WO2023161507A1

WO2023161507A1 - Method of brightening a texturized protein product

Info

Publication number: WO2023161507A1
Application number: PCT/EP2023/054931
Authority: WO
Inventors: Jan Engmann; Charlotte GANCEL; Fanny LAZZARO; Martin Michel; Stefan Palzer; Patrick Pibarot; Yu-Jie Wang; Hannes WENZEL; Yogesh HARSHE
Original assignee: Société des Produits Nestlé S.A.
Priority date: 2022-02-28
Filing date: 2023-02-28
Publication date: 2023-08-31

Abstract

The present invention relates to a method of brightening a meat analogue product, in particular a texturized protein product. More specifically, the invention relates to a method of making a texturized protein product which has been brightened by the incorporation of immiscible particles, wherein the immiscible particles are insoluble mineral and/or emulsified lipid.

Description

Method of brightening a texturized protein product

Background of the invention

The shift towards vegetarian and vegan diets has been driven by many factors including health and wellbeing, impact on the planet, and animal welfare. As a consequence, the demand for vegetarian and vegan analogues of animal food products is growing. An analogue can be defined as a product mimicking the sensorial and nutritional characteristics of a specific type of animal food product.

High and low moisture extrusion are the most widely applied technologies for manufacturing meat, poultry or seafood analogues, enabling functionalization and texturization of plant proteins. While key challenges remain with obtaining the desired taste and texture experience, efforts are also made to obtain adequate appearance, especially in terms of color. Plant-based analogues produced by extrusion technology generally carry shades of color from light beige or yellow to dark brown, distinguishing themselves significantly from the animalbased product they intend to replace.

Among the whole color spectra, white is of key importance when it comes to the development of chicken, pork and fish analogues. Today, the solutions enabling whitening of plant-based analogues relies mostly on addition of titanium dioxide (TiO2). However, TiO2 has regulatory and safety concerns due to the nano status of this ingredient. Various other ingredients provided for their coloring function, particularly in extruded applications, fail to provide the desired level of whiteness.

Embodiments of the invention

The present invention relates to a method of making a protein product which has been brightened by the incorporation of immiscible particles, wherein the immiscible particles are insoluble mineral, emulsified lipid and/or insoluble fiber.

In a first aspect, the invention relates to a method of making a protein product, said method comprising the steps of forming a protein product from a mixture of raw materials, wherein the raw materials include immiscible particles, preferably insoluble mineral, emulsified lipid and/or insoluble fiber.

In a second aspect, the invention relates to a method of making a texturized protein product, said method comprising the steps of a. Forming a mixture of raw materials; wherein the raw materials include immiscible particles, preferably insoluble mineral, emulsified lipid and/or insoluble fiber; b. Forming a texturized protein product from the mixture by a texturizing process; In a third aspect, the invention relates to a method of making a texturized protein product, said method comprising the steps of a. Forming a mixture of raw materials, wherein the raw materials include protein and immiscible particles, preferably insoluble mineral, emulsified lipid and/or insoluble fiber, more preferably insoluble mineral and/or emulsified lipid; b. Forming a texturized protein product from the mixture by a texturizing process, for example an extrusion process;

In a fourth aspect, the invention relates to a method of making a texturized protein product, said method comprising the steps of a. Forming a mixture of raw materials, wherein the raw materials include protein, immiscible particles, preferably insoluble mineral, emulsified lipid and/or insoluble fiber, more preferably insoluble mineral and/or emulsified lipid, and water; b. Forming a texturized protein product from the mixture by a texturizing process, for example an extrusion process; characterized in that between 0.1 wt% to 15 wt% immiscible particles on a wet basis is dispersed in the mixture.

In a fifth aspect, the invention relates to a method of making a texturized protein product, said method comprising the steps of a. Forming a mixture of raw materials, wherein the raw materials include protein, immiscible particles, preferably insoluble mineral, emulsified lipid and/or insoluble fiber, more preferably insoluble mineral and/or emulsified lipid, and water; b. Forming a texturized protein product from the mixture by a texturizing process, for example an extrusion process, a trigger process, a freeze-thaw process, a 3-D printing process, a shear cell technology process, a microwave tube heating process, an injection molding process, a freeze structuring or freeze alignment process, an electro-spinning process, or a high-pressure treatment process; characterized in that between 0.1 wt% to 15 wt% immiscible particles on a wet basis is dispersed in the mixture.

These aspects of the invention can be further described according to one or more of the following embodiments.

