Field

The present invention relates to a texturized vegetable protein particle. Further, the present invention relates to a process for manufacturing a texturized vegetable protein particle, and to the use of the texturized vegetable protein particle. Further the present invention relates to a method analogue product and a method for the preparation of a meat analogue product.

Background

Proteins are an essential element in animal and human nutrition. Meat, in the form of animal flesh and fish, are the most common sources of high protein food. The many disadvantages associated with the use of animal-derived protein for human consumption, ranging from acceptability of raising animals for consumption to the fact that such meat production is inefficient in terms of feed input to food output and carbon footprint, makes the ongoing search for improved meat alternatives one of the most active developments in present day society.

Historically, meat alternatives achieve a certain protein content using vegetable sources such as soy (e.g. tofu, tempeh) or gluten/wheat (e.g. seitan). Today, modern techniques are used to make meat alternatives with more meat-like texture, flavor and appearance. Soy and gluten are favorable sources for such meat alternatives because they are widely available, affordable, relatively high in protein and well processable. In combination with extrusion, the formation of fibrous character can be generated in soy or soy/gluten-based compositions, which is an aspect that is key for approaching the fibrous structure of animal meat. These properties as found when using soy or wheat or soy/gluten mixtures are generally not found in other plant-based proteins. Simultaneously however, there also is a strong driver to avoid soy and gluten for reasons of allergenicity and/or consumer trust. Manufacturers of meat alternatives turn to other proteins, for example like those derived from legumes, e.g. pea, fava bean, lupin, chickpea. However, use of these alternative protein sources is accompanied with new problems. The protein mixtures are often not as easily processible as the traditional soy or gluten or their combinations, and in many cases also lead to texturized food proteins that do not mimic the nutrition, texture, appearance, and/or the taste of animal-derived meat products. As a result, consumers typically consider such meat alternatives unappealing and unpalatable.

The majority of meat alternatives are made from solid plant-based materials produced by extrusion. Commonly, two types of extrusion are distinguished, high moisture (or wet) extrusion and dry extrusion.

The problem of poor texture, notably lack of fibrous texture, may be addressed by using high-moisture extrusion (HME), leading to products (called HMEs) with a highly fibrous nature, such as for example described in WO 2015/020873 for proteins that may be animal or plant-derived, or in WO 2019/143859 for compositions comprising two or more plant-based proteins that are not soy and do not contain gluten. Generally, in high-moisture extrusion, a water level of from 40 to 70% on the total extruder feed is used. In the process a blend of solids is fed into the extruder, water is added, and the material is kneaded into a homogeneous mass. A melt is formed at elevated temperature and pressure, which is fed into a cooling die where controlled cooling under flow leads to fibrous nature of the material. This material can be described as anisotropic, i.e. the properties and microstructure of the material are not the same in all three dimensions. Such anisotropic, fibrous material is clearly discernable while eating the product, and is generally linked to meat-like textures that are often also fibrous and anisotropic.

However, high-moisture extrusion is not a straightforward technology, but is costly and leads to materials that are high in water content, thus microbiologically vulnerable, which constrains the producer to frozen storage and complexifies further processing into consumer end products.

Dry extrusion is a cost effective, simple and robust technology, proven for decades and leads to material that can be used in meat alternatives with a relatively homogeneous texture, having a more isotropic character. Dry extrusion is used to make so-called texturized vegetable protein (TVP), which is material that forms the base of the largest categories of meat alternatives such as burgers, (“meat”) balls, minced beef, minced chicken, minced pork, minced veal, (stir-fry) pieces, sausages and the like. Advantageously, dry extrusion leads to products with a low moisture content that are less susceptible to microbiological contamination due to the low water activity.

A drawback of conventional TVPs is that the homogenous or isotropic texture limits the application of TVPs. For example, conventional TVPs are less applicable for preparing products with more flaky or fibrous texture such as a chicken-like, fish-like texture, as can be found in for instance chicken-style or beef style stir-fry pieces, breaded products such as nuggets alternatives or schnitzel alternatives, or fish finger alternatives, or meat/fish alternatives in cold applications like tuna-style salad or chicken-style salad, or applications where a ‘pulled-meat’ character is wished, pulled chicken or pulled pork.

Therefore, there is a need in the art for texturized vegetable protein material I particles that has the benefits of dry extrusion and at the same time provides an inhomogeneous, anisotropic, texture.

Detailed description

1 . The present invention relates to a texturized vegetable protein particle having a length within the range of 3 to 8 cm, preferably within the range of 3.5 to 6 cm, or having a width within the range of 1 to 3 cm, wherein the protein comprises a legume protein.

The present inventors found that TVPs can be produced that combine the processing benefits of TVPs (such as a low risk of contamination) with the texture benefits of HMEs.

The term ‘texturized vegetable protein’ or ‘textured vegetable protein’ as used herein refers to a composition obtainable by (dry) extrusion cooking of proteins. Dry extrusion cooking means wherein an amount of water is used in the process of less than 30 wt. % (of the composition being extruded).

