WO2025163063A1

WO2025163063A1 - A nutrition bar

Info

Publication number: WO2025163063A1
Application number: PCT/EP2025/052390
Authority: WO
Inventors: Margriet Sjanien GROENENBOOM; Cornelis KORNET; Silke Sterre RINSMA
Original assignee: Vivici BV
Current assignee: Vivici BV
Priority date: 2024-01-31
Filing date: 2025-01-30
Publication date: 2025-08-07
Anticipated expiration: 2026-07-31

Abstract

The present invention relates to a nutrition bar comprising a protein and extraneous β-lactoglobulin. The invention further relates to a process for the preparation of such nutrition bar. The invention further relates to the use of a β-lactoglobulin to modulate the hardness, firmness and/or cohesiveness of a nutrition bar.

Description

A nutrition bar

Field of the invention

The present invention relates to a nutrition bar comprising a protein and extraneous p-lactoglobulin. The invention further relates to a process for the preparation of such nutrition bar. The invention further relates to the use of a p-lactoglobulin to modulate the hardness, firmness and/or cohesiveness of a nutrition bar.

Background of the invention

Nutrition bars (sometimes referred to as nutritional bars) are food products that are formulated to provide a balanced mix of (essential) nutrients. These bars typically contain a combination of macronutrients such as protein, carbohydrates, and fats, as well as various vitamins and minerals. The exact recipe of nutrition bars can vary widely, and nutrition bars may be designed to meet specific dietary goals or nutritional requirements.

Nutrition bars may range from about 10 to about 150 grams in weight. Nutrition bars may range from about 0.5 cm to about 4 cm in height. Nutrition bars may range from about 4 cm to about 15 cm in length. Nutrition bars may range from about 1 cm to about 7 cm in width. The corners of a nutrition bar may be sharp or round. The person skilled in the art will comprehend that these features may vary without any impact on the essential features of a nutrition bar.

Protein bars are nutrition bars that are specifically designed to provide a convenient and concentrated source of protein. They come in a variety of flavors and formulations and typically contain a combination of protein sources, such as dairy protein e.g. whey or other plant-based proteins (e.g. pea, fava, oat, soy, rice, nuts, pumpkin seeds), carbohydrates, and fats, along with various vitamins and minerals. These bars are commonly consumed by individuals seeking to increase their protein intake, especially athletes, fitness enthusiasts, and those looking for a quick and portable snack option.

Meal replacement bars are nutrition bars that are specifically formulated to replace a full meal, for e.g. dietary goals.

Energy bars are nutrition bars that focus on providing a quick source of energy, often through carbohydrates and sometimes added caffeine or other energy-boosting ingredients. Usually, protein bars are lower in carbohydrates than energy bars, lower in vitamins and dietary minerals than meal replacement bars, and significantly higher in protein than either. Energy bars provide the larger part of their food energy (calories) in carbohydrate form. Meal replacement bars are intended to replace the variety of nutrients in a meal.

Nutrition bars usually are made of a fat, protein, and carbohydrate source. Flavoring components, stabilizers, fruits, and nuts may also be present. A common protein source for vegan protein bars are plant proteins such as pea protein. The fat source used is most often selected from the group consisting of vegetable shortening, nut butter, cocoa butter, or some type of oil (e.g., canola, coconut, or vegetable). The carbohydrate source used is most often selected from the group consisting of a sugar source (e.g., glucose, maltodextrin), a sugar alcohol (e.g. sorbitol or maltitol), a fiber syrup (e.g., inulin, fructose) or mixtures thereof. The above combination of ingredients is mixed to form a dough that is subsequently formed into bars. A drawback of state of the art nutrition bars is that they are relatively hard. This typically occurs already immediately after production but can also manifest upon storage. Many nutrition bars generally become too hard to make them difficult to consume, within only months from the production date. It is generally believed that the phenomenon of hardening becomes more severe with increasing amounts of protein. In traditional non-vegan high protein nutrition bars, the hardening effect is a well-known problem, see e.g. McMahon et al. (J. Food Sci. (2009) 74(6), E312-E321) wherein hardening was attempted to be reduced by applying hydrolyzed WPI or combining WPI with appropriate carbohydrates. Some manufacturers almost exclusively use hydrolyzed proteins, albeit that hydrolyzed proteins generally are more expensive and can create quality issues such as bitter off-flavor and negative textural changes. Furthermore, nutrition bars and dough containing hydrolyzed protein also tend to stick to equipment, making it more difficult to process the dough and bars. Alternatively, in WO 2022/015671 and US2022007681 , the hardening effect in protein bars containing WPI was reduced by acidifying the whey protein followed by extrusion to obtain puffed protein products. A drawback of acidification and extrusion is that the former requires additional chemicals and may impact flavour and taste while the latter is considered an energy-intensive process manufacturers may seek to replace by milder lower temperature mixing processes.

Malecki et al. (Int. J. Environ. Res. Public Health (2022) 19(7), 3923-3937) reported the reduction of hardness in protein bars by combining proteins with liquid syrups based on tapioca, oligofructose or maltitol. Malecki et al uses large amounts of these syrups (about 32% w/w), which may be undesired.. In addition, the addition of further carbohydrates to nutrition bars may be unwanted in view of nutritional and/or dietary requirements.

When nutrition bars comprise compounds from animal sources, such as lactose, this may cause problems since users may be intolerant to lactose. In addition, sugars such as lactose may crystalize and consequently attribute to the hardening problem.

Altogether, there remains a need to reduce product hardening in nutrition bars that are partly or fully based on plant proteins.

Summary of the invention

The invention relates to a nutrition bar comprising protein and extraneous p-lactoglobulin.

The invention further relates to a process for the preparation of a nutrition bar, comprising: mixing the desired ingredients comprising at least a protein and extraneous p-lactoglobulin to result in a dough, and forming a bar of the dough.

The invention further relates to the use of a p-lactoglobulin to modulate the hardness, firmness and/or cohesiveness of a nutrition bar.

Description of the invention

The inventors have established that when mixing protein with extraneous p-lactoglobulin in the preparation of nutrition bars, it resulted in nutrition bars that exhibited less hardening over time compared to nutrition bars prepared with the protein without extraneous p-lactoglobulin. This is a surprising finding since p-lactoglobulin is the main component of whey and nutrition bars containing whey protein are well-known to suffer from the hardening problem. In addition, it was established that the original softness of the nutrition bars is retained for a longer period compared to a nutrition bar prepared with the protein without extraneous p-lactoglobulin. Furthermore, it was established that, upon mixing protein with extraneous p-lactoglobulin, the resulting nutrition bars also display improved (higher) cohesiveness and improved (lower) firmness and that this improved (higher) cohesiveness and improved (lower) firmness was maintained over time.

Accordingly, in a first aspect, there is provided for a nutrition bar comprising protein and extraneous p-lactoglobulin. In the embodiments herein, the nutrition bar is referred to as the “nutrition bar according to the invention” and interchangeably the terms “nutritional bar according to the invention” or simply “nutrition bar” or “bar” are used. The person skilled in the art will comprehend that p-lactoglobulin is a protein itself, albeit it is not a plant protein. In the embodiments herein when the term protein is used, such as when discussing the ratio of protein to extraneous p-lactoglobulin, the protein is to be regarded as the protein fraction excluding the extraneous p-lactoglobulin, unless explicitly stated that the extraneous p-lactoglobulin is included. In the embodiments herein, the term “extraneous p-lactoglobulin” means that p-lactoglobulin is added as a separate fraction irrespective of a protein fraction of the nutrition bar that may comprise p-lactoglobulin as an intrinsic compound; the intrinsic p-lactoglobulin is thus part of the protein fraction excluding the extraneous p-lactoglobulin. In the embodiments herein, when the term p-lactoglobulin is used, it is to be construed as extraneous p-lactoglobulin; intrinsic p-lactoglobulin is only intended if the explicit term "intrinsic p-lactoglobulin” is used.

Preferably, the hardness of the nutrition bar at three months post production is at most 12.000 g, such as at most 8.000 g, 9.000 g, 10.000 g, 11 .000, or at most 12.000 g.

Preferably, the firmness of the nutrition bar at three months post production is at most 2000 kPa, such as 900 kPa, 1000 kPa, 1100 kPa, 1200 kPa, 1300 kPa, 1400k Pa, 1500 kPa, 1600 kPa, 1700kPa, 1800 kPa, 1900 kPa or at most 2000 kPa.

Preferably, the cohesiveness of the nutrition bar at three months post production is at least 0.09, such as 0.09, 0.1 , 0.11 , 0.12, 0.13, 0.14, 0.15, 0.16, 0.17, 0.18 or at least 0.2.

In the embodiments herein, at least part of the protein in the nutrition bar may be a plant protein. The part of the protein, excluding the extraneous p-lactoglobulin, that is plant protein may be at least 1 %, such as 1 %, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11 %, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21 %, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31 %, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41 %, 42%, 43%, 44%, 4%, 46%, 47%, 48%, 49%, 50%, 51 %, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61 %, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71 %, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%; 100% meaning that all protein in the nutrition bar, excluding the extraneous p-lactoglobulin, is plant protein.

In the embodiments herein, the protein may be any protein known to the person skilled in the art, such as but not limited to whey protein and casein protein. In the embodiments herein, the plant protein may be any plant protein known to the person skilled in the art. In the embodiments herein, the p-lactoglobulin may be any p-lactoglobulin known to the person skilled in the art.

In the embodiments herein, the nutrition bar may comprise at least 5% (w/w), 6%, 7%, 8%, 9%, 10%, 11 %, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21 %, 22%, 23%, 24%, 25%, 26%, 27%, 28%,

29%, 30%, 31 %, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41 %, 42%, 43%, 44%, 45%, 46%,

47%, 48%, 49%, 50%, 51 %, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61 %, 62%, 63%, 64%,

65%, 66%, 67%, 68%, 69%, 70%, 71 %, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, or at least 80%

(w/w) of total protein, including the extraneous p-lactoglobulin, in view of the total weight of the bar.