In some embodiments, the texturizing process is an extrusion process comprising the steps of a. Forming a mixture by metering raw materials into an extruder, wherein the raw materials include protein, immiscible particles, preferably insoluble mineral, emulsified lipid and/or insoluble fiber, more preferably insoluble mineral and/or emulsified lipid, and water; b. Passing the mixture through an extruder whilst heating the mixture to above the denaturation temperature of the protein, wherein the protein content of the mixture is in the range from 30 wt% to 95 wt% on a dry basis and the solids content of the mixture is in the range from 25 wt% to 90 wt% on a wet basis; c. Cooling the mixture to a temperature of less than 100°C to form a texturized protein product;

Additionally or alternatively, the raw materials are mixed before they are metered through the extruder. In some embodiments, the immiscible particles are metered separately into the extruder with a fraction of some of the other raw materials.

In some embodiments, the immiscible particles have a refractive index between 1.55 and 2.5. In some embodiments, the immiscible particles have a refractive index which is greater than the protein mixture, for example a refractive index between 1.55 and 2.5.

In some embodiments, the immiscible particles are selected from insoluble mineral, emulsified lipid, and insoluble fiber.

In some embodiments, the immiscible particles are insoluble mineral and/or emulsified lipid.

In some embodiments, the immiscible particle is an insoluble mineral having a D50 particle size of less than 50 microns, preferably between 0.10 microns to 10 microns.

In some embodiments, the insoluble mineral is a calcium salt, preferably calcium citrate.

In some embodiments, the immiscible particles are emulsified lipids. Typically, the emulsified lipids have been made by emulsifying lipids with protein, for example soy protein.

In some embodiments, the emulsified lipids have a D [3;2] particle size of less than 1 micron, preferably between 0.10 microns to 1 microns, preferably 0.3 microns to 0.7 microns.

In some embodiments, the insoluble fiber is microcrystalline cellulose.

In some embodiments, the microcrystalline cellulose has a D50 particle size of less than 50 microns, preferably between 0.5 microns to 5 microns.

In some embodiments, the emulsified lipid has a solid fat content at 20°C greater than 5 %, more preferably greater than 30%, more preferably greater than 50%, more preferably greater than 80%. The solid fat content can be determined using the method described in IUPAC 2.150(b) Solid content determination in fats by NMR.

In some embodiments, the emulsified lipid is shea stearin. In some embodiments, the emulsified lipid is emulsified canola oil.

In some embodiments, the protein ingredient has a protein content of greater than 40 wt% protein on a dry matter basis, preferably between 60 wt% to 95 wt%.

In some embodiments, the protein is plant protein, for example soy protein, gluten protein, or pea protein, or a combination thereof. Preferably, the plant protein is soy protein or gluten protein.

In some embodiments, the texturized protein product has an increased brightness (AL) value of greater than 2, compared to a product which does not comprise immiscible particles, preferably insoluble mineral, emulsified lipid and/or insoluble fiber.

In some embodiments, the texturized protein product is a plant-based meat analogue, for example plant-based chicken, plant-based fish, plant-based ham, plant-based burger, or plant-based seafood, preferably a plant-based chicken analogue or a plant-based fish analogue.

The invention further relates to a texturized protein product made by a method according to the invention.

The invention further relates to a method of making a protein product which has been brightened by the incorporation of insoluble mineral, emulsified lipid and/or insoluble fiber, more preferably insoluble mineral and/or emulsified lipid.

In one embodiment, said method comprises the steps of forming a protein product from a mixture of raw materials, wherein the raw materials include insoluble mineral, emulsified lipid and/or insoluble fiber, more preferably insoluble mineral and/or emulsified lipid.

In one embodiment, said method comprises the steps of a. Forming a mixture of raw materials; wherein the raw materials include insoluble mineral, emulsified lipid and/or insoluble fiber, more preferably insoluble mineral and/or emulsified lipid; and b. Forming a texturized protein product from the mixture by a texturizing process;

In a sixth aspect, the invention relates to a texturized protein product, said product comprising protein, immiscible particles, wherein said immiscible particles are preferably insoluble mineral, emulsified lipid and/or insoluble fiber, more preferably insoluble mineral and/or emulsified lipid, and water, wherein the protein content of the product is in the range from 30 wt% to 95 wt% on a dry basis and the solids content of the product is in the range from 25 wt% to 90 wt% on a wet basis, and wherein the immiscible particles are present at a concentration of between 0.1 wt% to 15 wt% on a wet basis in the product. In some embodiments, the immiscible particles are present at a concentration of between 1.5 wt% to 5 wt% on a wet basis in the product.

In some embodiments, the immiscible particles are selected from insoluble mineral, emulsified lipid, and insoluble fiber, more preferably insoluble mineral and/or emulsified lipid.

In some embodiments, the emulsified lipid is shea stearin.

In some embodiments, the protein is plant protein, for example soy protein, gluten protein, or pea protein, or a combination thereof.

In some embodiments, the texturized protein product has an increased brightness (AL) value of greater than 2, compared to a product which does not comprise immiscible particles. In some embodiments, the texturized protein product is a plant-based meat analogue, for example plant-based chicken, plant-based ham, plant-based burger, or plant-based seafood, preferably a plant-based chicken analogue.

