Meat substitute product, method for producing the same and twin-screw extruder

Abstract

In order to improve the mouthfeel of meat substitute production, improvements in meat substitute products and high moisture protein texturized extrusions are invented. The inventors have found that by appropriate selection of extrusion parameters and starting materials comprising mechanically processed starch-containing grains, the formation of an emulsion between starch and a protein melt forming a protein matrix can be prevented or reduced to such an extent that a large amount of unbound starch is present in the protein matrix. It has been observed that the presence of unbound starch in the protein matrix improves the mouthfeel and maintains an acceptable mouthfeel for a long time. This patent application contains a large number of independent claims for meat substitute products and methods.

Description

Technical Field

[0001] The present invention relates to meat substitute products and methods for their manufacture. Furthermore, the present invention relates to the use of starch in food products. Background Art

[0002] In recent years, many people have turned vegetarian or vegan, or at least increased the share of vegetables and vegetable products in their diet. While ecological concerns are the reason for some, it seems obvious that vegetables and products made from vegetables should be a central part of a healthy diet. Many consumers find it difficult to ensure a daily protein intake using vegetables or products made from vegetables, while some find it time-consuming to prepare protein-containing ingredients for steaming or baking.

[0003] Therefore, there is a market for producing vegetarian or vegan foods on an industrial basis by extrusion cooking. Extrusion cooking is a continuous process that enables the production of texturized proteins, which are unique products made by extrusion. Extrusion enables control of functional properties such as density, rate and time of rehydration, shape, product appearance and mouthfeel.

[0004] For the extrusion of meat substitute products (also called meat analogs) or texturized vegetable products, twin screw extruders are usually used.There are two main types of extrusion cooking processes for the preparation of meat substitute products.

[0005] A meat substitute product produced by texturizing extrusion with low moisture proteins. These products have a moisture content between 10% and 40% (15% to 40% during extrusion). They typically have a spongy texture and require rehydration before consumption. These products are often used as chopped meat substitutes or extenders in meat products, but do little to mimic fibrous whole muscle meat.

[0006] Another type of meat substitute product is made with high-moisture protein texturized extrusions. These products have a moisture content between 40% and 80%. They are generally more like muscle food than meat substitute products made with low-moisture texturized extrusions.

[0007] Meat substitute products are typically manufactured by mixing at least one protein matrix-forming ingredient such as a protein isolate or a protein concentrate (which are generally referred to as a protein component), possible starch-containing particles, possible oil, and extruding the mixed slurry of ingredients in an extruder configured to perform protein texturization extrusion.

[0008] In tests conducted by the inventors using high moisture protein texturized extrusions, we found that the mouthfeel of freshly extruded meat substitute products was generally very appealing. However, after a relatively short period of time (generally in the range of a few minutes, typically 5-10 minutes), as the meat substitute product cooled, the mouthfeel became unacceptable.

[0009] Currently, meat substitute products made from high-moisture protein texturized extrusion are often sold deep frozen. Alternatively, meat substitute products are sold chopped or torn into pieces, making the unacceptable mouthfeel less noticeable. Summary of the invention

[0010] The first object of the present invention is to improve the mouthfeel of a meat substitute product made by texturizing and extruding a high-moisture protein, so that the improved mouthfeel is comparable to that of steamed chicken thigh meat, and the improved mouthfeel is further maintained for a long time, such as overnight, or 24 hours, without the need to freeze the meat substitute product.

[0011] When the linear compressibility of the sample is relatively high and the cylindrical compressibility is relatively low, it can be assumed that the mouthfeel is comparable to that of a steamed chicken leg. When measured with a texture analyzer model TA.XTPlus (Stable Micro Systems, Inc., Surrey, UK) equipped with a 294.2 N (30 kg) load cell (detector sensor) and a sharp blade, the linear compressibility is preferably between 300 g and 1500 g. When measured with a texture analyzer TA.XTPlus (Stable Micro Systems, Inc.) equipped with a 294.2 N (30 kg) load cell (detector sensor) with a cylindrical probe (model "P / 36R", 36 mm radius edge cylindrical probe - aluminum - AACC standard probe for bread hardness), the cylindrical compressibility is preferably between 7000 g and 17500 g. For the measurement, a sample with a height between 7.0 and 12.0 mm should be used. The width and length of the sample are preferably selected to be 40 mm. Fig.11 The shear force and compression force analysis methods that should preferably be used are illustrated.

[0012] Alternatively, when the compressibility and chewing properties experienced are confirmed by a panel of testers to be similar to those of a cooked chicken leg, the mouthfeel of the meat substitute product can be said to be comparable to that of a cooked chicken leg.

[0013] This object can be achieved by a meat substitute product according to any of independent items 1, 3, 9, 10, 11, or 60, or any combination thereof comprising two, three, four or all of these independent items, and by a method for manufacturing a meat substitute product according to any of independent items 16, 19, 20, 21, 22, 24 or 53, or any combination thereof comprising two, three, four, five or all of these independent items.

[0014] A second object of the present invention is to increase starch solubility in meat substitute products made by high moisture protein texturization extrusion. This object can be achieved with a meat substitute product according to any of independent items 9 and 11 and with a method according to any of independent items 21 and 22.

[0015] A third object of the present invention is to control starch solubility in meat substitute products made by high moisture protein texturization extrusion. This object can be achieved with a meat substitute according to independent item 10 and with a method according to independent item 20.

[0016] A fourth object relates to the use of the novel starch ingredient in food products.

[0017] The fifth object relates to the improvement of a twin screw extruder. This object can be achieved with a twin screw extruder according to item 41.

[0018] The sixth object relates to improving the mouthfeel of meat substitute products made by high moisture protein texturization extrusion. This object can be achieved by a method according to parallel independent item 42 and by product item 52.

[0019] The dependent items describe advantageous aspects of the meat substitute products and methods for making the meat substitute products.

[0020] The above items are shown below.

[0021] Item 1. A meat substitute product comprising:

[0022] An extrudate made with high moisture protein texturized extrusion, i.e., such that the moisture content during extrusion is between 40% and 80%, and having a substantially linearly oriented continuous protein fiber matrix structure,

[0023] The matrix comprises a disruption in the matrix structure,

[0024] Some of the disruptions in the matrix structure are in the form of cavities having walls at least partially coated by gelatinized starch clusters formed from starch,

[0025] Such that when measured on the extrudate, at least 5.1%, preferably at least 5.2% of the starch is soluble starch, the soluble starch being located at the disruptions in the matrix structure and not emulsified by the matrix structure.

[0026] Item 2. The meat substitute product according to Item 1, wherein: the soluble starch is in the form of clusters and is phase-separated from the protein phase and is not emulsified by the protein.

[0027] Item 3. A meat substitute product, wherein: the meat substitute product comprises an extrudate, the extrudate being made by texturizing extrusion with a high moisture protein and having a substantially linearly oriented continuous protein fiber matrix structure, the extrudate comprising gelatinized starch clusters located at disruptions in the matrix structure and not emulsified by the matrix structure, such that when the extrudate is measured,

[0028] i) when the protein content of the extrudate is greater than 55% but less than 70% by weight, at least 10.5% of the starch in the extrudate is washable starch,

[0029] ii) when the protein content of the extrudate is at least 70 wt% but less than 90 wt%, at least 15% of the starch in the extrudate is washable starch,

[0030] iii) when the protein content of the extrudate is at least 90 wt% but equal to or less than 99 wt%, at least 16% of the starch in the extrudate is washable starch,

[0031] The weight % indicated therein is on a dry basis.

[0032] Item 4. The meat substitute product according to Item 3, wherein the washable starch is washable in water at a temperature of 50°C.

[0033] Item 5. The meat substitute product according to Item 3 or 4, wherein the washable starch is located at the destruction of the matrix structure and is not emulsified by the matrix structure.

[0034] Item 6. The meat substitute product of Item 5, wherein: some of the disruptions in the matrix structure are in the form of cavities having walls at least partially coated by gelatinized starch clusters formed from washable starch.

[0035] Item 7. The meat substitute product according to any one of the preceding items 1 and / or 6, wherein the starch clusters comprise washable starch that is washable in water at a temperature of 50°C.

[0036] Item 8. The meat substitute product according to any of the preceding items, comprising: starch clusters having a size (eg, length) greater than about 100 μm.

[0037] Item 9. A meat substitute product, wherein: the meat substitute product comprises an extrudate having a substantially linearly oriented continuous protein fiber matrix structure, the extrudate comprising starch,

[0038] And wherein: the extrudate is made using a high moisture protein texturized extrusion process in which the starchy grains are gelatinized and the proteins forming the protein matrix are melted so that the starchy grains are gelatinized before they are substantially powdered by the extruder screw.

[0039] Item 10. A meat substitute product, wherein: the meat substitute product comprises an extrudate having a substantially linearly oriented continuous protein fiber matrix structure, the extrudate comprising starch,

[0040] and wherein: the extrudate is made using a high moisture protein texturized extrusion process in which the starch-containing grains are gelatinized and the proteins forming the protein matrix are melted such that:

[0041] (a) before said gelatinized starch-containing grains are emulsified with said proteins of said protein matrix, and

[0042] (b) before the gelatinized starch forms a complete barrier that prevents the formation of a continuous protein fiber cross-linked matrix,

[0043] The protein melts.

[0044] Item 11. A meat substitute product comprising:

[0045] An extrudate having a substantially linearly oriented continuous protein fiber matrix structure, the extrudate being made by texturizing extrusion with a high moisture protein and comprising starch located at disruptions in the matrix structure and not emulsified by the matrix structure, wherein:

[0046] Some of the disruptions in the matrix structure are in the form of cavities having walls at least partially coated by clusters of gelatinized starch formed from the starch.

[0047] Item 12. The meat substitute product according to Item 11, wherein the gelatinized starch clusters formed from starch are formed from soluble starch or washable starch.

[0048] Item 13. The meat substitute product of any preceding item, wherein: the extrudate is an extrudate made using a high moisture protein texturizing extrusion process using a twin screw extruder with a long cooling die.

[0049] Item 14. The meat substitute product according to Item 13, wherein: the length of the long cooling mold is at least 300 mm, preferably at least 1000 mm, and most preferably between 1000 mm and 5000 mm.

[0050] Item 15. A meat substitute product according to any one of items 1-14, wherein: the meat substitute product is in the form of thick chunks, chops, nuggets, fillets, steaks or kebab-like slices, or in the form of kebab-like layered layers in yogurt or vegetarian yogurt and spices.

[0051] Item 16. A method for making a meat substitute product, the method comprising: producing the meat substitute product using an extruder configured to perform high moisture protein texturized extrusion, wherein the starch-containing grains are gelatinized and the proteins forming the protein matrix are melted,

[0052] The meat substitute product is an extrudate having a substantially linearly oriented continuous protein fiber matrix structure, the extrudate comprising gelatinized starch clusters located at disruptions in the matrix structure and not emulsified by the matrix structure,

[0053] The extrudate comprises starch, of which at least 5.1%, preferably at least 5.2%, when measured on the extrudate, is soluble starch.

[0054] Item 17. The method according to Item 16, wherein: the soluble starch is located at the destruction of the matrix structure and is not emulsified by the matrix structure.

[0055] Item 18. A method according to Item 16 or 17, wherein: some of the disruptions in the matrix structure are in the form of cavities having walls at least partially coated by gelatinized starch clusters formed from starch, preferably soluble starch.

[0056] Item 19. A method for making a meat substitute product, the method being characterized by: producing the meat substitute product using an extruder configured to perform high moisture protein texturized extrusion, wherein starch-containing grains are gelatinized and the protein forming the protein matrix is ​​melted, the meat substitute product being an extrudate having a continuous protein fiber matrix structure, the extrudate comprising gelatinized starch clusters formed from starch located at disruptions in the matrix structure and not emulsified by the matrix structure, such that when the extrudate is measured,

[0057] i) when the protein content of the extrudate is greater than 55% but less than 70% by weight, at least 10.5% of the starch is washable starch,

[0058] ii) when the protein content of the extrudate is at least 70 wt% but less than 90 wt%, at least 15% of the starch is washable starch,

[0059] iii) when the protein content of the extrudate is at least 90 wt% but equal to or less than 99 wt%, at least 16% of the starch is washable starch,

[0060] The weight % indicated therein is on a dry basis.

[0061] Item 20. A method for making a meat substitute product, the method characterized by: producing the meat substitute product using an extruder configured to perform high moisture protein texturized extrusion, wherein starch-containing grains are gelatinized and the proteins forming the protein matrix are melted, the meat substitute product being an extrudate having a continuous protein fiber matrix structure, the extrudate comprising starch such that:

[0062] (a) before gelatinizing the starch-containing grains and forming an emulsion with said protein of said protein matrix, and

[0063] (b) Before the gelatinized starch forms a complete barrier that prevents the formation of a continuous protein fiber cross-linked matrix,

[0064] The proteins forming the protein matrix melt.

[0065] Item 21. A method for making a meat substitute product, the method being characterized by: producing the meat substitute product using an extruder configured to perform high moisture protein texturized extrusion such that the moisture content during extrusion is between 40% and 80%, wherein during extrusion the starch-containing grains are gelatinized and the proteins forming the protein matrix are melted, the resulting meat substitute product being an extrudate having a continuous protein fiber matrix structure, the extrudate comprising starch, wherein: the step of heating the slurry in the extruder is performed such that:

[0066] The starchy grains are gelatinized before they are substantially pulverized by the extruder screw.

[0067] Item 22. A method for making a meat substitute product, the method being characterized in that the meat substitute product is produced using an extruder configured to perform high-moisture protein texturized extrusion, in which starch-containing grains are gelatinized and the protein forming the protein matrix is ​​melted, the meat substitute product being an extrudate having a continuous protein fiber matrix structure, the extrudate comprising gelatinized starch clusters formed from starch, located at locations where the matrix structure is disrupted and not emulsified by the matrix structure.

[0068] Item 23. A method according to Item 22, wherein: some of the disruptions in the matrix structure are in the form of cavities having walls at least partially coated by gelatinized starch clusters formed from starch, preferably soluble starch or washable starch.

[0069] Item 24. A method for manufacturing a meat substitute product, the method comprising the steps of:

[0070] a) feeding a mixture into an extruder configured to perform high moisture protein texturizing extrusion, the mixture comprising:

[0071] a1) at least one protein matrix-forming component, such as a protein isolate or a protein concentrate, and

[0072] a2) Mechanically processed starch-containing grains having a diameter of at least 0.125 mm 3 , preferably at least 1 mm 3 , most preferably at least 6mm 3 The particle volume;

[0073] b) feeding water into the extruder;

[0074] c) heating the mixture in the extruder to gelatinize the starch-containing grains;

[0075] d) after gelatinization of the starch is achieved, further heating the mixture in the extruder to melt at least one protein matrix-forming ingredient; and

[0076] e) extruding the mixture through an extrusion die at a temperature between 70°C and 100°C

[0077] in:

[0078] i) carrying out the heating step c) with impingement heating such that the starch-containing grains are gelatinized before they are substantially pulverized by the extruder screw; and

[0079] ii) carrying out the heating step d) by shock heating, such that:

[0080] (a) before the gelatinized starch and the protein matrix forming component form an emulsion,

[0081] as well as

[0082] (b) Before the gelatinized starch forms a complete barrier that prevents the formation of a continuous protein fiber cross-linked matrix,

[0083] The protein melting temperature of the protein matrix forming component is reached.

[0084] Item 25. The method of Item 24, wherein: the starch-containing grains are soaked before being fed into the extruder.

[0085] Item 26. The method according to Item 24 or 25, wherein: the starch-containing grains are treated before being fed into the extruder so that the starch is gelatinized before being fed into the extruder.

[0086] Item 27. The method of any one of Items 24-25, wherein: water is fed to the starch-containing grain at an elevated temperature.

[0087] Item 28. The method according to Item 27, wherein: the water temperature is higher than 60°C, preferably higher than 65°C.

[0088] Item 29. The method according to Item 27, wherein: the water temperature is higher than 75°C.

[0089] Item 30. The method according to any one of the preceding items 24-29, wherein the heating step d) is performed at a temperature between 140°C and 200°C.

[0090] Item 31. The method according to any one of the preceding items 24-30, wherein: the heating step d) is performed so that protein melting occurs between 1 s and 40 s, preferably between 10 s and 30 s, after step b).

[0091] Item 32. The method according to any one of the preceding items 24-31, wherein: the heating step c) is performed so that starch gelatinization occurs between 0 s and 18 s, preferably between 1 s and 15 s, after step b).

[0092] Item 33. The method according to any one of Items 24 to 32, wherein: after the starch-containing grains are ground by the extruder screw into particles with a volume of less than 5000 μm 3 Before, and preferably after, the starch-containing grains are ground by an extruder screw to a particle size of less than 0.001 mm 3 Prior to this, the heating step c) is carried out.

[0093] Item 34. A method according to any one of items 24 to 33 above, wherein: after the heating step d), the mixture is continued to be extruded for more than 5 s, preferably more than 10 s, at a temperature not higher than the temperature in the heating step d), preferably between 90°C and the temperature in the heating step d).

[0094] Item 35. A method according to any one of the preceding items 16-34, wherein: the mechanically processed starch-containing grains comprise one or more of the following or consist of one or more of the following: oat flakes (such as compressed, rolled or flaked), steel-cut grains, shelled and pearled grains, crushed grains, shelled but not pearled grains, but excluding: shelled but not pearled oat grains, shelled but not pearled rye grains, shelled but not pearled barley grains, shelled but not pearled corn grains.

[0095] Item 36. A method according to any one of the preceding items 16-35, wherein: the mechanically processed starch-containing grains comprise one or more of the following or consist of one or more of the following: oats, barley, rye, wheat, rice, corn, lentils, chickpeas, mung beans, broad beans, peas, quinoa, pigeon peas, sorghum, buckwheat, but excluding: shelled but unpealed oat grains, shelled but unpealed rye grains, shelled but unpealed barley grains, shelled but unpealed corn grains.

[0096] Item 37. The method according to any one of the preceding items 16-36, wherein the extrusion step is performed using an extrusion die having a length greater than 300 mm, preferably greater than 1000 mm, and most preferably between 1000 mm and 5000 mm.

[0097] Item 38. Use of insoluble washable starch in cluster form in meat substitute food products produced by high moisture protein texturized extrusion.

[0098] Item 39. Use of the insoluble washable starch according to Item 38, wherein the meat substitute food product is manufactured using the method according to any one of Items 16-37.

[0099] Item 40. Use of an insoluble washable starch according to Item 38 or 39, wherein: the insoluble washable starch is contained in the form of clusters, the clusters having a size greater than about 100 μm.

