconnected by pectin. In the secondary vascular tissue (xylem and phloem tissues), the cell wall is much thicker than that of soft tissue cells and is bound to lignin.

[0047] Polysaccharides usually constitute more than 90% of the primary plant cell wall, and cellulose, hemicellulose, and pectin are the main components. The exact form and (chemical) composition of the soft tissue cell wall can vary by species. The soft tissue cell wall material in the plant-derived particles provided in step (a) can be obtained from various plant sources containing soft tissue cell walls.

[0048] Preferably, the plant-derived or fungus-derived particles provided in step (a), preferably the plant-derived particles, contain at least 15% by weight, preferably 15 - 40% by weight, more preferably 15 - 35% by weight, and most preferably 15 - 30% by weight of cellulose based on the dry matter of the plant-derived or fungus-derived particles, preferably the plant-derived particles.

[0049] Preferably, the plant-derived or fungus-derived particles provided in step (a), preferably the plant-derived particles, contain at least 15% by weight, preferably 15 - 40% by weight, more preferably 15 - 35% by weight, and most preferably 15 - 30% by weight of pectin based on the dry matter of the plant-derived or fungus-derived particles, preferably the plant-derived particles.

[0050] Preferably, the plant-derived or fungus-derived particles provided in step (a), preferably the plant-derived particles, contain at least 15% by weight, preferably 15 - 40% by weight, more preferably 15 - 35% by weight, and most preferably 20 - 35% by weight of hemicellulose based on the dry matter of the plant-derived or fungus-derived particles, preferably the plant-derived particles.

[0051] Other typical insoluble or poorly soluble soft tissue cell (wall) components that may be present in the plant-derived or fungus-derived particles, preferably the plant-derived particles, provided in step (a) include proteins, lignin, residual sugars, fats, and ash.

[0052] Thus, in certain embodiments, the plant-derived or fungus-derived particles, preferably the plant-derived particles, provided in step (a) contain 1 - 25% by weight, preferably 3 - 15% by weight of protein based on the dry matter of the plant-derived or fungus-derived particles, preferably the plant-derived particles.

[0053] In certain embodiments, the plant-derived particles provided in step (a) contain 0 - 5% by weight, preferably 0 - 4% by weight, more preferably 0 - 3% by weight of lignin based on the dry matter of the plant-derived particles.

[0054] In certain embodiments, the plant-derived or fungus-derived particles, preferably plant-derived particles, provided in step (a) contain less than 10% by weight, preferably less than 6% by weight, more preferably less than 3% by weight of sugar, based on the dry matter of the plant-derived or fungus-derived particles, preferably plant-derived particles.

[0055] In certain embodiments, the plant-derived or fungus-derived particles, preferably plant-derived particles, provided in step (a) contain less than 2% by weight, preferably less than 1% by weight, more preferably less than 0.5% by weight of fat, based on the dry matter of the plant-derived or fungus-derived particles, preferably plant-derived particles.

[0056] In certain embodiments, the plant-derived or fungus-derived particles, preferably plant-derived particles, provided in step (a) contain less than 6% by weight, preferably less than 5% by weight, more preferably less than 4% by weight of ash, based on the dry matter of the plant-derived or fungus-derived particles, preferably plant-derived particles.

[0057] In certain preferred embodiments, the plant-derived particles provided in step (a) contain at least 70% by weight of soft tissue cell wall material and less than 6% by weight of lignin, based on the dry matter of the plant-derived particles, and are selected from the group consisting of used sugar beet pulp particles, pulp particles from citrus fruits, pulp particles from tomatoes, used pulp particles from chicory, pulp particles from potatoes, pulp particles from pineapples, pulp particles from apples, pulp particles from cranberries, pulp particles from grapes, and / or pulp particles from carrots (excluding the stems and leaves) and combinations thereof, more preferably selected from the group consisting of used sugar beet pulp particles, used chicory pulp particles, and combinations thereof, and most preferably used sugar beet pulp particles.

[0058] The term "spent" in "spent sugar beet pulp particles" and "spent pulp particles from chicory" is well-known to those skilled in the art and relates to pulp particles of either sugar beet or chicory from which sugar or inulin has been extracted, respectively. Preferably, the spent sugar beet pulp particles referred to herein can be obtained or are obtained after sucrose extraction by a warm water diffusion process carried out at a temperature of 60 to 80 °C, preferably 65 to 75 °C. The residence time of the sugar beet pulp particles in the warm water diffusion process is preferably 30 to 180 minutes. Such a sucrose extraction process is known to those skilled in the art and is typically carried out in a so-called diffusion tower.

[0059] In a very preferred embodiment of the present invention, the components of the plant-derived or fungus-derived particles, preferably plant-derived particles, provided in step (a) are similar to those of common spent sugar beet pulp, and based on the dry matter of the plant-derived or fungus-derived particles, preferably plant-derived particles, contain 15 to 35 wt%, preferably 15 to 30 wt%, more preferably 18 to 26 wt% of cellulose, 15 to 40 wt%, preferably 20 to 38 wt%, more preferably 22 to 35 wt% of hemicellulose, 15 to 35 wt%, more preferably 20 to 30 wt%, more preferably 21 to 27 wt% of pectin, 5 to 15 wt% of protein, less than 5 wt% of lignin, less than 5 wt% of sugar, less than 6 wt% of ash and less than 1 wt% of fat.

[0060] In a preferred embodiment, the plant-derived or fungus-derived particles, preferably plant-derived particles, provided in step (a) are obtained from raw plant-derived or fungus-derived materials that are subjected to one or more of the following steps: · Heating, preferably cooking; · Pulsed electric field treatment; · Cutting, slicing or grinding; · Sieving; · Removal of water by pressing, drying, sieving or combinations thereof; and · Freezing and thawing.

[0061] In a very preferred embodiment, the plant-derived or fungus-derived particles provided in step (a), preferably the plant-derived particles, are obtained from raw plant-derived or fungus-derived materials to be subjected to the following steps: · Heating, preferably cooking; · Freezing and thawing; · Preferably cutting, slicing or grinding; · Preferably pressing.

[0062] In a very preferred embodiment, the plant-derived or fungus-derived particles provided in step (a), preferably the plant-derived particles, are used sugar beet pulp that has been subjected to the following: (i) Heating at a temperature of at least 80°C, preferably at least 85°C; (ii) Preferably freezing and thawing; (iii) Optionally, pressing, drying, sieving or combinations thereof, more preferably water removal only by pressing and / or sieving.

[0063] Therefore, as will be understood by those skilled in the art from the previous paragraph, the plant-derived particles provided in step (a) are used sugar beet pulp obtained after sucrose extraction by a warm water diffusion process carried out at a temperature of 60 to 80°C, preferably 65 to 75°C. The residence time of the sugar beet pulp particles in the warm water diffusion process is preferably 30 to 180 minutes, and it is highly preferred that the used sugar beet pulp particles are then subjected to step (i) of heating at a temperature of at least 80°C, preferably at least 85°C. The inventors have surprisingly found that subjecting the used pulp to a simple heat treatment at a temperature higher than the normal sucrose extraction temperature activates the pulp and is sufficient to make it susceptible to the protein loading in step (d) of the process of the present invention. The optional freezing and thawing step (ii) may be carried out before or after the heating step (i). The inventors have found that there is no need to carry out the freezing and thawing step (ii) after the heating step (i) in order to sufficiently activate the used pulp for protein loading. The used pulp may be subjected to the freezing and thawing step (ii) before the heating step (i), which is useful for long-term storage of the pulp.

[0064] In a preferred embodiment, step (i) involves heating at a temperature of at least 90°C, more preferably at least 95°C, and even more preferably at least 100°C. Such temperatures have been found to result in a lighter-colored material, which is a desirable characteristic for consumers.

[0065] In one embodiment, the heating step (i) is carried out over a period of at least 1 minute, such as at least 5 minutes, at least 10 minutes, at least 15 minutes, at least 20 minutes, at least 30 minutes, at least 40 minutes, at least 50 minutes, at least 80 minutes or at least 120 minutes.

[0066] In a preferred embodiment, the heating step (i) is carried out over a period of 1 to 60 minutes, 2 to 60 minutes, 5 to 60 minutes, 10 to 60 minutes, 15 to 60 minutes, 20 to 60 minutes, 30 to 60 minutes or 40 to 60 minutes.

[0067] In another preferred embodiment, the heating step (i) is carried out over a period of 1 to 55 minutes, 2 to 50 minutes, 5 to 45 minutes, 10 to 40 minutes, 15 to 30 minutes, 17 to 25 minutes or 18 to 23 minutes.

[0068] In a preferred embodiment, step (i) involves heating at a temperature of at least 80 °C, preferably at least 85 °C, over a period of 1 to 120 minutes, 5 to 100 minutes, 8 to 90 minutes, 10 to 80 minutes, 15 to 75 minutes, 20 to 70 minutes, 30 to 65 minutes or 40 to 60 minutes.

[0069] In another preferred embodiment, step (i) involves heating at a temperature of at least 90 °C over a period of 10 to 60 minutes, 15 to 50 minutes, 17 to 40 minutes or 18 to 30 minutes.

[0070] In yet another preferred embodiment, step (i) involves heating at a temperature of at least 95 °C over a period of 15 to 30 minutes, 17 to 25 minutes or 18 to 23 minutes.

[0071] As will be understood by those skilled in the art, step (i) can also be carried out under superheated conditions, i.e., at increased pressure. In a preferred embodiment, step (i) involves superheating at a temperature of 85 to 120 °C, more preferably at a temperature of 90 to 115 °C, and even more preferably at a temperature of 95 to 110 °C.

[0072] Most preferably, step (i) involves superheating at a temperature of about 100 °C at atmospheric pressure.

[0073] In a highly preferred embodiment, the method further comprises cutting, slicing or grinding before and / or after any of the process steps (i) to (iii).

[0074] There are generally two methods for loading proteins onto plant-derived or fungus-derived particles, preferably plant-derived particles, namely "dry protein loading" and "wet protein loading". In dry protein loading, the plant-derived or fungus-derived particles, preferably plant-derived particles, are contacted with dry protein powder. The inventors have found that dry loading can be advantageously performed on plant-derived or fungus-derived particles, preferably plant-derived particles, that are fully hydrated. Therefore, when dry protein loading is carried out, the plant-derived or fungus-derived particles, preferably plant-derived particles, are typically not subjected to processes that remove water by pressing, drying, or a combination thereof. In wet protein loading, the plant-derived or fungus-derived particles, preferably plant-derived particles, are contacted with an aqueous protein solution. The inventors have found that wet loading can be advantageously carried out on plant-derived or fungus-derived particles, preferably plant-derived particles, that are not fully hydrated. Therefore, when wet protein loading is carried out, the plant-derived or fungus-derived particles, preferably plant-derived particles, are preferably subjected to a process that removes water by pressing, drying, or a combination thereof.