In a seventh aspect, the invention relates to the use of immiscible particle to improve the brightness of a texturized protein product, wherein the immiscible particle is selected from emulsified shea stearin, and calcium citrate.

In some embodiments, the texturized protein product is made by a method comprising the steps of a. Forming a mixture of raw materials, wherein the raw materials include protein, immiscible particles, and water; b. Forming a texturized protein product from the mixture by a texturizing process, for example an extrusion process, a trigger process, a freeze-thaw process, a 3-D printing process, a shear cell technology process, a microwave tube heating process, an injection molding process, a freeze structuring or freeze alignment process, an electro-spinning process, or a high-pressure treatment process; characterized in that between 0.1 wt% to 15 wt% immiscible particles on a wet basis is dispersed in the mixture.

In some embodiments, the texturizing process is an extrusion process comprising the steps of a. Forming a mixture by metering raw materials into an extruder, wherein the raw materials include protein, immiscible particles and water; b. Passing the mixture through an extruder whilst heating the mixture to above the denaturation temperature of the protein, wherein the protein content of the mixture is in the range from 30 wt% to 95 wt% on a dry basis and the solids content of the mixture is in the range from 25 wt% to 90 wt% on a wet basis; c. Cooling the mixture to a temperature of less than 100°C to form a texturized protein product;

In some embodiments, the immiscible particles have a refractive index between 1.55 and 2.5. In some embodiments, the immiscible particles have a refractive index which is greater than the protein mixture, for example a refractive index between 1.55 and 2.5. In some embodiments, the immiscible particles are selected from insoluble mineral, emulsified lipid, and insoluble fiber, more preferably insoluble mineral and/or emulsified lipid.

In some embodiments, the emulsified lipid is shea stearin.

In some embodiments, the texturized protein product has an increased brightness (AL) value of greater than 2, compared to a product which does not comprise immiscible particles.

In some embodiments, the texturized protein product is a plant-based meat analogue, for example plant-based chicken, plant-based ham, plant-based burger, or plant-based seafood, preferably a plant-based chicken analogue.

Detailed description of the invention

Extrusion process

The extrusion process may use a twin screw extruder. A cooling die may be attached at the end of the extruder. Typically, the dry raw materials are fed into the first barrel of the extruder. The liquid raw materials may be injected in the second extruder barrel. The flowrate of the liquid feed was may be adapted to reach a solid content of between 38.1 - 47.8 g/lOOg in the product. The temperature profile of the extruder from the first zone to the sixth zone may be between 20-25°C for the first two barrels, about 80°C for the third barrel and about 230 °C for the last three barrels. The cooling die can be tempered to about 60°C using recirculating water.

Emulsified lipids

Where the immiscible particles are emulsified lipids, these are typically introduced to the mixture as a liquid feed, typically as an emulsion. The emulsion can be prepared by mixing a suspension of soy protein concentrate together with lipids, for example at a protein to lipids ratio of about 0.1. For coconut oil and shea stearin, the protein suspension can be prepared by dispersing the soy protein concentrate in warm deionized water, for example at 70°C under high shear. The temperature of the mix should be maintained above 60°C. The previously melted lipid is typically incorporated into the warm protein suspension, in an amount to reach the desired emulsified lipids concentration in the product. A coarse emulsion is then typically formed by stirring. The emulsification is typically further enhanced by passing this coarse emulsion in a two-stage homogenizer.

Where the immiscible particle is emulsified canola oil, the same process can be followed with the exception that all steps were performed at room temperature, for example between 20 - 25°C. Where polysorbate 80 and hydrolyzed lecithin were used as emulsifiers, the suspension of soy protein concentrate can be replaced with a suspension of polysorbate 80 or hydrolyzed lecithin. The content of polysorbate 80 and hydrolyzed lecithin were adjusted to reach an emulsifier to lipids ratio of about 0.1 and about 0.3 in the liquid feed, respectively.

The immiscible particles can thus be droplets of canola oil emulsified with different types of emulsifiers, for example soy protein, polysorbate 80, or hydrolyzed lecithin, preferably soy protein.

The immiscible particles can be insoluble minerals. Preferably, the insoluble minerals are calcium sulphate anhydrous, calcium citrate tribasic, calcium citrate, or zinc oxide. Preferably, the insoluble minerals have a refractive index greater than 1.63. Preferably, the insoluble minerals have a D90 below 35 pm.

In one embodiment, the immiscible particles are calcium citrate tribasic. Where the insoluble mineral is calcium citrate tribasic, preferably the D50 particle size is below 150 pm.

The size distributions of the emulsified lipid droplets in the emulsions can be determined by laser light diffraction, for example as described herein. The dynamic, quantitative shape analyses can be conducted on powdered insoluble minerals and fibers to determine their particle size properties, for example by using a CamsizerXT, for example as described herein. During extrusion, immiscible particles can be mixed at the inlet, but also could be injected at different points during extrusion.