[0100] Item 41. A twin-screw extruder for high-moisture protein texturization extrusion, comprising: a screw barrel (138) for accommodating an extruder screw (126), wherein the extruder screw (126) defines the direction of movement of the material in the extruder (13) relative to the barrel (138), the barrel (138) also comprising a first inlet hole (139) for receiving solid ingredients into the extruder (13) and a second inlet hole (140) for receiving liquid into the extruder (13), wherein the second inlet hole (140) is located downstream from the first inlet hole (139) along the flow direction, and the extruder (13)

[0101] i) connected to a warm water supply having a temperature of at least 50°C,

[0102] or

[0103] ii) comprising a heating element (14) configured to heat water from the water supply to a temperature of at least 50°C before passing the water from the water supply into the second inlet aperture (140);

[0104] The extruder further comprises a long cooling die (125), which is longer than 300 mm, preferably between 300 mm and 5000 mm in length, most preferably between 1000 mm and 3000 mm in length.

[0105] Item 42. A method for making a meat substitute product using high moisture protein texturized extrusion, wherein the improvement comprises: selecting extrusion parameters and starting materials, the starting materials comprising at least: i) a protein component, which is preferably a protein isolate or protein concentrate or a mixture thereof; ii) mechanically processed starch-containing grains; and iii) cereal flour, such that the formation of an emulsion between the starch and the protein melt forming the protein matrix is ​​substantially prevented or reduced to the extent that a substantial amount of the starch in the extrudate will be in the form of gelatinized starch clusters and will not be bound by the protein matrix.

[0106] Item 43. The method of Item 42, wherein: the clusters have a size greater than about 100 μm.

[0107] Item 44. A method according to Item 42 or 43, wherein: the controlled extrusion parameters include, or are: water feed temperature and / or heating distribution, such as along the extrusion screw and in the cooling die, so that the starting material is impact heated in the extruder.

[0108] Item 45. A method according to any one of Items 42-44, wherein the hardness or compressibility of the meat substitute product is controlled by controlling the ratio of the amount of soluble starch to the total amount of starch and / or the weight % of the soluble starch in the meat substitute product.

[0109] Item 46. A method according to Item 45, wherein: the ratio of the amount of soluble starch to the total amount of starch and / or the weight % of the soluble starch is controlled so that the linear compressibility is between 300 g and 1500 g, and the cylindrical compressibility is between 7000 g and 17500 g.

[0110] Item 47. The method of Item 46, wherein: linear and cylindrical compressibility are measured at least 24 hours after extrusion.

[0111] Item 48. The method of any one of Items 42-47, wherein the amount of starch not bound by the protein matrix is ​​determined as soluble starch.

[0112] Item 49. A method according to Item 48, wherein: the compressibility is controlled by changing the extrusion parameters so that in the meat substitute product after extrusion, the ratio of the amount of soluble starch to the total amount of starch is between 3 weight % and 10 weight %, and / or the soluble starch content is between 0.03 weight % and 1.1 weight %.

[0113] Item 50. A method according to any one of items 42-49, wherein: the mechanically processed starch-containing grains comprise one or more of the following or consist of one or more of the following: oat flakes (such as compressed, rolled or flaked), steel-cut grains, shelled and pearled grains, crushed grains, shelled but not pearled grains, but excluding: shelled but not pearled oat grains, shelled but not pearled rye grains, shelled but not pearled barley grains, shelled but not pearled corn grains.

[0114] Item 51. A method according to any one of the preceding items 42-50, wherein: the mechanically processed starch-containing grains comprise one or more of the following or consist of one or more of the following: oats, barley, rye, wheat, rice, corn, lentils, chickpeas, mung beans, broad beans, peas, quinoa, pigeon peas, sorghum, buckwheat, but excluding: shelled but unpealed oat grains, shelled but unpealed rye grains, shelled but unpealed barley grains, shelled but unpealed corn grains.

[0115] Item 52. A meat substitute product, the meat substitute product being made using the method according to any one of Items 42-51.

[0116] Item 53. A method for manufacturing a meat substitute product, the method comprising the steps of:

[0117] a) feeding a mixture into an extruder configured to perform high moisture protein texturizing extrusion, the mixture comprising:

[0118] a1) at least one protein matrix-forming component, such as a protein isolate or a protein concentrate, and

[0119] a2) Mechanically processed starch-containing grains, which are steel-cut grains and have a diameter of at least 0.125 mm 3 , preferably at least 1mm 3 , most preferably at least 6 mm 3 The particle volume;

[0120] b) feeding water into the extruder so that the moisture content during extrusion is between 40% and 80%;

[0121] c) heating the mixture in the extruder to gelatinize the starch-containing grains;

[0122] d) after gelatinization of the starch is achieved, further heating the mixture in the extruder to melt at least one protein matrix-forming ingredient; and

[0123] e) extruding the mixture through an extrusion die at a temperature between 70° C. and 100° C.,

[0124] in:

[0125] i) carrying out the heating step c) with impingement heating such that the starch-containing grains are gelatinized before they are substantially pulverized by the extruder screw; and

[0126] ii) carrying out the heating step d) by shock heating, such that:

[0127] (a) before the gelatinized starch and the protein matrix forming component form an emulsion,

[0128] as well as

[0129] (b) Before the gelatinized starch forms a complete barrier that prevents the formation of a continuous protein fiber cross-linked matrix,

[0130] The protein melting temperature of the protein matrix forming component is reached.

[0131] Item 54. The method of Item 53, wherein: the steel cut grains are soaked in water before being fed into the extruder.

[0132] Item 55. The method of Item 53, wherein: the steel cut grains are not soaked when fed into the extruder.

[0133] Item 56. The method of any one of Items 53-55, wherein the steel cut grains comprise steel cut oats.

[0134] Item 57. A method according to any one of Items 53-56, wherein: steel cut oats are replaced with any of the following: steel cut barley, rice kernels, broken rice, pearled barley, pearled rye, pearled wheat, pearled oats, crushed pea seeds (e.g., with a particle size of 2 mm), crushed broad bean seeds, crushed chickpea seeds, lentil seeds or a mixture thereof.

[0135] Item 58. The method according to any one of items 53-57, wherein: in the method, a combination of (a) using an extrusion impact heating temperature setting and (b) using hot water as a liquid feed is used to increase starch solubility.

[0136] Item 59. The method of any one of Items 53-58, wherein: in the method, (a) grains and water are mixed, and (b) the grains combined with water are heated sufficiently early before starch in the grains is emulsified by the protein matrix.

[0137] Item 60. A meat substitute product made using the method of any one of Items 54-59.

[0138] Advantages of the invention

[0139] According to a first aspect, a meat substitute product showing an improved mouthfeel that is maintained for a prolonged period of time is produced using high moisture protein texturized extrusion and comprising an extrudate having a substantially linearly oriented continuous protein fiber matrix structure, the extrudate comprising starch,

[0140] Of these, at least 5.1%, preferably at least 5.2%, of the starch is soluble starch.

[0141] Accordingly, a meat substitute product showing an improved mouthfeel that is maintained for an extended period of time can be made using a manufacturing method that uses an extruder configured to perform high moisture protein texturized extrusion in which the starch-containing grains are gelatinized and the proteins that form the protein matrix are melted so that the meat substitute product is an extrudate having a continuous protein fiber matrix structure, the extrudate comprising starch, wherein at least 5.1%, preferably at least 5.2%, of the starch is soluble starch.

[0142] The soluble starch is preferably located at disruptions in the matrix structure and is not emulsified by the matrix structure. Most preferably, some of the disruptions in the matrix structure are in the form of cavities having walls at least partially coated by gelatinized starch clusters formed from starch, preferably soluble starch.

[0143] According to a second aspect, which replaces or supplements the first aspect, a meat substitute product shows an improved mouthfeel that is maintained for a long time, the meat substitute product is made with high moisture protein texturized extrusion and comprises an extrudate having a substantially linearly oriented continuous protein fiber matrix structure, the extrudate comprising starch such that:

[0144] i) when the protein content of the extrudate is greater than 55% but less than 70% by weight, at least 10.5% of the starch is washable starch,

[0145] ii) when the protein content of the extrudate is at least 70 wt% but less than 90 wt%, at least 15% of the starch is washable starch,

[0146] iii) when the protein content of the extrudate is at least 90 wt% but equal to or less than 99 wt%, at least 16% of the starch is washable starch,

[0147] The weight % indicated therein is on a dry basis.

[0148] Accordingly, a meat substitute product showing an improved mouthfeel that is maintained for an extended period of time can be made using a manufacturing method that uses an extruder configured to perform high moisture protein texturized extrusion in which the starch-containing grains are gelatinized and the proteins forming the protein matrix are melted, the meat substitute product being an extrudate having a continuous protein fiber matrix structure, the extrudate comprising starch such that:

[0149] i) when the protein content of the extrudate is greater than 55% but less than 70% by weight, at least 10.5% of the starch is washable starch,

[0150] ii) when the protein content of the extrudate is at least 70 wt% but less than 90 wt%, at least 15% of the starch is washable starch,

[0151] iii) when the protein content of the extrudate is at least 90 wt% but equal to or less than 99 wt%, at least 16% of the starch is washable starch,

[0152] The weight % indicated therein is on a dry basis.

[0153] Preferably, the washable starch is located at disruptions in the matrix structure and is not emulsified by the matrix structure. Most preferably, some of the disruptions in the matrix structure are in the form of cavities having walls at least partially coated by gelatinized starch clusters formed from the washable starch. The washable starch is washable in water at a temperature of 50°C, which is below the gelatinization temperature of the starch.

[0154] According to a third aspect, which replaces the first and second aspects or is in addition to one or both of them, a meat substitute product showing an improved mouthfeel that is maintained for a long time, the meat substitute product is made with high moisture protein texturized extrusion and comprises an extrudate having a substantially linearly oriented continuous protein fiber matrix structure, the extrudate comprising starch, and wherein the extrudate is made using a high moisture protein texturized extrusion process in which starch-containing grains are gelatinized and the proteins forming the protein matrix are melted such that:

[0155] The starchy grains are gelatinized before they are substantially pulverized by the extruder screw.

[0156] Accordingly, a meat substitute product showing an improved mouthfeel that is maintained for a long time can be manufactured by producing a meat substitute product using a manufacturing method that uses an extruder configured to perform high-moisture protein texturized extrusion, in which the starch-containing grains are gelatinized and the protein forming the protein matrix is ​​melted, the meat substitute product being an extrudate having a continuous protein fiber matrix structure, the extrudate containing starch, wherein: the step of heating the slurry in the extruder is performed in such a way that the starch-containing grains are gelatinized before they are substantially powdered by the extruder screw.

[0157] The method for manufacturing the meat substitute product increases starch solubility, and accordingly, the meat substitute product has increased starch solubility.

[0158] According to a fourth aspect, replacing the first, second and third aspects or supplementing one, two or all of them, a meat substitute product showing an improved mouthfeel that is maintained for a long time, the meat substitute product is made with high moisture protein texturized extrusion and comprises an extrudate having a substantially linearly oriented continuous protein fiber matrix structure, the extrudate comprising starch, and wherein: the extrudate is made using a high moisture protein texturized extrusion process in which starch-containing grains are gelatinized and the proteins forming the protein matrix are melted such that:

[0159] (a) before the gelatinized starch-containing grains are emulsified with the proteins of the protein matrix,

[0160] and

[0161] (b) before the gelatinized starch forms a complete barrier that prevents the formation of a continuous protein fiber cross-linked matrix,

[0162] Protein melting.

[0163] Accordingly, a meat substitute product showing an improved mouthfeel that is maintained for an extended period of time can be made by producing a meat substitute product using a manufacturing method that uses an extruder configured to perform high moisture protein texturized extrusion in which the starch-containing grains are gelatinized and the proteins forming the protein matrix are melted, the meat substitute product being an extrudate having a continuous protein fiber matrix structure, the extrudate comprising starch such that:

[0164] (a) before the gelatinized starch-containing grains are emulsified with proteins of the protein matrix,

[0165] and

[0166] (b) before the gelatinized starch forms a complete barrier that prevents the formation of a continuous protein fiber cross-linked matrix,

[0167] The proteins forming the protein matrix melt.

[0168] The manufacturing method of the meat substitute product enables the control of starch solubility, and accordingly, the meat substitute product can have controlled starch solubility.

[0169] According to a fifth aspect, which replaces the first, second, third and fourth aspects, or supplements one, two, three or all of them, a meat substitute product shows an improved mouthfeel that is maintained for a long time, the meat substitute product is made by texturizing and extruding a high moisture protein, and comprises:

[0170] An extrudate having a substantially linearly oriented matrix structure of continuous protein fibers, the extrudate comprising starch located at disruptions in the matrix structure and not emulsified by the matrix structure.

[0171] Accordingly, a meat substitute product showing an improved mouthfeel that is maintained for an extended period of time can be made using a manufacturing method that uses an extruder configured to perform high moisture protein texturized extrusion in which the starch-containing grains are gelatinized and the proteins that form the protein matrix are melted, the meat substitute product being an extrudate having a continuous protein fiber matrix structure,

[0172] The extrudate contains starch located at disruptions in the matrix structure and not emulsified by the matrix structure.

[0173] The method for manufacturing the meat substitute product increases starch solubility, and accordingly, the meat substitute product has increased starch solubility.

[0174] Particularly advantageously, some of the disruptions in the matrix structure may be in the form of cavities having walls at least partially coated by gelatinized starch clusters formed from starch, preferably soluble starch or washable starch.

[0175] The advantage generated in particular by the fifth aspect is that the damage and, in particular, the cavities coated at least partially (preferably completely) by the starch clusters (and the starch clusters separated from the phase) prevent the hardening of the extrudate (generated by the gel hardness reinforcement). The damage and at least partially coated cavities formed by the starch clusters (and the starch clusters separated from the phase) act as a new type of destructive complex, which prevents the further formation of protein-protein interactions between protein fibers after extrusion. They are different from and superior to other destructive particles known to the inventors, such as starch, cereal flour, insoluble salts, dietary fiber, pregelatinized starch, gas, which either (a) disappear after extrusion (e.g., gas), or (b) will be emulsified by the protein matrix during extrusion (e.g., insoluble salts, dietary fiber, cereal flour, starch), or (c) become factors that accelerate or worsen the deterioration (hardening) of the extrudate (e.g., starch retrogradation effect, starch gel aging refers to the rearrangement of starch amylose and amylopectin molecules and the resulting recrystallization, which usually leads to the leathery mouthfeel and hard texture of starch-containing foods such as bread. These phenomena occur most rapidly at temperatures slightly above freezing).

[0176] According to the sixth aspect, a meat substitute product showing an improved taste maintained for a long time can be manufactured by a manufacturing method as follows, replacing the first, second, third, fourth and fifth aspects, or supplementing one, two, three, four or all of them:

[0177] a) feeding a mixture into an extruder configured to perform high moisture protein texturizing extrusion, the mixture comprising:

[0178] a1) at least one protein matrix-forming component, such as a protein isolate or a protein concentrate, and

[0179] a2) has a minimum of 0.125 mm 3 , preferably at least 1 mm 3 , most preferably at least 6 mm 3 Mechanically processed starchy grains having a particle volume of

[0180] b) feeding water into the extruder;

[0181] c) heating the mixture in the extruder to gelatinize the starch-containing grains;

[0182] d) after starch gelatinization is achieved, further heating the mixture in the extruder to melt at least one protein matrix-forming ingredient; and

[0183] e) extruding the mixture through an extrusion die at a temperature between 70°C and 100°C

[0184] in:

[0185] i) carrying out the heating step c) with impingement heating such that the starch-containing grains are gelatinized before they are substantially pulverized by the extruder screw;

[0186] and

[0187] ii) carrying out the heating step d) by shock heating, such that:

[0188] a) before the gelatinized starch and the protein matrix forming component form an emulsion,

[0189] as well as

[0190] b) before the gelatinized starch forms a complete barrier that prevents the formation of a continuous protein fiber cross-linked matrix,

[0191] The protein melting temperature of the protein matrix forming component is reached.

[0192] "Particle volume" and "volume per particle" are terms that describe the size of a particle. They can be calculated based on the dimensions of the particle, for example:

[0193] - When the particles are mostly close to cubic shape, their particle volume can be calculated as length times width times thickness;

[0194] - When the particles are close to a sphere, the particle volume can be calculated using the diameter value of the particles. For example, the average particle size (diameter) can be calculated using the Dv0.5 value in a conventional particle size distribution analysis method.

[0195] Particle size is at least 0.125 mm 3 This indicates that the average volume of the particles is 0.125 mm 3 The particle size of typical commercial oatmeal flour measured by sieving is less than 0.300 mm, from which the average particle volume can be calculated to be no more than 0.014 mm 3 .

[0196] Traditionally, in high moisture protein texturizing extrusion, a heating temperature profile is used in which the temperature gradually increases from the material feed side to the other end of the screw chamber in the extruder, because protein melting is expected to occur at this end of the extruder, and the ingredients gradually absorb heat and increase their temperature. Under the concept of shock heating of the present invention, the materials to be heated to the target temperature in the extruder are heated significantly faster, preferably within a few seconds after they are fed into the extruder, which is before they are conveyed to the last part of the extruder screw chamber.

[0197] Preferably, water is fed to the starch-containing grains at an elevated temperature. The specific heat capacity of water is about 220% higher than that of protein powder and flour. So feeding water at an elevated temperature can heat the material in the extruder to reach the target temperature in a significantly shorter time.

[0198] Preferably, the starch-containing grains are treated before feeding to the extruder such that the starch is gelatinized before feeding to the extruder, in such a way that the size (particle volume) of the grains at least remains the same or even increases.

[0199] The inventors have observed a stable consistency of the first five aspects in the investigated samples with improved mouthfeel.Furthermore, the objects of the present invention can be solved with a method according to a sixth aspect.

[0200] According to either aspect, the meat substitute product and method have in common that the extrudate is an extrudate made using a high moisture protein texturized extrusion process, preferably a twin screw extruder with a long cooling die (the cooling die preferably has a length greater than 300 mm, most preferably greater than 1000 mm). In the extrusion, the machined starch-containing grains are processed with at least one protein isolate / concentrate / combination of the foregoing, oil and flavor to make a slurry which is then extruded.

[0201] The term "mechanically processed" refers to cereal flakes - such as compressed, rolled or flaked - steel cut kernels, shelled and pearled, crushed kernels or shelled but not pearled kernels, however excluding: shelled but not pearled oat kernels, shelled but not pearled rye kernels, shelled but not pearled barley kernels, shelled but not pearled corn kernels.

[0202] The mechanically processed starch-containing grains preferably comprise or consist of one or more of: oats, barley, rye, wheat, rice, corn, lentils, chickpeas, mung beans, broad beans, peas, quinoa, pigeon peas, sorghum, buckwheat, however excluding: hulled but not pearled oat grains, hulled but not pearled rye grains, hulled but not pearled barley grains, hulled but not pearled corn grains.