[0075] Step (b): Providing a soluble protein that can be immobilized In step (b) of the process as defined herein, a "soluble protein that can be immobilized" is provided. As is well known to those skilled in the art, many proteins isolated in their native state from their corresponding sources are water-soluble and can be used as techno-functional components in food preparations, for example, to provide (thermal) gelling, foaming, water-binding, or emulsification processes. This functionality is typically (reversibly or irreversibly) lost by subjecting the native protein to, for example, heat, extreme pH, changes in pH to the isoelectric point, or a combination thereof, resulting in at least partial denaturation, precipitation, and / or coagulation of the protein.

[0076] In a preferred embodiment, the water solubility of the soluble protein that can be immobilized in the plant-derived or fungus-derived particles, preferably plant-derived particles, provided in step (b) is at least 20%, more preferably at least 50%, even more preferably at least 70%, and still more preferably at least 90% at pH = 7.0 and T = 20°C when measured according to the protocol defined in the experimental section.

[0077] In another preferred embodiment, the water solubility of the soluble protein that can be immobilized in the plant-derived or fungus-derived particles, preferably plant-derived particles, provided in step (b) is 20 - 100%, more preferably 50 - 100%, even more preferably 70 - 100%, and still more preferably 90 - 100% at pH = 7.0 and T = 20°C when measured according to the protocol defined in the experimental section.

[0078] In a preferred embodiment, the water solubility of the soluble protein that can be immobilized in the plant-derived or fungus-derived particles, preferably plant-derived particles, provided in step (b) is at least 20%, more preferably at least 50%, even more preferably at least 70%, and still more preferably at least 90% at pH = 3.0 and T = 20°C when measured according to the protocol defined in the experimental section.

[0079] In another preferred embodiment, the water solubility of the soluble protein that can be immobilized in the plant-derived or fungus-derived particles, preferably plant-derived particles, provided in step (b) is 20 - 100%, more preferably 50 - 100%, even more preferably 70 - 100%, and still more preferably 90 - 100% at pH = 3.0 and T = 20°C when measured according to the protocol defined in the experimental section.

[0080] In a preferred embodiment, the soluble protein that can be immobilized in the plant-derived or fungus-derived particles, preferably plant-derived particles, provided in step (b) is a plant protein including proteins derived from legumes, leguminous plants, oil seeds, algae, and kelp; a microbial protein including proteins derived from yeast, mold, and fungi; an animal protein including whey protein, chicken egg protein, and insect-derived protein; their hydrolysates; and combinations thereof, which is selected from the group consisting of them.

[0081] In a highly preferred embodiment, the soluble protein that can be immobilized in the plant-derived or fungus-derived particles, preferably plant-derived particles, provided in step (b) is selected from the group consisting of potato protein, rubisco, lentil-derived protein, pea protein, wheat protein, barley-derived protein, rice-derived protein, soybean protein, broad bean protein, chickpea-derived protein, chicken egg protein, whey protein, canola protein, lupin bean protein, chickpea protein, almond protein, sunflower protein, their hydrolysates, and combinations thereof, and even more preferably, is selected from the group consisting of potato protein, broad bean protein, pea protein, lentil-derived protein, whey protein, their hydrolysates, and combinations thereof.

[0082] In an embodiment, the soluble protein that can be immobilized in the plant-derived or fungus-derived particles, preferably plant-derived particles, provided in step (b) has a molecular weight of 3 to 650 kDa, for example, 3 to 500 kDa, 3 to 450 kDa, 3 to 420 kDa, 3 to 400 kDa, 3 to 380 kDa, 50 to 650 kDa, 100 to 500 kDa, 150 to 450 kDa, 170 to 420 kDa, 180 to 400 kDa, or 190 to 380 kDa when measured using size exclusion chromatography (SEC).

[0083] Step (c): adding the soluble protein and the plant-derived or fungus-derived particles, preferably plant-derived particles In step (c) of the process as previously defined, the soluble protein provided in step (b) and the plant-derived or fungus-derived particles, preferably plant-derived particles, provided in step (a) are added and preferably subsequently mixed.

[0084] In certain embodiments, the soluble protein that can be immobilized in the plant-derived or fungus-derived particles, preferably plant-derived particles, and the plant-derived or fungus-derived particles, preferably plant-derived particles, are applied in step (c) at a weight ratio of 1:0.05 (protein: plant-derived or fungus-derived particles, preferably plant-derived particles) to 1:1, preferably at a weight ratio of 1:0.1 to 1:0.5, on a dry matter basis.

[0085] Step (c) preferably includes mixing of the components, such as gentle stirring of the components.

[0086] In certain embodiments, the plant-derived or fungus-derived particles, preferably plant-derived particles, are loaded with different soluble proteins that can be immobilized in the plant-derived or fungus-derived particles, preferably plant-derived particles.

[0087] In one embodiment, the soluble protein that can be immobilized in the plant-derived or fungus-derived particles, preferably plant-derived particles, provided in step (b) is added as an aqueous solution to the plant-derived or fungus-derived particles, preferably plant-derived particles, in step (c) (“wet protein loading”). The water solubility of the protein depends on temperature, pH, and the protein species. Selecting the optimal conditions is within the skill of those in the art. In this embodiment, the total amount of soluble protein based on the dry weight to be added to the plant-derived or fungus-derived particles, preferably plant-derived particles, is typically an amount in excess of the intended amount of soluble protein to be loaded into the plant-derived or fungus-derived particles, preferably plant-derived particles. In this embodiment, the protein that can be immobilized in the plant-derived or fungus-derived particles, preferably plant-derived particles, is preferably added as an aqueous solution to the plant-derived or fungus-derived particles, preferably plant-derived particles, at a weight ratio of 1:0.05 to 1:0.3 on a dry weight basis, preferably at a weight ratio of 1:0.1 to 1:0.2 on a dry weight basis.

[0088] As will be understood by those skilled in the art, loading the plant-derived or fungus-derived particles, preferably plant-derived particles, with the soluble protein by contacting the particles with an aqueous solution of the soluble protein also enables loading of additional soluble components via the same aqueous solution.

[0089] In one embodiment, the additional component is water-insoluble or slightly partially soluble in water and is loaded into the plant-derived or fungus-derived particles, preferably plant-derived particles, in the form of a water-in-oil (micro) emulsion.

[0090] Non-limiting examples of additional components are salts, flavorants, colorants, and preservatives. In one highly preferred embodiment, the additional component is a food-grade additional component.

[0091] The inventors have unexpectedly established that plant-derived or fungus-derived particles, preferably plant-derived particles as previously defined, can effectively load soluble proteins even at high protein loading levels when the soluble proteins are added as a dry powder to the plant-derived or fungus-derived particles, preferably plant-derived particles, in step (c) ("dry protein loading"). In one highly preferred embodiment, the soluble protein that can be immobilized in the plant-derived or fungus-derived particles, preferably plant-derived particles, provided in step (b) is added as a dry powder in step (c), preferably having a particle size characterized by a Sauter mean diameter (D[3,2]) of 10 to 100 μm when determined by a laser diffraction particle size analyzer.

[0092] In this embodiment, the soluble protein that can be immobilized in the plant-derived or fungus-derived particles, preferably plant-derived particles, is preferably added as a dry powder to the plant-derived or fungus-derived particles, preferably plant-derived particles, at a weight ratio of 1:0.1 to 1:1, preferably 1:0.2 to 1:0.5, on a dry weight basis in step (c).

[0093] Step (d): contacting the soluble protein with the plant-derived or fungus-derived particles, preferably plant-derived particles In step (d) of the process as previously defined, the plant-derived or fungus-derived particles, preferably plant-derived particles, are contacted with the soluble protein in the mixture provided in step (c), and the protein is loaded onto the plant-derived or fungus-derived particles, preferably plant-derived particles, under conditions where the protein remains soluble. It is important that the protein remains sufficiently soluble during loading because this allows the protein to penetrate as far as possible into the matrix of the plant-derived or fungus-derived particles, preferably plant-derived particles. Partially immobilized proteins cannot be loaded deep into the plant-derived or fungus-derived particles, preferably plant-derived particles. The definition of "soluble protein" with respect to immobilization (b) applies equally to step (d).

[0094] The aqueous solubility of proteins typically depends on pH and temperature. It is within the skill of the artisan to select the most appropriate conditions to achieve efficient and / or high loading for different soluble proteins. The pH can be adjusted, for example, by adding 1 M NaOH or by adding concentrated lactic acid.

[0095] In an embodiment in which "wet protein loading" is performed, the plant-derived or fungal-derived particles, preferably plant-derived particles, and the soluble proteins in the mixture provided in step (c) are preferably contacted for at least 1 minute, more preferably at least 5 minutes, even more preferably at least 30 minutes, still more preferably at least 60 minutes, even more preferably at least 120 minutes, such as from 1 minute to 12 hours or from 5 minutes to 240 minutes.

[0096] In an embodiment where "dry protein loading" is performed, the plant-derived or fungal-derived particles, preferably plant-derived particles, and the soluble proteins in the mixture provided in step (c) are preferably contacted for at least 1 minute, more preferably at least 5 minutes, even more preferably at least 1 hour, still more preferably at least 2 hours, such as from 5 minutes to 24 hours, from 1 hour to 24 hours or from 2 hours to 24 hours.

[0097] In another preferred embodiment, the plant-derived or fungal-derived particles, preferably plant-derived particles, and the soluble proteins in the mixture provided in step (c) are contacted at a temperature between 4 and 60°C, more preferably at a temperature between 4 and 40°C, even more preferably at a temperature between 4 and 20°C.

[0098] In another preferred embodiment, the plant-derived or fungal-derived particles, preferably plant-derived particles, and the soluble proteins in the mixture provided in step (c) are contacted at a pH of 6-9, such as a pH of 6.5-8 or a pH of 6.5-7.5.

[0099] In yet another preferred embodiment, the plant-derived or fungus-derived particles, preferably the plant-derived particles, are contacted with the soluble proteins in the mixture provided in step (c) at a pH of 2 to 4, for example, at a pH of 2.5 to 3.5 or 3 to 4.

[0100] Step (d) may include mixing of the components, for example, gentle stirring of the components.

[0101] Step (e): Immobilizing the protein The method as previously defined further includes step (e) of immobilizing at least a portion of the protein in the plant-derived or fungus-derived particles, preferably the plant-derived particles.

[0102] The immobilization of the protein in the plant-derived or fungus-derived particles, preferably the plant-derived particles, has the advantage that the plant-derived or fungus-derived particles, preferably the plant-derived particles loaded with the protein, can be rehydrated after an optional drying step without leakage of the protein from the plant-derived or fungus-derived particles, preferably the plant-derived particles.