In the embodiments herein, the nutrition bar may comprise at least 5% (w/w), 6%, 7%, 8%, 9%, 10%, 11 %, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21 %, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31 %, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41 %, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51 %, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61 %, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71 %, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, or at least 80% (w/w) of plant protein in view of the total weight of the bar.

In the embodiments herein, the nutrition bar may comprise, further to protein and extraneous p-lactoglobulin, a fat and a carbohydrate. The person skilled in the art knows which fats and carbohydrates are suitable for use in a nutrition bar and any of those may be used. The nutrition bar may further comprise compound that are regularly used in nutrition bars, such as vitamins, minerals, energy boosting compounds such as caffein, and flavoring compounds. The fat in the nutrition bar herein may be selected from the group consisting of vegetable shortening, nut butter, cocoa butter, and vegetable oil, such as canola oil, sunflower oil, and coconut oil. The carbohydrate in the a nutrition bar herein may be selected from the group consisting of a sugar source, such as glucose and maltodextrin, glycerin, a sugar alcohol, such as sorbitol and maltitol, and a fiber syrup such as inulin and fructose syrup.

In the embodiments herein, the plant protein in the nutrition bar may be any plant protein known to the person skilled in the art to be suitable for use in a nutrition bar. In the embodiments herein, the plant protein in the nutrition bar may be a legume protein, a cereal protein and/or a nut protein.

In the embodiments herein, the plant protein in the nutrition bar may comprise or may be soy protein, rice protein, lupine protein, chickpea protein, lentil protein, pea protein, fava protein and/or protein from nuts, such as almond and cashew.

In the embodiments herein, the extraneous p-lactoglobulin may be any p-lactoglobulin known the person skilled in the art. The p-lactoglobulin may be a non-recombinant or a recombinant p-lactoglobulin. p-lactoglobulin is the major whey protein in the milk of many mammals. In bovine milk it accounts for approximately 10 - 15% of total milk proteins and about 50 - 54% of whey protein. Bovine p-lactoglobulin is expressed as a precursor protein comprising a 16 amino acid N-terminal signal peptide (referred to herein and elsewhere as the "full-length" p-lactoglobulin protein), which is cleaved to form a mature 162 amino acid protein. There are two primary variants of bovine p-lactoglobulin - variants A and B and a less common variant - variant C. Sequences for both the mature and full-length forms of bovine p-lactoglobulin variants A, B and C, and wild type full length and mature forms of p- lactoglobulin from other species are presented in Table 1. Preferred p-lactoglobulins include proteins comprising an amino acid sequence having at least about 70% sequence identity to the sequence of a wild type (native) p-lactoglobulin (either full length or mature p-lactoglobulin lacking a signal sequence, but preferably the mature sequence), but particularly any wild type bovine, ovine, caprine, buffalo, equine, donkey or reindeer p-lactoglobulin sequence, including any sequence of SEQ ID NO: in Table 1 . In some embodiments the amino acid sequence of such variants may comprise a truncation or an elongation at the N-terminus and/or the C-terminus relative to the wildtype sequence, for example, truncations or elongations of from about 1 to about 20 amino acids, such as 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, 16, 17, 18, 19, or 20 amino acids. In some embodiments, variants or modified p-lactoglobulin proteins may contain from 1 to 20 amino acid insertions, deletions, and/or substitutions (collectively) with respect to the wild-type sequence. Such proteins may be referred to herein as "elongated p-lactoglobulin proteins". In some embodiments the variants or modified p- lactoglobulin proteins may comprise one or more post-translational modifications that differ to a wild type p-lactoglobulin protein, including glycosylation and or phosphorylation at one or more residues. An N-terminal elongation may have a sequence comprising or consisting of amino acid A. An N-terminal elongation may have a sequence comprising or consisting of EA, or two or more repeats of EA, for example three or more repeats of EA, four or more repeats of EA, or five or more repeats of EA. For example, the N-terminal elongation may have a sequence comprising or consisting of A, EA, EAEA (SEQ ID NO: 22), EAEAEA (SEQ ID NO: 23), EAEAEAEA (SEQ ID NO: 24), EAEAEAEAEA (SEQ ID NO: 25, REAEAM (SEQ ID NO: 26), REAEAEAM (SEQ ID NO: 27), REAEAEAEAM (SEQ ID NO: 28), KREAEAM (SEQ ID NO: 29), KREAEAEAM (SEQ ID NO: 30), or KREAEAEAEAM (SEQ ID NO: 31). In the embodiments herein, the p-lactoglobulin may be a mixture of heterogenous p-lactoglobulin proteins. In the embodiments herein, the p-lactoglobulin may be a plurality of recombinant p- lactoglobulin proteins heterogeneous in amino acid sequence, such as at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 14, 15, 16, 18 or 20 recombinant p -lactoglobulin proteins of differing amino acid sequence and/or of differing elongation. In a non-limiting embodiment, the p-lactoglobulin may comprise a native p- lactoglobulin and a p-lactoglobulin having an N-terminal elongation of the amino acid A. In a non-limiting embodiment, the p-lactoglobulin may comprise a native p-lactoglobulin and a p-lactoglobulin having an N-terminal elongation of the amino acids EAE. In a non-limiting embodiment, the p-lactoglobulin may comprise a p-lactoglobulin having an N-terminal elongation of the amino acid A and a p-lactoglobulin having an N-terminal elongation of the amino acids EAE.

The elongated p-lactoglobulins and mixes thereof as set forward in WO2022/269549 are preferred p- lactoglobulins and are herein incorporated by reference. Accordingly, the recombinant p-lactoglobulins comprising or consisting of an amino acid sequence having at least about 70% sequence identity to a sequence selected from the group consisting of: SEQ ID NO: 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, 35, 36, 37, 38, 39, 40, 41 , 42, 43, 44, 45, 46, 47, 48, 61 , 71 , 72, 73, and 74 as set forward in WO2022/269549 are preferred p- lactoglobulins herein. In the embodiments herein, the p-lactoglobulin may comprise non-native post- translational modification modulating e.g. the glycosylation and/ or phosphorylation of the p- lactoglobulin, as set forward in WQ2020219596A1 and US2022211061 , which are herein incorporated by reference. Accordingly, the p-lactoglobulin may be a recombinant p-lactoglobulin comprising an amino acid sequence that comprises one or more of amino acid residues selected from T4, T6, T18, S21 , S27, S30, S36, T49, T76, T97, S110, S116, T125, S150, N152, and T154 of Bos taurus p-lactoglobulin, and having non-native glycosylation on one or more of such amino acid residues. The p-lactoglobulin may be a recombinant p-lactoglobulin comprising an amino acid sequence that comprises amino acid residue N 152 of Bos taurus p-lactoglobulin, and having non-native N-glycosylation on such amino acid residue. The p-lactoglobulin may be a recombinant p-lactoglobulin comprising an amino acid sequence that comprises one or more of amino acid residues selected from T4, T6, T18, S21 , S27, S30, S36, T49, T97, SI 10, SI 16, T125, S150, and T154 of Bos ta urus p-lactoglobulin, and having non-native O- glycosylation on one or more of such amino acid residues. The p-lactoglobulin may be a recombinant p-lactoglobulin comprising an amino acid sequence that comprises one or more of amino acid residues selected from T4, T6, T18, Y20, S21 , S27, S30, S36, Y42, T49, T76, T97, Y99, Y102, SI 10, SI 16, T125, S150, and T154 of Bos taurus p-lactoglobulin, and having non native phosphorylation on one or more of such amino acid residues. The p-lactoglobulin may be a recombinant p-lactoglobulin comprising an amino acid sequence that comprises one or more of amino acid residues selected from K8, K14, R40, K47, K60, K69, K70, K75, K77, K83, K91 , K100, K101 , R124, K135, K138, K141 , and R148 of Bos taurus p-lactoglobulin, and having non native methylation on one or more of such amino acid residues. The p-lactoglobulin may be a recombinant p-lactoglobulin comprising an amino acid sequence that comprises one or more of amino acid residues selected from C66, C106, C119, C121 , and C160 of Bos taurus p-lactoglobulin, and having non-native palmitoylation on one or more of such amino acid residues. The p-lactoglobulin may be a recombinant p-lactoglobulin comprising an amino acid sequence that comprises one or more of amino acid residues selected from K8, K14, K47, K60, K69, K70, K75, K77, K83, K91 , K100, K101 , K135, K138, and K141 of Bos taurus p-lactoglobulin, and having non- native sumoylation on one or more of such amino acid residues. The p-lactoglobulin may be a recombinant p-lactoglobulin comprising an amino acid sequence that comprises one or more of amino acid residues selected from C66, C106, C119, C121 , and C160 of Bos taurus p-lactoglobulin, and having non-native nitrosylation on one or more of such amino acid residues. The p-lactoglobulin n may be a recombinant p-lactoglobulin comprising an amino acid sequence that comprises one or more of amino acid residues selected from Y20, Y42, Y99, and Y102 of Bos taurus p-lactoglobulin, and having non-native tyrosine nitration on one or more of such amino acid residues. The p-lactoglobulin may be a recombinant p-lactoglobulin comprising an amino acid sequence that comprises one or more of amino acid residues selected from F151 of Bos taurus p-lactoglobulin, and having non-native glypiation on such amino acid residue. The p-lactoglobulin may be a recombinant p-lactoglobulin comprising an amino acid sequence that comprises one or more of amino acid residues selected from C160 of Bos taurus p-lactoglobulin, and having non-native farnesylation on such amino acid residue. The p- lactoglobulin may be a recombinant p-lactoglobulin comprising an amino acid sequence that comprises one or more of amino acid residues selected from C160 of Bos taurus p-lactoglobulin, and having non- native geranylgeranylation on such amino acid residue.