The term ‘particle’ as used in the present content means a single piece of the texturized vegetable protein. The word ‘chunk’ can alternatively be used. Preferably, the present texturized vegetable protein particle has a length within the range of 3.1 to 7.9 cm, 3.2 to 7.8 cm, 3.3 to 7.7 cm, 3.4 to 7.6 cm, 3.5 to 7.5 cm. Such as 3.6 to 7 cm, or 4 to 6 cm. Preferably, the present texturized vegetable protein particle has a length within the range of 3.1 to 4.5 cm, 3.2 to 4.4 cm or 3.3 to 4.3 cm.

In a preferred embodiment, the present texturized vegetable protein particle has a width within the range of 1 to 3 cm. Preferably within the range of 1.1 to 2.9 cm, 1 .2 to 2.8 cm, 1 .3 to 2.7 cm, 1 .4 to 2.6 cm or 1 .5 to 2.5 cm.

The present inventors found that the present TVP particles can be used as such, without a need for further processing such as chopping. This is advantageous in industries for production of meat analogues, such as chicken fingers, nuggets etc. It is surprising that a legume based TVP particle can be provided that have a size a structure which is good enough to consume the TVP particle as a whole, i.e. without further processing (milling) steps.

Therefore, in a preferred embodiment, the present texturized vegetable protein particle has a length within the range of 3.1 to 7.9 cm and a width of 1 .1 to 2.9 cm, or a length within the range of 3.2 to 4.4 cm and a width within the range of 1 .5 to 2.5 cm.

Preferably the present TVP particles have a ratio length I width of 1 .5 to 6, such as 2 to 5.

The legume protein can be any protein derived from a legume, preferably the legume protein is from lupin, pea (yellow pea, green pea), bean (such as soybean, fava (faba) bean or broad bean, kidney bean, green bean, haricot bean, pinto bean, mung bean, adzuki bean), chickpea, lupin, lentil, and peanut, and the like. Fava bean and faba bean and broad bean can be used interchangeably. Advantageously, the legume-derived protein is non-allergenic.

In a preferred embodiment, the legume protein is selected from pea protein, chickpea protein and faba bean protein. More preferably from pea protein and faba bean protein.

As the present invention is about providing allergen free TVPs, in a preferred embodiment, the present texturized vegetable protein does not comprise soy protein and/or gluten (protein) and/or peanut protein. Gluten can be wheat gluten, or gliadin.

Pea protein, obtained from yellow pea Pisum sativum, is a mixture of various proteins (see for instance Lam et al. Food Rev. International 2018 34(2) p126-147), consisting of globulins (70- 80%) and albumins (10-20%). The globulin fraction consists of several proteins: Legumin (11 S, 300-400 kDa), vicilin (7S, 150-170 kDa) and convicilin (210 kDa as trimer), the water-soluble albumin fraction consists of proteins with molecular masses up to 80 kDa comprising enzymes protease- and amylase inhibitors and lectins. Furthermore, a small fraction consists of among others prolamins and glutenins. The method of extraction highly influences the composition of the protein concentrate or isolate, as well as its physico-chemical properties and its flavour. The general process for producing a pea protein isolate is known in the art and described for instance by Frederikson et al. (J. Agric. Food Chem. 2001 , 49, p1208-1212 Production Process for High-Quality Pea-Protein Isolate with Low Content of Oligosaccharides and Phytate). Several industrial methods to obtain isolates are described such as WO2020221978 (Gelling leguminous protein), US2020229462 (Pea protein composition having improved nutritional quality), EP3071045 B1 (Method for extracting pea proteins), US2020281224 (Product analogs or components of such analogs and processes for making same). Next to pea protein isolates and pea protein concentrates, usually obtained by a wet extraction process, also dry fractionation can be used to obtain a protein-enriched pea flour. Dry fractionation may occur by processes such as wind sifting or electrostatic separation. Preferably the pea protein is a pea protein concentrate, pea protein isolate or a combination of both.

The term isolate means that on a dry basis, 85 wt. % of the total weight of the isolate is protein. This is calculated using the Dumas method according to AOAC Official Method 991.20 Nitrogen (Total) in Milk, using a conversion factor of 6.25 was used to determine the amount of protein (% (w/w)). Typically, the non-protein content of the protein isolate includes non-protein compounds such as, fibre and/or other carbohydrates, minerals, anti-nutritional substances. Preferably the present protein isolate has a protein content of at least 90 wt.% (calculated as Dumas N x 6.25) on a dry weight basis, preferably at least 91 , 92, 93, 94, 95, 96, 97, 98, or at least 99 wt.% on a dry weight basis (calculated as Dumas N x 6.25).

In a preferred embodiment, the present texturized vegetable protein particle further comprises rapeseed protein or rapeseed protein isolate. The predominant storage proteins found in rapeseed protein are cruciferins and napins. Cruciferins are globulins and are the major storage protein in the seed. A cruciferin is composed of 6 subunits and has a total molecular weight of approximately 300 kDa. Napins are albumins and are low molecular weight storage proteins with a molecular weight of approximately 14 kDa. Napins are more easily solubilized and are primarily proposed for use in applications where solubility is key. Preferably the present rapeseed protein is a rapeseed protein isolate.

In a preferred embodiment, the present rapeseed protein (isolate) comprises 40 to 65 wt. % cruciferins and 35 to 60 wt. % napins (of the rapeseed protein). Preferably, the present rapeseed protein comprises 40 to 55 wt. % cruciferins and 45 to 60 wt. % napins.