[0203] The meat substitute product is preferably further processed so that it can be sold in the form of chunks, chops, nuggets, fillets, steaks or doner meat-like slices, or in the form of doner kebab-like layered layers in yogurt or vegetarian yogurt and spices.

[0204] The use of insoluble washable starch in the form of clusters in food products may present interesting possibilities for the food industry.

[0205] The inventors have observed using a microscope equipped with polarized light that the starch in the extruded product does not have the "Maltese cross" feature that it had before extrusion or before immersion in hot water. This shows that the starch in the extruded product is gelatinized.

[0206] The protein fiber matrix structure of the chopped extruded product remained insoluble and intact after testing with the Starch Washability Test. The protein fiber matrix structure of the meat substitute product also remained insoluble and intact after autoclaving in water at 110°C for 10 minutes. The cutting force of the autoclaved meat substitute product remained between 40% and 50% of the cutting force before autoclaving. These are important differences in the properties of products produced by other extrusion processes that are not high moisture protein texturized extrusion. Products produced by other extrusion processes substantially dissolve, soften or collapse after being cooked in water or soaked in warm water overnight.

[0207] According to another aspect, a method for making a meat substitute product using high moisture protein texturized extrusion can be improved by selecting extrusion parameters and starting materials, the starting materials comprising at least: i) a protein component, which is preferably a protein isolate or a protein concentrate or a mixture thereof, ii) a mechanically processed starch-containing grain, and iii) a cereal flour, such that the formation of an emulsion between the starch and the protein melt forming the protein matrix is ​​substantially prevented or reduced to the extent that a large amount of starch that is not bound to the protein matrix is ​​present in the extruded meat substitute product.

[0208] The extrusion parameters controlled preferably include water feed temperature and / or heating profile (eg along the extrusion screw and in the cooling die) such that the starting material is shock heated in the extruder.

[0209] Advantageously, the firmness or compressibility of the meat substitute product is controlled by controlling the starch solubility in the meat substitute product. Most advantageously, the starch solubility is controlled so that the linear compressibility is between 300 g and 1500 g, and the cylindrical compressibility is between 7000 g and 17500 g. Preferably, the linear and cylindrical compressibility are measured at least 24 hours after extrusion.

[0210] Advantageously, the amount of starch not bound to the protein matrix is ​​determined as soluble starch. The compressibility is preferably controlled by varying the extrusion parameters so that the ratio of the amount of soluble starch in the extruded meat substitute product to the total amount of starch (starch solubility) is between 3% and 10% by weight. In this case, the soluble starch content in the extruded meat substitute product is between 0.03% and 1.10% by weight. BRIEF DESCRIPTION OF THE DRAWINGS

[0211] The meat substitute product and the method of making the meat substitute product will be described in more detail below with reference to the accompanying drawings, in which:

[0212] Figure 1 These are photos of samples No. 5, No. 7, and No. 8;

[0213] Figure 2A This is an X-ray micro-tomography (Micro-CT) scan image of sample No. 5 taken after being immersed in water at 60°C for 24 hours and air-dried;

[0214] Figure 2B This is an X-ray micro-CT scan image of sample 8 taken after it was immersed in water at 60°C for 24 hours and air-dried. Figure 2A Cut in the same way.

[0215] Figure 3 The observed relationship between starch solubility and the compression force required to compress a meat substitute product is shown (exponential curve fit to the measurement points);

[0216] Figure 4 shows the particle weight distribution of the extruded material as affected by the component composition and extrusion heating temperature distribution for Experiments 1-6;

[0217] Figure 5 Shown are the results of compression testing of dry (unsoaked) steel cut oats versus soaked steel cut oats (soaked in hot water);

[0218] Fig. 6A and 6B is a microscope image (10x magnification) of a sample taken from sample No. 2;

[0219] Figure 6C and 6D is a microscope image (10x magnification) of a sample taken from sample No. 2;

[0220] Fig. 6E and 6F is a microscope image (10x magnification) of a sample taken from sample No. 6;

[0221] Figure 6G and 6H is a microscope image (20x magnification) of a sample taken from sample No. 6;

[0222] Fig. 7A is a microscope image of a sample of washable starch taken from sample No. 2 washed with water at 50°C;

[0223] Figure 7Bis a microscope image of a sample of washable starch taken from sample No. 2 washed with water at 50°C;

[0224] Figure 8 is an example of a food made from a meat substitute product (Sample No. 2) after being torn into pieces;

[0225] Fig. 9 Examples of food made from a meat substitute product (Sample No. 2) after tearing the extruded product into pieces, marinating the pieces (left side), coating the extruded product with batter, and crumb-coating the extruded product and deep-frying it in oil (right side);

[0226] Fig.10 The gelation of pea protein as affected by heating temperature is shown;

[0227] Fig.11 The cutting force and compression force analysis methods are illustrated;

[0228] Fig. 12A and B illustrate a schematic arrangement of the extrusion process;

[0229] Fig.13 The quantitative analysis methods of soluble starch and washable starch are illustrated;

[0230] Fig.14A Starch coated on the inner surface of the cavity of an extruded product is shown;

[0231] Fig. 14B shows the inner surface of the cavity of the extruded product observed by iodine staining;

[0232] Fig. 14C shows the inner surface of the cavity of the extruded product observed by iodine staining;

[0233] Fig.14D and Fig.14E shows the inner surface of the cavity of the extruded product observed by iodine staining; and

[0234] Fig.15 Photographs of sample No. 2 before (top photo) and after (bottom two photos) expansion are shown.

[0235] The same reference numerals refer to the same parts throughout the drawings. DETAILED DESCRIPTION

[0236] I: Current Status and Purpose

[0237] The mouthfeel of cooked chicken thighs is different from that of chicken breast fillets. The difference in mouthfeel is particularly related to tenderness. Cooked chicken breast fillets generally require a relatively high compression force at a compression ratio of 40%, which indicates that cooked chicken breast fillets generally have a relatively low compressibility.

[0238] As described in the introduction, the inventors have been working on meat substitute products made from high moisture protein texturized extrusion. Fig. 12A An extruder 12 configured to implement a conventional high moisture protein texturizing extrusion process is illustrated. In the extruder 12, the ingredients in powder form are mixed in a mixer 121, which is connected to a supply line 122, which leads to an inlet funnel 123. The extruder 12 has a liquid feed line 124 connected (preferably via a valve 130 and a collection tank 131, to achieve a constant water volume flow) to a conventional tap water supply (tap water typically has a temperature not higher than room temperature or, for example, 30°C). The extruder 12 has a long cooling die 125. Extrusion is carried out with two extruder screws, hence the name "twin screw extruder".

[0239] The research focuses on improving the mouthfeel and finding ways to produce meat substitute products made from high-moisture protein texturized extrusion, so that the meat substitute products have the right high compressibility and chewiness to make it taste as close to steamed chicken thighs as possible. In addition, in order to optimize the mouthfeel, the meat substitute products should have a long continuous fiber protein matrix structure.

[0240] In the market, there are meat substitute products made by extrusion of high-moisture protein texturization, which are sold in shreds or pieces, and when the meat substitute products are cooled after extrusion, they have a mouthfeel comparable to that of steamed chicken breast slices to a certain extent. Table I shows some data of selected existing meat substitute products compared with tofu, chicken breast and chicken thigh.

[0241] Table I: Physical properties of selected meat substitute products on the market

[0242]

[0243] [Oumph!® is a trademark of Food for Progress Scandinavia Ab of Sweden registered in at least the EU, the USA, New Zealand, Switzerland, Australia, the Islands and Norway. The product "chunks" has the composition of water, soy protein (23%) and salt.]

[0244] None of the products the inventors were able to test resembled the steamed chicken thighs, which were more tender, more compressible and had a more pliable structure than steamed chicken breast fillets.

[0245] The cooked chicken thigh meat has a chewy mouthfeel comparable to that of chicken breast fillet due to the long continuous fiber protein matrix structure of the cooked chicken thigh meat.

[0246] In tests conducted by the inventors using high moisture protein texturized extrusion, we found that the mouthfeel of freshly extruded meat substitute products made using high moisture protein texturized extrusion was generally very appealing.

[0247] However, after a relatively short time (usually in the range of a few minutes, typically 5 to 10 minutes), as the meat substitute product cools, the mouthfeel becomes unacceptable. The unacceptable mouthfeel stems from the meat substitute product losing its tenderness, becoming less compressible, and the structure of the meat substitute product becoming less pliable.

[0248] Currently, most meat substitute products made from high-moisture protein texturized extrusion are sold deep frozen. After thawing, these products will have a mouthfeel comparable to that of a steamed chicken breast fillet, which is far from being similar to a steamed chicken thigh.

[0249] In order to improve the mouthfeel of meat substitute products made with low-moisture extruded protein texturization, it is known to add particulates to the extrudate, for example, already including starch; cereal flour; soluble and insoluble polymer fibers such as pea fiber, cellulose, agar-agar, xanthan gum (such as in U.S. Patent Application Publication 2016 / 0205985 A1); insoluble salts such as gypsum (such as in U.S. Patent 5,922,392); and fats that disrupt protein fibers to tenderize the extruded product to produce a meat substitute product (such as in U.S. Patent Application Publication 2016 / 0205985 A1).

[0250] However, most of these compounds are small in size (less than 100 μm in each dimension) before extrusion, or will be broken into small parts (less than 100 μm in each dimension) during extrusion. In practice, they will all be homogenized by the extruder screw and emulsified by the protein material covering them.

[0251] Tolstoguzov [reference 1] has studied and described in detail different types of emulsions in protein extrusion, including emulsions of polysaccharides in proteins. Tolstoguzov found that the emulsion systems extruded under protein texturized extrusion conditions are different from typical water-in-water emulsions or oil-in-water emulsions that exist at temperatures below 140°C. The emulsion of polysaccharides in proteins can be considered as an emulsion of polysaccharide melt in protein melt. During the meat substitute product manufacturing process, i.e., in the high-moisture protein texturized extrusion process, protein is the main ingredient. On a dry basis, protein usually accounts for between 50% and 100% of the weight of the extruded raw material. Typically, in the extruder, vegetable proteins suitable for this type of extrusion process can be melted at a heating temperature between 140°C and 200°C. Therefore, the protein can form a continuous phase.

[0252] Therefore, the microparticles disclosed in US 2016 / 0205985 A1 and 5,922,392 will be dispersed in the protein and form a dispersed phase. The dispersed microparticles are stably captured or embedded in the continuous phase, evenly distributed throughout the continuous phase, and have a small particle size.

[0253] The spinneretless spinning effect in extrusion leads to the formation of anisotropic (fibrous or lamellar) structures in the flowing heterogeneous liquid system.

[0254] In the final stage of the extrusion process, the shape of the emulsion, liquid filaments and anisotropic structures is fixed by the rapid gelation of the protein phase with a gelation time that is shorter than the lifetime of the liquid filaments. After this, if the protein matrix structure or the protein layer covering the dispersed particles is not broken apart, the dispersed particles remain uniformly dispersed, firmly embedded, and can hardly be separated from the protein matrix by mechanical forces (e.g. centrifugation, gravity) or by extraction (e.g. water washing, water extraction).

[0255] When producing meat substitute products using protein texturized extrusion, it is known that known methods of including microparticles in the extrusion will tenderize the extruded product to a certain extent, especially when the extruded product is freshly produced and before being cooled and stored overnight. The microparticles can disrupt the protein fibers by being present in between the protein fibers or between adjacent protein fibers.

[0256] The addition of such microparticles can also dilute the concentration (ratio) of protein in the ingredients used for extrusion, which forms the protein fiber matrix and contributes to the strength of the extruded product. In this way, the addition of microparticles can soften the extruded product, especially when the product is fresh and warm before being stored overnight at a cooling temperature (e.g., between 0°C and 6°C). In low-moisture protein texturized extrusions (e.g., the moisture content of the material during extrusion is between 15% and 40%) for the production of meat substitute products, the extruded products mostly have a large amount of expansion and contain a large number of bubbles between the protein fibers. The expansion and bubbles are attributed to the large amount of water evaporation that occurs when the extruded material just leaves the extruder die at high temperatures (e.g., above 100°C). In such cases, the broken protein fibers are further separated by the bubbles and fixed in separate (distanced) positions from each other. Therefore, in low-moisture protein texturized extrusions for the production of meat substitute products, the breaking effect from these microparticles can be attractive to a certain extent.

[0257] However, in high moisture protein texturized extrusions used in the production of meat substitute products (e.g., moisture content of the material during extrusion is between 40% and 80%), the extruded material expands much less and the protein fibers are evenly distributed among them with far fewer bubbles that disrupt their cross-links between adjacent fibers.

[0258] Akdogan [reference 2] found that the reduced expansion level in high moisture protein texturized extrusions is caused by the increased concentration of water during extrusion. More specifically, the higher moisture content extrusion has a different distribution of shear (generally less shear forces are present in high moisture protein texturized extrusions), mixing, mechanical heat (generally less mechanical heat dissipation in high moisture protein texturized extrusions), and convective heat. The high moisture content extrusion has much less viscous dissipation of energy in the extruder barrel due to the greatly reduced melt viscosity and greatly reduced pressure buildup in the extruder barrel. The pressure along the die is greatly reduced, and this is therefore partly responsible for the minimal to non-existent expansion at the die. The use of a long cooling die during high moisture protein texturized extrusion cools the extruded material, and thus water evaporation is greatly reduced. It is also known from the background art that when the starch content of the extruded material is low, and when the starch gelatinization level is low, the expansion level of the extruded material leaving the extrusion die will be lower. The low viscosity of the extruded material associated with the high moisture content also results in a certain degree of inability to maintain (maintain) expansion stability from collapsing into a dense sheet.

[0259] Variations in moisture content during extrusion can also result in changes in the major contributing protein-protein forces that stabilize the protein matrix. Lin et al. [Reference 3] found that under high moisture extrusion (e.g., when the moisture content during extrusion was between 40% and 80%), a large portion of the protein was linked and stabilized by hydrogen bonds, while disulfide bonds and hydrophobic interactions were not the major forces stabilizing the protein. In contrast, under low moisture extrusion (e.g., when the moisture content during extrusion was between 30% and 40%), the major important protein matrix stabilizing forces were disulfide bonds and hydrophobic bonds. After extrusion, during the cooling process, hydrogen bonds in the protein matrix can significantly contribute to further increase the gel strength (firmness) of the extruded product. It is well known and disclosed by Sun and Arntfield [Reference 4] that low temperatures (e.g., between 0°C and 6°C) for storage and the cooling process after protein gel formation can favor the extensive and increasing formation of hydrogen bonds. In addition, it is also known that during the cooling process after starch is heated and gelatinized in water, the starch gel strength also mostly and significantly increases because of the extensive generation of hydrogen bonds between starch molecules during cooling. Starch retrogradation can occur after starch gelatinization. Longer storage time periods lead to further formation of hydrogen bonds and therefore to further tightness (firmness) of the structure, as well as lower water holding capacity. Thus, starch gelatinization and retrogradation are another factor leading to the problem of firm texture and loss of attractive mouthfeel of meat substitute products produced by high moisture protein texturized extrusion in the known methods of the background art.

[0260] In the context of baking bread, the adverse effects of retrogradation on crumb texture are well known: retrogradation significantly leads to increased crumb staling and firmness during storage time.

[0261] Hydrogen bonds are short-range chemical bonds, which means that crosslinks associated with hydrogen bonds occur primarily between adjacent compounds that are in close or direct contact with each other (e.g., protein-protein, protein-starch, starch-starch). Starch of the amylose type has a high ability to form starch-starch hydrogen bonds because it has many hydroxyl groups and linear polymer chains in its molecular structure. Starch before gelatinization cannot form a gel in water because the starch is embedded in the starch granule structure and is therefore insoluble. Starch gelatinization may occur more excessively during high-moisture extrusion than in low-moisture extrusion. During high-moisture protein-textured extrusion, the starch is heated sufficiently and leached into the water by heat and shear forces, causing the leached amylose molecules to be arranged linearly and in close contact with each other.

[0262] Since extruded products from high moisture protein texturized extrusions have higher compactness (less expansion, higher density) and more excessive formation of hydrogen bond type protein-protein crosslinks than those from low moisture extrusions, the addition of microparticles (such as starch powders, insoluble salts, fibers, fats, etc.) does not nearly as well to disrupt the excessive protein-protein crosslinks or interaction forces that occur during the cooling stage and after extrusion as in low moisture extrusions. Therefore, these extruded products with or without the addition of microparticles still suffer from structural hardening (firmness) and loss of acceptable mouthfeel (e.g., compressibility) during cooling and storage time. More specifically, the microparticles are easily homogenized, covered, and emulsified by the protein matrix either very quickly during extrusion or immediately after the microparticles are extruded together with the protein material. The microparticles are then unable to provide sufficiently large disruptive forces or barrier effects between protein fibers and may only be able to provide limited disruptive areas around each individual microparticle point without extending. More seriously, when starch is added in the form of starch powder (including or not including modified starch or pregelatinized starch) or cereal grain flour powder, they are also homogenized, covered and emulsified by protein matrix very soon after extruding with protein material. Then, emulsified starch is heated and gelatinized. In the whole extrusion process and in the final product, starch remains as small particles. So starch can hardly provide large destructive power or barrier effect between protein fibers, and may only be able to provide limited destructive area around each individual particle point, and will not extend. After extrusion, the protein matrix around the starch particles can continue to become firm, forming protein-protein interaction forces, such as more hydrogen bonds. In addition, starch becomes extremely easy to stand starch gelatinization, retrogradation, hardening, drying and form possible starch-protein interaction with hydrogen bonds after being sheared, gelatinized, distributed in (between) linearly arranged protein fibers and linearly arranged. Like this, extruded products stand the very important problem of structural hardening (firmness) and losing acceptable mouthfeel (for example compressibility) during cooling and storage time.

[0263] II: Processing machine (extruder system) for carrying out the tests described in the following examples

[0264] Fig. 12B An extruder 13 configured to implement a high moisture protein texturizing extrusion process for implementing the method described in accordance with the present invention is illustrated. The extruder 13 implements the technical features required for the new process.