In the embodiments herein, the p-lactoglobulin protein may have an attenuated or essentially eliminated allergenicity, such e.g. the p-lactoglobulin as set forward in WO2021168343 and US2023106635, which are herein incorporated by reference. Accordingly, the recombinant p-lactoglobulins comprising or consisting of an amino acid sequence having at least about 70% sequence identity to a sequence selected from the group consisting of: SEQ ID NO: 1 , 2, 3, 4, 5, 6, 7, 8, 9, and 10 of WO2021168343 and US2023106635 are preferred p-lactoglobulins herein.

It will be evident that the p-lactoglobulin may be a naturally occurring p-lactoglobulin, or may be a recombinant p-lactoglobulin. A natural p-lactoglobulin may be added as such or may be part of a composition distinct from the composition according to the invention, such as whey, provided that the whey comprises a high amount of p-lactoglobulin, such as e.g. in example 5 herein. A preferred p-lactoglobulin is a recombinant p-lactoglobulin as set forth herein. A preferred recombinant p-lactoglobulin is one selected from the group consisting of: a. a p-lactoglobulin having an amino acid sequence that has at least 90% sequence identity with the amino acid sequence as said forth in any one of SEQ ID NO: 1 to 21 and 32 and 33; b. a p-lactoglobulin having an amino acid sequence that has at least 90% sequence identity with the amino acid sequence as set forth in SEQ ID NO: 1 , wherein the p- lactoglobulin comprises at least one additional charged amino acid compared to p- lactoglobulin with the amino acid sequence as set forth in SEQ ID NO: 1 , and wherein, preferably, the additional charged amino acid is not an N-terminal EA, EAEA, EAEAEA, EAEAEAEA, or R; c. a p-lactoglobulin having an amino acid sequence that has at least 90% sequence identity with the amino acid sequence as set forth in SEQ ID NO: 3, wherein the p- lactoglobulin comprises at least one additional charged amino acid compared to p- lactoglobulin with the amino acid sequence as set forth in SEQ ID NO: 3, and wherein, preferably, the additional charged amino acid is not an N-terminal EA, EAEA, EAEAEA, EAEAEAEA, or R; d. a p-lactoglobulin having an amino acid sequence that has at least 90% sequence identity with the amino acid sequence as set forth in SEQ ID NO: 5, wherein the p- lactoglobulin comprises at least one additional charged amino acid compared to p- lactoglobulin with the amino acid sequence as set forth in SEQ ID NO: 5; e. a p-lactoglobulin having an amino acid sequence that has at least 90% sequence identity with the amino acid sequence as set forth in SEQ ID NO: 7, wherein the p- lactoglobulin comprises at least one additional charged amino acid compared to p- lactoglobulin with the amino acid sequence as set forth in SEQ ID NO: 7; f. a p-lactoglobulin having an amino acid sequence that has at least 90% sequence identity with the amino acid sequence as set forth in SEQ ID NO: 9, wherein the p- lactoglobulin comprises at least one additional charged amino acid compared to p- lactoglobulin with the amino acid sequence as set forth in SEQ ID NO: 9; g. a p-lactoglobulin having an amino acid sequence that has at least 90% sequence identity with the amino acid sequence as set forth in SEQ ID NO: 11 , wherein the p- lactoglobulin comprises at least one additional charged amino acid compared to p- lactoglobulin with the amino acid sequence as set forth in SEQ ID NO: 11 ; h. a p-lactoglobulin having an amino acid sequence that has at least 90% sequence identity with the amino acid sequence as set forth in SEQ ID NO: 13, wherein the p- lactoglobulin comprises at least one additional charged amino acid compared to p- lactoglobulin with the amino acid sequence as set forth in SEQ ID NO: 13; i. a p-lactoglobulin having an amino acid sequence that has at least 90% sequence identity with the amino acid sequence as set forth in SEQ ID NO: 15, wherein the p- lactoglobulin comprises at least one additional charged amino acid compared to p- lactoglobulin with the amino acid sequence as set forth in SEQ ID NO: 15; j. a p-lactoglobulin having an amino acid sequence that has at least 90% sequence identity with the amino acid sequence as set forth in SEQ ID NO: 17, wherein the p- lactoglobulin comprises at least one additional charged amino acid compared to p- lactoglobulin with the amino acid sequence as set forth in SEQ ID NO: 17; k. a p-lactoglobulin having an amino acid sequence that has at least 90% sequence identity with the amino acid sequence as set forth in SEQ ID NO: 19, wherein the p- lactoglobulin comprises at least one additional charged amino acid compared to p- lactoglobulin with the amino acid sequence as set forth in SEQ ID NO: 19; and l. a p-lactoglobulin having an amino acid sequence that has at least 90% sequence identity with the amino acid sequence as set forth in SEQ ID NO: 21 , wherein the p- lactoglobulin comprises at least one additional charged amino acid compared to p- lactoglobulin with the amino acid sequence as set forth in SEQ ID NO: 21 .

In the embodiments herein, the p-lactoglobulin comprising at least one additional charged amino acid may have a surface charge (^-potential) of at least 12mV at a pH of at most 4 and/or has a surface charge (^-potential) of at most -12mV at a pH of at least 6.

In the embodiments herein, the p-lactoglobulin comprising at least one additional charged amino acid may have a surface charge ( -potential)of at most -12mV at a pH of at least 5.5 and/or has a surface charge (^-potential) of at most -8mV at a pH of at least 5.

In the embodiments herein, the p-lactoglobulin comprising at least one additional charged amino acid may have enhanced physical stability compared to the p-lactoglobulin selected from the group consisting of SEQ ID NO: 1 , 3, 5, 7, 9, 11 , 13, 15, 17, 19, and 21 that has the highest sequence identity with the p-lactoglobulin with the at least one additional charged amino acid.

In the embodiments herein, in the p-lactoglobulin the additional charged amino acids may be selected from the group consisting of Glutamic acid, Aspartic acid, Arginine, Lysine and Histidine.

In the embodiments herein, in the p-lactoglobulin the additional charged amino acid may be located between the N-terminal amino acid and the C-terminal amino acid of the p-lactoglobulin.

In the embodiments herein, in the p-lactoglobulin at least one of the additional charged amino acid may be located at the surface of the mature p-lactoglobulin. In the embodiments herein, a more preferred p-lactoglobulin is a p-lactoglobulin that is not of an animal source, a p-lactoglobulin not isolated and/or purified from milk, whey or the like. Accordingly, a more preferred nutrition bar according to the invention is a nutrition bar that comprises a p-lactoglobulin that is not of an animal source. An even more preferred nutrition bar is a nutrition bar that does not comprise any compounds from an animal source, i.e. an animal-free nutrition bar. Such animal-free nutrition bar preferably comprises at most 0.1% (w/w) compounds from animal origin, more preferably at most 0.01% (w/w), at most 0.001 % (w/w), even more preferably at most 0.0001 % (w/w). Most preferably no compounds from animal origin are detectable in an animal-free nutrition bar. A specifically preferred nutrition bar according to the invention is devoid of lactose. Accordingly, in the embodiments herein, the nutrition bar may be devoid of, or may have low amounts of, compounds that occur in animal- derived p-lactoglobulin sources. Examples of such compounds are a-lactalbumin, bovine serum albumin, casein, glycomacropeptide, immunoglobulins, lactoferrin, and lactose. A nutrition bar that is devoid of compounds that occur in animal-derived p-lactoglobulin sources may be advantageous for applications wherein such components are undesirable or detrimental, for example for allergenic, health, or nutritional considerations. The amount of any of a-lactalbumin, bovine serum albumin, casein, glycomacropeptide, immunoglobulins, lactoferrin, or lactose in a nutrition bar according to the invention may be at most 1 % (w/w), such as between 0.0001 % and 0.5% (w/w), between 0.001 and 0.2% (w/w), or between 0.01 and 0.1 % (w/w), based on the total weight of the nutrition bar. The amount of any of a-lactalbumin, bovine serum albumin, casein, glycomacropeptide, immunoglobulins, lactoferrin, or lactose in a nutrition bas according to the invention may be at most 1 % (w/w), such as at most 0.1 % (w/w), at most 0.01 % (w/w), at most 0.001 % (w/w), or most preferably at most 0.0001 % (w/w).

In an embodiment, the ratio of p-lactoglobulin : a-lactalbumin (w/w) may be from 10,000 to 2, from 5,000 to 5, or from 1 ,000 to 10.

In an embodiment, the ratio of p-lactoglobulin : bovine serum albumin (w/w) may be from 10,000 to 5, from 5,000 to 10, or from 1 ,000 to 50.

In an embodiment, the ratio of p-lactoglobulin : glycomacropeptide (w/w) may be from 10,000 to 5, from 5,000 to 10, or from 1 ,000 to 50.

In an embodiment, the ratio of p-lactoglobulin : lactoferrin (w/w) may be from 100,000 to 10, from 10,000 to 50, or from 1 ,000 to 100.