In a preferred embodiment, the present rapeseed protein (isolate) comprises 50 to 95 wt. % or 50 to 90 wt. % cruciferins and 5 to 40 wt. % or 20 to 40 wt. % napins, preferably 60 to 80 wt. % cruciferins and 20 to 40 wt. % napins. Preferably, the present rapeseed protein comprises 65 to 75 wt. % cruciferins and 25 to 35 wt. % napins.

In a preferred embodiment, the present rapeseed protein (isolate) comprises 0 to 15 wt. % or 0 to 10 wt. % cruciferins and 85 to 100 wt. % or 90 to 100 wt. % napins. Preferably, the present rapeseed protein comprises 1 to 5 wt. % cruciferins and 95 to 100 wt. % napins.

Preferably, the amounts of cruciferins and napins calculated based on the total amount of canola protein (isolate). Or alternatively, the amounts of cruciferins and napins are calculated based on the sum of cruciferins and napins present in the canole protein (isolate). Preferably, the amounts of cruciferins and napins are determined by size exclusion chromatography (SEC).

Preferably, the amounts of cruciferins and napins are determined by size exclusion chromatography (SEC) using the following test: samples of protein isolate are dissolved in a 500 mM NaCI saline solution and analyzed by High Performance SEC using the same solution as the mobile phase, followed by detection using measuring UV absorbance at 280 nm, wherein the relative contribution of cruciferin and napin (wt. %) was calculated as the ratio of the peak area of each protein with respect to the sum of both peak areas.

Preferably, the present rapeseed protein (isolate) comprises 40 to 65 wt. % 12S and 35 to 60 wt. % 2S. Preferably, the present rapeseed protein comprises 40 to 55 wt. % 12S and 45 to 60 wt. % 2S.

In a preferred embodiment, the present rapeseed protein (isolate) comprises 45 to 80 wt. % 12S and 20 to 40 wt. % 2S, preferably 50 to 75 wt. 12S and 25 to 35 wt. 2S, preferably 60 to 80 wt. % 12S and 20 to 40 wt. % 2S. Preferably, the present rapeseed protein (isolate) comprises 65 to 75 wt. % 12S and 25 to 35 wt. % 2S.

In a preferred embodiment, the present rapeseed protein (isolate) comprises 0 to 30 wt. % 12S and 70 to 100 wt. % 2S. Preferably, the present rapeseed protein (isolate) comprises 0 to 20 wt. % 12S and 80 to 100 wt. % 2S, preferably 1 to 15 wt. % 12S and 85 to 99 wt. % 2S, such as 1 to 10 wt. 12S and 90 to 99 wt. % 2S. Or the present rapeseed protein (isolate) comprises 0 to 10 wt. % cruciferins and 90 to 100 wt. % napins. Preferably, the present rapeseed protein isolate) comprises 1 to 5 wt. % cruciferins and 95 to 100 wt. % napins.

In a preferred embodiment, the present rapeseed protein (isolate) comprises 10 to 50 wt. % 12S and 50 to 90 wt. % 2S. Preferably, the present rapeseed protein (isolate) comprises 20 to 40 wt. % 12S and 60 to 80 wt. % 2S, preferably 25 to 35 wt. % 12S and 65 to 75 wt. % 2S.

Preferably, the amounts of 12S and 2S is determined by sedimentation velocity analytical ultracentrifugation (SV-AUC) analysis. Preferably, the amounts of 12S and 2S is determined by sedimentation velocity analytical ultracentrifugation (SV-AUC) analysis using the following test: samples of protein isolate are dissolved in a 3.0% (or 500 mM) NaCI saline solution and amounts determined using interference optics.

In a preferred embodiment, the present rapeseed protein (isolate) comprises a conductivity in a 2 wt.% aqueous solution of less than 9000 pS/cm over a pH range of 2 to 12. More preferably the conductivity of the native rapeseed protein isolate in a 2 wt. % aqueous solution is less than 4000 pS/cm over a pH range of 2.5 to 11.5. For comparison the conductivity of a 5 g/l NaCI aqueous solution is around 9400 pS/cm. Preferably conductivity is measured with a conductivity meter, for example Hach senslON+ EC71.

In a preferred embodiment, the present rapeseed protein (isolate) comprises a solubility of at least 88 % when measured over a pH range from 3 to 10 at a temperature of 23 +1-2 °C. Preferably a solubility of at least 90, 91 , 92, 93, 94, 95, 96, 97, 98 or at least 99% over a pH range from 3 to 10 at a temperature of 23 +1-2 °C. This is also known as the soluble solids index (SSI).