Color analyses of the products

Products can be analyzed by color measurement after 4 h storage at room temperature in vacuum sealed plastic bags. Color measurements of the products can be performed using a Digi-Eye® system integrated with a digital camera D-90 Nikon. L* (0 = black, 100 = white), a* (+value = red, -value = green), and b* (+value = yellow, -value = blue) values according to the CIELAb system definition can be determined. The total color difference (AE) and the brightness difference (AL) can be calculated as described herein.

Definitions

As used herein, the singular forms "a," "an" and "the" include plural referents unless the context clearly dictates otherwise.

The words "comprise," "comprises" and "comprising" are to be interpreted inclusively rather than exclusively. Likewise, the terms "include," "including" and "or" should all be construed to be inclusive, unless such a construction is clearly prohibited from the context.

An immiscible particle is defined as a particle which is not capable of combining with another substance to form a homogeneous mixture. In other words, it is a small discrete quantity of matter that has an interface with the surrounding environment.

The compositions disclosed herein may lack any element that is not specifically disclosed. Thus, a disclosure of an embodiment using the term "comprising" includes a disclosure of embodiments "consisting essentially of and "consisting of the components identified. Similarly, the methods disclosed herein may lack any step that is not specifically disclosed herein. Thus, a disclosure of an embodiment using the term "comprising" includes a disclosure of embodiments "consisting essentially of" and "consisting of" the steps identified.

The term "and/or" used in the context of "X and/or Y" should be interpreted as "X," or "Y," or "X and Y." Where used herein, the terms "example" and "such as," particularly when followed by a listing of terms, are merely exemplary and illustrative and should not be deemed to be exclusive or comprehensive. Any embodiment disclosed herein can be combined with any other embodiment disclosed herein unless explicitly stated otherwise.

As used herein, "about" and "approximately" are understood to refer to numbers in a range of numerals, for example the range of -10% to +10% of the referenced number, preferably within -5% to +5% of the referenced number, more preferably within -1% to +1% of the referenced number, most preferably within -0.1 % to +0.1 % of the referenced number. As used herein, a product "substantially devoid" of an ingredient means that none of that ingredient is added as such to the product, and that any of the ingredient present originates from minor traces or impurities present in other ingredients.

A vegan product is defined as being devoid of animal products, for example devoid of dairy products and meat products. A product of the invention has the look, taste, and texture which is close to real its meat counterpart.

The invention will now be illustrated by way of examples, which should in no way be thought to limit the scope of the invention as herein described.

EXAMPLES

Example 1

Composition, manufacture, and analyses of the protein-based products

Protein-based products were made using soy concentrate, wheat gluten as protein sources. Sodium chloride, water and in some cases vinegar was also added. Immiscible particles, such as emulsified lipid droplets, insoluble minerals and insoluble fibers were introduced as part of the recipes. The recipe mixes were further textured through an extrusion process and analyzed to determine their color properties.

Tables 1, 2 and 3 show the composition and details regarding the manufacturing process of soy and wheat gluten protein-based products, that can be used as chicken meat replacer:

Table 1 shows a reference product without immiscible particles and without vinegar (reference A) and products with emulsified lipid droplets as immiscible particles.

Table 2 shows a reference product without immiscible particles (reference B) and products with insoluble minerals as immiscible particles.

Table 3 shows a reference product without immiscible particles (reference B) and products with insoluble fibers as immiscible particles.

The comparison of the color, and especially brightness differences (AL), of the products with immiscible particles to the reference products without are reported and described in the following examples. Table 1:

Ci_P: concentration immiscible particles ; Rl: refractive index

Table 2 :

Ci_P: concentration immiscible particles ; Rl: refractive index

Table 3 :

Ci_P: concentration immiscible particles ; Rl: refractive index ; MCC : microcrystalline cellulose ;

The powder ingredients specified in tables 1, 2 and 3 were dry mixed using a Kenwood Cooking Chef Major mixer (model KM080, Kenwood, Switzerland) equipped with its whisk at speed 2 for 10 min.

For the products with emulsified lipids (Table 1), the liquid feed was an emulsion. The emulsion was first prepared by mixing a suspension of soy protein concentrate together with lipids, at a protein to lipids ratio of 0.1. For coconut oil and shea stearin, the protein suspension was prepared by dispersing the soy protein concentrate in warm deionized water at 70°C using a high shear mixer (L5M-A, Silverson, United States). The temperature of the mix was maintained above 60°C by maintaining it into a warm water bath during stirring. The lipid ingredient, previously melted at 70°C, was incorporated into the warm protein suspension, in amount to reach the desired emulsified lipids concentration in the product (table 1), and further stirred to form a coarse emulsion. The emulsification was furthered enhanced by passing this coarse emulsion in a two-stage homogenizer (Lab Homogenizer Twin PANDA 600, GEA, Italy). The pressure of the two stages were set as indicated in table 1, depending on the product.