[0265] In the novel process, the machined starch-containing grains are mixed with the starch-containing grains in powder form (preferably flour), at least one (preferably vegetable or dairy) protein isolate / at least one (preferably vegetable or dairy) concentrate / a mixture of at least one such isolate and at least one such concentrate, possible oils and possible flavors and any other ingredients in a mixer 121 and fed into an extruder 13 via a feed line 122, for example via an inlet funnel 123. The extruder 13 has a liquid feed line 124 connected to a water heating element 14, which is configured to provide heated water (so that the heated water is significantly higher than the temperature of tap water, for example, has a temperature of at least 50° C.) and is preferably configured to provide water with a stable temperature (for this purpose, the heating element 14 preferably has a pump 132 and a heater tank 133, and the heater tank 133 preferably has a water heating element and a temperature probe). The extruder 13 also comprises a long cooling die 125. The pump 132 can be controlled so that the water fed to the tank 131 always has a target temperature, and the pump 130 can feed water into the extruder 13 at a target flow rate (e.g., kilograms of water per hour). If the tap water is directly connected to the tank 131, and an attempt is made to heat the water in the tank 131, it will be more difficult to accurately control the temperature of the water.

[0266] In the following examples, experiments performed by the inventors are described in more detail.

[0267] III: First Experiment (Examples 1 and 2)

[0268] In the following, and also throughout the description of the compositions of the samples in the other experiments and tests, the percentages of the compositions are given in % by weight on a dry basis.

[0269] Using Examples 1 and 2, we demonstrate exemplary parameters of the manufacturing process (ingredients, shock heating) and their impact on the quality of the resulting meat substitute product (e.g. in terms of specific physical properties such as compressibility, hardening, expansion, cavity structure).

[0270] Mechanically processed starchy grains Comprising or consisting of one or more of: flakes (such as compressed, rolled or flaked), steel cut kernels, shelled and pearled kernels, crushed kernels, shelled but not pearled kernels.

[0271] Mechanically processed starchy grains Contains or consists of one or more of the following: oats, barley, rye, wheat, rice, corn, lentils, chickpeas, mung beans, broad beans, peas, quinoa, pigeon peas, sorghum, buckwheat.

[0272] However, the following ingredients are excluded Mechanically processed starchy grains Others: hulled but not pearled oat kernels, hulled but not pearled rye kernels, hulled but not pearled barley kernels and hulled but not pearled corn kernels.

[0273] Although other extruder configurations can be used, the extruder 13 used to carry out the experiments is a twin-screw co-rotating extruder with a screw 126 having a diameter between 30 mm and 50 mm. The extruder 13 has a screw chamber 138 surrounding the screw 126. The configuration used has a screw chamber 138 with 6 zones (although another number of zones is possible), which can be numbered from the side where the solid ingredients are fed into the extruder and the extrusion starts as zone 1 to zone 6. Thus, there is an inlet hole 139 for feeding the solid ingredients (e.g. in zone 1). Zones 2, 3, 4, 5 and 6 are all equipped with heating, cooling and temperature sensing elements, which preferably can control the temperature of each zone individually between, for example, 10° C. and 220° C. In addition, there is an inlet hole 140 for feeding a liquid into the extruder 13 to be extruded together with the solid ingredients (e.g. in zone 2).

[0274] At a typical screw rotation speed (e.g., between 150 rpm and 300 rpm), the material can take about 45 s to 75 s to pass through the screw chamber 138. The inventors set to allow the liquid feed line 124 and the heating element 14 to feed water with water at different temperatures between 5°C and 99°C, for example, in some cases, the heated water is fed into the tank 131, and the heated water is pumped to the extruder 13 by the pump 130 of the liquid feeder. A test was conducted to stop the extruder and remove the screw after a continuous run of extruding dry oat flakes without water. It was observed that with a spiral milling time of 5-15 s (e.g., calculated by the conveying distance), at about zone 2, the oat flakes were mostly (more than 90%) and substantially pulverized into grain-like particles that were significantly smaller than their original size (e.g., their size was less than 200 μm).

[0275] Differently, the conventional liquid feed line 124 is connected to regular tap water and feeds tap water between 5°C and 25°C to the extruder (eg Fig. 12A The feed rates (eg kg / h) of the solid component and the liquid can be controlled individually.

[0276] After the last zone (e.g. zone 6), there is a long cooling die 125 connected to the extruder 13, which also has heating, cooling and temperature detection elements. The long cooling die 125 is longer than 300 mm, preferably its length is between 300 mm and 5000 mm, most preferably between 1000 mm and 3000 mm. Between the last zone (e.g. zone 6) and the cooling die 125, there are pressure detection sensors and temperature detection sensors. In addition, a tool can be connected after the long cooling die 125.

[0277] Those skilled in the art have sufficient knowledge from the background technology to know how to adjust or select the screw 126 diameter, screw 126 speed, cooling die 125 length and shape, tool type and cutting speed according to different types of customized requirements such as production stability, production speed, product size and shape.

[0278] Example 1 (Samples No. 1, 2, 3, 4) - Effect of ingredients on textural properties of extruded products

[0279] The inventor prepared 4 samples (No. 1, No. 2, No. 3, No. 4) using Fig. 12B The extruder 13 shown in FIG. 1 is used for texturizing extrusion processing with high moisture protein.

[0280] Sample No. 1 contained 90 wt% pea protein, 5 wt% oat flour, 4 wt% fiber, to which other ingredients (such as salt, spices, yeast extract, oil, oat malt extract, non-starchy grains - such as sunflower seeds) were added.

[0281] Sample No. 2 contained 90 wt% pea protein, 5 wt% steel cut oats, 4 wt% fiber, with other ingredients added (such as salt, spices, yeast extract, oil, oat malt extract, non-starchy grains - such as sunflower seeds).

[0282] Sample No. 3 contained 62 wt% pea protein, 20 wt% oat flour, 10 wt% fiber, to which other ingredients (such as salt, spices, yeast extract, oil, oat malt extract, non-starchy grains - such as sunflower seeds) were added.

[0283] Sample No. 4 contained 62 wt% pea protein, 1 wt% steel cut oats, 19 wt% oat flour, 10 wt% fiber, to which other ingredients (such as salt, spices, yeast extract, oil, oat malt extract, non-starchy grains - such as sunflower seeds) were added.

[0284] After production, samples No. 1, No. 2, No. 3, No. 4 were cooled and stored overnight. The next day their mechanical properties were measured to study the texture. The results of the measurements are shown in Table II.

[0285] The results in Table II show that samples No. 1 and No. 3 produced from ingredients comprising a starchy flour (oat flour) had a hard and rubbery texture and had a high resistance to cylindrical compression.

[0286] The results in Table II also show that Samples No. 2 and No. 4, in which the starchy flour (oat flour) was replaced or partially replaced by starchy grains (steel cut oats), were more flexible and compressible than Samples No. 1 and No. 3.

[0287] After cooking in water at 110°C in a pressure cooker (e.g. in a pressure cooker), sample No. 2 has a much higher thickness cooking expansion (265%) than sample No. 1 (143%). This difference is caused only by the change in the starch-containing ingredient (from flour to steel-cut grains). Other conditions such as extrusion parameters remain the same; and the ingredients have the same chemical (nutrient) composition.

[0288] Table II. Texture of samples No. 1, No. 2, No. 3, and No. 4

[0289]

[0290] As protein in Example 1, we used pea protein isolate. It can be replaced at least partially by pea protein concentrate, or by any other protein isolate or protein concentrate (such as bean, soy, chickpea, wheat gluten, oat), dairy (milk or whey) protein, or a mixture of at least one of these. The results are comparable.

[0291] The grain used in Example 1 is steel cut oats. It can be used as described above (please note the excluded species described above). Mechanically processed starchy grains Substitution, in particular, with steel cut barley, rice grains, cracked rice, pearled barley, pearled rye, pearled wheat, pearled oats, crushed pea seeds (e.g. with a particle size of 2 mm), crushed broad bean seeds, crushed chickpea seeds, lentil seeds, etc. and mixtures thereof. The results are comparable.

[0292] In this embodiment, before extrusion, Mechanically processed starchy grains Soak in hot water. Soaking is done by gently mixing the grains with hot water (e.g. 90°C) 1:2 and then keeping at a warm temperature (e.g. 75°C) for 2 hours. After soaking, the grains absorb all the water and become softer and larger.

[0293] The flour in Example 1 is oat flour. It can be replaced by barley flour, wheat flour, rice flour, pea flour, chickpea flour, bean flour, lentil flour, etc. and mixtures thereof. The results are comparable.

[0294] The fiber in Example 1 is pea fiber. It can be replaced by oat fiber, oat bran, potato fiber, broad bean fiber, etc. and mixtures thereof. The results are comparable.

[0295] Other ingredients in Example 1 include all of the following: salt, spices, yeast extract, oil, oat malt extract, non-starchy grains (such as sunflower seeds), etc. Some of these may be omitted or replaced with other ingredients as desired.

[0296] The resistance to cutting with a sharp blade was measured as the cutting force in Example 1. The measurement was carried out using the texture analyzer described above.

[0297] The resistance to compression with a cylinder was measured as the compression force in Example 1. The measurement was carried out using the texture analyzer described above.

[0298] As for the texture observations in Example 1 in Table II, the texture property observations recorded were analyzed by an expert panel performing sensory evaluation.

[0299] Extrusion parameters used in Example 1:

[0300] (1) Liquid feed: hot water (e.g., with an elevated temperature of 65° C.);

[0301] (2) The moisture content of the slurry (extruded material) during extrusion is about 50%. Depending on the desired properties of the extruded product (e.g., moisture content, color, etc.) and composition changes (e.g., different proteins may have different melting requirements, and different starches may have different gelatinization requirements), the moisture content of the slurry can be adjusted between 40% and 80%;

[0302] (3) Extruder heating profile: an impact heating profile with a temperature of 80-125-160-145-130 (° C.) in zones 2-3-4-5-6. The cooling die temperature is 90° C. Depending on the change in ingredients (e.g., different proteins may have different melting temperatures, and different starches may have different gelatinization temperatures), the temperature may be adjusted within the range described in the method item;

[0303] (4) Productivity: about 18 kg of product per hour. Pressure at the end of the screw: between 1.0 mPa and 3.0 mPa.

[0304] (5) After extrusion, the extruded product is immediately immersed in water (e.g., 20° C.) for 2 hours to cool and prevent drying. Then it is taken out of the water. After being stored in a cold room (e.g., 5° C.) for 24 hours, the samples are analyzed for cutting force, compression force, texture observation, and thickness cooking expansion rate.

[0305] NA stands for not analyzed.

[0306] The expansion in Example 1 represents the thickness cooking expansion rate analyzed by the cooking test method, which will be described below. Unless otherwise specified such as "extrusion expansion rate", throughout this application, "expansion" or "expansion rate" always refers to thickness cooking expansion rate.

[0307] In further experiments, the ingredients of sample No. 1 (90 wt% pea protein + 5 wt% oat flour + 4 wt% fiber, with other ingredients added) were processed with different extrusion parameters, such as different liquid feed water temperatures (15°C-90°C), extruder heating profiles ("shock heating" such as (in 2-3-4-5-6 zones) 80-125-160-145-130°C), "overheating" such as 80-125-160-160-160°C, "slow heating" such as 40-75-100-140-165°C, all produced unacceptable products (similar to sample No. 1) with a hard and rubbery structure and mouthfeel, cutting forces between 500 g and 1100 g, compression forces between 18 200 g and 44 000 g, and cooking expansion ratios between 125% and 149%. The results from these experiments were not satisfactory. The taste is totally incomparable to that of steamed chicken thighs.

[0308] Replacing oat flour with other starchy flours such as oat starch, potato starch, rice starch, chickpea starch, wheat starch, pea starch, etc. also produced unacceptable results similar to Sample No. 1. The inventors performed a large number of tests.

[0309] Substituting non-starchy grains such as sunflower seeds, peanut flakes, almond seed flakes, coconut particles, chia seeds for the oatmeal also resulted in an unacceptable product similar to Sample No. 1.

[0310] Replacing oat grist with starchy grains with intact husks or intact, thick and strong seed coat (also known as pericarp layer, bran layer) or intact husks, such as whole oat seeds, whole barley seeds, whole rye seeds, also produced an unacceptable product similar to Sample No. 1.

[0311] However, adding between 0% and 20% (preferably between 0% and 10%) of these microparticles (e.g., sunflower seeds, chia seeds, whole oat seeds) to the ingredients to partially replace the protein in acceptable samples such as Sample No. 2 will not adversely affect the quality of the extruded product.

[0312] Adding additives such as calcium chloride, calcium carbonate, gypsum powder (calcium sulfide dihydrate), baking powder, psyllium, alginates, ascorbic acid, xanthan gum, agar-agar, etc. to the ingredients of sample No. 1 did not produce the desired properties observed in acceptable samples such as sample No. 2.

[0313] However, it is still possible to add some of these additives (such as baking powder, gypsum powder, ascorbic acid) between 0% and 5% to the other ingredients of acceptable samples such as Sample No. 2 without causing serious adverse effects on the quality (compression characteristics and mouthfeel) of the extruded product.

[0314] Example 2 (Samples No. 5, 6, 7, 8, 9) - Effect of extrusion composition and extrusion heating profile on the texture and expansion properties of the extruded product.

[0315] The inventor prepared 5 samples (No. 5, No. 6, No. 7, No. 8, No. 9), which were Fig. 12B The extruder 13 shown in FIG. 1 is used for texturizing extrusion processing with high moisture protein.

[0316] Sample No. 5 contained 70 wt% pea protein and 30 wt% oat flour.

[0317] Like sample No. 5, sample No. 6 contains 70 wt% pea protein and 30 wt% oat flour.

[0318] Sample No. 7 contained 70 wt% pea protein, 10 wt% oat flakes, and 20 wt% oat flour.

[0319] Like sample No. 7, sample No. 8 comprises 70 wt% pea protein, 10 wt% oat flakes, and 20 wt% oat flour.

[0320] Sample No. 9 contained 70 wt% pea protein, 20 wt% oat flakes, and 10 wt% oat flour.

[0321] After production, samples No. 5, No. 6, No. 7, No. 8, No. 9 were cooled and stored overnight. The next day their mechanical properties were measured to study the texture. The results of the measurements are shown in Table III.

[0322] Table III. Texture of samples No. 5, No. 6, No. 7, No. 8, and No. 9

[0323]

[0324] Table III shows that the oat flakes containing extruded products Sample No. 8 and Sample No. 9 produced by extrusion with shock heating temperature profile (hot water liquid feed used with temperature profile 80-125-160-145-130° C. in the 2-3-4-5-6 zone) have a more flexible and compressible texture, which produces a very good mouthfeel and is pleasant to eat. After cooking in water, it also has a high cooking expansion ratio (189% - 206%), which is consistent with its nature of having a flexible and extensible structure and texture.

[0325] When oat flakes were completely replaced by oatmeal flour with the same chemical composition but much smaller particle size (sample 6), the extruded product became hard, rubbery and had a lower cooking expansion (129%). The mouthfeel was not comparable to that of cooked chicken thighs. The shock heat extrusion condition did not result in a large difference between the products without oat flakes (between samples 5 and 6).

[0326] When oat flakes were used in extrusion without shock heating settings (e.g. if the liquid feed water temperature was 25°C and the zone 2 temperature was set to 40°C), the product (Sample No. 7) had a hard and rubbery texture and low overrun (164%). The mouthfeel was not comparable to that of steamed chicken thighs.

[0327] The protein in Example 2 is pea protein isolate. It can be replaced by other proteins in the manner described above and below in Example 1.

[0328] Oat flakes were used as the mechanically processed starch-containing grain in Example 2. Oat flakes may be replaced by other mechanically processed starch-containing grains in the manner described above and in the context of Example 1. In particular, barley flakes, steel cut oats, steel cut barley, rice grains, cracked rice, pearled barley, pearled rye, pearled wheat, etc. and mixtures thereof may be used. The results are comparable.

[0329] In Example 2, the machined starch-containing grains were not immersed in hot water prior to extrusion.

[0330] The flour in Example 2 is oat flour. It can be replaced by barley flour, wheat flour, rice flour, pea flour, chickpea flour, broad bean flour, quinoa, pigeon pea, sorghum, buckwheat, etc. or a mixture thereof. The results are comparable.

[0331] The expansion in Example 2 represents the thickness retort expansion ratio analyzed by the retort test method, which will be described below.

[0332] The visible air cavities in Example 2 represent visible air cavities in an extruded product analyzed by a visual inspection method, which is described below.

[0333] The texture observations in Example 2 represent textural property observations recorded by an expert panel of sensory evaluations.

[0334] Extrusion parameters:

[0335] (1) The moisture content of the slurry (material being extruded) during extrusion is about 50%;

[0336] (2) After extrusion, the extruded products were immediately immersed in water (20°C) for 2 hours to cool and prevent drying. They were then taken out of the water. After storage in a cold room (e.g., 5°C) for 24 hours, the samples were analyzed for texture observation, visible air cavities, thickness, and cooking expansion.

[0337] (3) Productivity: About 18 kg of product per hour. The cooling mold temperature is 90°C.

[0338] Examples of air cavities can be found in Figure 1 As seen in sample No. 8 in Figure 2.

[0339] IV: Results of the First Experiment

[0340] Figure 1 The following are photos of samples No. 5, No. 7, and No. 8 (from bottom to top) after being immersed in water at 60°C for 24 hours: On the right, the sample was cut parallel to the fiber direction so that the fiber, length, and thickness of the sample are visible. On the left, the sample was cut across the fiber direction so that the cross-section (width and thickness) of the sample is visible. Sample No. 8 has significantly more visible air cavities than Samples No. 7 and No. 5. The air bubbles in Sample No. 8 are more evenly distributed in the protein fiber matrix, with a larger total volume and a larger average size than the air bubbles in Samples No. 5 and No. 7. Figure 1 The presence of white particles in sample No. 7 included intact oat flake particles within the protein matrix. The inclusion of particles did not solve the problem of a rubbery, hard and difficult to compress product. The visible particles were not powdered by the extruder, mainly due to the fact that a very small portion (e.g., less than 5%) of the particles slipped through the narrow gap between the screw and the screw chamber. They remained largely intact throughout the extrusion and were not effectively mixed with the other ingredients. The degree of gelatinization of these particles was insufficient and much lower than that of other particles that were effectively mixed by the screw (e.g., those that were powdered in sample No. 7). At the end of the process, they were covered with other materials. They were unable to disrupt the overall formation of the protein fiber structure or the growth of the inter-fiber interaction forces. These and Figure 1 These are confirmed by microscopic studies (although no photographs are provided in this application) and textural (compressibility) studies.

[0341] Figure 2AThis is an X-ray micro-tomography (Micro-CT) scan image of sample No. 5 taken after being immersed in water at 60°C for 24 hours and air-dried. The sample was cut parallel to the fiber direction so that the fiber, length and thickness of the sample are visible.