Preferably, solubility is calculated by:

Protein solubility (%) = (concentration of protein in supernatant (in g/l) I concentration of protein in total dispersion (in g/l)) x 100. Preferably, the solubility is measured using the following test:

-sufficient protein to supply 0.8 g of protein is weighed into a beaker;

-a small amount of demineralized water is added to the powder and the mixture is stirred until a smooth paste is formed;

-additional demineralized water is then added to make a total weight of 40 g (yielding a 2 % w/w protein dispersion);

-the dispersion is slowly stirred for at least 30 min using a magnetic stirrer;

-afterwards the pH is determined and adjusted to the desired level (2, 3, 4, etc.) with NaOH or HCI; -the pH of the dispersion is measured and corrected periodically during 60 minutes stirring;

-after 60 minutes of stirring, an aliquot of the protein dispersion is reserved for protein concentration determination (Dumas analysis; Dumas N x 6.25), another portion of the sample is centrifuged at 20,000 G for 2 min;

-the supernatant and pellet are separated after centrifugation;

-the protein concentration of the supernatant is also determined by Dumas analysis (Dumas N x 6.25);

- and protein solubility is calculated by:

Protein solubility (%) = (concentration of protein in supernatant (in g/l) I concentration of protein in total dispersion (in g/l)) x 100.

For use in human food consumption the removal of phytates, phenolics (or polyphenolics) and glucosinolates prevents unattractive flavour and coloration and prevents decreased nutritional value of the protein isolate. At the same time this removal enhances the protein content of the protein isolate.

In a preferred embodiment, the present rapeseed protein (isolate) has a phytate level less than 5 wt.%, preferably less than, 4, 3, 2, 1 , 0.5, 0.4, 0.3, 0.2. 0.1 or less than 0.01 wt. %. Alternatively, the present rapeseed protein (isolate) has a phytate level of 0.01 to 4, 0.05 to 3, 0.1 to 1 wt.%. Preferably the phytate level is measured using method QD495, based on Ellis et al, Analytical Biochemistry Vol. 77:536-539 (1977).

In a preferred embodiment, the present rapeseed protein (isolate) has a phenolic content of less than 1 wt.% on dry matter expressed as sinapic acid equivalents. Preferably less than 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1 , 0.05 or less than 0.01 wt.% on dry matter expressed as sinapic acid equivalents.

In a preferred embodiment, the present rapeseed protein (isolate) comprises < 10 ppm gliadin. Preferably the rapeseed protein (isolate) comprises less than 5 ppm gliadin and most preferably no gliadin can be detected. Preferably, gliadin content is determined using sandwich ELISA from R-Biopharm (cat no R7001 , lot 14434) used according to the manufacturer’s instructions to determine the gliadin ppm in extracts.

In a preferred embodiment, the present texturized vegetable protein particle has a protein content between 55 and 80 wt. %, preferably between 60 and 70 wt. % (on dry weight of the particle). Preferably the protein content is 61 , 62, 63, 64, 65, 66, 67, 68 or 69 wt. % (on dry weight of the particle).

In a preferred embodiment, the present texturized vegetable protein particle has a dry matter of greater than 80 wt. %, preferably of greater than 90 wt. %. Preferably a dry matter of greater than 91 , 92, 93, 94, 95, 96, 97, 98 or 99 wt. %.

In a preferred embodiment, the present texturized vegetable protein particle has a density within the range of 80 to 170 g/l, preferably within the range of 110 to 150 g/l. Preferably the density is within the range of 115 to 135 g/l or 120 to 130 g/l. Preferably the density is determined by using the density test as shown in example 1 .

In a preferred embodiment, the present texturized vegetable protein particle has a water holding capacity of greater than 3 gram water per gram texturized vegetable protein particle. Preferably the water holding capacity is from 3 to 8 gram, preferably 3.1 to 7 gram, preferably from 3.5 to 6 gram water per gram texturized vegetable protein particle. Preferably, the water holding capacity is determined by using the water holding capacity test as shown in example 1 .

In a preferred embodiment, the present texturized vegetable protein particle further comprises a dietary fiber, preferably in an amount of 5 to 20 wt. % (on dry weight of the particle). Preferably the amount of dietary fiber is from 6 to 19, 7 to 18, 8 to 17, 9 to 16 or 10 to 15 wt. % (on dry weight of the particle).

Dietary fibers may be added to improve the texture, and/or firmness, and/or consistency, and/or the nutritional value and/or as a filler. Examples of dietary fiber are pea fiber, fava bean fiber, lupin fiber, oil seed fiber (such as sunflower seed fiber or cotton seed fiber), fruit fiber (such as apple fiber), cereal fiber (such as oat fiber, maize fiber, rice fiber), bamboo fiber, potato fiber, inulin, or combinations thereof. Fibers are commonly present in plant-based foods and cannot (completely) be broken down by the human digestive enzymes, are either water-soluble or water-insoluble fibers. They may consist of (mixtures of) cellulose, hemicellulose, pectins and other non-starch polysaccharides or plant cell-wall biopolymers. Fiber fractions are materials that also can comprise protein, starch, lignin and/or ash. Preferably the dietary fiber is pea fiber or fava bean fiber.

In a preferred embodiment, the present texturized vegetable protein particle further comprises a flavor. A flavor is a compound that provides a flavourto the texturized vegetable protein particle. Examples of flavours are meat flavors, like chicken flavor. Alternatively, the flavour is a fish flavor.