[0103] The immobilization conditions are not the same for all types of proteins. Selecting the optimal conditions to achieve efficient immobilization is within the skill of those in the art.

[0104] In one embodiment, step (e) of immobilizing at least a portion of the protein in the plant-derived or fungus-derived particles, preferably the plant-derived particles, is carried out as follows: (I) The plant-derived or fungus-derived particles, preferably the plant-derived particles loaded with the protein provided in step (d) are superheated to a temperature of at least 85°C, preferably 85 to 100°C, preferably 90 to 100°C, or (II) The plant-derived or fungus-derived particles, preferably the plant-derived particles loaded with the protein provided in step (d) are exposed to an acidic solution, or (III) exposing the plant-derived or fungus-derived particles, preferably plant-derived particles, loaded with the protein provided in step (d) to a pH equal to the isoelectric point, or (IV) drying the plant-derived or fungus-derived particles, preferably plant-derived particles, loaded with the protein provided in step (d), or (V) performing the combination of (I) and (II), or (VI) performing the combination of (I) and (III).

[0105] In certain preferred embodiments, the protein is completely immobilized in step (e). In another preferred embodiment, the protein is completely denatured in step (e). In yet another preferred embodiment, the protein is completely precipitated in step (e). In yet another preferred embodiment, the protein is completely soft-aggregated in step (e). In yet another preferred embodiment, the protein is completely coagulated in step (e).

[0106] Option (I) or (IV) is most preferred because it does not require the addition of chemicals that result in fewer processed products, which is preferred for food applications.

[0107] Option (I) is preferably carried out in the presence of a salt, preferably a food-grade salt such as NaCl or a calcium salt. The heating in option (I) is preferably carried out for at least 1 minute, for example, 1 to 5 minutes.

[0108] The acidic solution of option (II) preferably comprises a solution containing an acid selected from the group consisting of organic acids, carbonic acid, phosphoric acid, and combinations thereof, more preferably a solution containing a C 2 ~C 6 organic acid, more preferably a solution containing a C 2 ~C 6 organic carboxylic acid. In some embodiments, the acidic solution of option (II) is a solution containing an acid selected from the group consisting of citric acid, acetic acid, lactic acid, phosphoric acid, malic acid, tartaric acid, carbonic acid, fumaric acid, salts thereof, and combinations thereof.

[0109] The acidic solution of alternative (II) has a pH of less than 6, more preferably less than 5. In an exemplary embodiment, the acidic solution of alternative (II) is an acidic solution having a pH of 4 - 5, preferably a pH of 4.2 - 4.8. Alternative (II) preferably comprises contacting plant-derived or fungus-derived particles, preferably plant-derived particles, loaded with the protein provided in step (d) with the acidic solution for at least 1 minute, for example 1 - 5 minutes.

[0110] The inventors have found that immobilizing a protein in plant-derived or fungus-derived particles, preferably plant-derived particles, can also be carried out by simply drying the material as specified in alternative (IV). This results in a satisfactory immobilization of the protein and does not result in significant leakage when the dried material is rehydrated or washed. Drying may be carried out by any method known to those skilled in the art, but is preferably a simple air drying using hot air that results in dehydration (i.e., a decrease in water content) of the plant-derived or fungus-derived particles, preferably plant-derived particles, loaded with the protein. The hot air preferably has a temperature in the range of 70 - 95 °C, preferably in the range of 80 - 95 °C. The inventors have found that above 95 °C, the particles tend to blacken. Drying is preferably carried out to obtain a product having a water activity (AW) of 0.10 - 0.80, more preferably 0.20 - 0.76, even more preferably 0.30 - 0.60, when measured at 25 °C using a Lab Master-aw neo water activity measuring device (Novasina AG).

[0111] Step (f): Drying or freezing If the plant-derived or fungus-derived particles, preferably plant-derived particles, loaded with the protein obtained in step (e) are not used immediately, the inventors have found it advantageous to dry and / or freeze them so that they can be conveniently stored in a stable manner for future use.

[0112] Thus, in certain embodiments, the method as previously defined further comprises step (f) of drying and / or freezing the plant-derived or fungus-derived particles, preferably plant-derived particles, loaded with the protein obtained in step (e). Drying and / or freezing can help increase the microbiological stability of the product, which is preferred for food applications. Drying also reduces the transportation costs of the product.

[0113] If the immobilization of step (e) is carried out by drying, step (f) will typically not include drying the protein obtained in step (e). If the immobilization of step (e) is carried out by drying, the dried material obtained in step (e) may be further frozen to extend its shelf life. It is also possible to carry out a two-stage drying process, where the first drying step is applied in step (e) to immobilize at least a portion of the protein in the plant-derived or fungus-derived particles, preferably plant-derived particles, resulting in a dried material obtained in step (e) having a first water activity level, and subsequently the second drying step (f) results in a dried material obtained in step (f) having a second water activity level lower than the first water activity level.

[0114] In certain preferred embodiments, step (f) is carried out, which comprises drying the plant-derived or fungus-derived particles, preferably plant-derived particles, loaded with the protein obtained in step (e) to obtain a product having a water activity (AW) of 0.10 to 0.80, more preferably 0.20 to 0.76, even more preferably 0.30 to 0.60, when measured at 25 °C using a Lab Master-aw neo water activity measuring device (Novasina AG).

[0115] Drying may be carried out by any method known to those skilled in the art, but is preferably a simple air drying using hot air that results in dehydration (i.e., a decrease in water content) of plant-derived or fungus-derived particles loaded with protein, preferably plant-derived particles. The hot air preferably has a temperature in the range of 70 to 95 °C, preferably in the range of 80 to 95 °C. The inventors have found that when exceeding 95 °C, the particles are prone to blackening.

[0116] In a preferred embodiment, step (f) is carried out, which includes freezing plant-derived or fungus-derived particles loaded with the protein obtained in step (e), preferably plant-derived particles, preferably including freezing at a temperature below -15 °C for at least 12 hours.

[0117] The inventors have found that plant-derived or fungus-derived particles loaded with the protein according to the present invention, preferably plant-derived particles, can be thawed without significantly losing protein immobilization.

[0118] As previously described, the preferred average size or average maximum dimension of the plant-derived or fungus-derived particles provided in step (a), i.e., before loading the protein, preferably plant-derived particles, is not necessarily the preferred average size or average maximum dimension of the plant-derived or fungus-derived particles loaded with the protein that will ultimately be in food or applied as food. In other words, plant-derived or fungus-derived particles loaded with the protein obtained via the method as previously defined, preferably plant-derived particles, can be subjected to cutting, slicing or grinding in a further step. This step can be carried out after step (e) and before step (f), or after step (f), or in combination thereof.

[0119] Step (g): Rehydration In certain embodiments, the plant-derived or fungus-derived particles, preferably plant-derived particles, loaded with the protein obtained in step (e), or the dried plant-derived or fungus-derived particles, preferably plant-derived particles, loaded with the protein obtained in step (f) are subsequently rehydrated, for example, for application as or in a food product or for application as a food ingredient. In embodiments, the time between step (e) or (f) and the rehydration step (g) can be up to 1 day, up to 15 days, up to 1 month or up to 5 months.

[0120] As shown in the attached examples, the inventors have unexpectedly found that plant-derived or fungus-derived particles, preferably plant-derived particles, loaded with the protein according to the invention can be rehydrated without substantial loss of the protein. Furthermore, the rehydration of the protein-loaded particles does not result in cellulose particles sticking to each other, and the shape of the protein-loaded particles is maintained upon rehydration.

[0121] Plant-derived or fungus-derived particles, preferably plant-derived particles, loaded with a protein In a second aspect, the invention relates to plant-derived or fungus-derived particles, preferably plant-derived particles, loaded with a protein obtainable or obtained by a process as defined herein.

[0122] To the best of the inventors' knowledge, plant-derived or fungus-derived particles, preferably plant-derived particles, obtainable or obtained by a process as defined herein are novel over the prior art because they contain a high loading of immobilized protein, at least a portion of which is based on the weight of the plant-derived or fungus-derived particles, preferably plant-derived particles, loaded with the protein obtained in step (e).

[0123] In a third aspect, the present invention relates to protein-loaded plant-derived or fungus-derived particles, preferably plant-derived particles from vegetables, fruits, and edible fungus fruiting bodies, comprising a continuous matrix of a plant-derived or fungus-derived material having proteins distributed throughout the matrix, wherein the protein-loaded plant-derived or fungus-derived particles, preferably plant-derived particles, (i) based on the total weight of the protein-loaded plant-derived or fungus-derived particles, preferably plant-derived particles, 3 to 97% by weight of water and 97 to 3% by weight of dry matter (the amount of water and the amount of dry matter together constitute 100% by weight of the protein-loaded plant-derived or fungus-derived particles, preferably plant-derived particles), and (ii) based on the dry weight of the protein-loaded plant-derived or fungus-derived particles, preferably plant-derived particles, 20 to 65% by weight of the plant-derived or fungus-derived material, and (iii) based on the dry weight of the protein-loaded plant-derived or fungus-derived particles, preferably plant-derived particles, at least 35% by weight of protein, and (iv) based on the dry weight of the protein-loaded plant-derived or fungus-derived particles, preferably plant-derived particles, 0 to 20% by weight of additional components, and consisting of, the amounts of the plant-derived or fungus-derived material, protein, and additional components together constitute 100% by weight of the dry matter of the protein-loaded plant-derived or fungus-derived particles, preferably plant-derived particles, at least a portion of the protein is immobilized in the continuous matrix of the plant-derived or fungus-derived material.

[0124] In a preferred embodiment, the plant-derived or fungus-derived particles, preferably plant-derived particles, loaded with the protein according to the third aspect are rehydratable, i.e., they can absorb and retain an amount of water that is at least 2 times, more preferably at least 2.5 times, even more preferably at least 3 times, such as at least 3.5 times, at least 4 times, at least 4.5 times, at least 5 times, at least 5.5 times, at least 6 times or at least 7 times their own dry weight.

[0125] In another preferred embodiment, the plant-derived or fungus-derived particles, preferably plant-derived particles, loaded with the protein according to the third aspect are rehydratable, i.e., they can absorb and retain an amount of water that is 2 to 25 times, more preferably 2.5 to 22 times, even more preferably at least 3 to 20 times, such as 3.5 to 18 times or at least 4 to 16 times their own dry weight.

[0126] The expression "at least xx weight % based on the dry weight of the plant-derived or fungus-derived particles, preferably plant-derived particles, loaded with the protein" refers to the weight of the protein determined using the Kjeldahl method with a conversion factor of 6.25 divided by the dry weight of the plant-derived or fungus-derived particles, preferably plant-derived particles, loaded with the protein.