In the case of emulsified canola oil, the same process was followed with the exception that all steps were performed at room temperature (20 - 25°C). In the cases where polysorbate 80 and hydrolyzed lecithin were used as emulsifiers, i.e., E-Can_F and E-Can-G respectively, the suspension of soy protein concentrate was replaced with a suspension of polysorbate 80 or hydrolyzed lecithin. The content of polysobarte 80 and hydrolyzed lecithin were adjusted to reach an emulsifier to lipids ratio of 0.1 and 0.3 in the liquid feed, respectively.

For the references A and B, the insoluble minerals and the fibers products, the liquid feed consisted in water or water mixed manually with vinegar to ensure its homogeneity prior injection in the extruder.

All blends of raw materials were extruded in similar extrusion conditions using a twin screw extruder (EuroLab, ThermoFischer, Germany). It was equipped with two screws consisting solely of conveying elements and a cooling die was attached at the end of the extruder.

This dry blend was then fed into the first barrel of the extruder. The liquid feed was injected in the second extruder barrel. The flowrate of the liquid feed was adapted as described in table 1, 2 and 3 to reach a solid content of 38,1 - 47,8 g/lOOg in the product.

The temperature profile of the extruder from the first zone to the sixth zone was 20-25°C (room temperature) for the first two barrels, 80°C for the third barrel and 230 °C for the last three barrels. The cooling die was tempered to 60°C using recirculating water.

The products were subsequently packed in sealed plastic bags under vacuum right at the extruder die output.

Size properties of the immiscible particles

Emulsified lipid droplets

The size distributions of the emulsified lipid droplets in the emulsions were determined by laser light diffraction immediately after their preparation. A Mastersizer 2000 (Malvern Instruments, Worcestershire, UK) equipped with a He/Ne laser (X = 633 nm) and an electroluminescent diode (X = 466 nm) was used. The refractive indices reported in table 1 were indicated as refractive index of the dispersed phase for each lipid source, and the refractive index of water, i.e. 1.33, was indicated for the dispersing phase. The surface area moment mean, or Sauter mean diameter, D[3;2] was extracted from all the particle size distributions of the emulsions and used as their size descriptor.

Insoluble minerals and fibers

Dynamic, quantitative shape analyses were conducted on the following powdered ingredients to determine their particle size properties using a CamsizerXT (Retsch Technology, Haan, Germany):

Insoluble minerals: different calcium citrate ingredients, calcium carbonate, calcium sulphate, Insoluble fibers: different microcrystalline cellulose ingredients.

The analysis involved the loading of the powder ingredients into a stainless-steel feeder allowing the particles to be transported from a hopper to an analyzing zone by vibration. The particles circulated in front of two light sources, and images of their shadows were captured by the basic or zoomed cameras depending on their size. Pictures were taken at a frame rate of 277 images/second and the software program examined each particle shadow to determine its size and shape features. The particle size distribution of each ingredient was determined by the software: for each particle detected, the software determined the mean diameter equivalent to the sphere of same area. The D50 parameter, that corresponds to the particle diameter at 50% in the cumulative distribution, was extracted for each ingredient and used as its size descriptor.

Color analyses of the products

After 4 h storage at room temperature in their vacuum sealed plastic bags, the products were analyzed by color measurement. The color measurements of the products were performed using a Digi-Eye® system (Carl von Gehlen Spezialmaschinen und Zubehbr GmbH & Co. KG, Mbnchengladbach, Germany) integrated with a digital camera D-90 Nikon (Tokyo, Japan). L* (0 = black, 100 = white), a* (+value = red, -value = green), and b* (+value = yellow, -value = blue) values according to the CIELAb system definition were determined. The total color difference (AE) and the brightness difference (AL) were calculated as follow:

AL — L L_re/ where L_ref, a_ref and b_ref represent the color values of the reference product that does not contain added immiscible particles and L, a and b the color values of the products containing immiscible particles. The standard deviation for the total color difference (AE) and the brightness difference (AL) were determined on 17 and 7 repetitions of references A and B and used for all products according to which reference product they are compared to.

Example 2: Impact of the type of emulsified lipid

Figure 1 and 2 shows a comparison of products which have 3 g of immiscible particles per 100 g of product consisting in emulsified lipid droplet of canola oil, coconut oil and shea stearin. Figure 2 represents these same products, plotted according to the solid fat content at 20°C of the lipids composing the emulsified droplets (IUPAC 2.150(b) Solid content determination in fats by NMR).

The products were prepared and analyzed by color measurements according to the method described in example 1. The results show that all type of emulsified lipid enabled to increase the brightness of the product by more than 2 units compared to the reference A product that does not contain immiscible particles (AL). The results shows that the higher the solid fat content of the lipids at 20°C, the brighter the product.