According to another aspect, the present invention relates to a process for manufacturing a texturized vegetable protein particle as defined herein, comprising the steps of:

(a) mixing a legume protein (composition) and from 5-30% (w/w) water in an extruder;

(b) heating the mixture obtained in step (a) to a temperature of from 100-180°C;

(c) extruding the mixture obtained in step (b) through an extrusion die; and/or (d) cutting the extruded mixture into particles resulting in a length within the range of 3 to 8 cm, preferably within the range of 3.5 to 6 cm, to provide the texturized vegetable protein particle.

Preferably, the present process further comprises drying the texturized vegetable protein particle to a dry matter of greater than 80 wt. %, preferably of greater than 90 wt. %.

Preferably, the legume protein is selected from pea protein, chickpea protein and faba bean protein.

Preferably, the legume protein is as defined above. More preferably, step (a) comprises mixing a legume protein (composition) and from 5-30% (w/w) water in an extruder, without the presence of soy protein, gluten and/or peanut protein.

In an embodiment the protein may be in the form of a flour, a concentrated flour (obtained for example by wind sifting), a concentrate (>50% protein) or an isolate (>80% protein), or a combination thereof. Preferably the amount of protein is between 55 and 80 wt. %, preferably between 60 and 70 wt. % (on dry weight of the legume protein composition).

Preferably, step (d) of cutting the extruded mixture into particles results in a particle having a length within the range of 3.1 to 7.9 cm, 3.2 to 7.8 cm, 3.3 to 7.7 cm, 3.4 to 7.6 cm, 3.5 to 7.5 cm, such as 3.6 to 7 cm, or 4 to 6 cm. Preferably, resulting in a length within the range of 3.1 to 4.5 cm, 3.2 to 4.4 cm or 3.3 to 4.3 cm.

Preferably, step (d) of cutting the extruded mixture into particles results in a particle having a has a width within the range of 1 to 3 cm. Preferably within the range of 1 .1 to 2.9 cm, 1 .2 to 2.8 cm, 1 .3 to 2.7 cm, 1 .4 to 2.6 cm or 1 .5 to 2.5 cm.

Preferably, step (d) of cutting the extruded mixture into particles results in a particle having a length within the range of 3.1 to 7.9 cm and a width of 1.1 to 2.9 cm, or a length within the range of 3.2 to 4.4 cm and a width within the range of 1 .5 to 2.5 cm.

Dietary fibers may be added to the mixture (in step a) to improve the texture, and/or firmness, and/or consistency, and/or the nutritional value and/or as a filler. Examples of plant-based fiber are pea fiber fava bean fiber, lupin fiber, oil seed fiber (such as sunflower seed fiber or cotton seed fiber), fruit fiber (such as apple fiber), cereal fiber (such as oat fiber, maize fiber, rice fiber), bamboo fiber, potato fiber, inulin, or combinations thereof. Fibers are commonly present in plantbased foods and cannot (completely) be broken down by the human digestive enzymes, are either water-soluble or water-insoluble fibers. They may consist of (mixtures of) cellulose, hemicellulose, pectins and other non-starch polysaccharides or plant cell-wall biopolymers. Fiber fractions are materials that also can comprise protein, starch, lignin and/or ash.

Other components that can be added in present step (a) or to the material are starch, either native or modified (chemically or physically), from any source such as tapioca, corn, potato, pea or other legume, wheat, rice or other cereal. Next to normal salt (sodium chloride), other salts can be added, like potassium salts, calcium salts. This can be soluble or insoluble salts and minerals. Insoluble salts can also act as inert fillers and ways to change the colour of the end product. Soluble salts such as sodium bicarbonate can also be added to increase the pH during processing or in the end product. Alternatively, acids can be used to reduce the pH in during the process or in the end product. Any type of acid can be used, such as malic acid, citric acid, lactic acid, phosphoric acid, tartaric acid. Such soluble salts and pH modulators can be added as a solid to the powder premix or dissolved in the water stream. The modulation of the pH during extrusion can be used advantageously as a means to modify the texture, flavour and appearance of the TVP, it can impact on density, and on mechanical properties (dry and after hydration) such as resilience or elasticity.

Preferably, the pH of the present mix of protein powder and water is within the range of pH 6 to 10, preferably pH 6 to 7, preferably pH 7 to 9, preferably pH 7 to 8, preferably pH 6 to 8.

In an embodiment, the mix of protein powder and water is brought into the extruder, either separately (through different feeders) or (partially) combined. In the extruder, the screws knead the resulting mixture into a paste. The temperature at which this takes place may be from 40-200°C, 90-190°C, or from 110-180°C, or from 120-170°C or from 130-140°C or at 140±30°C. Preferably, the temperature at the beginning of the extruder is 100 to 120°C, and 160 to 180°C at the end of the extruder.

In an embodiment, the process takes place at elevated pressure such as from 5-80 bar, or from 20-60 bar or at 40±30 bar. The skilled person understands that the choice of pressure is related to the scale of the extrusion process. Preferably, the process is carried out in a continuous mode.