[0127] In the protein-loaded plant-derived or fungus-derived particles according to the third aspect, preferably the protein in the plant-derived particles is preferably completely immobilized. In another preferred embodiment, the protein in the plant-derived or fungus-derived particles loaded with the protein according to the third aspect, preferably the plant-derived particles, is completely denatured. In yet another preferred embodiment, the protein in the plant-derived or fungus-derived particles loaded with the protein according to the third aspect, preferably the plant-derived particles, is completely precipitated. In yet another preferred embodiment, the protein in the plant-derived or fungus-derived particles loaded with the protein according to the third aspect, preferably the plant-derived particles, is completely soft-aggregated. In yet another preferred embodiment, the protein in the plant-derived or fungus-derived particles loaded with the protein according to the third aspect, preferably the plant-derived particles, is completely coagulated.

[0128] The plant-derived or fungus-derived material is preferably the material previously disclosed with respect to the plant or fungus-derived particles of the first aspect, preferably the plant-derived particles.

[0129] The protein-loaded plant-derived or fungus-derived particles, preferably the plant-derived particles, can be freshly protein-loaded wet particles, dried particles, or wet particles rehydrated after drying. Thus, the protein-loaded plant-derived or fungus-derived particles, preferably the plant-derived particles, according to the third aspect can have different moisture contents.

[0130] In one embodiment, the protein-loaded plant-derived or fungus-derived particles, preferably the plant-derived particles, according to the third aspect contain 70 to 97% by weight of water, more preferably 80 to 97% by weight of water, based on the total weight of the protein-loaded plant-derived or fungus-derived particles, preferably the plant-derived particles.

[0131] In a preferred embodiment, the protein-loaded plant-derived or fungus-derived particles, preferably plant-derived particles according to the third aspect, contain 3 to 20% by weight of water, preferably 4 to 17% by weight of water, based on the total weight of the protein-loaded plant-derived or fungus-derived particles, preferably plant-derived particles. In another preferred embodiment, the protein-loaded plant-derived or fungus-derived particles, preferably plant-derived particles according to the third aspect, have a water activity (AW) of 0.1 to 0.8, more preferably 0.2 to 0.76, even more preferably 0.30 to 0.60 when measured at 25 °C using a Lab Master-aw neo water activity measuring device (Novasina AG).

[0132] The protein-loaded plant-derived or fungus-derived particles, preferably plant-derived particles according to the third aspect, contain at least 38% by weight, more preferably at least 40% by weight, even more preferably at least 42% by weight, such as at least 44% by weight, at least 46% by weight, at least 48% by weight, at least 50% by weight, at least 52% by weight, at least 54% by weight, at least 56% by weight, at least 58% by weight, at least 60% by weight or at least 62% by weight of protein, based on the dry weight of the protein-loaded plant-derived or fungus-derived particles, preferably plant-derived particles.

[0133] In another preferred embodiment, the protein-loaded plant-derived or fungus-derived particles, preferably plant-derived particles according to the third aspect, contain 35 to 80% by weight, more preferably 38 to 80% by weight, even more preferably 40 to 80% by weight, such as 42 to 80% by weight, 44 to 80% by weight or 46 to 80% by weight of protein, based on the dry weight of the protein-loaded plant-derived or fungus-derived particles, preferably plant-derived particles.

[0134] In yet another preferred embodiment, the protein-loaded plant-derived or fungus-derived particles, preferably plant-derived particles according to the third aspect, contain 35 to 75% by weight, such as 35 to 65% by weight, 35 to 60% by weight, 35 to 56% by weight, 35 to 50% by weight or 35 to 46% by weight of protein, based on the dry weight of the protein-loaded plant-derived or fungus-derived particles, preferably plant-derived particles.

[0135] In a preferred embodiment, the protein is distributed throughout the voids of the matrix of the plant-derived or fungus-derived material.

[0136] In a preferred embodiment, the protein in the protein-loaded plant-derived or fungus-derived particles, preferably plant-derived particles according to the third aspect, includes plant proteins including proteins derived from legumes (seeds), leguminous plants, oilseed grains, algae, and kelp; microbial proteins including proteins derived from yeast, molds, and fungi; animal proteins including whey protein, chicken egg protein, and insect-derived proteins; their hydrolysates; and proteins selected from the group consisting of combinations thereof.

[0137] In a highly preferred embodiment, the protein in the protein-loaded plant-derived or fungus-derived particles, preferably plant-derived particles according to the third aspect, is selected from the group consisting of potato protein, rubisco, lentil-derived protein, pea protein, wheat protein, barley-derived protein, rice-derived protein, soybean protein, broad bean protein, chickpea-derived protein, chicken egg protein, whey protein, canola protein, lupin bean protein, chickpea protein, almond protein, sunflower protein, their hydrolysates, and combinations thereof, and even more preferably, proteins selected from the group consisting of potato protein, broad bean protein, pea protein, lentil-derived protein, whey protein, their hydrolysates, and combinations thereof.

[0138] In certain embodiments, the protein loaded in the plant-derived or fungus-derived particles, preferably the plant-derived particles, according to the third aspect, has a molecular weight of 3 to 650 kDa, such as 3 to 500 kDa, 3 to 450 kDa, 3 to 420 kDa, 3 to 400 kDa, 3 to 380 kDa, 50 to 650 kDa, 100 to 500 kDa, 150 to 450 kDa, 170 to 420 kDa, 180 to 400 kDa or 190 to 380 kDa when measured using size exclusion chromatography (SEC).

[0139] As will be appreciated by those skilled in the art, different proteins can be loaded into the plant-derived or fungus-derived particles, preferably the plant-derived particles. The plant-derived or fungus-derived particles, preferably the plant-derived particles, loaded with the protein according to the third aspect contain different proteins, i.e., proteins specific to the plant-derived or fungus-derived material and one or more proteins from different sources loaded into the plant-derived or fungus-derived particles, preferably the plant-derived particles.

[0140] A protein loaded into a plant-derived or fungus-derived particle, preferably a plant-derived particle, means a protein that was not present in the plant-derived or fungus-derived particle, preferably the plant-derived particle, before loading. Such protein loading is described elsewhere in this specification. In some embodiments of the invention, the proteins from different sources loaded into the plant-derived or fungus-derived particle, preferably the plant-derived particle, are proteins originating from the same species as the plant-derived or fungus-derived particle, preferably the plant-derived particle. In other embodiments of the invention, the proteins from different sources loaded into the plant-derived or fungus-derived particle, preferably the plant-derived particle, are proteins originating from different species. According to a preferred embodiment of the invention, the plant-derived or fungus-derived particle, preferably the plant-derived particle, loaded with protein has a protein content that does not occur naturally, because the total concentration of the protein is higher than that which occurs naturally, and / or because the identity of one or more of the proteins contained in the plant-derived or fungus-derived particle, preferably the plant-derived particle, loaded with protein does not occur naturally for the species of the plant-derived or fungus-derived particle, preferably the plant-derived particle, and / or because the concentration profile of the different proteins contained in the plant-derived or fungus-derived particle, preferably the plant-derived particle, loaded with protein does not occur naturally.

[0141] In a preferred embodiment of the invention, plant-derived or fungus-derived particles loaded with protein, preferably plant-derived particles from vegetables, fruits and fruiting bodies of edible fungi, are provided as such, and the proteins from different sources than the plant-derived or fungus-derived particle, preferably the plant-derived particle, comprise a continuous matrix of plant-derived or fungus-derived material having proteins from different sources than the plant-derived or fungus-derived particle, preferably the plant-derived particle, distributed throughout the matrix, and the plant-derived or fungus-derived particle, preferably the plant-derived particle, loaded with the protein (i) Based on the total weight of the protein-loaded plant-derived or fungus-derived particles, preferably plant-derived particles, 3 to 97% by weight of water and 97 to 3% by weight of dry matter (the amount of water and the amount of dry matter together constitute 100% by weight of the protein-loaded plant-derived or fungus-derived particles, preferably plant-derived particles), and (ii) Based on the dry weight of the protein-loaded plant-derived or fungus-derived particles, preferably plant-derived particles, 20 to 65% by weight of the plant-derived or fungus-derived material, and (iii) Based on the dry weight of the protein-loaded plant-derived or fungus-derived particles, preferably plant-derived particles, at least 35% by weight of the protein from a source different from the plant-derived or fungus-derived particles, preferably plant-derived particles, and (iv) Based on the dry weight of the protein-loaded plant-derived or fungus-derived particles, preferably plant-derived particles, 0 to 20% by weight of additional components, consisting of The plant-derived or fungus-derived material, the plant-derived or fungus-derived particles, preferably the protein from a source different from the plant-derived or fungus-derived particles, and the additional components together constitute 100% by weight of the dry matter of the protein-loaded plant-derived or fungus-derived particles, preferably plant-derived particles. At least a part of the protein from a source different from the plant-derived or fungus-derived particles, preferably plant-derived particles, is immobilized in the continuous matrix of the plant-derived or fungus-derived material.

[0142] In some embodiments, the protein-loaded plant-derived or fungus-derived particles, preferably plant-derived particles, according to the third aspect contain proteins from a source different from the plant-derived or fungus-derived particles, preferably plant-derived particles, loaded with the protein, based on the dry weight of the protein-loaded plant-derived or fungus-derived particles, preferably plant-derived particles, of at least 38% by weight, more preferably at least 40% by weight, even more preferably at least 42% by weight, such as at least 44% by weight, at least 46% by weight, at least 48% by weight, at least 50% by weight, at least 52% by weight, at least 54% by weight, at least 56% by weight, at least 58% by weight, at least 60% by weight or at least 62% by weight.

[0143] In another embodiment, the protein-loaded plant-derived or fungus-derived particles, preferably plant-derived particles, according to the third aspect contain proteins from a source different from the plant-derived or fungus-derived particles, preferably plant-derived particles, loaded with the protein, based on the dry weight of the protein-loaded plant-derived or fungus-derived particles, preferably plant-derived particles, of 35% to 80% by weight, more preferably 38% to 80% by weight, even more preferably 40% to 80% by weight, such as 42% to 80% by weight, 44% to 80% by weight or 46% to 80% by weight.

[0144] In yet another preferred embodiment, the protein-loaded plant-derived or fungus-derived particles, preferably plant-derived particles, according to the third aspect contain proteins from a source different from the plant-derived or fungus-derived particles, preferably plant-derived particles, loaded with the protein, based on the dry weight of the protein-loaded plant-derived or fungus-derived particles, preferably plant-derived particles, of 35% to 75% by weight, such as 35% to 65% by weight, 35% to 60% by weight, 35% to 56% by weight, 35% to 50% by weight or 35% to 46% by weight.