In the embodiments herein, the ratio of protein to extraneous p-lactoglobulin may be within any useful range. The ratio may be within the range from 99 : 1 (w/w) to 5 : 95 (w/w), such as from 98 : 2(w/w), from 97 : 3(w/w), from 96 : 4(w/w), from 95 : 5 (w/w), from 90 : 10 (w/w), from 80 : 20 (w/w), from 75 : 25, from 70 : 30 (w/w), from 65 : 35, from 60 : 40 (w/w), from 50 : 50 (w/w), from 40 : 60 (w/w), from 30 : 70 (w/w), from 20 : 80 (w/w), from 10 : 90 (w/w) and from 5 : 95 (w/w). A preferred ratio is within the range from 95 : 5 (w/w) to 60 : 40 (w/w). In the embodiments herein, the ratio of plant protein to extraneous p-lactoglobulin may be within any useful range. The ratio may be within the range from 99 : 1 (w/w) to 5 : 95 (w/w), such as from 98 : 2 (w/w), from 97 : 3 (w/w), from 96 : 4 (w/w), from 95 : 5 (w/w), from 90 : 10 (w/w), from 80 : 20 (w/w), from 75 : 25, from 70 : 30 (w/w), from 65 : 35, from 60 : 40 (w/w), from 50 : 50 (w/w), from 40 : 60 (w/w), from 30 : 70 (w/w), from 20 : 80 (w/w), from 10 : 90 (w/w) and from 5 : 95 (w/w). A preferred ratio is within the range from 95 : 5 (w/w) to 60 : 40 (w/w). A specific preferred ratio in all embodiments of the invention is within the range from 80 : 20 (w/w) to 5 : 95 (w/w). Accordingly, there is provided for a nutrition bar comprising protein and extraneous p-lactoglobulin, wherein the ratio of protein to extraneous p-lactoglobulin is within the range from 80 : 20 (w/w) to 5 : 95 (w/w). A preferred ratio is within the range from 80 : 20 (w/w) to 20 : 80 (w/w). A further preferred ratio is within the range from 80 : 20 (w/w) to 50 : 50 (w/w).

In the embodiments herein, in the nutrition bar, the amount of plant protein plus extraneous p-lactoglobulin in view of the total weight of the nutrition bar may be from 5% (w/w) to 80% (w/w), such as 5%, 6%, 7%, 8%, 9%, 10%, 11 %, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21 %, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31 %, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41 %, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51 %, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61 %, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71 %, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, or 80% (w/w).

In the embodiments herein, the nutrition bar may be a coated nutrition bar. The coating may be any coating known to the useful for a nutrition bar, such as a dairy coating such a yoghurt, or chocolate. In a coated bar, protein, plant protein and/or intrinsic and/or extraneous p-lactoglobulin may or may not be present in the coating. In the embodiments herein, the nutrition bar may have a homogenous content or a non-homogenous content, such as a bar that contains different layers. Each different part of such nutrition bar with non-homogenous content may comprise plant protein and/or extraneous p-lactoglobulin. In the embodiments herein, the nutrition bar may comprise further ingredients such as chocolate, nougat, jam, yoghurt, caramel, dried fruit and/or nuts.

In a second aspect, there is provided for a process for the preparation of nutrition bar according to the invention, comprising: a. mixing the desired ingredients comprising at least a protein and extraneous p-lactoglobulin to result in a dough, and b. forming a bar of the dough.

In this aspect, the features are preferably those of the first aspect.

In a third aspect, there is provided for the use of a p-lactoglobulin to modulate the hardness, firmness and/or cohesiveness of a nutrition bar, preferably after storage. For the sake of clarity for all embodiments herein, this means that the p-lactoglobulin that is added during production maintains its modulative effects during a prolonged storage time. In this aspect, the features are preferably those of the first aspect.

Accordingly, there is provided for the use of a p-lactoglobulin to modulate the hardness of a nutrition bar, preferably after storage. Preferably, the hardness of the nutrition bar is significantly improved after a period of at least 10, 20, 30, 40, 50, 60, 70, 80, or most preferably after 90 days from the production date of the nutrition bar compared to the hardness of a corresponding nutrition bar that does not comprise p-lactoglobulin. In the embodiments herein, the corresponding nutrition bar that does not comprise p-lactoglobulin is a nutrition bar that comprises all corresponding components, except for the added p-lactoglobulin.

In addition, there is provided for the use of a p-lactoglobulin to modulate the firmness of a nutrition bar, preferably after storage, with the proviso that in a nutrition bar comprising 30% protein, the ratio of pea protein and p-lactoglobulin is not 90 : 10. Preferably, the firmness of the nutrition bar is significantly improved after a period of at least 10, 20, 30, 40, 50, 60, 70, 80, or most preferably after 90 days from the production date of the nutrition bar compared to the firmness of a corresponding nutrition bar that does not comprise p-lactoglobulin.

In addition, there is provided for the use of a p-lactoglobulin to modulate the cohesiveness of a nutrition bar, preferably after storage. Preferably, the the cohesiveness of the nutrition bar is significantly improved after a period of at least 10 , 20, 30, 40, 50, 60, 70, 80, or most preferably after 90days from the production date of the nutrition bar compared to the cohesiveness of a corresponding nutrition bar that does not comprise p-lactoglobulin.

In the embodiments herein, the modulation is preferably effective after storage of at least 1 day, 2 days, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, 35, 36, 37, 38, 39, 40, 41 , 42, 43, 44, 45, 46, 47, 48, 49, 50, 55, 60, 70, 80, 90 days, or 4 months, 5 months, 6, 7, 8, 9, 10, 11 months or at least a year of storage. Preferably, the storage period is between 10 and 90 days from the date of production of the nutrition bar.

In this aspect, the p-lactoglobulin may be an extraneous p-lactoglobulin.

In this aspect, the p-lactoglobulin may be a recombinant p-lactoglobulin.

In this aspect, the p-lactoglobulin may be a non-recombinant p-lactoglobulin.

In this aspect, the amount of p-lactoglobulin in view of the total amount of protein in the nutrition bar is preferably at least 15% w/w, 20% w/w, 25%w/w, 30% w/w, 40% w/w, 50% w/w, 60% w/w or at least 70% w/w. In this aspect, the amount of p-lactoglobulin in view of the total amount of protein in the nutrition bar is preferably at least 20% w/w and at most 80% w/w.

In this aspect, modulation of hardness of a nutrition bar preferably means a decrease of hardness, preferably after storage. Such decrease in hardness may be a decrease of at least 10%, 20%, 30%, 40% 50%, 60%, 70%, 80%, 90%, 100%, 110%, 120%, 130%, 140%, 150%, 160%, 170%, 180%, 190%, 200%, 300%, 400%, or at least 500%. Such decrease in hardness may be a decrease within the range of between 10% to 200%, between 20% to 100%, between 25% to 70% or between 30% to 50%. Preferably, the hardness of the nutrition bar at three months post production is at most 12.000 g, such as at most 8.000 g, 9.000 g, 10.000 g, 11 .000, or at most 12.000 g.

In this aspect, modulation of firmness of a nutrition bar preferably means a decrease of firmness, preferably after storage. Such decrease in firmness may be a decrease of at least 10%, 20%, 30%, 40% 50%, 60%, 70%, 80%, 90%, 100%, 110%, 120%, 130%, 140%, 150%, 160%, 170%, 180%, 190%, 200%, 300%, 400%, or at least 500%. Such decrease in firmness may be a decrease within the range of between 10% to 200%, between 20% to 100%, between 25% to 70% or between 30% to 50%. Preferably, the firmness of the nutrition bar at three months post production is at most 2000 kPa, such as 900 kPa, 1000 kPa, 1100 kPa, 1200 kPa, 1300 kPa, 1400k Pa, 1500 kPa, 1600 kPa, 1700kPa, 1800 kPa, 1900 kPa or at most 2000 kPa. In this aspect, modulation of cohesiveness of a nutrition bar preferably means an increase of cohesiveness, preferably after storage. Such increase in cohesiveness may be an increase of at least 10%, 20%, 30%, 40% 50%, 60%, 70%, 80%, 90%, 100%, 110%, 120%, 130%, 140%, 150%, 160%, 170%, 180%, 190%, 200%, 300%, 400%, or at least 500%. Such increase in cohesiveness may be a decrease within the range of between 10% to 200%, between 20% to 100%, between 25% to 70% or between 30% to 50%. Preferably, the cohesiveness of the nutrition bar at three months post production is at least 0.09, such as 0.09, 0.1 , 0.11 , 0.12, 0.13, 0.14, 0.15, 0.16, 0.17, 0.18 or at least 0.2.

Table 1 : Overview of sequences Figure legends

Figure 1 depicts the effect of p-lactoglobulin on hardness (y-axes, in g) in pea-based nutrition bars (1A) or soy-based nutrition bars (1 B). The percentage of p-lactoglobulin relative to the total of p-lactoglobulin plus pea or p-lactoglobulin plus soy, respectively, is on the x-axes.

Figure 2 depicts the effect of p-lactoglobulin on cohesiveness (y-axes) in pea-based protein bars (2A) or soy-based protein bars (2B). The percentage of p-lactoglobulin relative to the total of p-lactoglobulin plus pea or p-lactoglobulin plus soy, respectively, is on the x-axes.

Figure 3 depicts the effect of p-lactoglobulin on firmness (y-axes, in kPa) in pea-based protein bars (3A) or soy-based protein bars (3B). The percentage of p-lactoglobulin relative to the total of p-lactoglobulin plus pea or p-lactoglobulin plus soy, respectively, is on the x-axes.

Figure 4 depicts the effect of p-lactoglobulin on (4A) hardness (y-axes, in g), (4B) cohesiveness (y- axes, and (4C) firmness (y-axes, in kPa) in soy based protein bars. The percentage of p-lactoglobulin relative to the total of p-lactoglobulin plus soy protein is on the x-axes.

Figure 5 depicts the effect of p-lactoglobulin on (5A) hardness (y-axes, in g), (5B) cohesiveness (y- axes, and (5C) firmness (y-axes, in kPa) in pea based protein bars. The percentage of p-lactoglobulin relative to the total of p-lactoglobulin plus pea protein is on the x-axes.