[0342] Figure 2B This is an X-ray micro-CT scan image of sample No. 8 taken after it was immersed in water at 60°C for 24 hours and air-dried. Figure 2A The samples were cut in the same manner. Figure 2A and Figure 2B The difference between the two is clear, and it can be seen that sample No. 8 has more air bubbles (black cavities between white fibers), which are widely and evenly distributed in the protein fiber matrix, with a larger total volume and a larger average size than sample No. 5. In addition, sample No. 8 obviously has a long continuous fiber structure. The fibers of sample No. 8 are thinner and have a more uniform thickness than the fibers of sample No. 5. Most of the fibers are parallel to each other. This shows that although protein fibers tend to stick to each other and form larger bundles or groups, the protein fibers are well broken and separated in sample No. 8. The thinner fiber structure of sample No. 8 contributes to a good, chewy and compressible texture, which is close to the chicken thigh meat of steaming. The aggregation and layered structure of sample No. 5 makes it have an unfavorable, hard, leather-like and rubber-like texture.

[0343] Fig. 6A is a microscope image of a sample taken from sample number 2. The sample was stained with a protein stain (Thermo Scientific Pierce Coomassie Brilliant Blue R-250). The sample was observed using an optical microscope (Zeiss AxioLab.A1 laboratory microscope) at 10x magnification. The protein fibers are stained black. The protein fibers are continuous throughout the image and are much larger than 1 mm in length. The protein fibers are mostly arranged parallel to each other. Crosslinking is low, with only a few connections between adjacent fibers.

[0344] Figure 6B is a microscope image of a sample taken from sample No. 2. The sample was stained with a dilute iodine solution, such as a 1:5 diluted Sigma-Aldrich Lugol's solution stabilized with polyvinyl pyrrolidone for Gram staining. The sample was observed with an optical microscope at 10X magnification. The dark black material (lumps) indicated starch-rich material, which formed a dark blue iodine-starch complex with the iodine stain. Figure 6BThe protein fiber matrix is ​​also shown in gray, which is lighter in color than the starch material and more transparent than the starch material, but not completely transparent. The starch-rich material appears rounded or random in shape, is not tightly embedded in the protein fiber matrix, and is not evenly distributed throughout the structure. These findings indicate that the starch is in the form of clusters, a phase separated from the protein phase, and is not emulsified by the protein.

[0345] Figure 6C is a microscope image of a sample taken from sample No. 2. Fig. 6A The samples were stained with a protein dye in and observed at 20x magnification. The protein fibers were mostly arranged parallel to each other. Cross-linking was low, with only a few connections between adjacent fibers.

[0346] Fig.6D is a microscope image of a sample taken from sample No. 2. The sample was stained with a dilute iodine solution, such as 1:5 diluted Sigma-Aldrich Lugol's solution stabilized with polyvinyl pyrrolidone for Gram staining, and observed at 20X magnification. The dark black material (lumps) indicate starch-rich material, which forms a dark blue iodine-starch complex with the iodine stain. Fig.6D The protein fiber matrix is ​​also shown in gray, which is lighter in color than the starch material, more transparent than the starch material, but not completely transparent. The starch-rich material presents a round or random shape, is not tightly embedded in the protein fiber matrix, and is not evenly distributed throughout the structure. These findings show that starch is in the form of clusters, is the phase separated from the protein phase, and is not emulsified by protein. There are starch clusters (shown as dark spots) greater than 30 μm in size (e.g., length).

[0347] Fig. 6E is a microscope image of a sample taken from sample number 6. The sample was stained with a protein stain such as ThermoScientific Pierce Coomassie Brilliant Blue R-250 and viewed at 10x magnification. The protein fibers are stained black. The protein fibers are continuous throughout the image and are much larger than 1 mm in length. The protein fibers are mostly aligned parallel to each other. Cross-linking is high: Fig. 6E The connections between adjacent fibers are significantly Fig. 6A The richer in. Fig. 6E The interstitial space between adjacent fibers is significantly larger than Fig. 6A There are two rows of bright white spaces between the three protein fibers. They are the empty spaces between two protein fibers.

[0348] Fig. 6Fis a microscope image of a sample taken from sample No. 6. The sample was stained with a dilute iodine solution, such as 1:5 diluted Sigma-Aldrich Lugol's solution stabilized with polyvinyl pyrrolidone for Gram staining, and observed at a magnification of 10. The dark black material (lumps) indicate starch-rich material, which forms a dark blue iodine-starch complex with the iodine stain. Fig. 6F The protein fiber matrix is ​​also shown in gray, which is lighter in color than the starch material, more transparent than the starch material, but not completely transparent. The starch-rich material is in a narrow linear shape and is tightly embedded in the protein fiber matrix, and is obviously substantially evenly distributed between and along the entire structure of the protein fibers, and the distribution, shape and distribution of the starch-rich material are highly ordered. These indicate that the starch is emulsified by the protein.

[0349] Figure 6G is a microscope image of a sample taken from sample number 6. The sample was stained with a protein stain such as ThermoScientific Pierce Coomassie Brilliant Blue R-250 and viewed at 20x magnification. The protein fibers are stained black. The protein fibers are mostly arranged parallel to each other. Cross-linking is high: Figure 6G The connections between adjacent fibers are significantly Figure 6C The richer in. Figure 6G The interstitial space between adjacent fibers is significantly larger than Figure 6C The one in is narrower and smaller.

[0350] Figure 6H is a microscope image of a sample taken from sample No. 6. The sample was stained with a dilute iodine solution, such as 1:5 diluted Sigma-Aldrich Lugol's solution stabilized with polyvinyl pyrrolidone for Gram staining, and viewed at 20X magnification. The dark black material (lumps) indicate starch-rich material, which forms a dark blue iodine-starch complex with the iodine stain. Figure 6H The protein fiber matrix is ​​also shown in gray, which is lighter in color than the starch material, more transparent than the starch material, but not completely transparent. The starch-rich material presents a narrow linear shape, is tightly embedded in the protein fiber matrix, and is obviously evenly distributed between the protein fibers and along the entire structure of the protein fibers. The distribution, shape and distribution of the starch-rich material are highly ordered. These indicate that the starch is emulsified by protein.

[0351] Fig. 7A is a microscope image of a sample of washable starch taken from sample No. 2 washed with water at 50°C. Fig. 7AInsoluble washable starch (black material in the image) in the form of clusters is shown, with sizes ranging from 50 μm to 800 μm. Each cluster contains more than 5 individual starch granules (circles) therein. Within each cluster, the individual starch granules are tightly bonded to each other. The samples were observed under an optical microscope at a magnification of 5 times.

[0352] Figure 7B is a microscope image of a sample of washable starch taken from sample No. 2 washed with water at 50°C. Figure 7B Insoluble washable starch (black material in the image) is shown to be present in the form of clusters, approximately 100 μm in size. Each cluster contains more than 5 individual starch granules (circles) therein. Within each cluster, the individual starch granules are tightly bonded to each other. There is starch leaching out of the clusters of aggregated starch granules into the water. This leached starch allows these clusters to be "washed" by 50°C water. The starch embedded in such clusters is insoluble in 50°C water, but soluble in 110°C water. The samples were observed under an optical microscope at a magnification of 20 times.

[0353] Fig.10 The gelation of pea protein affected by heating temperature is shown. In order to see how the heating temperature can affect the gelation of pea protein, pea protein was mixed with water in a 1:1 ratio, then packaged into vacuum bags, and then heated at different temperatures (50°C to 110°C). The texture of the gel / block was then measured. As can be seen from the results in the table, the samples heated to 90°C and above have significantly higher hardness. These show that when heated to 90°C or above, a significantly firmer gel is formed.

[0354] Fig.14A The starch coating on the inner surface of the cavity of the extruded product as observed by iodine staining and visual inspection is shown. Left: Slice of sample No. 2. Right: Slice of a sample produced under similar conditions to sample No. 2, but using whole oat kernels that were hulled but not pearled instead of the steel cut oats used in sample No. 2. The sample on the right had an unacceptable texture: for example, the compression force exceeded 20 000 g.

[0355] The two samples were cut into slices about 1 mm thick, about 10 mm wide and 40 mm long. The length direction was mainly parallel to the direction of fiber orientation. One slice of each sample was stained for 45 minutes with a diluted Lugol's solution (iodine solution for staining) ranging from 1 mL to 3 mL to cover the sample in all directions. The stained sample was then gently moved and immersed in 50 ml of water for 5 minutes. We then placed the slices on white paper for visual observation.

[0356] Fig.14AThe gray blocks in the photos refer to the overall structure (protein matrix structure and all other materials embedded in the protein matrix structure). The darker colors (black) indicate starch-enriched materials.

[0357] The slice of sample No. 2 (ie, the left side) has obvious dark coating material on the inner wall and the outer wall (surface) of the cavity of the extruded product.

[0358] The slice of another sample (i.e., on the right) has dark colors within the structure as large spots (e.g., 1 mm dots). The dark spots should be uncracked oat seeds. The sample contains visible uncracked seeds as inclusion particles, but it has an unacceptable texture.

[0359] No obvious dark coating material was found in No. 1, No. 3, No. 5, No. 6 and No. 7.

[0360] Fig. 14B Shown is the inner surface of the cavity of the extruded product observed by iodine staining and microscopy (5 times magnification, using a stereo microscope, such as a Zeiss Stemi 305 stereo microscope). The sample specimen is taken from sample No. 2. The sample was stained for 30 minutes with a diluted Lugol's solution (iodine solution for staining) before observation. The gray blocks in the photo refer to the overall structure (protein matrix structure and all other materials embedded in the protein matrix structure). Dark (black) indicates the material enriched in starch content. When observed via a microscope, the color view is blue or dark blue or black.

[0361] Fig. 14C The inner surface of the cavity of the extruded product observed by a microscope with 20 times magnification by iodine staining is shown. The sample sample is taken from sample No. 2. The sample was dyed for 30 minutes with a diluted Lugol's solution (iodine solution for dyeing) before observation. The dark gray block with a specific fiber (anisotropic) structure in the figure (from the left to the middle of the picture) refers to the overall structure (protein matrix structure and all other structures embedded in the protein matrix). There is a cluster of black dots on the left side of the picture, indicating a starch cluster of gelatinization. The light gray blocks (on the right side of the picture) adjacent to very bright white and blank areas indicate the material enriched in starch content. Because the wall is more directly exposed to the microscope light, the starch at the cavity wall observed with this magnification and angle has a lighter color than the protein matrix structure. When observed via a microscope, the starch at the cavity wall observed with this magnification and angle is light blue.

[0362] Fig.14D and Fig.14EThe inner surface of the cavity of the extruded product observed by iodine staining and microscopic examination (40 times magnification) is shown. The sample sample is taken from sample No. 2. The sample was stained for 30 minutes with a diluted Lugol's solution before observation. The dark gray block with a specific fiber (anisotropic) structure in the figure refers to the overall structure (protein matrix structure and all other structures embedded in the protein matrix). The light gray block without fiber structure adjacent to the very bright white and blank area (the middle of the picture) indicates the material enriched in starch content. The starch at the cavity wall observed with this magnification and angle has a lighter color than the protein matrix structure. When observed via a microscope, the starch at the cavity wall observed with this magnification and angle is light blue.

[0363] Fig.15 are photographs of sample No. 2 (reference number 1) before (top photo) and after (bottom two photos, reference number 2) expansion by cooking in water at 110°C for 10 minutes in an autoclave.

[0364] V: Further Experiments (Examples 3 and 4)

[0365] Using Examples 3 and 4, we further demonstrate exemplary parameters of the manufacturing process (shock heating) and their impact on the quality of the resulting meat substitute product (e.g. in terms of specific physical properties such as compressibility, hardening, expansion, cavity structure).

[0366] Example 3 (Samples No. 10, 11, 12, 13) - Hardening and compressibility of extruded products as affected by extrusion temperature setting

[0367] Samples No. 10, No. 11, No. 12, and No. 13 contain 70 wt% pea protein, 5 wt% steel cut oats, 24 wt% oat flour, and 1 wt% salt. Samples No. 10, No. 11, No. 12, and No. 13 are processed in the extruder 13 using different extrusion temperature settings.

[0368] Table IV shows that when mechanically processed starch-containing grains (such as steel cut oats) are used in the ingredients, the impact heating temperature setting of the extrusion conditions leads to good compressibility (compression force 10 234 g) and moderate hardening (129%) of the produced product (Sample No. 13).

[0369] However, when the liquid feed water temperature is low (such as 25°C commonly used in the known extruder 12), and / or when the temperature in the extruder is not subjected to shock heating distribution (the temperature of zone 2 is below 100°C, and / or the temperature of zone 4 is below 160°C), the products thus produced (samples 10, 11, and 12) have more serious hardening problems (186%-232%) and poor compressibility (compression force 17 803 g - 20 844 g). Although they have lower hardness (lower than sample 13) when fresh (5 minutes after extrusion), they have much higher hardness (higher than sample 13) after 5 hours of storage.

[0370] Table IV. Texture of samples No. 10, No. 11, No. 12, and No. 13

[0371]

[0372] The protein in Example 3 is pea protein isolate. It can be replaced by other proteins in the manner described in the context of Example 1.

[0373] Steel cut oats were used as the mechanically processed starchy grain in Example 3. Oat grits were used as the flour. Steel cut oats and oats can be replaced by other mechanically processed starchy grains and flours in the manner described above and in the context of Example 1.

[0374] In particular, steel cut oats can be replaced with steel cut barley, rice grains, cracked rice, pearled barley, pearled rye, pearled wheat, pearled oats, etc., or mixtures thereof. The results are comparable. Oat flour can be replaced with barley flour, wheat flour, rice flour, pea flour, chickpea flour, fava bean flour, quinoa, pigeon pea, sorghum, buckwheat, etc., and mixtures thereof. The results are comparable.

[0375] In this example, the steel cut oats were not soaked in hot water prior to extrusion.

[0376] Extrusion parameters:

[0377] (1) The moisture content of the slurry (material being extruded) during extrusion is about 50%;

[0378] (2) After extrusion, some of the extruded products were immediately immersed in water (e.g., 20°C) for 2 hours to cool and prevent drying. They were then taken out of the water. After being stored at 5°C for 24 hours, their compression force was analyzed;

[0379] (3) Some of the extruded products were immediately packaged in closed plastic bags to prevent drying, stored at room temperature, and analyzed for hardness and hardening;

[0380] (4) Extrusion productivity: about 18 kg of product per hour. The cooling die temperature is 90°C.

[0381] The compression force in Example 3 represents the resistance to compression with a cylinder as analyzed by the texture analysis method described above.

[0382] The texture observations in this example represent the observation records of texture properties analyzed by sensory evaluation by an expert panel.

[0383] The hardness in this example represents the hardness of an unsoaked extruded product analyzed by a texture analyzer using a cylindrical compression method to be described below.

[0384] Hardening refers to the hardening rate after 5 hours of storage, which can be calculated as:

[0385] Hardening rate = 100% x hardness (5 hours) / hardness (5 minutes).

[0386] Example 4 (Samples 14, 15, 16, 17) - Structure and compressibility of extruded products as affected by extrusion temperature setting

[0387] The ingredients used in samples No. 14, No. 15, No. 16, and No. 17 were: 90 wt% pea protein isolate, 5 wt% steel cut oats, 4 wt% pea fiber, and 1 wt% salt.

[0388] Table V shows that the functional combination (of sample No. 16) of (a) using an extrusion impact heating temperature setting and (b) using hot water as liquid feed, results in good compressibility of the resulting product (compression force 16 290 g) when mechanically processed starchy grains (now: steel cut oats) are used in the ingredients.

[0389] When the extrusion temperature was changed to a slower heating profile (temperature reduction in zone 4, 130°C; and temperature increase in zone 6, 160°C), the resulting product (Sample No. 15) was much less compressible (26 484 g).

[0390] When the extrusion heating temperature becomes an "excessive" heating profile as in the production of sample No. 17, where the temperatures of zones 5 and 6 are increased (160° C. and 160° C.), the produced product (sample No. 17) no longer has the desired continuous or complete structure. So its compression force is not measurable. And the product does not have the ideal chewiness similar to sample No. 16. These make it impossible for sample No. 17 to produce a meat substitute product similar to chicken legs or chicken nuggets.

[0391] When the extrusion temperature is changed to a "very slow" heating profile as in the production of sample No. 14, where the temperature of zone 2 is below 80°C, the temperature of zone 4 is below 160°C, and the liquid feed water is cold (25°C), the product produced (sample No. 14) no longer has the desired continuous or complete structure. So the compression force is not measurable. And the product does not have the ideal chewiness similar to sample No. 16. These make it impossible for sample No. 14 to produce a meat substitute product similar to chicken legs or chicken nuggets.

[0392] Table V. Texture of samples No. 14, No. 15, No. 16, and No. 17

[0393]

[0394] The protein in Example 4 is pea protein isolate. It can be replaced with other proteins in the manner described in the context of Example 1. The results will be comparable.

[0395] • For the possibility of replacing steel cut oats and oat grits, the same considerations as in Example 3 apply.

[0396] In Example 4, the steel cut oats were not soaked in hot water prior to extrusion.

[0397] Extrusion parameters:

[0398] (1) The moisture content of the slurry (material being extruded) during extrusion is about 50%;

[0399] (2) The extruded products were immediately immersed in water (e.g., 20°C) for 2 hours to cool and prevent drying. They were then taken out of the water. After being stored at 5°C for 24 hours, their compression force was analyzed;

[0400] (3) Extrusion productivity: about 18 kg of product per hour. The cooling die temperature is 90°C.

[0401] The compression force in this example represents the resistance to compression with a cylinder as analyzed using the texture analysis method described above.

[0402] VI - Advanced Experiments (Examples 5 and 6)

[0403] With Examples 5 and 6, we demonstrate the effect of extrusion conditions and ingredients on the formation of cavities with gelatinized starch coatings, which is closer to the mechanism of how these processing methods can lead to quality improvement. Some of the samples used in Examples 5 and 6 are the same as in Example 1.

[0404] Table VI shows that the functional combination of Sample No. 13 (a) using an extrusion shock heating temperature setting and (b) using hot water as the liquid feed resulted in increased starch solubility when steel cut oats were used in the ingredients.

[0405] The presence of soluble starch in the extruded product results from the combined effects of (a) mixing the grains and water, and (b) heating the grains with water early enough before the starch in the grains is emulsified by the protein matrix.

[0406] During extrusion, soluble starch can cause phase separation between protein gel and protein fiber, preventing the formation of a strong, completely isotropic (three-dimensional) cross-linked network structure. Soluble starch can also form a coating material between the gaps in the protein matrix, which will later become the cavity inside the extruded product. The coating material strengthens the cavity and prevents it from being sealed by protein cross-linking.

[0407] Table VI.

[0408]

[0409] The ingredients and extrusion parameters used in this example are the same as those described in Example 3

[0410] • Total starch in Example 5 represents the total amount of starch in the extruded product, which can be analyzed by any standard starch analysis method or by the hot water extraction method. The hot water analysis method is described below.