[0145] The plant-derived or fungus-derived particles loaded with the protein according to the third aspect, preferably the plant-derived particles, contain additional components in an amount of 0 to 20% by weight, preferably 0.01 to 10% by weight, such as 0.01 to 5% by weight, 0.01 to 3% by weight, or 0.05 to 1% by weight, based on the dry weight of the plant-derived or fungus-derived particles loaded with the protein, preferably the plant-derived particles.

[0146] In certain preferred embodiments, the additional component is a water-soluble additional component. In certain embodiments, the additional component is water-insoluble or slightly partially soluble in water and is loaded into the plant-derived or fungus-derived particles, preferably the plant-derived particles, in the form of a water-in-oil (micro) emulsion. In certain highly preferred embodiments, the additional component is a food-grade additional component.

[0147] Non-limiting examples of water-soluble additional components are salts, flavoring agents, coloring agents, and preservatives.

[0148] Food The fourth aspect relates to a food comprising or consisting of plant-derived or fungus-derived particles loaded with a protein, preferably plant-derived particles, obtainable as defined previously or via a method as defined previously.

[0149] In certain preferred embodiments, the food contains plant-derived or fungus-derived particles loaded with a protein, preferably plant-derived particles, in an amount of 0.05 to 99.9% by weight, more preferably 0.05 to 95% by weight, such as 0.05 to 90% by weight, 0.05 to 80% by weight, 0.05 to 70% by weight, 0.05 to 60% by weight, 0.05 to 50% by weight, 0.05 to 40% by weight, or 0.05 to 30% by weight, based on the weight of the food.

[0150] In another preferred embodiment, the food product comprises plant-derived or fungus-derived particles loaded with 5 to 99.9% by weight, more preferably 10 to 99.9% by weight, such as 20 to 99.9% by weight, 30 to 99.9% by weight, 40 to 99.9% by weight, 50 to 99.9% by weight, 60 to 99.9% by weight or 70 to 99.9% by weight of protein, preferably plant-derived particles, based on the weight of the food product.

[0151] The food product can be in any form known in the art, provided that the form contains plant-derived or fungus-derived particles loaded with protein, preferably plant-derived particles. Examples include liquids such as dispersions, creams, emulsions and solutions, and solids such as granules, flakes, foams, gels or powders.

[0152] Preferred but non-limiting examples of the food product are selected from the group consisting of meat substitutes or alternatives, fish substitutes or alternatives, breakfast cereals, cereal bars, pastries, snacks and salads. The snack is preferably selected from the group consisting of plant-based meat snacks, vegan meat sticks, pizza bites and vegan protein bites.

[0153] In other embodiments, the food product is a vegetarian or vegan food product, preferably a vegetarian or vegan meat substitute or alternative, fish substitute or alternative, breakfast cereal, cereal bar, pastry, snack or salad. In other embodiments, the food product does not contain animal-derived ingredients.

[0154] In a preferred embodiment, the food product is a burger, more preferably a vegetarian or vegan burger. In one embodiment, the raw burger, i.e., the burger before cooking, grilling and / or frying, consists of the following components based on the total weight of the burger: · 40 to 70% by weight of water; · Based on dry weight, 5 to 25% by weight of protein that can be obtained as previously defined or via a method as previously defined, loaded onto plant-derived or fungus-derived particles, preferably plant-derived particles; · 0.5 to 2% by weight of salt; · 5 to 20% by weight of fat or oil; · 1 to 6% by weight of a techno-functional protein; and · 1 to 15% by weight of additional components.

[0155] Based on the present disclosure, as will be understood by those skilled in the art, plant-derived or fungus-derived particles, preferably plant-derived particles, loaded with protein that can be obtained as previously defined or via a method as previously defined, contain at least some water. However, in the above recipe, for clarity, water and the dry weight of plant-derived or fungus-derived particles loaded with protein, preferably plant-derived particles, are defined separately, but they may be added as one component or may be added.

[0156] Use The fifth aspect of the present invention relates to the use of plant-derived or fungus-derived particles, preferably plant-derived particles, loaded with protein that can be obtained as previously defined or via a method as previously defined, as a food ingredient.

[0157] Dry loading In a sixth aspect, the present invention relates to a method for loading protein onto plant-derived or fungus-derived particles, preferably plant-derived particles from vegetables, fruits, and fruiting bodies of edible fungi, the method comprising (aa) providing plant-derived or fungus-derived particles, preferably plant-derived particles; (bb) providing a soluble protein that can be immobilized in plant-derived or fungus-derived particles, preferably plant-derived particles, in the form of a dry powder; The step of adding the soluble protein in the form of a dry powder provided in step (cc) to plant-derived or fungus-derived particles, preferably plant-derived particles, provided in step (aa), and preferably subsequently mixing them; The step of contacting the plant-derived or fungus-derived particles, preferably plant-derived particles, and the protein in the mixture provided in step (cc) to load the protein onto the plant-derived or fungus-derived particles, preferably plant-derived particles, under conditions where the protein remains soluble; The step of immobilizing at least a part of the protein in the plant-derived or fungus-derived particles, preferably plant-derived particles, is included.

[0158] Therefore, in this sixth aspect, the plant-derived or fungus-derived particles, preferably plant-derived particles, are loaded with the protein in a dry form. No additional water or solvent is applied.

[0159] In a preferred embodiment, the plant-derived or fungus-derived particles, preferably plant-derived particles, loaded with the protein obtained in step (ee) contain at least 2% by weight, more preferably at least 5% by weight, even more preferably at least 10% by weight, for example at least 15% by weight, at least 20% by weight, at least 25% by weight, at least 30% by weight, at least 35% by weight, at least 40% by weight, at least 45% by weight, at least 50% by weight, at least 55% by weight or at least 60% by weight of the protein, based on the dry weight of the plant-derived or fungus-derived particles, preferably plant-derived particles, loaded with the protein obtained in step (ee).

[0160] In another preferred embodiment, the plant-derived or fungus-derived particles, preferably plant-derived particles, loaded with the protein obtained in step (ee) contain 2 to 80% by weight, more preferably 5 to 80% by weight, even more preferably 10 to 80% by weight, such as 15 to 80% by weight, 20 to 80% by weight, 25 to 80% by weight, 30 to 80% by weight or 35 to 80% by weight of the protein, based on the dry weight of the plant-derived or fungus-derived particles, preferably plant-derived particles, loaded with the protein obtained in step (ee).

[0161] In yet another preferred embodiment, the plant-derived or fungus-derived particles, preferably plant-derived particles, loaded with the protein obtained in step (ee) contain 2 to 75% by weight, such as 2 to 65% by weight, 2 to 60% by weight, 2 to 56% by weight, 2 to 50% by weight or 2 to 46% by weight of the protein, based on the dry weight of the plant-derived or fungus-derived particles, preferably plant-derived particles, loaded with the protein obtained in step (e).

[0162] In a seventh aspect, the present invention relates to plant-derived or fungus-derived particles, preferably plant-derived particles, loaded with the protein obtainable or obtained by the process according to the sixth aspect.

[0163] An eighth aspect relates to a food product comprising or consisting of plant-derived or fungus-derived particles, preferably plant-derived particles, loaded with the protein according to the seventh aspect.

[0164] A ninth aspect relates to the use of plant-derived or fungus-derived particles, preferably plant-derived particles, loaded with the protein according to the seventh aspect as a food ingredient.

[0165] Other preferred embodiments defined with respect to the first, second, fourth and fifth aspects of the present invention are equally applicable to the sixth to ninth aspects of the present invention.

Examples

[0166] Examples Method for measuring the water solubility at pH = 7.0 or pH = 3.0 and T = 20 °C The water solubility of the protein at pH = 7.0 (or pH = 3.0) and a temperature of 20 °C was tested using the following protocol: (a) A sample of the protein was added to deionized water in an amount of 5% by weight, based on the total weight of all components, (b) The composition of step (a) was stirred at a temperature of 20 °C for 1 hour, (c) The pH of the composition obtained in step (b) was measured, (d) If the pH measured in step (c) is different from 7.0 (or 3.0), the pH was adjusted to 7.0 (or 3.0) with 1 M HCl or 1 M NaOH, (e) A first subsample of the composition obtained in step (d) was taken, and the total protein content (A) was determined using the Kjeldahl method with a conversion factor of 6.25 (g / L), (f) A second subsample of the composition obtained in step (d) was taken, centrifuged at 4000 G for 10 minutes (Beckman Coulter Avanti J-E centrifuge), the resulting supernatant was isolated, and its total protein content (B) was determined using the Kjeldahl method with a conversion factor of 6.25 (g / L), (g) The solubility of the protein was calculated at pH = 7.0 (or 3.0) and T = 20 °C from the following: % Solubility = (B) / (A)·100%.

[0167] Method for measuring the dry matter and moisture content A sample having a first "wet weight" was subjected to drying in a hot air oven at a temperature of 80 °C for 20 hours, followed by drying at a temperature of 105 °C for 2 hours to determine the dry matter content and moisture content of the sample. Then, the dry matter content and moisture content were determined from the weight loss.

[0168] Method for measuring the protein content The protein content of the sample was determined using the Kjeldahl method with a conversion factor of 6.25.

[0169] Method for measuring the water uptake during rehydration A sample of protein-loaded plant-derived or fungus-derived particles was dried in a hot air oven at a temperature of 90 °C until no further weight loss could be observed. The sample thus dried was rehydrated in an excess amount of water at room temperature. The weight of the protein-loaded plant-derived or fungus-derived particles during rehydration was measured several times during the rehydration process. After about 1 hour, the protein-loaded plant-derived or fungus-derived particles reached a constant weight. The total water uptake during rehydration [grams of water per gram of dry matter] was calculated from the total weight increase of the sieved sample during rehydration and the moisture content of the protein-loaded plant-derived or fungus-derived particles before rehydration.

[0170] Method for measuring the water activity (AW) of a dried sample A sample of protein-loaded plant-derived or fungus-derived particles was dried in a hot air oven at 90 °C for 6 hours. The water activity (AW) of the sample thus dried was measured at 25 °C using a Lab Master-aw neo water activity measuring device (Novasina AG).

[0171] Example 1: Loading of proteins in pretreated used sugar beet pulp Materials Used sugar beet pulp was sampled from the diffusion tower of Cosun Beet Company, Dinteloord, the Netherlands. The sugar beet was washed, sliced into so-called "cossettes", and the sugar beet cossettes were subjected to thermal cell disruption and extraction in a diffusion tower to obtain used sugar beet pulp. In the diffusion tower, other water-soluble components were extracted from the heat-treated sugar beet cossettes together with sucrose by a warm water diffusion process to obtain a so-called "raw juice" or "diffusion juice" at a temperature of 65 - 75 °C and a residence time of 30 - 180 minutes. This heat treatment resulted in the denaturation of the cell membranes and partial destruction of the cell wall structure of the remaining used sugar beet pulp.