Example 3: Impact of the concentration of lipid

Figure 3 shows a comparison of products which have between 1 to 10 g of immiscible particles per 100 g of product, where the immiscible particles are emulsified lipid droplets of canola oil.

The products were prepared and analyzed by color measurements according to the method described in example 1. The results show that all concentrations of immiscible particles increased the brightness of the product by more than 2 units compared to the reference A product that does not contain immiscible particles (AL).

Example 4: Impact of the emulsion size

Figure 4 shows a comparison of products containing 3 g of immiscible particles per 100 g of product, where the immiscible particles are emulsified lipid droplets of shea stearin of different Sauter mean diameter (D[3;2]).

The products were prepared and analyzed by color measurements according to the method described in example 1. The results show that the immiscible particles increased the brightness of the product by more than 2 units for a Sauter mean diameter (D[3;2]) below 1 pm compared to the reference A product that does not contain immiscible particles (AL).

Example 5: Impact of the type of emulsifier

Figure 5 shows a comparison of products containing 3 g of immiscible particles per 100 g of product, where the immiscible particles are droplets of canola oil emulsified with different types of emulsifiers, i.e. soy proteins, polysorbate 80 and hydrolyzed lecithin.

The products were prepared and analyzed by color measurements according to the method described in example 1. The results show that the type of emulsifier impacted the color, and the brightness of the product. Only soy proteins, when used as emulsifier, increased the brightness of the product by more than 2 units compared to the reference A product that does not have immiscible particles (AL).

Example 6: Impact of the type of insoluble minerals

Figure 5 shows a comparison of products containing 1 g of immiscible particles per 100 g of product, where the immiscible particles are insoluble minerals of a different nature.

The products were prepared and analyzed by color measurements according to the method described in example 1. The results show that the immiscible particles increased the brightness of the product by more than 2 units, compared to the reference B product that does not have immiscible particles (AL), especially for calcium sulphate anhydrous (CaSO4), calcium citrate tribasic (CaCit - A), and Zinc Oxide (ZnO). Example 7: Impact of the refractive index of the insoluble minerals

Figure 6 shows a comparison of products containing 1 g of immiscible particles per 100g of product, where the immiscible particles are insoluble minerals with different refractive index.

The products were prepared and analyzed by color measurements according to the method described in example 1. The results show that the immiscible particles increased the brightness of the product by more than 2 units compared to the reference B product that does not have immiscible particles (AL), especially for a refractive index superior to 1.63.

Example 8: Impact of the size of the particles of insoluble minerals

Figure 7 shows a comparison of products containing 1 g of immiscible particles per 100 g of product, where the immiscible particles are insoluble minerals with different particle size (D90).

The products were prepared and analyzed by color measurements according to the method described in example 1. The results show that the immiscible particles increased the brightness of the product by more than 2 units, especially for a D90 below 35 pm, compared to the reference B product that does not have immiscible particles (AL).

Example 9: Impact of concentration of calcium citrate tribasic

Figure 9 shows a comparison of products containing from 1 to 3.6 g of immiscible particles per 100 g of product, consisting in insoluble particles of calcium citrate.

The products were prepared and analyzed by color measurements according to the method described in example 1. The results show that all concentrations of immiscible particles enable to increase the brightness of the products by more than 2 units compared to the reference B product that does not contain immiscible particles (AL).

Example 10: Impact of particle size of calcium citrate tribasic on the color and brightness of the products compared to a product which does not comprise immiscible particles.

Figure 10 shows a comparison of products containing 1.9 g of immiscible particles per 100 g of product, consisting in insoluble particles of calcium citrate with different particle size (D90).

The products were prepared and analyzed by color measurements according to the method described in example 1. The results show that the immiscible particles enabled to increase the brightness of the products by more than 2 units compared to the reference B product that does not contain immiscible particles (AL), especially for a D50 below 150 pm.

Example 11: Impact of the type of fibers on the color and brightness of the product compared to a product which does not comprise immiscible particles

Figure 11 shows a comparison of products containing 5 g of immiscible particles per 100 g of product, consisting in insoluble fibers of different nature. The products were prepared and analyzed by color measurements according to the method described in example 1. The results show that only bamboo fibers and microcrystalline cellulose (MCC) enabled to increase the brightness of the products by more than 2 units compared to the reference B product that does not contain immiscible particles (AL).

Example 12: Impact of the concentration of microcrystalline cellulose on the color and brightness of the products compared to a product which does not comprise immiscible particles

Figure 12 shows a comparison of products containing from 1 to 7 g of immiscible particles per 100 g of product, consisting in microcrystalline cellulose.

The products were prepared and analyzed by color measurements according to the method described in example 1. The results show that all concentrations of immiscible particles enable to increase the brightness of the products by more than 2 units compared to the reference product that does not contain immiscible particles (AL).