[0411] The washable starch in Example 5 (g of washable starch in 100 g of product) represents the amount of starch that can be washed out by 50°C water from a slice cut from the extruded product, as analyzed by the water washing test. The analytical method is described separately in another paragraph. Fig. 7A and Figure 7B There are microscope images of washable starch.

[0412] The soluble starch (g of soluble starch in 100 g of product) in Example 5 represents the amount of starch that can be dissolved into 50°C water from a slice cut from the extruded product, as analyzed by the water solubility test. The analytical method is described separately in another paragraph.

[0413] In this example, starch solubility represents the ratio between soluble starch and total starch.

[0414] Starch solubility = 100% x soluble starch / total starch.

[0415] Example 6. Starch that can be washed out by warm water and starch that can be dissolved by warm water from extruded products as affected by ingredients

[0416] Table VII shows that the use of oat flour (Sample No. 1) in the ingredients resulted in an extruded product with very low starch solubility (3.4%) and little washable starch (0.08 g / 100 g). However, when the oat flour was replaced with steel cut oats of the same chemical composition but larger size, the resulting product (Sample No. 2) had much higher starch solubility (8.4%) and more washable starch (0.41 g / 100 g).

[0417] like Figure 3 As shown in the Figure 1, and as shown in Example 1, Sample No. 2 has a more flexible and compressible texture than Sample No. 1. This is attributed to the higher amount of soluble starch and washable starch. This is consistent with the results of Example 5.

[0418] Table VII. Analysis of washable starch and soluble starch

[0419]

[0420] The ingredients and extrusion parameters used in this example are the same as those described in Example 1.

[0421] The washable starch in Example 6 (g of soluble starch in 100 g of product) represents the amount of starch that can be washed out by 50°C water from the cut slices of the extruded product.

[0422] Example 6 Starch solubility represents the "ratio between soluble starch and total starch".

[0423] Figure 3 A mathematical model is shown where an exponential curve is fitted to the measured values. It shows that there is a relationship between starch solubility and the compression force required to compress a meat substitute product made with high moisture protein texturized extrusion.

[0424] VII: Manufacturing Examples (Examples 8 and 9)

[0425] The following steps are used to produce a (preferably vegan) meat replacement product in the form of chunks (imitating chicken chunks). Figure 8 The results are shown in , which is an example of a food made from a meat substitute product (Sample No. 2) after being shredded into pieces having a size of more than 5 cm in length, 1 cm in width, and 0.8 cm in thickness, marinated, and pan-fried. The food simulates a thick chunk or slice of chicken thigh meat.

[0426] Step 1) Produce a meat substitute product, such as sample No. 2 or No. 13.

[0427] Step 2) tearing the extruded product into thin strips (e.g., about 2 cm - 4 cm long, 1 cm - 3 cm wide, 0.8 cm thick) so that the fiber direction is along the length direction. The tearing can be done manually or by a shredder.

[0428] Step 3) Soaking the shredded / chopped squeezed product in a marinade sauce (eg, containing water, oil, lemon juice, balsamic vinegar, sugar, salt and other spices) for a suitable time (eg, 2 hours), preferably immediately after squeezing.

[0429] Step 4) Remove the extruded product from the marinade and pan fry it preferably for 2 to 3 minutes until warm and golden and crispy on the surface.

[0430] The extruded product may be frozen or cooled after step 3). Step 4) may be performed just before consumption, such as at home or at work, or after purchasing the product in a restaurant.

[0431] Example 8 - Manufacturing a meat substitute product in the form of (preferably vegan) chunks

[0432] Fig. 9 An example food made of a meat substitute product such as sample No. 2 or sample No. 13 is shown after tearing the extruded product into pieces having a size preferably exceeding 3 cm in length, 2 cm in width, 0.8 cm in thickness, marinating the pieces (left side), battering the extruded product, crumb coating the extruded product and deep frying it in oil (right side). The food simulates chicken nuggets.

[0433] A meat substitute product in (preferably vegan) bite-sized form can be produced using the following steps:

[0434] Step 1) Produce a meat substitute product, such as sample No. 2 or No. 13. Soak the extruded product in water or a marinating sauce (e.g., containing water, oil, lemon juice, balsamic vinegar, sugar, salt and other spices) for a suitable time (e.g., 24 hours) after extrusion;

[0435] Step 2) Cut the soaked extruded product into a size and shape similar to regular or typical commercial pieces (eg at least 3 cm long, 2 cm wide, 0.8 cm thick).

[0436] Step 3) preparing a batter by mixing the ingredients, such as a formula of 40% by weight chickpea flour and 60% by weight water;

[0437] Step 4) covering the cut extruded product with batter liquid;

[0438] Step 5) coating the battered extruded product with a breading component (such as a commercial wheat-based fried breading component), breadcrumbs, or with a commercial gluten-free breading component;

[0439] Step 6) Deep fry the crumbed extruded product, preferably in oil, at, for example, 170°C for a suitable time, for example 3 minutes.

[0440] VIII: Advanced Analytical Methods

[0441] The analytical methods used to analyze different properties such as compression force, swelling ratio, starch solubility are described below.

[0442] Method for measuring thickness cooking expansion

[0443] The extruded product is cut into chunks by cutting through the protein fiber direction perpendicular to the direction in which the extruded product exits the extruder die. The length of this chunk is equal to the original width of the extruded product. The thickness of this chunk is equal to the original thickness of the extruded product. The width of this chunk is 20 mm. The width is measured in the direction parallel to the fiber direction.

[0444] The chunks were placed in a beaker-shaped container. Water was then added to the container to submerge the chunks. The water and chunks were then boiled in a pressure cooker (autoclave) at 110°C for 10 minutes.

[0445] After cooking, the chunks are removed from the water and allowed to rest on a kitchen sieve to drain. The thickness of the chunks is measured and compared before and after cooking. The expansion ratio is calculated as: the thickness after cooking divided by the thickness before cooking. The thickness of the chunk is measured at the center of the length of the chunk. Unless otherwise expressly stated, such as "extrusion expansion ratio", the thickness cooking expansion ratio is expressed as "expansion" or "expansion ratio" throughout this application.

[0446] Expansion rate = 100% x thickness (after cooking) / thickness (before cooking).

[0447] Methods for observing visible air cavities in extruded products:

[0448] The extruded product was cut into chunks (chunk A) by cutting through in a direction perpendicular to the direction of the protein fibers (the direction in which the extruded product exits the extruder die). The length of this chunk was equal to the original width of the extruded product. The thickness of this chunk was equal to the original thickness of the extruded product. The width of this chunk was 20 mm. The width was measured in the direction parallel to the fiber direction.

[0449] The extruded product is cut into a thick block (thick block B) by cutting the extruded product and taking the middle part (in the middle of the width of the extruded product), so the thick block has a thickness of its original thickness, a length of 40 mm in a direction parallel to the fiber direction of the extruded product, and a width of 20 mm in a direction parallel to the width of the extruded product.

[0450] Chunk A and Chunk B were placed in a beaker-shaped container. Water was then added to the container to submerge the chunks. The water and chunks were then heated at 60°C for 24 hours.

[0451] After heating, the chunks were removed from the water and allowed to rest on a kitchen sieve to drain. The cut cross-sections (length x thickness) of Chunk A and Chunk B were then observed by visual inspection and photographing.

[0452] The slabs were then air dried at room temperature for 7 days. The dried slabs were analyzed by X-ray micro-tomography (Micro-CT) scanning.

[0453] Method for soluble starch concentration measurement

[0454] This method adopts modifications of [Ref 10] and [Ref 11].

[0455] A solution containing soluble starch (1 mL) was mixed with diluted Lugol's solution* (1 mL) and water (4 mL). The mixture was shaken by hand for about 10 seconds, and then the mixture was allowed to stand for 10 minutes. The absorbance of the mixture solution at a wavelength of 600 nm (the wavelength of the light beam used in the spectrophotometer measurement) was then measured**.

[0456] * Prepare a diluted Lugol's solution by mixing 1 part of Lugol's solution (synonyms: iodine / potassium iodide solution, an aqueous solution of potassium iodide and iodine, with an iodine concentration between 3% and 10%) or a stabilized Lugol's solution (a complex of iodine-polyvinylpyrrolidone (PVP) (a homopolymer derived from 1-vinyl-2-pyrrolidone, complexed with iodine at a concentration between 3% and 10%)) and 5 parts of water. An example of the final concentration after dilution: an iodine concentration of 0.0100 mol / L and a potassium iodide concentration of 0.0260 mol / L.

[0457] **Absorbance is measured by UV / Vis spectrophotometer (one example UV / Vis spectrophotometer may be UV-1600PC from Supplier VWR Collection).

[0458] Prepare a standard curve of absorbance and soluble starch concentration as follows:

[0459] Potato starch (0.05 g, 0.1 g and 0.2 g) was dispersed in 200 mL of cold water by hand shaking for 1 minute. The dispersion was then cooked twice in an autoclave (10 minutes at 110°C each time and hand shaking for 1 minute after each cooking when the mixture was still above 60°C). In this way, the potato starch was completely dissolved in water. The potato starch dispersion was centrifuged at 644 g (g is the unit of RCF = relative centrifugal force) at room temperature. The supernatant was then taken as the starch solution for further analysis. Centrifugation can be performed by the centrifuge used in this study, namely the Heraeus™ Megafuge™ 8 small tabletop centrifuge equipped with a 50 mL conical bucket rotor (supplier's product number 75005703).

[0460] Based on the standard curve and the absorbance value at a wavelength of 600 nm, the concentration of soluble starch in the starch solution can be calculated.

[0461] Cited by McGrance (1998) [reference 10], "The reaction between starch and iodine has been known for over a hundred years. Over fifty years ago, Rundle and Baldwin proposed that the iodine component of the complex exists as a one-dimensional array within the amylose helix with six glucose residues per turn. Two important aspects of the colorimetric method using the iodine reaction are its versatility and simplicity. It can be used with starch from a variety of plant sources and requires no special equipment other than a simple spectrophotometer capable of measuring absorbance near 600 nm. Samples high in amylose and low in amylose can be analyzed and require only that the volume of the aliquot be varied to obtain the best result. The sensitivity of the iodine-starch reaction is quite high." Although the iodine colorimetric method is not routinely used as an official analytical method, it is reliable for starch quantification and is known to those skilled in the art.

[0462] Method for the analysis of soluble and washable starch from extruded products

[0463] The method for extracting and defining soluble starch and washable starch adopts a modification of [reference 12]. Soluble starch is starch that can be extracted (extracted = washed out) from the product by water at 50°C, passes through a sieve with a pore size of 1200 μm, and is soluble in water. Washable starch is starch and starch-containing material that can be extracted (extracted = washed out) from the product by water at 50°C and passes through a sieve with a pore size of 1200 μm. Soluble starch is part of washable starch, in other words, soluble starch is a synonym of "soluble washable starch". Washable starch includes soluble washable starch and insoluble washable starch. Insoluble washable starch can be dissolved in water when it is cooked above its gelatinization temperature, preferably at about 100°C. Soluble components are components in solution, which disperse well in the liquid and do not precipitate during centrifugation at 644 g (g is the unit of RCF = relative centrifugal force).

[0464] Fig.13 The method for analyzing soluble starch and washable starch from an extruded product 61 is illustrated:

[0465] (Step 62) Cutting away from the edges (5% of the width) to take a sample 63 approximately from the middle of the extruded product 62;

[0466] (Step 64) Slicing the sample 63 into thin slices 65, the thin slices 65 of the extruded product having a size of about 1 mm x 10 mm x 40 mm, wherein the length direction (40 mm) of the slice is parallel to the fiber orientation direction of the extruded product;

[0467] (Step 66) Immerse the thin slice 65 in 50°C water for 24 hours and shake it manually for 2 minutes;

[0468] (Step 67) sieve with 1.2 mm pore size;

[0469] Reference numeral 68 refers to the insoluble washable components in the wash extract;

[0470] (Step 69) Centrifuge at 644 g (RCF) for 30 min;

[0471] Reference numeral 70 refers to the supernatant obtained by centrifugation, which contains soluble starch;

[0472] (Step 71) Cook in an autoclave at 110° C. for 10 minutes and shake by hand;

[0473] (Step 72) Centrifuge at 644 g (RCF) for 30 min;

[0474] Reference numeral 73 refers to the supernatant resulting from the centrifugation, which contains washable starch.

[0475] Measurements were made on 20 g of the slice extrudate, which was soaked (step 66) in 200 mL of water and maintained at 50°C for 24 hours.

[0476] g is the unit of RCF = relative centrifugal force.

[0477] Starch solubility of extruded product = (soluble starch content / total starch content in extruded product) x 100%

[0478] Starch washability of extruded product = (washable starch content / total starch content in extruded product) x 100%.

[0479] Method for measuring total starch in extruded products

[0480] The total amount of starch in the extruded product can be analyzed by standard starch analysis methods such as AACCI Method 76-13.01 "Total Starch Determination Procedure" (Megazyme Amyloglucosidase / α-amylase Method). It can also be measured by a hot water analysis method having the following steps: (1) Cut the extruded product into approximately 1 mm 3 cubes; (2) cook 4 g of the extrudate in 200 mL of water in an autoclave at 110°C for 10 minutes; (3) shake the extrudate-water mixture by hand when it is removed from the autoclave oven and above 70°C. (4) repeat step (3) cooking and shaking once more. With this treatment, it can be assumed that all the starch is dissolved in the water. (5) centrifuge the extrudate-water mixture at 644 g (RCF) for 30 minutes, and (6) measure the soluble starch concentration of the supernatant. The total amount of starch in the supernatant is equal to the total starch content of the extrudate, which can be calculated using the volume of water and the soluble starch concentration value.

[0481] Methods for measuring cutting and compression forces

[0482] For cutting force measurement, we measured the resistance of the sample during the compression test with a blade. The measurement was carried out so that the TA.XTPlus texture analyzer (supplier Stable Micro Systems) was equipped with a 294.2 N (30 kg) load cell (detector sensor) and a sharp blade. The knife was a "double bevel (grinding) Scandi" type. The knife had a blade with a total wedge angle of about 16 degrees at the sharpest part (edge), which means that the main angle of the bevel of the knife was about 8 degrees. The knife had a flat part (ridge) with a thickness of 0.6 mm higher than the blade part. The height of the sample was between 7.0 and 12.0 mm. The width of the sample was 20 mm. The sample was stabilized and placed horizontally on a plate, and the direction of the sample was adjusted so that the blade was compressed (i.e. cut) in the cross-sectional direction (length direction of the fiber) of the elongated fiber. The downward speed before the blade contacted the fiber was 4 mm / s (speed before the test). The speed of compression when the blade contacted the fiber was 20 mm / s (test speed), and the compression was carried out until a cutting depth of 90% of the sample height was reached. For samples with heights above 9.0 mm, compression was performed to a cutting depth of 8.0 mm. For this study, the peak positive force (peak positive force is a term used in the equipment software that refers to the maximum force detected during the measurement) was taken as the cutting force.

[0483] For the compression force measurement, we measured the resistance of the sample during the compression test with a cylindrical probe (model "P / 36R", 36 mm radius edge cylindrical probe - aluminum - AACC standard probe for bread hardness, supplier Stable Micro Systems). The measurement was carried out so that the TA.XTPlus texture analyzer was equipped with a 294.2 N (30 kg) load cell (detector sensor) and a cylindrical probe. The height of the sample was between 7.0 and 12.0 mm. The width and length of the sample were 40 mm. The sample was stabilized and placed horizontally on a plate, and the sample was oriented so that the cylindrical compression was towards the center of the sample. The downward speed before the blade contacted the fiber was 2 mm / s (pre-test speed). The speed of compression when the blade contacted the fiber was 0.5 mm / second (test speed), and the compression was carried out until a cutting depth of 40% of the sample height was reached. For this study, the peak positive force (peak positive force is a term used in the equipment software, which refers to the maximum force detected during the measurement) was taken as the compression force. There is a "trigger force" setting, which was set to 1000 g in this study. The trigger force is set to control the machine (texture analyzer). When the detected resistance is lower than the trigger force, the probe is not at the position where the top surface of the sample is contacted, and the probe moves downward at a pre-test speed of 2 mm / s. When the detected resistance is not less than the trigger force, the probe reaches the sample and the probe moves downward at a test speed of 0.5 mm / s.

[0484] Methods used to measure hardness

[0485] For the hardness measurement, we measured the resistance of the sample during the compression test with a cylindrical probe (model "P / 36R", 36 mm radius edge cylindrical probe - aluminum - AACC standard probe for bread hardness, supplier Stable Micro Systems). The measurement was carried out using a TA.XTPlus texture analyzer equipped with a 294.2 N (30 kg) load cell (detector sensor) and a cylindrical probe. The height of the sample was between 7.0 and 12.0 mm. The width and length of the sample were 40 mm. The sample was stabilized and placed horizontally on a plate and the sample was oriented so that the cylindrical compression was towards the center of the sample.

[0486] The test procedure uses a standard TPA measurement protocol (quote from the manual of the measuring equipment "Texture Profile Analysis (TPA) is an objective method of sensory analysis pioneered in 1963 by Szczesniak [Ref 6], who defined the texture parameters used for the first time in this analytical method. Later in 1978 Bourne [Ref 7] adapted the Instron to perform TPA by twice compressing a standard sized food sample. TPA is based on the recognition of texture as a multi-parameter property. For research purposes, it may be desirable to determine the texture profile with respect to several parameters on a small homogeneous sample. The test consists of twice compressing a bite-sized piece of food with a reciprocating motion that mimics the action of the jaws, and extracting from the resulting force-time curves a number of texture parameters that correlate well with the sensory evaluation of those parameters [Ref 8]. The mechanical texture properties of foods, which largely govern the choice of rheological procedure and equipment, can be divided into primary parameters of hardness, cohesion, elasticity (springiness) and adhesion, and secondary (derived) parameters of breakability (brittleness), chewiness and stickiness [Ref 9].

[0487] The downward speed before the blade contacts the fiber was 5 mm / s (pre-test speed). The speed of compression when the blade contacts the fiber was 2 mm / s (test speed), and compression was performed until a cutting depth of 30% of the sample height was reached. For this study, the peak positive force (peak positive force is a term used in the equipment software, which refers to the maximum force detected during the measurement) was taken as the compression force. There is a "trigger force" setting, which was set to 5000 g in this study. The waiting time between the first compression and the second compression was 1 second, and the hardness was calculated by the software of the measuring equipment. The hardness is equal to the peak positive force during the first compression.