[0172] The whey protein isolate (BiPro) was obtained from Davisco Foods Int. When the water solubility of this protein was measured according to the previously defined analytical protocol at pH = 7.0 and T = 20 °C, it was 93.5%.

[0173] The potato protein isolate (Solanic® 200) was obtained from Avebe B.V., the Netherlands. When the water solubility of this protein was measured according to the previously defined analytical protocol at pH = 7.0 and T = 20 °C, it was 97.7%.

[0174] The broad bean protein isolate (HQ isolate) was obtained from Cosun, Dinteloord, the Netherlands. When the water solubility of this protein was measured according to the previously defined analytical protocol at pH = 7.0 and T = 20 °C, it was 93.0%. This broad bean protein isolate has the following composition: Protein (Nx6.25): 88%, Carbohydrates: 4.0%, Ash: 4.6%, Fat: <1%, Moisture: 2.9%.

[0175] The soy isolate (Clarisoy 100) was obtained from ADM, US. The water solubility of this soy protein isolate at pH = 7.0 and T = 20 °C is very low. However, it has a significant water solubility at pH = 3.0 and T = 20 °C when measured according to the previously defined analytical protocol.

[0176] The soy TVP (Response 4410) was obtained from DuPont Nutrition & Biosciences.

[0177] First pretreatment of used sugar beet pulp The used sugar beet pulp defined above was pretreated by washing with tap water, cooking at 100 °C for 5 minutes, and freezing (-18 °C for 24 hours). The frozen used sugar beet pulp was thawed in a microwave oven and pulverized with a meat grinder to obtain used sugar beet pulp having an average size of about 5×5×5 mm. The used sugar beet pulp thus obtained was cooked again in an excess amount of water at 100 °C for 5 minutes. As used herein, the term "excess amount of water" means that the amount of water was more than the amount that could be absorbed by the sugar beet pulp treated in this way. When the obtained wet used sugar beet particles were separated from "free water" by sieving, they had a moisture content of 95% by weight based on the weight of the wet used sugar beet pulp. Because an excess amount of water was used during cooking, the wet used sugar beet particles were saturated (hydrated) with water up to the maximum extent.

[0178] Dry protein loading The wet used sugar beet pulp obtained after the first pretreatment step as described above, having a moisture content of 95% by weight based on the weight of the wet used sugar beet particles, was mixed with dry protein powder in a weight ratio of 6:1 (wet used sugar beet pulp):(dry protein powder) without further processing steps. This is a weight ratio of 1:3.3 on a dry matter basis. The mixing of both components was carried out manually using a spatula until a homogeneous mixture was obtained. This procedure was carried out for (i) whey protein isolate, (ii) pea protein isolate, and (iii) potato protein isolate. For (iv) soy protein isolate, a small amount of concentrated lactic acid (88%) was added dropwise during mixing until a pH of 3.0 was obtained. The obtained homogeneous mixtures (i) to (iv) were stored at a temperature of 5 °C for at least 12 hours to enable the loading of the protein onto the pretreated used sugar beet particles.

[0179] Second pretreatment of used sugar beet pulp The used sugar beet particles in the wet state obtained after the first pretreatment step as described above, having a moisture content of 95% by weight based on the weight of the used sugar beet particles in the wet state, were subjected to a second pretreatment by subjecting them to a pulp press in a Gezang (the Netherlands) press at 9 bar for 5 minutes, obtaining pre-pressed and pre-treated used sugar beet particles having a moisture content of 90% by weight based on the weight of the pre-pressed and pre-treated used sugar beet particles.

[0180] Wet protein loading (i) Whey protein isolate, (ii) pea protein isolate, (iii) potato protein isolate and (iv) soybean protein isolate were used to prepare four concentrated (20% by weight) protein solutions in demineralized water. Subsequently, 200 g of pre-pressed used sugar beet particles obtained after the second pretreatment step as described above, having a moisture content of 90% by weight based on the weight of the pre-pressed used sugar beet pulp particles, were dispersed in 1 kg of each of the concentrated protein solutions and mixed for 1 hour using an overhead stirrer. For the soybean protein isolate, a small amount of concentrated lactic acid (88%) was added dropwise during mixing until a pH of 3.0 was obtained. Thus, the pre-pressed used sugar beet particles obtained after the second pretreatment step as described above were mixed with the concentrated protein solution at a weight ratio of 1:5. This corresponds to a weight ratio of 1:1 (pre-pressed used sugar beet particles in the wet state):(dry protein). On a dry matter basis, this is a weight ratio of 1:10. The resulting dispersion was stored at a temperature of 5 °C for at least 12 hours to enable the loading of the protein onto the pre-treated used sugar beet particles. The sugar beet particles loaded with the protein in the subsequent step were removed from the liquid phase by sieving.

[0181] Immobilization using thermal coagulation The protein loaded onto the pre-treated used sugar beet particles was immobilized in the pre-treated used sugar beet particles using thermal coagulation, i.e., denaturation and precipitation using heat.

[0182] In the first step, the pretreated used sugar beet particles loaded with protein were placed on a sieve and subjected to a washing step using water (from Quooker) having a temperature of 100 °C. This washing step partially removes the protein present on the outer surface of the sugar beet particles. This is an optional step because valuable proteins that can be immobilized on the pretreated used sugar beet particles may be removed from the outer surface. In the second step, the washed and pretreated used sugar beet particles loaded with protein were immersed in an excess amount of boiling water containing 0.2 wt% NaCl. After a residence time of about 1 minute, the sugar beet particles loaded with protein were taken out of the boiling water.

[0183] Immobilization using precipitation at the isoelectric point The protein loaded on the pretreated used sugar beet particles was immobilized in the pretreated used sugar beet particles using precipitation at the isoelectric point.

[0184] In the first step, the pretreated used sugar beet particles loaded with protein were dispersed in an excess amount of water. For the sugar beet particles loaded with soy protein isolate, the pH was adjusted to 4.7 using 1M NaOH. For the sugar beet particles loaded with whey protein isolate, pea protein isolate, and potato protein isolate, the pH was adjusted to 4.4 using concentrated lactic acid (88%).

[0185] Analysis The dry matter content of the sugar beet particles without protein loading (reference) and those after loading / immobilization were measured according to the analysis protocol specified above.

[0186] A reference sample (reference) consisting of used sugar beet particles in a wet state (subjected to the first pretreatment step) having a moisture content of 95% by weight was then placed on a sieve and subjected to a washing step using water (from Quooker) having a temperature of 100 °C, immersed in an excess of boiling water having 0.2% by weight of NaCl for a residence time of about 1 minute, and removed from the boiling water.

[0187] The Kjeldahl protein content of the reference sample (reference) was determined according to the measurement protocol specified above. The Kjeldahl protein content of the sugar beet particles after loading / immobilization was also measured according to the measurement protocol specified above. From these measurements, the Kjeldahl protein content [% by weight] based on the total wet matter and dry matter content of the sugar beet particles loaded with protein can be determined. The results are presented in Table 1.

[0188]

Table 1

[0189] Sugar beet particles loaded with whey protein isolate were dried in a hot air oven at a temperature of 90 °C via a wet method in which the protein is immobilized using heat (thermal coagulation) until no further weight loss could be observed. Similarly, sugar beet particles loaded with potato protein isolate were dried in a hot air oven at a temperature of 90 °C via a wet method in which the protein is immobilized using heat (thermal coagulation) until no further weight loss could be observed. The moisture content and water activity (AW) of these dried products were measured according to the protocol specified above and presented in Table 2. As a reference, the moisture content and water activity (AW) of soy TVP Response 4410 were measured.

[0190] The total water uptake (g of water per g of dry matter) upon rehydration of dry sugar beet particles loaded with whey protein isolate was measured according to the protocol specified above. Similarly, the total water uptake (g of water per g of dry matter) upon rehydration of dry sugar beet particles loaded with potato protein isolate was measured according to the protocol specified above. As a reference, the total water uptake (g of water per g of dry matter) upon rehydration of soy TVP Response 4410 was measured. The results are presented in Table 2. From the moisture content of the "dry" product before water uptake and its own water uptake, the total moisture content after rehydration can be calculated. The results are also presented in Table 2.

[0191]

Table 2

[0192] It was concluded that the protein-loaded plant-derived or fungus-derived particles according to the present invention showed increased water uptake and moisture content upon rehydration compared to the reference soy TVP. The dried plant-derived particles loaded with the protein according to the present invention were able to absorb an amount of water about 10 to 13 times their own dry weight. For example, 1 g of dry sugar beet pulp loaded with whey protein having a moisture content of 16.6 wt% was able to absorb 10.8 g of water, resulting in a water uptake of 12.9 g of water / [g of dry matter] and a total moisture content of 13.1 g of water / [g of dry matter].

[0193] Example 2: Water Uptake and Release with or without Different Pretreatment Steps Fresh sugar beet was sliced into particles having a size of 4×4×6 mm. Different samples (Samples 2 - 8) of fresh sugar beet particles were subjected to different pretreatment steps as shown in Table 3. Sample 1 was not subjected to any pretreatment (reference). Using a Dil, Elcrack HVP 30, bath TB 140 device, it was subjected to pulsed electric field (PEF) treatment (electric field strength: 1 kV / cm, conductivity of the treatment water: 1700 μS / cm, treatment water temperature: 25 °C, belt speed 0.04 m / s). Heat treatment at 70 °C for 120 minutes was carried out with an excess amount of water to mimic the conditions in the diffusion tower. The treatment at 100 °C for 10 minutes was directly connected to the heat treatment at 70 °C for 120 minutes with the same excess amount of water. The samples pretreated with an excess amount of water were sieved to remove "free water". The pretreated sugar beet particles had a moisture content based on the weight of the pretreated sugar beet particles as shown in Table 3.

[0194] Subsequently, the influence of different pretreatments on water uptake and the amount of water released during the subsequent pressing step (Chinkchar press, Gezang, the Netherlands; operating at a pressure of 9 bar for 5 minutes) was investigated. The amount of water (in g) released from 200 grams of (pretreated) used sugar beet particles is shown in Table 3. The percentage of water released from 200 grams of sugar beet particles during pressing, based on the total amount of water present before pressing, is also shown in Table 3.

[0195]

Table 3

[0196] It was concluded that increasing the amount of heat applied during pretreatment had a clear effect on the water uptake of sugar beet particles. Pretreatment with PEF alone or followed by membrane disruption by freezing / thawing also had a clear effect on the water uptake of raw sugar beet particles, even without heat treatment.