During the above processing, a melt is formed in the extruder, which, in an embodiment, is released through holes in the end plate at the end of the extruder, where immediate expansion occurs. This expansion may be caused by water flashing off, causing next to expansion also an immediate temperature reduction, converting the melt into a ‘glassier’ type of material. The number and size of the holes in the end plate may vary, and can be any shape such as circular, elongated, ellipsoidal, or even more complex. In an embodiment, the stream leaving the extruder may be cut into pieces using methods known to the skilled person. Such methods may for example be a rotating knife directly at the exit of the extruder. This leads to particles of various sizes and shapes depending on the cutting mode. The latter refers to the rotation speed of the knife (which may be from 50-5000 rpm, orfrom 100-3000 rpm or at 1000±500 rpm), the distance between extruder head and rotating knife and the dimensions of the holes. This may lead to particles where 95% of the particles has a size of from 3 to cm 8 cm.

According to another aspect, the present invention relates to the use of a texturized vegetable protein particle as defined herein in a meat (or fish) analogue, preferably a meat analogue selected from the group of chicken, nuggets, schnitzels and fish. Preferably use in a chicken-like, fish-like texture, as can be found in for instance chicken-style or beef style stir-fry pieces, breaded products such as nuggets alternatives or schnitzel alternatives or fish finger alternatives, or meat/fish alternatives in cold applications like tuna-style salad or chicken-style salad, or applications where a ‘pulled-meat’ character is wished, pulled chicken or pulled pork. According to another aspect, the present invention relates to a meat analogue product comprising the present texturized vegetable protein particles, water, a binder and oil and/or fat, preferably wherein the texturized vegetable protein particles are positioned in longitudinal direction of the meat analogue product. Preferably wherein the length of the texturized vegetable protein particles is in substantially the same direction as the length of the meat analogue product.

The present inventors found that the texturized vegetable protein particles can be shaped into a meat analogue product having a whole cut meat appearance, such as a beef steak, tenderloin, chicken breast or filet mignon. Without wishing to be bound by any theory, it is expected that the length and texture of the present texturized vegetable protein particles enables the formation of a whole cut meat analogue product.

In a preferred embodiment, the present meat analogue product has an elongated shape, preferably the present meat analogue has a cylindrical, rectangular, tapered or irregular shape, preferably wherein the length is considerably longer than the width. More preferably an elongated shape wherein the texturized vegetable protein particles are positioned in longitudinal direction of the meat analogue product.

Preferably, one or more texturized vegetable protein particles are placed (substantially) parallel to each other, preferably (substantially) parallel in view of the longitudinal direction (or length) of the texturized vegetable particle.

In a preferred embodiment, the present meat analogue product is raw and requires cooking before consumption. The advantage of a raw product is that the meat analogue product provides a real raw meat like appearance.

In a preferred embodiment, the amount of texturized vegetable protein particles is within the range of 10 to 40 wt. % of the meat analogue product, preferably within the range of 12 to 25 wt. % of the meat analogue product.

In a preferred embodiment, the present meat analogue product has a visual marbled aspect, preferably due to the presence of fat, emulsified fat or fat-free emulsions such a fat mimetics. The term "marbled" means that the product has markings and colorings suggestive of marble, and which are marked herein by the intermixture of textured plant protein/binder composition, and a vegetable fat composition. This visual aspect reflects the fat and fat distribution as found in corresponding real meat products.

In one embodiment of the present invention, the fat is a vegetable fat selected from coconut fat, palm fat, shea butter, or a combination thereof.

In a preferred embodiment of the present invention, the fat has an average particle size of 0.5-4.5mm. In a more preferred embodiment of the present invention, the particles have an average particle size of 1-3mm. The presence of those particles significantly help to improve the forming of the visual aspect of the marbled meat analogue product.

In one embodiment of the present invention, the meat analogue product comprises between 1 to 12 wt.% fat (weight percent of the total meat analogue product), preferably between 1 to 7 wt. %, preferably between 1 to 5 wt. %, preferably between 2 to 5 wt.% (weight percent of the total meat analogue product).

The vegetable oil can be an algal oil, a fungal oil, corn oil, olive oil, soy oil, peanut oil, walnut oil, almond oil, sesame oil, cottonseed oil, rapeseed oil, canola oil, safflower oil, sunflower oil, flax seed oil, palm oil, palm kernel oil, coconut oil, babassu oil, wheat germ oil, borage oil, black currant oil, sea-buckhorn oil, macadamia oil, saw palmetto oil, conjugated linoleic oil, arachidonic acid enriched oil, docosahexaenoic acid (DHA) enriched oil, eicosapentaenoic acid (EPA) enriched oil, palm stearic acid, sea-buckhorn berry oil, macadamia oil, saw palmetto oil, or rice bran oil; or margarine or other hydrogenated fats. In some embodiments, for example, the oil is algal oil. In a preferred embodiment, the present plant oil is sunflower oil and/or the present plant fat is coconut fat. In one embodiment of the present invention, the meat analogue product comprises between 1 to 15 wt. % oil (weight percent of the total composition), preferably between 2 to 10 wt. % such as

3 to 8 wt. % oil.