[0488] IX: Advanced Mechanistic Studies

[0489] Mechanistic Study 1 shows the effect of processing methods (ingredients, impact heating) on ​​the properties (particle size distribution) of the test extrusion (extrusion without cooling die) material, which reveals the mechanism of how these processing methods affect the extruded product. This can also be used as an evaluation method for selecting processing parameters.

[0490] Further mechanistic studies revealed relevant knowledge on the differences between the properties of grains and flours, and between grains processed with cold water and grains processed with warm water.

[0491] Mechanism study-1-Effects of composition and extrusion temperature distribution on particle weight distribution

[0492] In order to study the effect of composition and extrusion temperature on the results, the inventors conducted a number of further experiments. Table VIII lists the composition and test extrusion parameters. Test extrusion means that during these tests the extruder was not equipped with any die, but only the composition was processed by a screw running in a heated chamber. A summary of the results and findings can be found in Table IX. Figure 4 The measured particle weight distribution of the extruded material as affected by the component composition and the extrusion heating temperature distribution for Experiments 1 to 6 is shown.

[0493] Table VIII. Sample preparation for test extrusion

[0494]

[0495] Rolled oats were used as the machined starchy grain in Experiments 2 and 3. Steel cut oats were used in Experiments 4, 5 and 6. The steel cut oats were not soaked prior to testing extrusion.

[0496] The test extrusions did not form a thick mass with a long continuous fiber matrix. Instead, the resulting material was agglomerates of varying sizes (thus each particle weighed in the range of 0.1 g to 10 g). The agglomerates (i.e. particles) were divided into different size (weight) groups (small, medium, large, etc.), each size group was then weighed, and its percentage relative to the total weight of the resulting agglomerate was calculated. Figure 4 The particle weight distribution curve is shown in .

[0497] Table IX: Test extrusion results and findings

[0498]

[0499] Comparisons should mainly be made between samples with the same chemical composition (protein content, starch content, etc.), for example between Experiment 1, Experiment 2 and Experiment 3, or between Experiment 4, Experiment 5 and Experiment 6, respectively.

[0500] In addition, there are similarities between Experiments 1 and 4, which have parameters that can produce products with good compressibility and flexibility. They both produce a percentage of medium-sized particles (0.5 g - 4 g) between 26%-30%; a percentage of large particles (> 4 g) between 0%-5%.

[0501] Mechanistic Study 2. Comparison of Particle Size, Seed Coat, Seed Structural Integrity, and Starch Extractability among Oat Flour, Oat Flakes, Steel Cut Oats, and Whole Oat Seeds

[0502] The measurements in Table X show that the starch extractability in water of oat flakes, steel cut oats, and whole oat seeds (9-26 g / 100 g) is much lower than that of oat flour (40 g / 100 g) due to the better integrity of the seed structure and seed coat. The starch extractability of whole oat seeds is very low (9 g / 100 g) due to the intact seed coat.

[0503] When the water is hot, steel cut oats can absorb water much more and faster (375%, 110°C, 10 minutes) than when the water temperature is lower (136%, 50°C, 12 hours). These explain why shock heating and soaking in hot water can change the behavior and effectiveness of oat flakes and steel cut oats in high moisture extrusion. Hot water can allow the starchy grains to absorb water faster and more completely and become gelatinized and more soluble.

[0504] Whole oat seeds are not as functional / substitutable as the oat flakes and steel cut oats in the above disclosed embodiments. At the time of writing, the inventors are still testing other treatments to make whole oat seeds functional. For example, boiling thoroughly in excess water.

[0505] Table X: Oat-based starting materials, starch extractability in water

[0506]

[0507] To measure extractable starch, 10 g of the starting material was cooked in 100 g of water in an autoclave for 10 minutes and the cooked mixture was centrifuged at 644 g (RCF) for 30 minutes. The soluble starch concentration of the supernatant was calculated as:

[0508] Extractable starch = 100% x soluble starch in supernatant / weight of starting material.

[0509] To measure water absorption at 50°C, 20 g of the starting material was immersed in 200 g of water, then kept immersed at 50°C for 24 hours, and then sieved to remove water that was not absorbed by the material. The weight of the material before and after the 24-hour immersion was recorded.

[0510] Water absorption = 100% x (weight after immersion - weight before immersion) / weight before immersion.

[0511] To measure water absorption at 50°C, 20 g of the starting material was added to 200 g of water and then cooked in this water at 110°C in an autoclave for 10 minutes and then sieved to remove the water not absorbed by the material. The weight of the material before and after cooking was recorded.

[0512] Water absorption = 100% x (weight after cooking - weight before cooking) / weight before soaking.

[0513] Steel cut oats can be produced in different sizes, ranging from 6 mm per particle 3 Up to 15 mm 3 In this mechanism study 2, 8 mm per particle was used. 3 Those.

[0514] Mechanistic Study 3: Effect of Soaking Steel-Cut Oats on Their Mechanical Properties

[0515] The inventors studied the effects of soaking steel cut oats. Figure 5 The results of compression testing of dry (unsoaked) steel cut oats versus soaked steel cut oats (soaked in hot water) are shown in the table.

[0516] Available from Figure 5 As can be seen in the graph, the steel cut oats that were not soaked in water were significantly more brittle and less compressible than the steel cut oats soaked in hot water. When the compression rate reached 27% (1.78 mm thick steel cut oats were compressed to 0.47 mm deep), the steel cut oats that were not soaked had cracked and split. On the other hand, the steel cut oats that were soaked in hot water (80°C, 2 hours) became softer, more sticky and mushy. The soaked steel cut oats did not crack or split throughout the compression (compression between 0%-90% during the test).

[0517] This revealed that the starchy grains could be broken into smaller pieces by the compression force, which was sufficient during the extrusion process.

[0518] Treating the starchy grains with hot water can soften the grains and help prevent the grains from being broken into smaller pieces by compression or pressing.

[0519] Table XI: Effect of soaking of steel cut oats on its mechanical properties

[0520]

[0521] As an overview of comparing the soluble starch content, washable starch content, starch solubility and starch washability properties when the protein content is the same, the inventors reviewed and classified the results and calculated the changes in these values. In Table XII, S1, S3, S4, S5 and S6 have the same composition and extrusion conditions as in Sample No. 1, Sample No. 2, Sample No. 6, Sample No. 11 and Sample No. 13. S2 has the same composition as Sample No. 2, but it has different extrusion conditions. In S2, the steel cut oats were not soaked in hot water before extrusion, and shock heating was achieved by using hot water (60°C) liquid feed and an extruder temperature profile of 100-125-160-145-130 (°C) at zones 2-3-4-5-6.

[0522] Table XII shows that S2 has 52% higher starch solubility and 63% higher starch washability than S1. These differences are attributed to shock heating and ingredient differences (e.g., using steel cut oats). The starch solubility and starch washability of S3, which uses steel cut oats, soaking, and shock heating, are even higher. When the pea protein content is reduced from 90% to 70%, the impact of ingredients (e.g., using steel cut oats) and shock heating is even greater. S6 has 261% higher starch solubility and 58% higher starch washability than S4. Due to the difference in shock heating, the starch solubility and starch washability of S5 are not as high as S6.

[0523] Table XII: Effect of extrusion conditions and ingredients on soluble starch content, washable starch content, starch solubility and starch washability properties

[0524]

[0525] X: Conclusion

[0526] The inventors have surprisingly found that starch added in the form of a farinaceous powder or flour can indeed cause the separate protein matrix parts to glue together to form even larger pieces or more complete structures during the extrusion process with or without a long cooling die.

[0527] The addition of starch-containing powders also produces extruded products with much higher isotropic properties and much lower anisotropic properties (anisotropic fiber structure, anisotropic texture).

[0528] The inventors also found that small-sized starch can be emulsified into and / or between protein fibers to become a filler material in a protein-based emulsion gel system, which can improve the uniformity and coverage (area, space, volume) of the protein material distribution. As a result, during the entire extrusion process, the proteins can form more isotropic interactions with each other. Starch gelation can also combine materials of different parts to be connected to each other.

[0529] The inventors have also found that when a long cooling die is used in extrusion, adding a higher amount of this type of material containing starch powder can form a thicker, denser and more isotropic block with a specific fiber structure. When a cooling die is not used in extrusion, adding a higher amount of this type of material containing starch powder can form a larger extruded product connection mass (sheet) without a fiber structure.

[0530] The inventors have also found that the protein matrix hardening problem can be prevented or at least further delayed when starch-containing grains are added to the protein material and extruded as described in the method claim.

[0531] Without wishing to be bound by any theory, and in view of the very limited amount of knowledge in the art, the inventors have discovered and have a possible explanation that when the particle size of the starch-containing grains is larger than conventional starch-containing powders, the starch-containing grains break into smaller parts at a much slower rate. In addition, the broken grain parts are not easily emulsified by the protein matrix. The broken grain parts can still be gelatinized by sufficient heat, shear and water. In addition, the naturally occurring grain cell wall structure and materials can limit the complete leaching, alignment and retrogradation of starch molecules.

[0532] The naturally occurring grain cell wall structure and the gelatinization effect of gelatinized starch can also prevent the grain from being completely powdered into small particles (e.g., particle size below 100 μm). As a result, a large number of gelatinized starch clusters are formed and remain throughout the extrusion process and in the final product.

[0533] The inventors were surprised to find that at least some of these clusters can be washed out of the extruded product by warm water (50° C.) without further gelatinization of the starch when the extruded product is cut into thin slices but without necessarily completely destroying the protein fibers. These starch clusters have a much larger particle size than the starch in conventional processes, in which the starch in conventional processes is homogenized and emulsified individuals in a protein matrix. These starch clusters are typically larger than 100 μm in at least one of their dimensions. As a result, these starch clusters can behave like large particles that can separate the protein fibers away from each other and thus prevent the formation of hydrogen bond type protein-protein interactions and texture hardening.

[0534] Large starch clusters as large particles also often lead to the formation of holes (cavities) or empty spaces next to them. This may be due to the flow behavior of the extruded material and the strength of the protein fibers during extrusion, allowing the protein fibers to flow away from each other after encountering the large particle barrier formed by the starch clusters. Then, after a while of continuous flow away from each other, the bundles of protein materials (protein fibers) approach and interact with each other again. During this period when the proteins flow away from each other, empty spaces are formed behind the large particles of starch clusters. Protein fibers separated by empty spaces cannot form hydrogen bonds. The inventors believe that this may help the improved taste to last longer, even in cooled or frozen meat substitute products.

[0535] In addition, the inventor has found that the earlier the starch in the grain is gelatinized before being emulsified by the protein matrix, the higher the concentration of the gelatinized starch clusters is, the higher the degree of the formation of the continuous protein matrix can be prevented. Without wishing to be bound by any theory, the inventor has a kind of explanation, that is, the gelatinized starch clusters that are not emulsified by the protein matrix are immiscible with the protein phase, and therefore can make the phase and the protein phase separate, therefore can form a considerable connection phase, and can destroy the protein-protein interaction formation, so they can prevent the formation of the continuous protein fiber matrix to a certain extent. This explanation is very consistent with the test results in the mechanism research experiment in the selected embodiment described below. The observed difference between the sample quantity checked by the inventor seems to also support this explanation.

[0536] After the formation of gelatinized starch clusters, the melting, cross-linking and gelatinization of the protein material should be initiated within a certain short time window. If this occurs too late, there may be two types of unacceptable results, namely (1) the gelatinized starch clusters are ultimately homogenized, broken up and emulsified by the protein matrix, which is particularly likely to be the case when the amount of starch-containing grains added is small, or the starch-containing grains are relatively easy to break up and the content of starch-containing powder in the ingredients is high; (2) by excessively separating and covering the protein material into individual clusters, the gelatinized starch clusters completely prevent the formation of long continuous protein fiber structures and prevent protein-protein coagulation, aggregation and gelatinization, which is particularly likely to be the case when the amount of starch-containing grains added is large and the content of starch-containing powder in the ingredients is low.

[0537] In addition, the inventors have found that when the starch-containing grains added to the extruder are not soaked in hot water or are added to the extruder without mixing with hot water at a very early stage (e.g. between 0 seconds and 15 seconds, preferably between 1 second and 15 seconds after feeding into the extruder), the starch-containing grains are more easily ground into powder in the extruder. In this way, the starch-containing grains behave similarly to their flour, which has the same chemical composition but a smaller particle size and a disrupted cell wall structure.

[0538] In contrast, starch-containing grains immersed in hot water prior to extrusion and mixed with hot water very early in the extrusion (e.g. between 0 and 15 seconds, preferably between 1 and 15 seconds after feeding into the extruder) will be less friable, more extensible, and therefore less prone to being emulsified by the protein matrix, and more prone to remain as large particles throughout the extrusion. This is therefore part of the reason why it is important and necessary to use extrusion conditions with an impact heating setting while using starch-containing grains in the ingredients for extrusion in order to produce an extruded product of acceptable quality.

[0539] The inventors have also surprisingly discovered that meat substitute products made by texturizing extrusion with high moisture proteins can have significantly higher levels of extrusion overrun shortly after the extruded product leaves the long cooling die of the extruder when produced by the method described in said method item.

[0540] One second after the extruded product comes out of the long cooling die of the extruder, the high extrusion expansion rate during extrusion is clearly visible, the extruded product clearly has air bubbles inside the expanded structure, and its thickness is much greater (e.g. 200% - 600% higher) than its original thickness just before leaving the long cooling die of the extruder (the original thickness is roughly the same as the opening height of the long cooling die of the extruder). After the extruded product cools, the expanded structure may collapse for the most part. However, there are still more cavity (in other words, air pocket) structural units remaining in the cooled extruded product. This difference may be an advantage of forming gelatinized starch clusters that are not emulsified by the protein matrix, which is produced by the method described in the method item.

[0541] Gelatinized starch can lead to greater expansion ratios in high moisture extrusion. The increase in expansion ratio can be attributed to the reduction in structural firmness and the decrease in viscosity of the extruded material.

[0542] In contrast, such extrusion expansion phenomena are essentially absent, or in other words, cannot be detected, in such tested processing methods that do not use starch-containing grains or do not have impact heating settings in the extrusion conditions. It was found that these processing methods that failed to produce products with a texture close to that of boiled chicken thighs tended to produce extruded products with a denser and tighter structure (0% to 199% higher thickness one second after coming out of the extruder long cooling die than before leaving the extruder long cooling die), and retained significantly fewer cavity structural units (in other words, air pockets) after cooking. During high-moisture extrusion, starch-containing flour can lead to higher amounts of starch leached out, more water absorption, and higher viscosity increases compared to starch-containing grains. These were found to be consistent with observations during extrusion testing and consistent with experiments investigating the mechanism of cooking starch-containing materials in water in a pressure cooker.

[0543] The inventors have surprisingly found that for extruded products produced by the method described in the method item, more starch molecules can be dissolved out of the extruded product by warm water (50°C) when the extruded product is cut into thin slices but the protein fibers are not necessarily completely destroyed. The temperature of 50°C is below the gelatinization temperature of starch. Generally, native (non-gelatinized) starch is insoluble in 50°C water. Pregelatinized starch and some modified starches can be dissolved in 50°C water before they are used in meat substitute production by high-moisture protein texturization extrusion, but they lose solubility after the extrusion process because they are emulsified by the protein matrix quickly after being extruded with the protein material.

[0544] The starch dissolved in the extruded product described here and below is soluble washable starch, which is a part of washable starch. Compared with insoluble washable starch, soluble starch (soluble washable starch) gelatinization is more complete, more from starch granule shell and grain cell wall structure (released from its restriction), has higher affinity to water, and their molecules have more expanded structure (such as volume and surface area). Soluble starch has even less affinity to protein matrix, and is even less tightly embedded in or captured by long continuous protein fiber structure. Soluble starch is more immiscible with protein phase, so it is more completely separated from protein phase by phase separation. Soluble starch is the main component of the inner wall of the cavity (cavitation) of the acceptable extruded product. Soluble starch compound is the main component and main site of extrusion expansion and cavity generation. After being dyed with diluted iodine solution, the coating material of the inner wall of the cavity in the extruded product of acceptable quality can be seen by visual observation and microscopic observation. After staining, the coated material turned dark blue or black, indicating a high concentration of starch. The cavities coated by the gelatinized starch clusters also acted as a new type of disruptive compound that prevented further formation of protein-protein interactions (eg, hydrogen bonds) between the protein fibers after extrusion. use The gelatinized starch cluster-coated cavities are different from and outperform other known destructive microparticles. For example, starch, cereal flour, insoluble salts, dietary fiber, obviously because starch clusters keep protein fibers away from each other in a volume larger than the size of individual particles.

[0545] There is no background art teaching about the role and impact of soluble starch, washable starch, insoluble washable starch, starch solubility, starch washability in producing meat substitute products with long continuous protein fiber structures by high moisture protein texturized extrusion or in low moisture protein texturized extrusion. There may be some research on starch solubility in starch extrusion processes, which mainly process the starch component of starchy foods and have a very different configuration from protein texturized extrusion. However, starch solubility is highly correlated with crumb aging and texture quality. For example, Boyacioglu and D'Appolonia [Reference 5] reported that breadcrumbs aged (stored, aged) for more than 4 days may have a continuous, gradual and significant decrease in starch solubility, while the firmness value continues to increase significantly; it is recommended that the soluble starch content be used to measure the rate and extent of aging, because a decrease in soluble starch content indicates an increase in crumb aging and firmness; the rate of increase in firmness value of aged breadcrumb samples with a higher amount of soluble starch is lower. In breadcrumbs, a decrease in starch solubility indicates an increase in the regeneration rate of starch molecules. Starch retrogradation is a well-known factor that is often responsible for the leathery mouthfeel and tough texture of starch-containing foods such as bread. It occurs most rapidly at temperatures slightly above freezing (e.g., between 0°C and 6°C). Starch retrogradation is caused in part by recrystallization of starch amylose and amylopectin molecules and is a result of increased starch-starch hydrogen bond formation and decreased starch-water affinity. It is possible but not obvious to think about the correlation between the knowledge about starch solubility behavior in meat products produced by high-moisture protein texturized extrusion and the knowledge about breadcrumbs. Meat substitute products produced by high-moisture protein texturized extrusion have completely different ingredient formulations, structures, and microstructures than breadcrumbs. Although the processes and structure formation mechanisms of protein texturized extrusion and bread baking are also completely different.

[0546] The inventors were surprised to find that most of the starch in meat substitute products made by high-moisture protein texturization extrusion and having low starch solubility and low starch washability was uniformly homogenized and emulsified by the protein matrix. Microscopic observation revealed that the emulsified starch in the product was arranged linearly so that the starch particles were parallel to each other. The protein fibers tightly covered and captured the starch compounds. The starch compounds were completely drained out. The original starch granule structure had basically disappeared. Therefore, the starch may have undergone severe retrogradation. These findings are consistent with the results that the starch solubility of those samples was low, the hardening was more serious during the 5-hour storage period, the compressibility was worse after storage overnight, and the ability to expand by cooking in water in a pressure cooker was worse. On the contrary, it was found that the meat substitute products with significantly high starch solubility and starch washability had better texture properties (good compressibility, good expansion properties, and a mouthfeel close to chicken thigh meat).