[0197] It was further concluded that increasing the amount of heat applied during the pretreatment had a pronounced effect on water release during pressing. Pretreatments involving PEF and / or freeze / thaw membrane disruption also had a pronounced effect on water release during pressing of raw sugar beet particles without heat treatment, i.e., on the untreated sugar beet particles.

[0198] The combination of heat treatment followed by freeze / thaw had the most pronounced effect on water uptake and water release during pressing.

[0199] Example 3: Protein loading of sugar beet particles Whey protein isolate was loaded onto sugar beet particles subjected to the different pretreatment steps shown in Table 3 using wet and dry protein loading followed by immobilization using heat coagulation as described in Example 1. The Kjeldahl protein content [wt%] based on the total dry matter content of the protein-loaded sugar beet particles was determined. The results are presented in Table 4.

[0200]

Table 4

[0201] It can be concluded from Table 4 that the freeze / thaw step significantly increased the protein loading. Furthermore, additional heating at 100 °C in particular increased the protein loading. Pretreatments to which the heating step at 100 °C and the freeze / thaw step were applied resulted in the highest protein loadings for both wet and dry loading. The PEF treatment had a limited effect on protein loading without a heating step and / or without a freeze / thaw step.

[0202] Figure 1a shows an image taken by a confocal scanning laser microscope of sugar beet particles loaded with whey protein using immobilization with wet protein loading followed by heat coagulation. Figure 1b is a copy of Figure 1a with slightly changed colors to emphasize the immobilized whey protein in the matrix of the sugar beet particles. The white areas in Figure 1b correspond to the whey protein, the gray areas correspond to the matrix of the sugar beet particles, and the black areas correspond to the background.

[0203] Figure 2a shows an image taken by a confocal scanning laser microscope of sugar beet particles loaded with whey protein using immobilization with dry protein loading followed by heat coagulation. Figure 2b is a copy of Figure 2a with slightly changed colors to emphasize the immobilized whey protein in the matrix of the sugar beet particles. The white areas in Figure 2b correspond to the whey protein, the gray areas correspond to the matrix of the sugar beet pulp, and the black areas correspond to the background.

[0204] Sugar beet particles pretreated according to Sample 8, which were loaded with an isolate of whey protein immobilized using dry loading and heat coagulation, were dispersed in an excess amount of tap water at a temperature of 20 °C for 6 hours. As a reference example, sugar beet particles pretreated according to Sample 8, which were loaded with an isolate of whey protein using dry loading without a subsequent immobilization step, were dispersed in an excess amount of tap water at a temperature of 20 °C for 6 hours.

[0205] Subsequently, the Kjeldahl protein content [% by weight] based on the total dry matter content of the sugar beet particles loaded with protein was determined for both samples. The sample prepared without immobilization had a Kjeldahl protein content of 17.2% by weight based on the total dry matter content of the sugar beet particles loaded with protein. The sample prepared with the immobilization process had a Kjeldahl protein content of 59.7% by weight based on the total dry matter content of the sugar beet particles loaded with protein, that is, 93% of the protein still remained in the sugar beet particles in water after 6 hours. It was concluded that the immobilization process effectively prevented the leakage of protein from the sugar beet particles.

[0206] Example 4: Loading of Proteins onto Apple, Mushroom, and Carrot Particles Particles of apple, mushroom, and carrot (approx. 5×5×5 mm) were heated in water at 70 °C for 1 minute and subsequently the "free water" was removed by sieving. The thus pretreated plant-derived or fungus-derived particles had a moisture content of approximately 85 - 90% by weight based on the weight of the pretreated plant-derived or fungus-derived particles, as shown in Table 5. The thus pretreated plant-derived or fungus-derived particles were subjected to dry protein loading using whey protein isolate. The whey protein isolate in dry powder form was added to the pretreated plant-derived or fungus-derived particles at a weight ratio of 1:3.3 (pretreated plant-derived or fungus-derived particles: protein) based on the dry weight. The protein was immobilized using heat coagulation.

[0207] The Kjeldahl protein content [% by weight] based on the total dry matter content of the pretreated plant-derived or fungus-derived particles before and after protein loading was determined. The results are presented in Table 5.

[0208]

Table 5

[0209] Figure 3a shows an image taken by a confocal scanning laser microscope of apple, mushroom, and carrot particles loaded with whey protein using immobilization with dry protein loading followed by heat coagulation. Figure 3b is a copy of Figure 3a with the colors slightly changed to emphasize the immobilized whey protein in a plant-derived or fungus-derived matrix. The white areas in Figure 3b correspond to the whey protein, the gray areas correspond to the matrix of apple, mushroom, and carrot particles, and the black areas correspond to the background.

[0210] Example 5: Production of Vegetarian Burger A vegetarian burger was produced using sugar beet particles loaded with protein produced as follows. Used sugar beet particles sampled from the diffusion tower of Cosun Beet Company, Dinteloord, the Netherlands were washed with tap water, cooked with an excess amount of water (5 minutes, 100 °C), and frozen (-18 °C). The frozen used sugar beet particles were thawed in a microwave oven and ground with a meat grinder to obtain used sugar beet particles having an average size of about 5×5×5 mm. The used sugar beet particles thus obtained were cooked again in an excess amount of water at 100 °C for 5 minutes. The used sugar beet particles thus obtained were loaded with a whey protein isolate (BiPro; obtained from Davisco Foods Int.; see Example 1) or a soy protein isolate (HQ isolate; Cosun, Dinteloord; see Example 1) using immobilization with dry loading and heat coagulation to provide sugar beet particles loaded with protein. A general recipe for a vegetarian burger containing sugar beet particles loaded with protein is provided in Table 6.

[0211] [Table 6]

[0212] As a reference, a vegetarian burger based on soybean TVP was manufactured. The recipe for this vegetarian burger is provided in Table 7.

[0213] [[Table 7]]

[0214] The vegetarian burger was manufactured using the following order of steps: (i) Mix all ingredients except coconut oil in a Hobart mixer to obtain a homogeneous dough, (ii) Heat coconut oil in a microwave oven to a temperature of about 50 °C and add it to the dough obtained in step (i), (iii) Using a burger press (Sammic S.L., Φ10 cm), form about 110 g of burgers from the dough obtained in step (ii) at about 20 °C, (iv) Pre-cook the raw burgers obtained in step (iii) in a steam oven at 100 °C for 3 minutes, (v) Freeze the pre-cooked burgers obtained in step (iv) in a freezer (-18 °C), and (vi) Thaw the frozen burgers in a microwave oven and cook them on a frying pan on an induction cooker.

[0215] The vegetarian burger was sensory evaluated by a panel of 4 trained individuals, who scored it for mouthfeel, flavor, and juiciness.

[0216] The vegetarian burger based on soybean TVP had a crumbly mouthfeel and was not juicy. The vegetarian burger based on sugar beet pulp loaded with protein was juicier than the vegetarian burger based on soybean TVP.

[0217] None of the vegetarian burgers had sand contamination. The vegetarian burger with soy protein isolate had a somewhat better flavor than the vegetarian burger with whey protein. Neither of the vegetarian burgers had the typical sugar beet taste.

[0218] Example 6: Immobilization of Protein by Drying Used sugar beet pulp was sampled from the diffusion tower of Cosun Beet Company, Dinteloord, the Netherlands. In the diffusion tower, other water-soluble components together with sucrose were extracted from heat-treated sugar beet cossettes by a warm water diffusion process to obtain a so-called "raw juice" or "diffusion juice" at a temperature of 65 - 75 °C and a residence time of 30 - 180 minutes. The used sugar beet pulp was washed with tap water, cooked at 90 °C for 20 minutes, and frozen (-18 °C). The frozen used sugar beet pulp was thawed in a magnetron and heated in an excessive amount of water at 95 °C for 2 minutes. When the obtained wet used sugar beet particles were separated from "free water" by sieving, they had a moisture content of 95% by weight based on the weight of the wet used sugar beet pulp.

[0219] The obtained wet used sugar beet particles were mixed with a dry protein powder (Isopro 510B soy protein isolate, Sinoglory Health Food, 90% protein by weight) at a weight ratio of 6:1 (wet used sugar beet particles):(dry protein powder). This is a weight ratio of 1:3.3 on a dry matter basis. The mixing of both components was carried out in a rotating drum until a homogeneous mixture was obtained. The obtained homogeneous mixture was stored at a temperature of 5 °C for at least 12 hours to enable the loading of protein into the pretreated used sugar beet particles.

[0220] The pulp particles loaded with protein were subjected to simultaneous protein immobilization and drying in a hot air oven using the following temperature profile: 75 °C (60 minutes) - 80 °C (80 minutes) - 95 °C (180 minutes). The dry matter content, water content, water activity and protein content of the oven-dried pulp particles were determined (see Table 8 below).

[0221] The oven-dried protein-loaded particles were rehydrated in an excess of water at room temperature. After 24 hours, the rehydrated pulp particles were separated from free water by sieving. The dry matter content, water content and protein content of the rehydrated particles were determined (see the table below).

[0222] The protein content of the pulp particles after oven drying and rehydration was 73 wt% based on the total dry matter content, i.e., 100% of the protein still remained in the pulp particles after oven drying and 24 hours of rehydration in an excess of water. It was concluded that oven drying effectively immobilized the protein in the pulp particles.

[0223]

Table 8

[0224] Example 7: Preparation of Particles Loaded with Broad Bean Protein Used sugar beet pulp was sampled from the diffusion tower of Cosun Beet Company, Dinteloord, the Netherlands. In the diffusion tower, other water-soluble components together with sucrose were extracted from heat-treated sugar beet cossettes by a warm water diffusion process to obtain a so-called "raw juice" or "diffusion juice" at a temperature of 65 - 75 °C and a residence time of 30 - 180 minutes. The used sugar beet pulp was washed with tap water, cooked at 90 °C for 20 minutes, and frozen (-18 °C). 1 kg of frozen used sugar beet pulp was divided into two parts of 500 g each, and each was thawed with a magnetron by heating at 600 watts for 2 minutes in a household microwave oven. 75 g of soy protein isolate powder (Cosun Protein product Tendra) was mixed into each part using a tumbler (Tenon engineering LTD, bowl size diameter 26 cm). The tumbler was set to the first set value (about 76 rpm). The product was placed in a refrigerator at 5 °C for 3 hours. Every 30 minutes, the product was manually mixed using a spoon. The obtained protein-loaded sugar beet particles were solidified. One part was placed in the refrigerator until application (non-dried version), and the other part was dried in a hot air oven at 80 °C until 87% of its weight was reduced. This took 2 hours and 45 minutes. The measured water activity was 0.28 (dried version).