Definitions

"Sequence identity" is herein defined as a relationship between two or more amino acid (peptide, polypeptide, or protein) sequences or two or more nucleic acid (nucleotide, polynucleotide) sequences, as determined by comparing the sequences. In the art, "identity" also means the degree of sequence relatedness between amino acid or nucleotide sequences, as the case may be, as determined by the match between strings of such sequences. "Similarity" between two amino acid sequences is determined by comparing the amino acid sequence and its conserved amino acid substitutes of one peptide or polypeptide to the sequence of a second peptide or polypeptide. In a preferred embodiment, identity or similarity is calculated over the whole SEQ ID NO as identified herein. "Identity" and "similarity" can be readily calculated by known methods, including but not limited to those described in Computational Molecular Biology, Lesk, A. M., ed., Oxford University Press, New York, 1988; Biocomputing: Informatics and Genome Projects, Smith, D. W., ed., Academic Press, New York, 1993; Computer Analysis of Sequence Data, Part I, Griffin, A. M., and Griffin, H. G., eds., Humana Press, New Jersey, 1994; Sequence Analysis in Molecular Biology, von Heine, G., Academic Press, 1987; and Sequence Analysis Primer, Gribskov, M. and Devereux, J., eds., M Stockton Press, New York, 1991 ; and Carillo, H., and Lipman, D., SIAM J. Applied Math., 48:1073 (1988).

Preferred methods to determine identity are designed to give the largest match between the sequences tested. Methods to determine identity and similarity are codified in publicly available computer programs. Preferred computer program methods to determine identity and similarity between two sequences include e.g. the GCG program package (Devereux, J., et al., NucleicAcids Research 12 (1): 387 (1984)), BestFit, BLASTP, BLASTN, and FASTA (Altschul, S. F. et al., J. Mol. Biol. 215:403-410 (1990). The BLAST X program is publicly available from NCBI and other sources (BLAST Manual, Altschul, S., et al., NCBI NLM NIH Bethesda, MD 20894; Altschul, S„ et al., J. Mol. Biol. 215:403-410 (1990). The well-known Smith Waterman algorithm may also be used to determine identity.

Preferred parameters for polypeptide sequence comparison include the following: Algorithm: Needleman and Wunsch, J. Mol. Biol. 48:443-453 (1970); Comparison matrix: BLOSUM62 from Hentikoff and Hentikoff, Proc. Natl. Acad. Sci. USA. 89:10915-10919 (1992); Gap Penalty: 12; and Gap Length Penalty: 4. A program useful with these parameters is publicly available as the "Ogap" program from Genetics Computer Group, located in Madison, Wl. The aforementioned parameters are the default parameters for amino acid comparisons (along with no penalty for end gaps).

Preferred parameters for nucleic acid comparison include the following: Algorithm: Needleman and Wunsch, J. Mol. Biol. 48:443-453 (1970); Comparison matrix: matches=+10, mismatch=0; Gap Penalty: 50; Gap Length Penalty: 3. Available as the Gap program from Genetics Computer Group, located in Madison, Wis. Given above are the default parameters for nucleic acid comparisons.

Optionally, in determining the degree of amino acid similarity, the skilled person may also take into account so-called "conservative" amino acid substitutions, as will be clear to the skilled person. Conservative amino acid substitutions refer to the interchangeability of residues having similar side chains. For example, a group of amino acids having aliphatic side chains is glycine, alanine, valine, leucine, and isoleucine; a group of amino acids having aliphatic-hydroxyl side chains is serine and threonine; a group of amino acids having amide-containing side chains is asparagine and glutamine; a group of amino acids having aromatic side chains is phenylalanine, tyrosine, and tryptophan; a group of amino acids having basic side chains is lysine, arginine, and histidine; and a group of amino acids having sulphur-containing side chains is cysteine and methionine. Preferred conservative amino acids substitution groups are: valine-leucine-isoleucine, phenylalanine-tyrosine, lysine-arginine, alaninevaline, and asparagine-glutamine. Substitutional variants of the amino acid sequence disclosed herein are those in which at least one residue in the disclosed sequences has been removed and a different residue inserted in its place. Preferably, the amino acid change is conservative. Preferred conservative substitutions for each of the naturally occurring amino acids are as follows: Ala to ser; Arg to lys; Asn to gin or his; Asp to glu; Cys to ser or ala; Gin to asn; Glu to asp; Gly to pro; His to asn or gin; He to leu or val; Leu to ile or val; Lys to arg; gin or glu; Met to leu or ile; Phe to met, leu or tyr; Ser to thr; Thr to ser; Trp to tyr; Tyr to trp or phe; and, Val to ile or leu.

A “nucleic acid molecule” or “polynucleotide” (the terms are used interchangeably herein) is represented by a nucleotide sequence. A “polypeptide” is represented by an amino acid sequence. A “nucleic acid construct” is defined as a nucleic acid molecule which is isolated from a naturally occurring gene or which has been modified to contain segments of nucleic acids which are combined or juxtaposed in a manner which would not otherwise exist in nature. A nucleic acid molecule is represented by a nucleotide sequence. Optionally, a nucleotide sequence present in a nucleic acid construct is operably linked to one or more control sequences, which direct the production or expression of the peptide or polypeptide in a cell or in a subject.

“Operably linked” is defined herein as a configuration in which a control sequence is appropriately placed at a position relative to the nucleotide sequence coding for the polypeptide of the invention such that the control sequence directs the production/expression of the peptide or polypeptide of the invention in a cell and/or in a subject. “Operably linked” may also be used for defining a configuration in which a sequence is appropriately placed at a position relative to another sequence coding for a functional domain such that a chimeric polypeptide is encoded in a cell and/or in a subject.

“Expression” is construed as to include any step involved in the production of the peptide or polypeptide including, but not limited to, transcription, post-transcriptional modification, translation, post- translational modification and secretion.

A “control sequence” is defined herein to include all components which are necessary or advantageous for the expression of a polypeptide. At a minimum, the control sequences include a promoter and transcriptional and translational stop signals. Optionally, a promoter represented by a nucleotide sequence present in a nucleic acid construct is operably linked to another nucleotide sequence encoding a peptide or polypeptide as identified herein.

The term "transformation" refers to a permanent or transient genetic change induced in a cell following the incorporation of new DNA (i.e. DNA exogenous to the cell). When the cell is a bacterial cell, as is intended in the present invention, the term usually refers to an extrachromosomal, self-replicating vector which harbors a selectable antibiotic resistance.

An “expression vector” may be any vector which can be conveniently subjected to recombinant DNA procedures and can bring about the expression of a nucleotide sequence encoding a polypeptide of the invention in a cell and/or in a subject. As used herein, the term "promoter" refers to a nucleic acid fragment that functions to control the transcription of one or more genes or nucleic acids, located upstream with respect to the direction of transcription of the transcription initiation site of the gene. It is related to the binding site identified by the presence of a binding site for DNA-dependent RNA polymerase, transcription initiation sites, and any other DNA sequences, including, but not limited to, transcription factor binding sites, repressor and activator protein binding sites, and any other sequences of nucleotides known to one skilled in the art to act directly or indirectly to regulate the amount of transcription from the promoter. Within the context of the invention, a promoter preferably ends at nucleotide -1 of the transcription start site (TSS).

A “polypeptide” or “protein" as used herein refers to any peptide, oligopeptide, polypeptide, gene product, expression product, or protein. A polypeptide is comprised of consecutive amino acids. The term "polypeptide" encompasses naturally occurring or synthetic molecules.

The term “recombinant polypeptide” or “recombinant protein” as used herein refers to a polypeptide that is produced in a cell of a different species or type as compared to the species or type of cell that produces the polypeptide in nature, or that is produced in a cell at a level at which it is not produced in nature.

The term "heterogeneous" as used herein with reference to a plurality of recombinant proteins means that the plurality of recombinant proteins comprises at least two or two or more, three or more, four or more, five or more, six or more, or seven or more proteins of differing amino acid sequence.

The term "mature" as used herein with reference to a protein refers to the protein, or amino acid sequence of the protein, after cleavage of the signal sequence. The term "full length" as used herein with reference to a protein refers to the protein, or amino acid sequence of the protein, comprising the signal sequence. Examples of mature and full-length proteins are provided in Table 1 herein. The term "wild-type" as used herein with reference to proteins or polynucleotides refers to a protein or polynucleotide having an amino acid or nucleotide sequences that is the same as that expressed naturally. This term includes all naturally occurring variants of a particular protein, for example, all naturally occurring variants of p-lactoglobulin. Furthermore, this term includes both full length proteins and mature proteins and polynucleotides that encode wild-type full length and mature protein. The term is generally synonymous with the term "native".

Sequence identity herein of a polynucleotide, polynucleotide construct or of a polypeptide is preferably at least 70%. Preferably at least 70% is defined as preferably at least 70%, more preferably at least 71 %, more preferably at least 72%, more preferably at least 73%, more preferably at least 74%, more preferably at least 75%, more preferably at least 76%, more preferably at least 77%, more preferably at least 78%, more preferably at least 79%, more preferably at least 80%, more preferably at least 81 %, more preferably at least 82%, more preferably at least 83%, more preferably at least 84%, more preferably at least 85%, more preferably at least 86%, more preferably at least 87%, more preferably at least 88%, more preferably at least 89%, more preferably at least 90%, more preferably at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, more preferably at least 98%, more preferably at least 99%, or most preferably 100% sequence identity. In case of 100% sequence identity, the polynucleotide or polypeptide has exactly the sequence of the depicted SEQ ID NO:. Sequence identity is preferably determined over the entire length of the subject sequence.

The sequence information as provided herein should not be so narrowly construed as to require inclusion of erroneously identified bases. The skilled person is capable of identifying such erroneously identified bases and knows how to correct for such errors.

In this document and in its claims, the verbs "to comprise", “to contain”, and its conjugations are used in its non-limiting sense to mean that items following the word are included, but items not specifically mentioned are not excluded. In addition, the verb “to consist of’ may be replaced by “to consist essentially of’ meaning that a product or a composition or a nucleic acid molecule or a peptide or polypeptide of a nucleic acid construct or vector or cell as defined herein may comprise additional component(s) than the ones specifically identified; the additional component(s) not altering the unique characteristic of the invention. In addition, reference to an element by the indefinite article "a" or "an" does not exclude the possibility that more than one of the elements is present, unless the context clearly requires that there be one and only one of the elements. The indefinite article "a" or "an" thus usually means "at least one". The word “about” or “approximately” when used in association with a numerical value (e.g. about 10) preferably means that the value may be the given value (of 10) more or less 10% of the value.