[0547] Starch solubility and starch washability are even more important than soluble starch content and washable starch content. Starch solubility and starch washability are calculated as the ratio of soluble starch content and washable starch content to the total amount of starch in the extruded product. Soluble starch and washable starch contribute positively to the quality (e.g., mouthfeel) of the extruded product. On the contrary, the higher the percentage and amount of insoluble starch and non-washable starch, the worse the quality (e.g., mouthfeel) of the extruded product may be, because the insoluble starch and non-washable starch are emulsified, captured, embedded relatively more completely in the protein matrix, and have more retrogradation.

[0548] With this background and the inventors' novel discoveries, there are reasons to believe it is important to monitor and control the levels of soluble starch content, washable starch content, starch solubility and starch washability in meat substitute products made with high moisture protein texturized extrusion.

[0549] Methods for controlling and improving starch solubility and starch washability in meat substitute products produced by high moisture protein texturized extrusion are not found in the background art but are disclosed in the following description.

[0550] The inventors have discovered that when a meat substitute product manufactured in an extruder configured to perform high-moisture protein texturized extrusion comprises a continuous protein fiber matrix structure that is substantially linearly oriented and has breaks that form cavities having walls that are at least partially coated with gelatinized starch clusters, the mouthfeel tends to remain acceptable for a longer period of time.

[0551] It is known that the reduction of starch solubility (e.g. in water at 50°C) and the increase of starch retrogradation are important factors in causing the texture firmness of foods (e.g. bread crumbs containing starch gel structures). See references (a) SOHOCH, TJ; FRENCH, D. 1947. Studies on bread staling. 1. The role of starch. CerealChemistry, 24: 231-249; (b) T. Inagaki and PA 1992. Firming of Bread Crumbwith Cross-Linked Waxy Barley Starch Substituted for Wheat Starch. CerealChem 69:321-325; (c) K. Ghiasi, RC Hoseney, and DR Lineback. 1979. Characterization of Soluble Starch from Bread Crumb. Cereal Chem 56:485–490.

[0552] Alternatively or additionally, the gelatinized starch clusters include starch that is not emulsified by the protein fiber matrix structure (non-emulsified starch). Advantages resulting therefrom are: (1) an increase in the percentage of non-emulsified starch results in a decrease in the percentage of emulsified starch. Non-emulsified starch does not behave like a filler that fills the gaps between the protein fibers and reinforces the overall extrudate structure, whereas emulsified starch does; (2) non-emulsified starch is less arranged (less disordered or less molecular) than emulsified starch and thus has less and / or delayed starch retrogradation and improved softness over long storage times at temperatures above freezing (e.g., between 0°C and 6°C); (3) non-emulsified starch interferes with the arrangement of the protein fiber matrix structure and thus improves its softness over long storage times at temperatures above freezing (e.g., between 0°C and 6°C) by reducing and / or delaying the formation of intermolecular (e.g., protein-protein, starch-starch) hydrogen bonds in the extrudate.

[0553] Alternatively or additionally, a meat substitute product may be made using a high moisture protein texturized extrusion process wherein the starchy grains are gelatinized and:

[0554] (a) before the gelatinized starch-containing grains are emulsified with the proteins of the protein matrix; and

[0555] (b) Before the gelatinized starch forms a complete barrier that prevents the formation of a continuous protein fiber cross-linked matrix,

[0556] The protein that forms the protein matrix melts. The advantage of this is that: in this way, the extruded material is controlled to be in a good balance between the following two: (a) sufficient formation of protein-protein crosslinks to form continuous protein fibers; and (b) crosslink formation is prevented by gelatinized starch. As a result, the extrudate can have a chewiness within a specific threshold range (cutting force is higher than 300g), and at the same time have a compressibility within a specific threshold range (compression force is lower than 17500g). If the protein melt is not achieved before the emulsion is formed between the gelatinized starch-containing grains and the protein material, the emulsification can still be achieved by continuous shearing, tearing and homogenization of the protein starch mixture, and then the starch becomes emulsified, and the increase of undesirable interaction forces (such as hydrogen bonds) and the hardening of the extrudate (such as the compression force becomes higher than 17500g) cannot be prevented. On the other hand, if the protein melt is not achieved before the gelatinized starch forms a complete barrier that prevents the formation of a continuous protein fiber crosslinked matrix, the protein-protein crosslink will be lacking. As a result, the chewiness will be too low and not within the threshold range (cutting force is higher than 300g).

[0557] The extrusion step can be performed with an extrusion die having a length greater than 300 mm, preferably greater than 1000 mm. This is advantageous in that such a die is a typical setting for implementing high moisture protein texturization extrusion. This die allows the extruder to handle extrusion cooking of materials having a moisture content greater than 40% to form a texturized (cross-linked) structure before the material leaves the extruder. This die also allows the molten protein material to be arranged into a long continuous fiber structure.

[0558] Preferably, the heating step d) is preferably carried out at a temperature between 140°C and 200°C. This provides the advantage that this temperature allows the proteins to melt, denature, form gels and form the protein-protein crosslinks required to form long continuous fiber structures.

[0559] Preferably, the mechanically processed starch-containing grains comprise or consist of one or more of oats, barley, rye, wheat, rice, corn, lentils, chickpeas, mung beans, broad beans, peas, quinoa, pigeon peas, sorghum, buckwheat. Advantages arising therefrom are that these grains are commercially available, contain high amounts of starch, are known to be tasty and nutritious, and are widely used in various other food applications.

[0560] Alternatively or additionally, the heating step d) is preferably carried out so that the protein melting occurs between 1 s and 40 s, preferably between 10 s and 30 s, after step b). The resulting advantage is that: in this way,

[0561] (a) before the gelatinized starch-containing grains are emulsified with the proteins of the protein matrix; and

[0562] (b) Before the gelatinized starch forms a complete barrier that prevents the formation of a continuous protein fiber cross-linked matrix,

[0563] The proteins forming the protein matrix melt.

[0564] In the tests, the time required for an extruder to break grains (eg rolled oats, steel cut oats, rice) into powder was observed.

[0565] Alternatively or additionally, heating step c) is performed such that starch gelatinization occurs between 0 s and 18 s, preferably between 1 s and 15 s. The resulting advantage is that, in this way, the starch-containing grains can be preferably ground by the extruder screw to a particle size of less than 5000 μm. 3 Before and preferably after the starch-containing grains are ground by the extruder screw to a particle size of less than 0.001 mm 3 Prior to this, a heating step c) is performed. Each particle has a volume greater than 5000 μm 3 The gelatinized starch clusters are non-emulsified starches, are larger than those that are emulsified, and can provide much greater disruptive forces to prevent the formation of too excessive protein-protein interactions and, therefore, can prevent hardening of the extrudate during storage.

[0566] Preferably, after heating step d), the mixture is further extruded for more than 5 s, preferably more than 10 s, at a temperature not higher than that of heating step c), preferably between 90°C and the temperature in heating step d). The resulting advantage is that such a heating level can result in a good balance between: (a) sufficient formation of protein-protein cross-linked structure (force) to provide acceptable chewiness (cutting force above 300 g); and (b) having acceptable compressibility (compression force below 17500 g). Higher temperatures may result in excessive cross-linking and, therefore, poor compressibility. Temperatures below 90°C may result in too weak a structure lacking a co-aligned long fiber structure and poor chewiness.

[0567] XI-Summary

[0568] In order to improve the mouthfeel of meat substitute products, improvements in meat substitute products and high moisture protein texturized extrusion have been invented. The inventors have found that by appropriate selection of extrusion parameters and starting materials comprising mechanically processed starch-containing grains, the formation of an emulsion between starch and a protein melt forming a protein matrix can be prevented or reduced to such an extent that a large amount of unbound starch is present in the protein matrix. It was observed that the presence of unbound starch in the protein matrix improves the mouthfeel and maintains an acceptable mouthfeel for a long time. This patent application contains a large number of independent claims for meat substitute products and methods.

[0569] It is obvious to the skilled person that as technology advances, the basic idea of ​​the invention can be implemented in many ways. The invention and its embodiments are therefore not limited to the examples and samples described above, but they can vary within the content of the patent claims and their legal equivalents.

[0570] In the foregoing description of the present invention, unless the context requires, due to expressive language or necessary implication, the word "comprise" or variations such as "comprises" or "comprising" are used in an inclusive sense, that is, to specify the presence of the features described in each embodiment of the present invention, but not to exclude the presence or addition of more features.

[0571] List of referenced publications:

[0572] [Reference 1] Tolstoguzov, VB (1993), Thermoplastic extrusion—themechanism of the formation of extrudate structure and properties. J Am OilChem Soc, 70: 417-424. doi:10.1007 / BF02552717

[0573] [Reference 2] Akdogan, H. (1999), High moisture food extrusion. International Journal of Food Science & Technology, 34: 195-207. doi:10.1046 / j.1365-2621.1999.00256.x

[0574] [Reference 3] Lin, S., Huff, H. and Hsieh, F. (2000), Texture andChemical Characteristics of Soy Protein Meat Analog Extruded at HighMoisture. Journal of Food Science, 65: 264-269. doi:10.1111 / j.1365-2621.2000.tb15991.x

[0575] [Reference 4] Xiang Dong Sun, Susan D. Arntfield. (2010) Gelationproperties of salt-extracted pea protein induced by heat treatment. FoodResearch International. Volume 43, Issue 2, 2010, Pages 509-515.

[0576] [Reference 5] MH Boyacioglu and BL D'Appolonia. (1994)Characterization and utilization of durum wheat for breadmaking III. Stalingproperties of bread baked from bread wheat flours and durum wheat flours.Cereal Chemistry. 71:34-41

[0577] [Reference 6] SZCZESNIAK, AS (1963). Classification of textural characteristics. J. Food Sci, 28, 385-389.

[0578] [Reference 7] BOURNE, MC (1978). Texture Profile Analysis. FoodTechnol., 32 (7), 62-66, 72.

[0579] [Reference 8] BOURNE, MC (1988). Basic Principles of Food TextureMeasurement. Lecture text of Dough Rheology and Baked Products TextureWorkshop - Chicago.

[0580] [Reference 9] SZCZESNIAK, AS (1966). Texture Measurements. FoodTechnol., 20, 50, 55-58.)

[0581] [Reference 10] McGrance, S. J., Cornell, H. J. and Rix, C. J. (1998),A Simple and Rapid Colorimetric Method for the Determination of Amylose inStarch Products. Starch / Stärke, 50: 158-163. doi:10.1002 / (SICI)1521-379X(199804)50:4<158::AID-STAR158>3.0.CO;2-7.

[0582] [Reference 11] Adedeji, O. E., Oyinloye, O. D., & Ocheme, O. B.(2014). Effects of germination time on the functional properties of maizeflour and the degree of gelatinization of its cookies. African Journal ofFood Science, 8(1), 42-47.

[0583] [Reference 12] Azarfar, A., Williams, B. A., Boer, H. and Tamminga,S. (2007) In vitro gas production profile and the formation of end productsfrom non‐ washable, insoluble washable and soluble washable fractions in someconcentrate ingredients. Journal of the Science of Food and Agriculture. 87:1345-1355

Claims

1. A method for producing a meat substitute product, the method comprising the following steps: a) feeding a mixture into an extruder configured to perform high moisture protein texturizing extrusion, the mixture comprising: a1) at least one proteinaceous matrix-forming component, and a2) Mechanically processed starch-containing grains having a diameter of at least 0.125 mm 3 The particle volume; b) feeding water into the extruder; c) heating the mixture in the extruder to gelatinize the starch-containing grains; d) after gelatinization of the starch-containing grains is achieved, further heating the mixture in the extruder to melt at least one protein matrix-forming ingredient; and e) extruding the mixture through an extrusion die at a temperature between 70°C and 100°C in: i) carrying out the heating step c) with impingement heating such that the starch-containing grains are gelatinized before they are substantially pulverized by the extruder screw; and ii) carrying out the heating step d) by shock heating, such that: (a) before the gelatinized starch-containing grains are emulsified with a protein matrix-forming component, as well as (b) the protein melting temperature of the protein matrix forming component is reached before the gelatinized starch-containing grains form a complete barrier that prevents the formation of a continuous protein fiber cross-linked matrix.

2. The method according to claim 1, wherein: The protein matrix-forming component is a protein isolate or a protein concentrate.

3. The method according to claim 1, wherein: The mechanically processed starch-containing grains have a thickness of at least 1 mm. 3 of particle volume.

4. The method according to claim 3, wherein: The starch-containing grains have a diameter of at least 6 mm 3 of particle volume.

5. The method according to claim 1, wherein: The starch-containing grains are soaked before being fed into the extruder.

6. The method according to claim 1 or 5, wherein: The starch-containing grains are treated before being fed into the extruder such that the starch-containing grains are gelatinized before being fed into the extruder.

7. The method according to any one of claims 1 to 5, wherein: In step b), water is fed to the starch-containing grains at elevated temperature.

8. The method according to claim 7, wherein: The water temperature is above 60°C.

9. The method according to claim 8, wherein: The water temperature is above 65°C.

10. The method according to claim 7, wherein: The water temperature is above 75°C.

11. The method according to any one of the preceding claims 1 to 5, wherein: Said heating step d) is carried out at a temperature between 140°C and 200°C.

12. The method according to any one of the preceding claims 1 to 5, wherein: The heating step d) is performed so that protein melting occurs between 1 s and 40 s after step b).

13. The method according to claim 12, wherein: The heating step d) is performed such that protein melting occurs between 10 s and 30 s after step b).

14. The method according to any one of the preceding claims 1 to 5, wherein: The heating step c) is performed such that gelatinization of the starch-containing grains occurs between 0 and 18 seconds after step b).

15. The method according to claim 14, wherein: The heating step c) is performed such that gelatinization of the starch-containing grains occurs between 1 s and 15 s after step b).

16. The method according to any one of the preceding claims 1 to 5, wherein: The starchy grains are ground into particles with a volume of less than 5000 μm by the extruder screw. 3 The heating step c) is performed previously.

17. The method of claim 16, wherein: The starch-containing grains are ground into particles with a volume of less than 0.001 mm by the extruder screw. 3 The heating step c) is performed previously.

18. The method according to any one of the preceding claims 1 to 5, wherein: After said heating step d), the mixture is extruded for more than 5 seconds at a temperature not higher than the temperature in the heating step d).

19. The method of claim 18, wherein: After said heating step d), the mixture is extruded for more than 5 seconds at a temperature between 90°C and the temperature in heating step d).

20. The method according to any one of the preceding claims 1 to 5, wherein: The mechanically processed starch-containing grains comprise one or more of the following or consist of one or more of the following: oat flakes, steel-cut grains, shelled and pearled grains, crushed grains, shelled but not pearled grains, however excluding: shelled but not pearled oat grains, shelled but not pearled rye grains, shelled but not pearled barley grains, shelled but not pearled corn grains.

21. The method of claim 20, wherein: The oatmeal is compressed, flattened or fragmented oatmeal.

22. The method according to any one of the preceding claims 1 to 5, wherein: The mechanically processed starch-containing grains comprise or consist of one or more of the following: oats, barley, rye, wheat, rice, corn, lentils, chickpeas, mung beans, broad beans, peas, quinoa, pigeon peas, sorghum, buckwheat, but exclude: hulled but not pearled oat grains, hulled but not pearled rye grains, hulled but not pearled barley grains, hulled but not pearled corn grains.

23. The method according to any one of the preceding claims 1 to 5, wherein: The extrusion step is carried out with an extrusion die having a length greater than 300 mm.

24. The method of claim 23, wherein: The extrusion step is performed using an extrusion die having a length greater than 1000 mm.

25. The method of claim 24, wherein: The extrusion step is performed using an extrusion die having a length between 1000 mm and 5000 mm.

26. A method for producing a meat substitute product, the method comprising the steps of: a) feeding a mixture into an extruder configured to perform high moisture protein texturizing extrusion, the mixture comprising: a1) at least one proteinaceous matrix-forming component, and a2) Mechanically processed starch-containing grains, which are steel-cut grains and have a diameter of at least 0.125 mm 3 The particle volume; b) feeding water into the extruder so that the moisture content during extrusion is between 40% and 80%; c) heating the mixture in the extruder to gelatinize the starch-containing grains; d) after gelatinization of the starch-containing grains is achieved, further heating the mixture in the extruder to melt at least one protein matrix-forming ingredient; and e) extruding the mixture through an extrusion die at a temperature between 70° C. and 100° C., in: i) carrying out the heating step c) with impingement heating such that the starch-containing grains are gelatinized before they are substantially pulverized by the extruder screw; and ii) carrying out the heating step d) by shock heating, such that: (a) before the gelatinized starch-containing grains are emulsified with a protein matrix-forming component, as well as (b) the protein melting temperature of the protein matrix forming component is reached before the gelatinized starch-containing grains form a complete barrier that prevents the formation of a continuous protein fiber cross-linked matrix.

27. The method according to claim 26, wherein: The protein matrix-forming component is a protein isolate or a protein concentrate.

28. The method of claim 26, wherein: The mechanically processed starch-containing grains have a thickness of at least 1 mm. 3 of particle volume.

29. The method of claim 27, wherein: The mechanically processed starch-containing grains have a diameter of at least 6 mm. 3 of particle volume.

30. The method of claim 26, wherein: The steel cut kernels were soaked in water before being fed into the extruder.

31. The method of claim 26, wherein: The steel cut kernels were not soaked when fed into the extruder.

32. The method according to any one of claims 26 to 31, wherein: The steel cut grains comprise steel cut oats.

33. The method according to any one of claims 26 to 31, wherein: The steel cut oats may be replaced with any of the following: steel cut barley, rice kernels, cracked rice, pearled barley, pearled rye, pearled wheat, pearled oats, cracked pea seeds, cracked fava bean seeds, cracked chickpea seeds, cracked lentil seeds or a mixture thereof.

34. The method of claim 33, wherein: The particle size of the crushed pea seeds is 2 mm.

35. The method according to any one of claims 26 to 31, wherein: In the method, the use of extrusion shock heating temperature setting and the use of hot water as liquid feed are combined to increase starch solubility.

36. The method according to any one of claims 26 to 31, wherein: In the method, grains and water are mixed, and the grains combined with the water are heated early enough before the starch of the grains is emulsified by the protein matrix.

37. A meat substitute product produced using the method according to any one of claims 26 to 36.

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