Example 13: Impact of the size of the microcrystalline cellulose on the color and brightness of the product compared to a product which does not comprise immiscible particles

Figure 13 shows a comparison of products containing 5 g of immiscible particles per 100 g of product, consisting in insoluble fibers of microcrystalline cellulose with different particle size (D90).

The products were prepared and analyzed by color measurements according to the method described in example 1. The results show that the immiscible particles enabled to increase the brightness of the product by more than 2 units compared to the reference B product that does not contain immiscible particles (AL), especially for a D50 below 50 pm.

Example 14: Composition, manufacture, and analysis of protein-based products textured by freezethaw process

Protein-based products were made using soy isolate as protein source. Vegetable oil and water were also added. Immiscible particles, such as emulsified lipid droplets of shea stearin and canola oil, and insoluble minerals such as calcium citrate tribasic were introduced as part of the recipes. The recipes were further textured in part through a freezing process and analyzed to determine their color properties.

Table 4 shows the composition and details regarding the manufacturing process of soy protein-based products that can be used as white fish replacer. It shows the recipes of a reference product without immiscible particles, products with insoluble calcium citrate tribasic, emulsified canola oil or shea stearin droplets as immiscible particles.

The comparison of the color, and especially brightness differences (AL), of the products with immiscible particles to the reference products without are reported and described in subsequent examples.

The products were manufactured by rehydrating the soy protein isolate in water together with sunflower oil and the immiscible particles. The calcium citrate tribasic was integrated as part of the mix in the form of a powder ingredient, while for the emulsified shea stearin and canola oil droplets, 10 % lipid emulsions were used. These emulsions were produced following the procedure described in Example 1. The ingredients were mixed with a Thermomix for 10 min at room temperature. The homogeneous mixes were molded under the form of fish analog filet and frozen at -15 °C for 72 h.

Table 4:

Color analyses was performed on the pieces which were defrosted at room temperature for 4 hours. The color measurements were performed as described in example 1 with the exception that the standard deviation for the total color difference (AE) and the brightness difference (AL) were determined by considering the standard deviation obtained through the measurement of the color components L, a and b of the reference and the samples.

Example 15: Impact of the concentration of calcium citrate tribasic and emulsified shea stearin and canola oil droplets on the color of protein-based products textured by freeze-thaw process

Figure 14 shows a comparison of products containing from 1.5 to 5 g of immiscible particles per 100 g of product, consisting of insoluble particles of calcium citrate tribasic or emulsified shea stearin or canola oil droplets.

The products were prepared and analyzed by color measurements according to the method described in example 1. The results show that all concentrations of immiscible particles enable an increase in the brightness of the products by more than 2 units compared to the reference fish analog products that do not contain immiscible particles.

Example 16 Composition, manufacture and analysis of the textured vegetable protein (TVP)-based products

Textured vegetable protein (TVP)-based products were made using TVP ingredients from soy and wheat and soy concentrate, and wheat protein isolate as protein sources. Water, vinegar, sodium chloride, flavors, garlic and onion powders, bamboo fibers, vegetable oil and methyl cellulose were also part of the composition of the products. Immiscible particles of calcium citrate tribasic, were introduced as part of the recipes.

A methyl cellulose gel was formed by mixing water, sunflower oil and methyl cellulose. The TVPs were hydrated in water and vinegar and further chopped to produce a homogeneous dough. The remaining ingredients were further added in the TVP dough followed by the incorporation of the methyl cellulose gel. The immiscible particle of calcium citrate tribasic were added on top of the dough, to reach a final concentration of between 0 and 2.9 %.

The dough was manually molded into nuggets or schnitzel pieces that were subsequently frozen. Color analyses was performed on the defrosted pieces (room temperature, 2 h) or on the cooked pieces (convection oven, at 180 °C for 14 min for the schnitzel and frying, at 170°C for 2 min 30 sec for the nuggets).

Tables 5 and 6 show the composition of soy and wheat gluten TVP based products, that can be used as white meat replacer.

The comparison of the color, and especially brightness differences (AL), of the products with immiscible particles of calcium citrate tribasic to the reference products without are reported and described in example 1.

Table 5:

Table 6:

Color analyses was performed on the defrosted pieces (room temperature, 2 h) or on the cooked pieces (convection oven, at 180 °C for 14 min for the schnitzel and frying, at 170°C for 2 min 30 sec for the nuggets). The color measurements were performed as described in example 1 with the exception that the standard deviation for the total color difference (AE) and the brightness difference (AL) were determined by considering the standard deviation obtained through the measurement of the color components L, a and b of the reference and the sample.

Example 17: Impact of the concentration of calcium citrate tribasic on the color of TVP-based products

Figure 15 shows a comparison of products containing from 1.5 to 2.9 g of immiscible particles per 100 g of product, wherein the insoluble particles are calcium citrate tribasic.