In a preferred embodiment, the present meat analogue product comprises a flavouring. The term "flavourings" according to this invention means ingredients selected from the group consisting of salt, sugar, vinegar, yeast extract, vegetable powder, bacterial extract, vegetable extract, reaction flavour, hydrolysed plant protein, acid, garnishes, herbs, spices or combinations of these. The meat analogue product according to the invention comprise 0 to 10 wt. % flavourings, preferably 0.5 to 10 wt. %, preferably 0.5 to 7 wt. %, preferably 0.5 to 5%, preferably 1 to 10 wt. %, preferably 1 to 5 wt. % (by weight of the meat analogue product). Vegetable powder means at least one ingredient of onion powder, garlic powder, tomato powder, celery root powder or a combination thereof. Garnishes, herbs, spices or a combination thereof are selected from the group comprising pieces of parsley, celery, fenugreek, lovage, rosemary, marjoram, dill, tarragon, coriander, ginger, lemongrass, curcuma, chili, ginger, paprika, mustard, garlic, onion, turmeric, tomato, coconut milk, cheese, oregano, thyme, basil, chillies, paprika, pimento, jalapeno pepper, white pepper powder and black pepper or combinations of these.

The term "binder" or "binding agent" as used herein relates to a substance for holding together particles and/or fibres in a cohesive mass. It is an edible substance that in the final product is used to trap components of the foodstuff with a matrix for the purpose of forming a cohesive product and/or for thickening the product. Binding agents of the invention may contribute to a smoother product texture, add body to a product, help retain moisture and/or assist in maintaining cohesive product shape; for example by aiding particles to agglomerate. Preferably the present meat analogue product comprises an amount of binder from 0.5 to 5% wt. %, preferably from 1 to

4 wt. %, such as from 2 to 3 wt. % (of the meat analogue product).

The binder may be or may comprise methyl cellulose. The methyl cellulose might be present in an amount of 0.5 to 2 wt. % such as from 1 to 2 wt. % (of the meat analogue product).

In a preferred embodiment the binder is gellan gum and the present gellan gum is high acyl gellan gum. Preferably the high acyl gellan gum is a polymer comprising various monosaccharides linked together to form a linear primary structure and the gum gels at temperatures of greater than 60 degrees centigrade. In some high acyl gellan gums, the gel temperature may be approximately 70 degrees centigrade or greater. In some high acyl gellan gums, the gel temperature may be approximately between 70 degrees centigrade and 80 degrees centigrade The properties of the high acyl gellan gum polymer may vary depending at least in part on its source, how it was processed, and/or the number and type of acyl groups present on the polymer.

Preferably, the amount of gellan gum in the present meat analogue product is within the range of 0.1 to 4 wt. %, preferably 0.2 to 3 wt. %, more preferably 0.5 to 1.0 wt. % (of the meat analogue product).

Preferably, the present gellan gum, or the present high acyl gellan gum, has a single gel setting temperature that is within the range of 70°C to 90°C. The advantage of high acyl gellan gum is that it forms soft and flexible gels, beneficial in providing a good texture of a vegetarian emulsified meat product, without introducing off flavors to the product. Preferably, the present gellan gum, or the present high acyl gellan gum, has more than 40% acetyl and more than 45% glyceryl residual substitutions per repeating unit.

In a preferred embodiment, the present meat analogue product further comprises a nutrient, preferably wherein the nutrient comprise both vitamins and minerals, preferably vitamins chosen from the group consisting of B2, B3, B6 and B12, preferably minerals chosen from the group consisting of iron, selenium and zinc. The term “nutrient” as used herein relates to a substances that provide nutritional value to the present meat analogue product, such as vitamins, minerals, trace elements and antioxidants for example. The advantage of adding these nutrients is that the present meat analogue product more closely resembles the nutritional value of a real meat hamburger, without introducing off flavors to the meat analogue product.

In another aspect, the present invention relates to a method for the preparation of a meat analogue product comprising the present texturized vegetable protein particles, water, a binder and oil and/or fat, the process comprises the steps of:

(i) Optionally hydrating the texturized vegetable protein particles with water;

(ii) providing a layer of hydrated texturized vegetable protein particles;

(iii) adding the binder with the oil and/or fat to the (layer of) hydrated texturized vegetable protein particles;

(iv) forming the hydrated texturized vegetable protein particles with the binder into the meat analogue product;

(v) optionally vacuumizing and/or freezing the formed meat analogue product.

Preferably, step (ii) comprising providing a layer of hydrated texturized vegetable protein particles wherein the hydrated texturized vegetable protein particles are adjacent to each other and/or wherein the texturized vegetable protein particles are positioned in a longitudinal direction.

Preferably, step (ii) comprising providing a layer of hydrated texturized vegetable protein particles wherein the layer has an elongated shape, preferably the layer has a rectangular, tapered or irregular shape, preferably wherein the length is considerably longer than the width. More preferably an elongated shape wherein the texturized vegetable protein particles are positioned in longitudinal direction of the layer.

Preferably wherein step (iv) comprises forming the hydrated texturized vegetable protein particles with the binder into the meat analogue product having a shape that is elongated, cylindrical, tapered or irregular shape, preferably wherein the length is of the meat analogue product is longer than the width. This can be achieved for example by rolling up the layer of hydrated texturized vegetable protein particles with the binder. Hence, preferably, present step (iv) comprises rolling or folding the hydrated texturized vegetable protein particles with the binder into the meat analogue product.

Present step (v) of vacuumizing and/or freezing the formed meat analogue product is advantageous to give the meat analogue product its shape. Vacuum can be used to provide a further improved coherent meat analogue product. Freezing can be used to conserve the shape of the meat analogue product.