All patent and literature references cited in the present specification are hereby incorporated by reference in their entirety.

Unless otherwise indicated each embodiment as described herein may be combined with another embodiment as described herein.

Unless stated otherwise, reference to such sources of information is not to be construed, in any jurisdiction, as an admission that such sources of information are prior art or form part of the common general knowledge in the art. Further embodiments of the invention

Further embodiments of the invention are listed here below.

1 . A nutrition bar comprising protein and extraneous p-lactoglobulin, wherein the ratio of protein to extraneous p-lactoglobulin is within the range from 80 : 20 (w/w) to 5 : 95 (w/w).

2. A nutrition bar according to embodiment 1 , wherein part of the protein excluding the extraneous p-lactoglobulin, is plant protein

3. A nutrition bar according to embodiment 1 or 2, wherein the nutrition bar comprises at least 5% (w/w) of total protein including the extraneous p-lactoglobulin in view of the total weight of the nutrition bar.

4. A nutrition bar according to any one of embodiments 1 to 3, wherein the nutrition bar comprises, further to protein and extraneous p-lactoglobulin, a fat and a carbohydrate.

5. A nutrition bar according to any one of embodiments 2 to 4, wherein the plant protein is a legume protein, a cereal protein and/or a nut protein.

6. A nutrition bar according to any one of embodiments 2 to 5, wherein the plant protein comprises soy protein, rice protein, lupine protein, chickpea protein, lentil protein, pea protein, fava protein, and/or protein from nuts, such as almond and cashew.

7. A nutrition bar according to any one of embodiments 1 to 6, wherein the extraneous p-lactoglobulin is a recombinant p-lactoglobulin.

8. A nutrition bar according to any one of embodiments 1 to 7, wherein the ratio of protein to extraneous p-lactoglobulin is within the range from 80 : 20 (w/w) to 20 : 80 (w/w).

9. A nutrition bar according to any one of embodiments 1 to 8, wherein the ratio of protein to p-lactoglobulin is within the range from 80 : 20 (w/w) to 50 : 50 (w/w).

10. A nutrition bar according to any one of embodiments 2 to 9, wherein the amount of plant protein plus extraneous p-lactoglobulin in view of the total weight of the nutrition bar is from 5% (w/w) to 70% (w/w).

11 . A nutrition bar according to any one of embodiments 1 to 10, wherein the nutrition bar is a coated nutrition bar.

12. A nutrition bar according to any one of embodiments 1 to 11 , wherein the nutrition bar is an animal-free nutrition bar. 13. A nutrition bar according to any one of embodiments 1 to 12, wherein the hardness of the nutrition bar at three months post production is at most 12.000 g, such as at most 8.000 g, 9.000 g, 10.000 g, 11 .000, or at most 12.000 g.

14. A nutrition bar according to any one of embodiments 1 to 13, wherein the firmness of the nutrition bar at three months post production is at most 2000 kPa, such as 900 kPa, 1000 kPa, 1100 kPa, 1200 kPa, 1300 kPa, 1400k Pa, 1500 kPa, 1600 kPa, 1700kPa, 1800 kPa, 1900 kPa or at most 2000 kPa.

15. A nutrition bar according to any one of embodiments 1 to 14, wherein the cohesiveness of the nutrition bar at three months post production is at least 0.09, such as 0.09, 0.1 , 0.11 , 0.12, 0.13, 0.14, 0.15, 0.16, 0.17, 0.18 or at least 0.2.

16. A process for the preparation of a nutrition bar according to any one of embodiments 1 to 15, comprising: a. mixing the desired ingredients comprising at least a protein and extraneous p-lactoglobulin to result in a dough, and b. forming a bar of the dough.

17. Use of a p-lactoglobulin to modulate the hardness, firmness and/or cohesiveness of a nutrition bar after storage.

18. Use according to embodiment 17, wherein the hardness of the nutrition bar is significantly improved after a period of at least 10 days from the production date of the nutrition bar compared to the hardness of a corresponding nutrition bar that does not comprise p-lactoglobulin.

19. Use according to embodiment 18, wherein the hardness of the nutrition bar at three months post production is at most 12.000 g, such as at most 8.000 g, 9.000 g, 10.000 g, 11.000, or at most 12.000 g.

20. Use according to any one of embodiments 17 to 19, wherein the firmness of the nutrition bar is significantly improved after a period of at least 10 days from the production date of the nutrition bar compared to the firmness of a corresponding nutrition bar that does not comprise p-lactoglobulin, with the proviso that in a nutrition bar comprising 30% protein, the ratio of pea protein and p-lactoglobulin is not 90 : 10.

21 . Use according to embodiment 20, wherein the firmness of the nutrition bar at three months post production is at most 2000 kPa, such as 900 kPa, 1000 kPa, 1100 kPa, 1200 kPa, 1300 kPa, 1400k Pa, 1500 kPa, 1600 kPa, 1700kPa, 1800 kPa, 1900 kPa or at most 2000 kPa. Use according to any one of embodiments 17 to 21 , wherein the cohesiveness of the nutrition bar is significantly improved after a period of at least 10 days from the production date of the nutrition bar compared to the cohesiveness of a corresponding nutrition bar that does not comprise p-lactoglobulin. Use according to embodiment 22, wherein the cohesiveness of the nutrition bar at three months post production is at least 0.09, such as 0.09, 0.1 , 0.11 , 0.12, 0.13, 0.14, 0.15, 0.16, 0.17, 0.18 or at least 0.2. Use according to any one of embodiments 17 to 23, wherein the period is between 10 and 90 days from the date of production of the nutrition bar. Use according to anyone of embodiments 17 to 24, wherein the p-lactoglobulin is an extraneous p-lactoglobulin. Use according to anyone of embodiments 17 to 25, wherein the p-lactoglobulin is a recombinant p-lactoglobulin. Use according to anyone of embodiments 17 to 26, wherein the p-lactoglobulin is a nonrecombinant p-lactoglobulin. A nutrition bar according to any one of embodiments 1 to 15, a process according to embodiment 16, and a use according to any one of embodiments 17 to 27, wherein the p-lactoglobulin is one or more selected from the group consisting of: a. a p-lactoglobulin having an amino acid sequence that has at least 90% sequence identity with the amino acid sequence as said forth in any one of SEQ ID NO: 1 to 21 and 32 and 33; b. a p-lactoglobulin having an amino acid sequence that has at least 90% sequence identity with the amino acid sequence as set forth in SEQ ID NO: 1 , wherein the p- lactoglobulin comprises at least one additional charged amino acid compared to p- lactoglobulin with the amino acid sequence as set forth in SEQ ID NO: 1 , and wherein, preferably, the additional charged amino acid is not an N-terminal EA, EAEA, EAEAEA, EAEAEAEA, or R; c. a p-lactoglobulin having an amino acid sequence that has at least 90% sequence identity with the amino acid sequence as set forth in SEQ ID NO: 3, wherein the p- lactoglobulin comprises at least one additional charged amino acid compared to p- lactoglobulin with the amino acid sequence as set forth in SEQ ID NO: 3, and wherein, preferably, the additional charged amino acid is not an N-terminal EA, EAEA, EAEAEA, EAEAEAEA, or R; d. a p-lactoglobulin having an amino acid sequence that has at least 90% sequence identity with the amino acid sequence as set forth in SEQ ID NO: 5, wherein the p- lactoglobulin comprises at least one additional charged amino acid compared to p- lactoglobulin with the amino acid sequence as set forth in SEQ ID NO: 5; e. a p-lactoglobulin having an amino acid sequence that has at least 90% sequence identity with the amino acid sequence as set forth in SEQ ID NO: 7, wherein the p- lactoglobulin comprises at least one additional charged amino acid compared to p- lactoglobulin with the amino acid sequence as set forth in SEQ ID NO: 7; f. a p-lactoglobulin having an amino acid sequence that has at least 90% sequence identity with the amino acid sequence as set forth in SEQ ID NO: 9, wherein the p- lactoglobulin comprises at least one additional charged amino acid compared to p- lactoglobulin with the amino acid sequence as set forth in SEQ ID NO: 9; g. a p-lactoglobulin having an amino acid sequence that has at least 90% sequence identity with the amino acid sequence as set forth in SEQ ID NO: 11 , wherein the p- lactoglobulin comprises at least one additional charged amino acid compared to p- lactoglobulin with the amino acid sequence as set forth in SEQ ID NO: 11 ; h. a p-lactoglobulin having an amino acid sequence that has at least 90% sequence identity with the amino acid sequence as set forth in SEQ ID NO: 13, wherein the p- lactoglobulin comprises at least one additional charged amino acid compared to p- lactoglobulin with the amino acid sequence as set forth in SEQ ID NO: 13; i. a p-lactoglobulin having an amino acid sequence that has at least 90% sequence identity with the amino acid sequence as set forth in SEQ ID NO: 15, wherein the p- lactoglobulin comprises at least one additional charged amino acid compared to p- lactoglobulin with the amino acid sequence as set forth in SEQ ID NO: 15; j. a p-lactoglobulin having an amino acid sequence that has at least 90% sequence identity with the amino acid sequence as set forth in SEQ ID NO: 17, wherein the p- lactoglobulin comprises at least one additional charged amino acid compared to p- lactoglobulin with the amino acid sequence as set forth in SEQ ID NO: 17; k. a p-lactoglobulin having an amino acid sequence that has at least 90% sequence identity with the amino acid sequence as set forth in SEQ ID NO: 19, wherein the p- lactoglobulin comprises at least one additional charged amino acid compared to p- lactoglobulin with the amino acid sequence as set forth in SEQ ID NO: 19; and l. a p-lactoglobulin having an amino acid sequence that has at least 90% sequence identity with the amino acid sequence as set forth in SEQ ID NO: 21 , wherein the p- lactoglobulin comprises at least one additional charged amino acid compared to p- lactoglobulin with the amino acid sequence as set forth in SEQ ID NO: 21 . 29. A nutrition bar according to any one of embodiments 1 to 15, a process according to embodiment 16, and a use according to any one of embodiments 17 to 27, wherein the p-lactoglobulin has a surface charge (^-potential) of at least 12mV at a pH of at most 4 and/or has a surface charge ( - potential) of at most -12mV at a pH of at least 6.