The products were prepared and analyzed by color measurements according to the method described in example 1. The results show that all concentrations of immiscible particles enable an increase in the brightness of the products by more than 2 units compared to the reference schnitzel and nugget products that does not contain immiscible particles.

Example 18: Role of droplet size on the whiteness impact of different oil emulsions

It was found that both optical contrast (difference between the refractive indices) of the material and whitening agent and also the particle / droplet size of the whitening agent are important. Thus, for a specific refractive index of a whitening agent, there is an optimum size or particle / droplet leading to highest impact on the whiteness. The meat analogue is found to have a refractive index of around 1.4. Whereas canola oil has a refractive index of 1.465. In figure 16 the scattering efficiency (Ssca), normalized by the maximum scattering efficiency possible, of droplets of canola oil with different size is demonstrated. As can be seen in the figure, there is an optimum droplet size (in this case dpopt = 3.8 micron) at which the scattering and whiteness efficiency of the whitening agent is the maximum. As the move is made away from the optimum droplet size, the whitening efficiency of the whitening agent diminishes significantly.

Similar to canola oil with Rl of 1.465, in Figure 17 the impact of droplet size of coconut oil with Rl of 1.44 on the scattering efficiency is shown. The optimum droplet size for coconut oil was found as 6.6 micron.

Thus, for each whitening agent, there is an optimum droplet / particle size where the whitening efficiency is expected to be the highest. Additionally, when the whitening agent is changed, optimum particle / droplet size changes as well. It was found that the optimum size increases as the optical contrast between the whitening agent and matrix decreases and vice versa. Thus, for a specific combination of matrix and whitening agent there exists an optimum size of the whitening agent and the range around this optimum size where the whitening efficiency is highest.

Claims

1. A method of making a texturized protein product, said method comprising the steps of a. Forming a mixture of raw materials, wherein the raw materials include protein, immiscible particles, wherein the immiscible particles are insoluble mineral and/or emulsified lipid, and water; and b. Forming a texturized protein product from the mixture by a texturizing process, for example an extrusion process, a trigger process, a freeze-thaw process, a 3-D printing process, a shear cell technology process, a microwave tube heating process, an injection molding process, a freeze structuring or freeze alignment process, an electro-spinning process, or a high-pressure treatment process; characterized in that between 0.1 wt% to 15 wt% immiscible particles on a wet basis is dispersed in the mixture, and that the immiscible particle has a refractive index between 1.55 and 2.5.

2. The method according to claim 1, wherein said texturizing process is an extrusion process comprising the steps of a. Forming a mixture by metering raw materials into an extruder, wherein the raw materials include protein, immiscible particles and water; b. Passing the mixture through an extruder whilst heating the mixture to above the denaturation temperature of the protein, wherein the protein content of the mixture is in the range from 30 wt% to 95 wt% on a dry basis and the solids content of the mixture is in the range from 25 wt% to 90 wt% on a wet basis; c. Cooling the mixture to a temperature of less than 100°C to form a texturized protein product.

3. The method according to any one of claims 1 or 2, wherein the immiscible particle is an insoluble mineral and/or emulsified lipid.

4. The method according to any one of claims 1 to 3, wherein the immiscible particle is an insoluble mineral having a D50 particle size of less than 50 microns, preferably between 0.10 microns to 10 microns.

5. The method according to claim 4, wherein the insoluble mineral is calcium citrate tribasic.

6. The method according to any one of claims 1 to 3, wherein the immiscible particles are emulsified lipids, wherein said emulsified lipids have been made by emulsifying lipids with soy protein.

7. The method according to claim 6, wherein the emulsified lipid has a D [3;2] particle size of between 0.3 microns to 0.7 microns.

8. The method according to any one of claims 6 to 7, wherein the emulsified lipid has a solid fat content at 20°C greater than 80%.

9. The method according to any one of claims 6 to 8, wherein the emulsified lipid is shea stearin.

10. The method according to any one of claims 1 to 11, the protein ingredient has a protein content between 60 wt% to 95 wt% on a dry matter basis.

11. The method according to any one of claims 1 to 12, wherein the protein is plant protein, for example soy protein, gluten protein, or pea protein, or a combination thereof.

12. The method according to any one of claims 1 to 13, wherein the texturized protein product is a plant-based chicken analogue.

13. A texturized protein product, said product comprising protein, immiscible particle, wherein said immiscible particle is preferably insoluble mineral and/or emulsified lipid, and water, wherein the protein content of the product is in the range from 30 wt% to 95 wt% on a dry basis and the solids content of the product is in the range from 25 wt% to 90 wt% on a wet basis, and wherein the immiscible particles are present at a concentration of between 0.1 wt% to 15 wt% on a wet basis in the product.

14. Use of insoluble mineral and/or emulsified lipid to improve the brightness of a texturized protein product, wherein the emulsified lipid is emulsified shea stearin, and the insoluble mineral is calcium citrate.