The inventor is further illustrated in the non-limiting examples below. In the examples, reference is made to the following figures.

Figure 1 shows the present TVP particles produced under conditions given in example 1 , showing particles of 5-8 cm and 1-3 cm in diameter.

Figure 2 shows the present TVP particles produced under conditions given in example 1 , showing particles of 3-5 cm and 1-3 cm in diameter.

Figure 3 shows the present TVP particles after hydration and cutting through perpendicular to the flow direction and in half in the flow direction. In the latter the air cells elongated in the flow direction can be seen.

Figure 4 shows the present TVP particles after frying.

Example 1

TVPs were made by adding to an extruder (Buhler PolyTwin 62 twin-screw extruder with a Priotherm preconditioner) a dry blend of 60 wt. % pea protein concentrate (Vestkorn Pea Protein F55X, Vestkorn, Norway, with 55 wt. % protein) and 40 wt. % pea protein isolate (HYPP-B80, Jianyuan, China with 80 wt.% protein). Water was added with an amount of 15 wt.% of the dry blend. The throughput was 500 kg/hr with a screw speed of 400 rpm using 4 inserts having a diameter of 8 mm. The temperature in the extruder was ranging from 100 to 180°C (first barrel to end plate). A cutter with 3 knives was used at a cutting speed adapted to the required length of the products, for a product with particle with a length of 3-5 cm, a rotation rate of 390 rpm was used, for particles of 5-8 cm, a slower cutting speed was used. After cutting, the TVPs were dried to approximately a moisture level of 8-9 wt. %. Thereafter, the density, particle size and water holding capacity was analysed as follows: Density:

A 1000 mL cylinder was tarred, then filled with TVPs just beyond 1000 mL mark, the cylinder was tapped 10 times at the table, then checked that it was exactly filled to 1000 mL and was weighed again. The measured weight was used as density in g/L.

Estimate of particle size:

Produced material was distributed over paper, and photographs were taken after which particle size was determined visually.

Water holding capacity:

The water holding capacity was determined by hydrating material: 30 gram TVP, on 120 gram cold water (1 :5), and allowed to hydrate for >1 hr. This 150 gram hydrated material was drained over a sieve and the drained water retrieved was weighed. From this, the water holding capacity in gram of water per gram of TVP was calculated, using the residual water levels by:

[weight drained water + weight water in 30g TVP] /[dry matter weight of 30g TVP]

Water holding time:

TVP was added to an excess of water, a panel of three persons assessed at least every minute the particles by sgueezing between the fingers, to feel if there are still hard pieces (domains in the TVP particle) in it. Once no hard pieces are felt, the particle was considered fully hydrated.

Firmness:

The particles were analysed on a TA.XT.PIusC texture analyser (50 kg load cell), in a heavy duty stand, using a Warner Bratzler knife blade, cutting the particle perpendicular to the direction of flow. The cutting force is recorded, in grams.

Results

Table 1

The products are represented in figure 1 and 2. Figure 1 shows the larger particles of 5-8 cm long and 1-3 cm diameter, figure 2 shows the particles of 3-4 cm and 1-3 cm diameter. Thereafter, the TVPs were hydrated in water, and cut through perpendicular to the flow direction and in half in the flow direction and pictures are made to study the structure. Figure 3 shows a picture of hydrated TPVs. Figure 3 clearly shows that the air cells are shaped / elongated in the length of the TVP (in the flow direction) and that the structure is inhomogeneous.

Example 2

5 The TVPs made in example 1 were tested in a meat strip application using the ingredients listed in table 1 .

Table 2

First, the TVP particles were hydrated with lukewarm water (water 1) for 15 minutes. The w dry rub was prepared by blending the ingredients shown in table 2 and mixing it with water 2.

Table 3

After hydration of the TVP particles the rub was applied on the TVP particles. Thereafter the flavoured TVP particles were fried in a dry pan. Figure 4 shows that after frying the TVP particles

15 remain in one piece. Upon testing it showed that the texture of the TVP particles remained after hydration and cooking. The texture had meat like fibrous structure. Example 3

The TVPs made in example 1 were tested in a whole cut filet mignon meat analogue using the ingredients listed in table 4.

Table 4

First, the flavors and colors where added to lukewarm water (water 1) and subsequently the TVP particles were hydrated in the water for 45 minutes. The methylcellulose and gellan gum were blended and thereafter mixed with the sunflower oil. Subsequently, (cold tap) water 2 was added together with the frozen coconut flakes. Thereafter, a layer of the hydrated and flavored TVP w particles was formed on a piece of plastic on a workbench, adjacent to each other, all in the same direction. Then a layer of the binder / oil / water / coconut mix was added to TVP particles followed by another layer of TVP particles. Subsequently the plastic was rolled to provide a cylindrical elongated shaped meat analogue product. Thereafter the meat analogue product was vacuumized in a vacuum bag and frozen until a hard product was obtained. This is shown in Figure 5.

15 To test the meat analogue product, the frozen product was cut into slices of 3 cm thick and directly cooked in a pan as shown in Figure 6 and analysed on appearance. Figure 7 shows the resulting meat analogue product, having a cohesive meaty like structure, marbled with fat.