30. A nutrition bar according to any one of embodiments 1 to 15, a process according to embodiment 16, and a use according to any one of embodiments 17 to 27, wherein the p-lactoglobulin has a surface charge ( -potential)of at most -12mV at a pH of at least 5.5 and/or has a surface charge (^-potential) of at most -8mV at a pH of at least 5.

31 . A nutrition bar according to any one of embodiments 1 to 15, a process according to embodiment 16, and a use according to any one of embodiments 17 to 27, wherein p-lactoglobulin has enhanced physical stability compared to the p-lactoglobulin selected from the group consisting of SEQ ID NO: 1 , 3, 5, 7, 9, 11 , 13, 15, 17, 19, and 21 that has the highest sequence identity with the p-lactoglobulin with the at least one additional charged amino acid.

Examples

The following examples are offered for illustrative purposes only, and are not intended to limit the scope of the present invention in any way. In this specification, where reference has been made to external sources of information, including patent specifications and other documents, this is generally for the purpose of providing a context for discussing the features of the present invention.

Materials and Methods

Pea protein concentrate (NUTRALYS_S85XF) and maltitol syrup (Lycasin 75/75) were obtained from Roquette. Soy protein was obtained from Buxtrade. Inulin (Frutalose L85) was obtained from Sensus. Glycerin 99,5% was obtained from Dokter-Klaus. Almond butter was obtained from Bulk. Sunflower oil was obtained from Reddy. Soy lecithin was obtained from Now Foods, p-lactoglobulin was prepared as described in example 1 of WO 2022/269549, which is herein incorporated by reference. Blending was carried out in a Kenwood mixer Titanium Chef Baker XL (Kenwood Limited, United Kingdom).

Texture analysis

After preparation, the nutrition bars were stored in a dark environment at ambient temperature. On the day of analysis, cylindrical pieces with a diameter of 21 mm were pressed out of the nutrition bars. A TA-XT plus Texture Analyzer (Stable Micro Systems, Surrey, United Kingdom) equipped with a 30 kg load cell was calibrated on height and weight. The nutrition bars were transferred to the texture analyzer for a TPA measurement, and uniaxially compressed twice by a cylindrical probe (50 mm diameter) with a pre-test speed of 1 mm/sec, a trigger force of 10 g, a strain deformation of 50%, using a testing speed of 1 mm/sec. There was a holding period of 5 sec between the first and second compression. The firmness was defined as the Young’s modulus (Pa) or the slope of the initial linear part of the stressstrain curve (Eq. 1). Hardness was defined as the maximum force required for 50% compression. Cohesiveness was defined as the area under the second peak of compression divided by the area under the first peak of compression.

E 77 = - a)

8

Where o is the compressive stress (Pa) and s the strain (-) in the linear elastic region.

All samples were measured in duplicate.

Example 1 ; Protein bars comprising soy protein and B-lactoqlobulin

Nutrition bars were prepared with ingredients as described in table 2 here below. This resulted in four different bars (trials 1 , 2, 3 and 4) wherein the amount of p-lactoglobulin relative to the total amount of protein including p-lactoglobulin was 0%, 10%, 30%, and 50%, respectively.

The liquid ingredients were blended in a saucepan and heated to 50±2°C. Separately, the proteins were blended in a mixing bowl. The liquid ingredients and almond butter were added to the mixing bowl and where mixed for one minute on the lowest speed (1). If needed, the dough was scraped from the K- blade and from the bottom of the bowl and mixing was continued at medium speed (2) until a homogeneous dough was obtained after approximately 2 minutes. The dough was transferred into a frame (~19 mm deep), spread evenly, covered with plastic film, and pressed down with a rolling pin to form bars. The dough was left overnight and heat-sealed in foil laminate sachets that were stored at 20±2°C).

Subsequently, after 1 day, 1 month and 3 months, hardness, cohesiveness and firmness of the bars was determined using the assay as set forward in the Materials and Methods section here above. The results are depicted in Tables 2 to 6 here below and in Figures 1 B, 2B, and 3B.

Results and discussion

The results indicate that incorporation of extraneous p-lactoglobulin in a nutrition bar comprising soy protein results in a nutrition bar with improved properties, namely a substantial reduction of hardness and firmness and an increase in cohesiveness. These effects are maintained over time.

Table 2: Ingredients of nutrition bars comprising soy protein

Table 3: Nutritional value of nutrition bars comprising soy protein

Table 4: Hardness of nutrition bars comprising soy protein

Table 5: Cohesiveness of nutrition bars comprising soy protein

Table 6: Firmness of nutrition bars comprising soy protein

Example 2; Protein bars comprising pea protein and B-lactoqlobulin

Nutrition bars were prepared according to Example 1 here above, with ingredients as set forward in table 7 here below. This resulted in six different bars wherein the amount of p-lactoglobulin relative to the total amount of protein was 0%, 10%, 20%, 30%, 40%, and 50%, respectively. Additionally, in a similar trial, a bar wherein the amount of p-lactoglobulin relative to the total amount of protein was 30%, was prepared.

At day 1 and day 34, hardness, cohesiveness and firmness of the bars was determined using the assay as set forward in the Materials and Methods section here above. The results are depicted in Tables 7 to 11 here below and in Figures 1A, 2A, and 3A.

Results and discussion

The results indicate that incorporation of extraneous p-lactoglobulin in a nutrition bar comprising pea protein results in a nutrition bar with improved properties, namely a substantial reduction of hardness and firmness and an increase in cohesiveness. These effects are maintained over time. Moreover, there appears to be an optimum concentration of extraneous p-lactoglobulin between 10% and 50% of p-lactoglobulin relative to the total amount of protein since the reduction of hardness and firmness and the increase in cohesiveness were even more pronounced in the nutrition bars comprising 30% extraneous p-lactoglobulin compared to the bars comprising 10% and 50% p-lactoglobulin, respectively. Table 7: Ingredients of nutrition bars comprising pea protein

Table 8: Nutritional value of nutrition bars comprising pea protein Table 9: Hardness of nutrition bars comprising pea protein

Table 10: Cohesiveness of nutrition bars comprising pea protein

Table 11 : Firmness of nutrition bars comprising pea protein

Example 3; Protein bars comprising soy protein and B-lactoqlobulin

Example 3 was conducted according to Example 1 here above with the difference that more timepoints were measured. The results are depicted in Tables 12 to 14 and Figures 4 A - 4C).

The results corroborate that incorporation of extraneous p-lactoglobulin in a nutrition bar comprising soy protein results in a nutrition bar with improved properties, namely a substantial reduction of hardness and firmness and an increase in cohesiveness. These effects are maintained over time Table 12: Hardness of nutrition bars comprising soy protein

Table 13: Cohesiveness of nutrition bars comprising soy protein

Table 14: Firmness of nutrition bars comprising soy protein Example 4; Protein bars comprising pea protein and B-lactoqlobulin

Example 4 was conducted according to Example 2 here above with the difference that more ratio’s of pean and p-lactoglobulin were prepared and more timepoints were measured. The results are depicted in Tables 15 to 17 and Figures 5 A - 5C).

The results corroborate that incorporation of extraneous p-lactoglobulin in a nutrition bar comprising pea protein results in a nutrition bar with improved properties, namely a substantial reduction of hardness and firmness and an increase in cohesiveness. These effects are maintained over time. Moreover, there appears to be an optimum concentration of extraneous p-lactoglobulin between 10% and 50% of p-lactoglobulin relative to the total amount of pea protein since the reduction of hardness and firmness and the increase in cohesiveness were even more pronounced in the nutrition bars comprising 30% extraneous p-lactoglobulin compared to the bars comprising 10% and 50% p-lactoglobulin, respectively. Table 15: Hardness of nutrition bars comprising pea protein

Table 16: Cohesiveness of nutrition bars comprising pea protein

Table 17: Firmness of nutrition bars comprising pea protein

Example 5; Protein bar comprising pea protein and high B-lactoglobulin whey protein isolate

A nutrition bar was prepared according to the method described in Example 2 herein above, with similar ingredients. However, the p-lactoglobulin was replaced with a whey protein isolate comprising 69% p- lactoglobulin. The amount of this high p-lactoglobulin whey protein isolate relative to the total amount of protein was 30%. Accordingly, the amount of p-lactoglobulin relative to the total amount of protein was 20.7% and the ratio of pea protein : p-lactoglobulin was 79.3 : 20.7. Cohesiveness, firmness, and hardness were measured after 1 day, 35 days, and 90 days using the assay as set forth in the Materials and Methods section herein above. The results are depicted in Table 18 here below.

Table 18: Hardness of nutrition bars comprising pea protein

It can be concluded that p-lactoglobulin from a whey protein isolate can have a similar effect on cohesiveness, firmness, and hardness of a nutrition bar comprising plant protein as does recombinant p-lactoglobulin. Without being bound by theory, it is likely that the amount of p-lactoglobulin has to be sufficiently high in the nutrition bar. Accordingly, it has herein been demonstrated that a p-lactoglobulin can be used to modulate cohesiveness, firmness, and/or hardness of a nutrition bar over time as measured after storage. The p-lactoglobulin can be added as an extraneous compound a such, but can also be added as an extraneous compound present in a composition that is added to a nutrition bar.

Claims

2. A nutrition bar according to claim 1 , wherein part of the protein excluding the extraneous p-lactoglobulin, is plant protein

3. A nutrition bar according to claim 1 or 2, wherein the nutrition bar comprises at least 5% (w/w) of total protein including the extraneous p-lactoglobulin in view of the total weight of the nutrition bar.