[0225] Example 8: Production of low-fat cake To demonstrate the suitability of the protein-loaded particles of the present invention for preparing cakes, a low-fat cake was prepared in which a part of the butter portion of the reference cake was replaced with the protein-loaded particles of Example 7 (non-dried version).

[0226] [Table 9]

[0227] Before use, the protein-loaded beat particles were passed through a meat grinder (Wolfmill: Kenwood Pro 2000 Excel). A cake was prepared using a Hobart N50 mixer according to the following procedure. - Mix butter and sugar in Hobart N50 at position 1 for 1 minute. - Slowly add whole eggs over 2 minutes and mix at position 1. - Lightly mix the butter at stand 2 for 3 minutes. - Add half of the flour and half of the milk and mix at setting 1 for 15 seconds. - Add the remaining flour and milk and mix again at position 1 for 15 seconds. - Lightly mix everything at stand 2 for 1 minute. - Use a spoon to mix in the protein-loaded beat particles. - Fill a 30 cm cake pan greased with 700 g of butter. - Bake the cake at 160 °C for 65 minutes. (Both recipes are baked simultaneously in the same baking group. Miwe Condor electric baking oven.) - Remove the cake from the pan and let it cool for 1 hour. - Wrap it in plastic.

[0228] It was found that the protein-loaded beat particles were well mixed with the butter and hardly visible as several dark parts. The cake was kept in the refrigerator for 5 days and then tasted. The tasting group consisted of 6 people. After evaluating the appearance, scoring was done individually. The scoring was done on a scale of 1 - 5 (very bad - very good). The average score for appearance is shown in Table 10.

[0229]

Table 10

[0230] Example 9: Production of Protein Muesli Bar To demonstrate the suitability for preparing the muesli bar of the protein-loaded particles of the present invention, a muesli bar provided by the protein-loaded particles (dry version) of Example 7 was prepared using a dry protein isolate of the reference bar.

[0231]

Table 11

[0232] The bar was prepared according to the following method: - Thoroughly mix the dry ingredients together (soybean protein isolate / protein-loaded particles; coarse-ground oatmeal flakes; rice crispies) - Prepare the syrup mixture: Mix Frutafit CLR and SP70 with water Add Frutalose L85 Heat until completely dissolved Add BR60 and glycerol Heat until 12.7% of the mixture evaporates. - Immediately add the resulting syrup to the dry mix and mix the combination well until the syrup spreads evenly in the dry ingredients. - Place the mass on baking paper and spread it on a flat plate with a width of 9 cm and a height of 1.5 cm between two metal bars. - Cover the bar with baking paper and let it cool overnight. Then, cut and package the bar.

[0233] The high-protein muesli bar was packaged in a polyethylene bag and kept at ambient temperature for 6 days. The tasting group consisted of 6 people. After evaluating the appearance, scoring was done individually. The scoring was on a scale of 1 - 5 (very bad - very good). The average score regarding the appearance is shown in Table 12.

[0234]

Table 12

Claims

1. A method for loading a protein onto plant-derived or fungus-derived particles from vegetables, fruits, and fruiting bodies of edible fungi, the method comprising: (a) providing the plant-derived or fungus-derived particles; (b) providing a soluble protein that can be immobilized in the plant-derived or fungus-derived particles; (c) adding the soluble protein provided in step (b) to the plant-derived or fungus-derived particles provided in step (a) and preferably mixing them thereafter; (d) contacting the plant-derived or fungus-derived particles with the protein in the mixture provided in step (c) and loading the protein onto the plant-derived or fungus-derived particles under conditions such that the protein remains soluble; (e) immobilizing at least a portion of the protein in the plant-derived or fungus-derived particles, wherein the plant-derived or fungus-derived particles loaded with the protein provided in step (e) contain at least 35% by weight of protein based on the dry weight of the plant-derived or fungus-derived particles loaded with the protein provided in step (e).

2. The step (e) of immobilizing at least a portion of the protein in the plant-derived or fungus-derived particles is carried out by: (I) heating the plant-derived or fungus-derived particles loaded with the protein provided in step (d) to a temperature of at least 85°C, or (II) exposing the plant-derived or fungus-derived particles loaded with the protein provided in step (d) to an acidic solution, or (III) exposing the plant-derived or fungus-derived particles loaded with the protein provided in step (d) to a pH equal to the isoelectric point, or (IV) drying the plant-derived or fungus-derived particles loaded with the protein provided in step (d), or (V) performing a combination of (I) and (II), or (VI) performing a combination of (I) and (III), according to the method of Claim 1.

3. The method according to Claim 1 or 2, further comprising a step (f) of drying and / or freezing the plant-derived or fungus-derived particles loaded with the protein obtained in step (e).

4. The method according to Claim 3, further comprising a step (g) of rehydrating the plant-derived or fungus-derived particles loaded with the protein obtained in step (e) or step (f).

5. The method according to any one of claims 1 to 4, wherein the soluble protein provided in step (b) is added to the plant-derived or fungus-derived particles in step (c) as a dry powder, preferably having a particle size distribution characterized by a Sauter mean diameter (D[3,2]) of 10 to 100 μm as determined by a laser diffraction particle size analyzer.

6. The method according to any one of claims 1 to 4, wherein the soluble protein provided in step (b) is added to the plant-derived or fungus-derived particles in step (c) as an aqueous solution.

7. The method according to any one of claims 1 to 6, wherein the soluble protein and the plant-derived or fungus-derived particles are applied in step (c) at a weight ratio of 1:0.05 to 1:1 on a dry matter basis, preferably 1:0.1 to 1:0.5 on a dry matter basis.

8. The method according to any one of claims 1 to 7, wherein the plant-derived or fungus-derived particles provided in step (a) are particles from vegetables, fruits, and fruiting bodies of edible fungi selected from the group consisting of sugar beet, carrot, chicory, potato, apple, pineapple, citrus fruits, cranberry, grape, mushroom, and combinations thereof.

9. The plant-derived or fungus-derived particles provided in step (a) are obtained from raw plant-derived or fungus-derived materials that have been subjected to one or more of the following steps: ・ Heating, preferably cooking; ・ Pulsed electric field treatment; ・ Cutting, slicing, or grinding; ・ Pressing; ・ Drying; and ・ Freezing and thawing. The method according to any one of claims 1 to 8.

10. The plant-derived or fungus-derived particles provided in step (a) are (i) heated at a temperature of at least 80°C, (ii) preferably frozen and thawed, (iii) optionally subjected to pressing, drying, sieving, or combinations thereof, more preferably only pressing and / or sieving for water removal. The method according to any one of claims 1 to 9, and are used sugar beet pulp particles.

11. The soluble protein that can be immobilized in the plant-derived or fungus-derived particles provided in step (b) is a plant protein including proteins derived from legumes (seeds), leguminous plants, oilseed grains, algae, and kelp; a microorganism-derived protein including proteins derived from yeast, mold, and fungi; an animal protein including whey protein, chicken egg protein, and insect-derived protein; their hydrolysates; and combinations thereof, preferably selected from the group consisting of potato protein, rubisco, lentil-derived protein, pea protein, wheat protein, barley-derived protein, rice-derived protein, soybean protein, broad bean protein, chickpea-derived protein, chicken egg protein, whey protein, canola protein, lupin bean protein, chickpea protein, almond protein, sunflower protein, their hydrolysates, and combinations thereof, more preferably selected from the group consisting of potato protein, broad bean protein, pea protein, lentil-derived protein, whey protein, their hydrolysates, and combinations thereof, the method according to any one of claims 1 to 10.

12. Plant-derived or fungus-derived particles loaded with protein, obtainable via a process according to any one of claims 1 to 11.

13. Plant-derived or fungus-derived particles from protein-loaded vegetables, fruits, and edible fungal fruiting bodies, comprising a continuous matrix of plant-derived or fungus-derived material with protein distributed throughout the matrix, wherein the protein-loaded plant-derived or fungus-derived particles (i) based on the total weight of the protein-loaded plant-derived or fungus-derived particles, 3 to 97% by weight of water and 97 to 3% by weight of dry matter (the amount of water and the amount of dry matter together constituting 100% by weight of the protein-loaded plant-derived or fungus-derived particles), and (ii) based on the dry weight of the protein-loaded plant-derived or fungus-derived particles, 20 to 65% by weight of the plant-derived or fungus-derived material, and (iii) based on the dry weight of the protein-loaded plant-derived or fungus-derived particles, at least 35% by weight of protein, and (iv) It consists of 0 to 20% by weight of a further component based on the dry weight of the plant-derived or fungus-derived particles loaded with the protein. The amounts of the plant-derived or fungus-derived material, the protein, and the further component together constitute 100% by weight of the dry matter of the plant-derived or fungus-derived particles loaded with the protein. A protein-loaded plant-derived or fungus-derived particle in which at least a part of the protein is immobilized in the continuous matrix of the plant-derived or fungus-derived material.

14. The protein is a plant protein containing proteins derived from legumes (seeds), leguminous plants, oilseed grains, algae, kelp; a microorganism-derived protein containing proteins derived from yeast, mold, and fungi; an animal protein containing whey protein, chicken egg protein, and insect-derived protein; their hydrolysates; and a combination thereof, preferably selected from the group consisting of potato protein, rubisco, lentil-derived protein, pea protein, wheat protein, barley-derived protein, rice-derived protein, soybean protein, broad bean protein, chickpea-derived protein, chicken egg protein, whey protein, canola protein, lupin bean protein, chickpea protein, almond protein, sunflower protein, their hydrolysates, and a combination thereof, more preferably selected from the group consisting of potato protein, broad bean protein, pea protein, lentil-derived protein, whey protein, their hydrolysates, and a combination thereof, the protein-loaded plant-derived or fungus-derived particle according to claim 13.

15. The protein-loaded plant-derived or fungus-derived particle according to claim 13 or 14, wherein the protein is distributed across the voids of the matrix of the plant-derived or fungus-derived material.

16. The plant-derived or fungus-derived material is from vegetables, fruits, and fruiting bodies of edible fungi selected from the group consisting of sugar beet, carrot, chicory, potato, apple, pineapple, citrus fruits, cranberry, grape, mushroom, and combinations thereof, the protein-loaded plant-derived or fungus-derived particle according to any one of claims 13 to 15.

17. A food comprising or consisting of plant-derived or fungus-derived particles loaded with the protein according to any one of claims 12 to 16, wherein the food is preferably selected from the group consisting of meat substitutes, fish substitutes, breakfast cereals, cereal bars, pastries, snacks and salads.

18. Use of the plant-derived or fungus-derived particles loaded with the protein according to any one of claims 12 to 16 as a food ingredient.

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