4. A nutrition bar according to any one of claims 1 to 3, wherein the nutrition bar comprises, further to protein and extraneous p-lactoglobulin, a fat and a carbohydrate.

5. A nutrition bar according to any one of claims 2 to 4, wherein the plant protein is a legume protein, a cereal protein and/or a nut protein.

6. A nutrition bar according to any one of claims 2 to 5, wherein the plant protein comprises soy protein, rice protein, lupine protein, chickpea protein, lentil protein, pea protein, fava protein, and/or protein from nuts, such as almond and cashew.

7. A nutrition bar according to any one of claims 1 to 6, wherein the extraneous p-lactoglobulin is a recombinant p-lactoglobulin.

8. A nutrition bar according to any one of claims 1 to 7, wherein the ratio of protein to extraneous p-lactoglobulin is within the range from 80 : 20 (w/w) to 20 : 80 (w/w).

9. A nutrition bar according to any one of claims 1 to 8, wherein the ratio of protein to p-lactoglobulin is within the range from 80 : 20 (w/w) to 50 : 50 (w/w).

10. A nutrition bar according to any one of claims 2 to 9, wherein the amount of plant protein plus extraneous p-lactoglobulin in view of the total weight of the nutrition bar is from 5% (w/w) to 70% (w/w).

11 . A nutrition bar according to any one of claims 1 to 10, wherein the nutrition bar is a coated nutrition bar.

12. A nutrition bar according to any one of claims 1 to 11 , wherein the nutrition bar is an animal-free nutrition bar.

13. A nutrition bar according to any one of claims 1 to 12, wherein the hardness of the nutrition bar at three months post production is at most 12.000 g, such as at most 8.000 g, 9.000 g, 10.000 g, 11 .000, or at most 12.000 g.

14. A nutrition bar according to any one of claims 1 to 13, wherein the firmness of the nutrition bar at three months post production is at most 2000 kPa, such as 900 kPa, 1000 kPa, 1100 kPa, 1200 kPa, 1300 kPa, 1400k Pa, 1500 kPa, 1600 kPa, 1700kPa, 1800 kPa, 1900 kPa or at most 2000 kPa.

15. A nutrition bar according to any one of claims 1 to 14, wherein the cohesiveness of the nutrition bar at three months post production is at least 0.09, such as 0.09, 0.1 , 0.1 1 , 0.12, 0.13, 0.14, 0.15, 0.16, 0.17, 0.18 or at least 0.2.

16. A process for the preparation of a nutrition bar according to any one of claims 1 to 15, comprising: a. mixing the desired ingredients comprising at least a protein and extraneous p-lactoglobulin to result in a dough, and b. forming a bar of the dough.

18. Use according to claim 17, wherein the hardness of the nutrition bar is significantly improved after a period of at least 10 days from the production date of the nutrition bar compared to the hardness of a corresponding nutrition bar that does not comprise p-lactoglobulin.

19. Use according to claim 18, wherein the hardness of the nutrition bar at three months post production is at most 12.000 g, such as at most 8.000 g, 9.000 g, 10.000 g, 11.000, or at most 12.000 g.

20. Use according to any one of claims 17 to 19, wherein the firmness of the nutrition bar is significantly improved after a period of at least 10 days from the production date of the nutrition bar compared to the firmness of a corresponding nutrition bar that does not comprise p-lactoglobulin, with the proviso that in a nutrition bar comprising 30% protein, the ratio of pea protein and p-lactoglobulin is not 90 : 10.

21 . Use according to claim 20, wherein the firmness of the nutrition bar at three months post production is at most 2000 kPa, such as 900 kPa, 1000 kPa, 1100 kPa, 1200 kPa, 1300 kPa, 1400k Pa, 1500 kPa, 1600 kPa, 1700kPa, 1800 kPa, 1900 kPa or at most 2000 kPa.

22. Use according to any one of claims 17 to 21 , wherein the cohesiveness of the nutrition bar is significantly improved after a period of at least 10 days from the production date of the nutrition bar compared to the cohesiveness of a corresponding nutrition bar that does not comprise p-lactoglobulin.

23. Use according to claim 22, wherein the cohesiveness of the nutrition bar at three months post production is at least 0.09, such as 0.09, 0.1 , 0.11 , 0.12, 0.13, 0.14, 0.15, 0.16, 0.17, 0.18 or at least 0.2.

24. Use according to any one of claims 17 to 23, wherein the period is between 10 and 90 days from the date of production of the nutrition bar.

25. Use according to anyone of claims 17 to 24, wherein the p-lactoglobulin is an extraneous p-lactoglobulin.

26. Use according to anyone of claims 17 to 25, wherein the p-lactoglobulin is a recombinant p-lactoglobulin.

27. Use according to anyone of claims 17 to 26, wherein the p-lactoglobulin is a non-recombinant p-lactoglobulin.

28. A nutrition bar according to any one of claims 1 to 15, a process according to claim 16, and a use according to any one of claims 17 to 27, wherein the p-lactoglobulin is one or more selected from the group consisting of: a. a p-lactoglobulin having an amino acid sequence that has at least 90% sequence identity with the amino acid sequence as said forth in any one of SEQ ID NO: 1 to 21 and 32 and 33; b. a p-lactoglobulin having an amino acid sequence that has at least 90% sequence identity with the amino acid sequence as set forth in SEQ ID NO: 1 , wherein the p- lactoglobulin comprises at least one additional charged amino acid compared to p- lactoglobulin with the amino acid sequence as set forth in SEQ ID NO: 1 , and wherein, preferably, the additional charged amino acid is not an N-terminal EA, EAEA, EAEAEA, EAEAEAEA, or R; c. a p-lactoglobulin having an amino acid sequence that has at least 90% sequence identity with the amino acid sequence as set forth in SEQ ID NO: 3, wherein the p- lactoglobulin comprises at least one additional charged amino acid compared to p- lactoglobulin with the amino acid sequence as set forth in SEQ ID NO: 3, and wherein, preferably, the additional charged amino acid is not an N-terminal EA, EAEA, EAEAEA, EAEAEAEA, or R; d. a p-lactoglobulin having an amino acid sequence that has at least 90% sequence identity with the amino acid sequence as set forth in SEQ ID NO: 5, wherein the p- lactoglobulin comprises at least one additional charged amino acid compared to p- lactoglobulin with the amino acid sequence as set forth in SEQ ID NO: 5; e. a p-lactoglobulin having an amino acid sequence that has at least 90% sequence identity with the amino acid sequence as set forth in SEQ ID NO: 7, wherein the p- lactoglobulin comprises at least one additional charged amino acid compared to p- lactoglobulin with the amino acid sequence as set forth in SEQ ID NO: 7; f. a p-lactoglobulin having an amino acid sequence that has at least 90% sequence identity with the amino acid sequence as set forth in SEQ ID NO: 9, wherein the p- lactoglobulin comprises at least one additional charged amino acid compared to p- lactoglobulin with the amino acid sequence as set forth in SEQ ID NO: 9; g. a p-lactoglobulin having an amino acid sequence that has at least 90% sequence identity with the amino acid sequence as set forth in SEQ ID NO: 11 , wherein the p- lactoglobulin comprises at least one additional charged amino acid compared to p- lactoglobulin with the amino acid sequence as set forth in SEQ ID NO: 11 ; h. a p-lactoglobulin having an amino acid sequence that has at least 90% sequence identity with the amino acid sequence as set forth in SEQ ID NO: 13, wherein the p- lactoglobulin comprises at least one additional charged amino acid compared to p- lactoglobulin with the amino acid sequence as set forth in SEQ ID NO: 13; i. a p-lactoglobulin having an amino acid sequence that has at least 90% sequence identity with the amino acid sequence as set forth in SEQ ID NO: 15, wherein the p- lactoglobulin comprises at least one additional charged amino acid compared to p- lactoglobulin with the amino acid sequence as set forth in SEQ ID NO: 15; j. a p-lactoglobulin having an amino acid sequence that has at least 90% sequence identity with the amino acid sequence as set forth in SEQ ID NO: 17, wherein the p- lactoglobulin comprises at least one additional charged amino acid compared to p- lactoglobulin with the amino acid sequence as set forth in SEQ ID NO: 17; k. a p-lactoglobulin having an amino acid sequence that has at least 90% sequence identity with the amino acid sequence as set forth in SEQ ID NO: 19, wherein the p- lactoglobulin comprises at least one additional charged amino acid compared to p- lactoglobulin with the amino acid sequence as set forth in SEQ ID NO: 19; and l. a p-lactoglobulin having an amino acid sequence that has at least 90% sequence identity with the amino acid sequence as set forth in SEQ ID NO: 21 , wherein the p- lactoglobulin comprises at least one additional charged amino acid compared to p- lactoglobulin with the amino acid sequence as set forth in SEQ ID NO: 21 .

29. A nutrition bar according to any one of claims 1 to 15, a process according to claim 16, and a use according to any one of claims 17 to 27, wherein the p-lactoglobulin has a surface charge ( - potential) of at least 12mV at a pH of at most 4 and/or has a surface charge (^-potential) of at most -12mV at a pH of at least 6.

30. A nutrition bar according to any one of claims 1 to 15, a process according to claim 16, and a use according to any one of claims 17 to 27, wherein the p-lactoglobulin has a surface charge ( - potential)of at most -12mV at a pH of at least 5.5 and/or has a surface charge (^-potential) of at most -8mV at a pH of at least 5.

31. A nutrition bar according to any one of claims 1 to 15, a process according to claim 16, and a use according to any one of claims 17 to 27, wherein p-lactoglobulin has enhanced physical stability compared to the p-lactoglobulin selected from the group consisting of SEQ ID NO: 1 , 3, 5, 7, 9, 11 , 13, 15, 17, 19, and 21 that has the highest sequence identity with the p-lactoglobulin with the at least one additional charged amino acid.