Hybrid substitute dairy products

Abstract

Provided herein are edible compositions comprising; i. a non-animal derived globular protein, wherein the non-animal derived globular protein is at least partially denatured; and ii. at least one functionalised plant protein; as well as processes for the preparation thereof.

Description

HYBRID SUBSTITUTE DAIRY PRODUCTSTECHNICAL FIELD

[0001] The technology described herein resides in the field of protein-based food products and dairy substitutes. The present invention relates to edible compositions that are substitute dairy products, and processes for the manufacture thereof.BACKGROUND ART

[0002] As of 5 July 2022, the global dairy market was valued at approximately 830 billion U.S. dollars, and it was predicted to grow to about 1130 billion U.S. dollars by 2026. Bovine milk holds the most significant share of this market, whilst plant-derived dairy alternatives, lactose- intolerant milk, milk products that are reduced in carbohydrates and enriched in proteins, and other modified milk products such as A1 p-casein free milk (A2 milk) are increasingly prominent in an increasingly informed market of diet-conscious consumers.

[0003] Mammalian-derived milk is a highly complex liquid composition comprising, aside from water, thousands of different compounds, from lipids, triglycerides, carbohydrates, saccharides, peptides, inorganic salts and other molecular entities. Although many consider mammalian- derived milk, including bovine milk, to be an ideal nutrition source, various alternatives to mammalian-derived milk are now successfully on the market, including plant- or nut-based milks, such as soy, almond, or coconut milk, and are accepted by consumers for reasons related to mammalian-derived milk's allergenicity, lactose intolerance of certain components, personal preference, or the perception of adverse environmental impacts arising from the dairy industry.

[0004] For example, the majority of mammalian-derived milk is sourced from ruminant animals including cows, buffalos, yaks, goats and sheep, as well as pseudo-ruminants such as camels, alpacas and llamas. Cattle-rearing and ruminant livestock agriculture in general produces more global warming greenhouse gases, as measured in carbon dioxide (CO2) equivalents, than transportation, according to a recent UN assessment. Ruminants are estimated to account for 10% of total greenhouse gas emissions in Australia. Ruminants produce methane (CH4) as a by-product of digestion via anaerobic microbial feed fermentation in the rumen and, to a lesser extent, the large intestine. This process is referred to as methanogenesis.

[0005] Methane absorbs solar infrared radiation efficiently, and it has a global warming potential 25 times that of CO2. The ruminal microbial population is made up of bacteria, protozoa, fungi, and bacteriophages, all of which work together to digest ingested organic matter and produce CO2, H2, volatile fatty acids, and formates. These end-products are used by methanogenic archaea in the rumen, which produces CH4. Although the generation of CH4lowers the partialpressure of H2, this has the potential to cause problems as it also limits the amount of energy and carbon available for the synthesis of volatile fatty acids, which are critical for ruminant nutrition and could otherwise restrict rumen fermentation. The majority of CH4 generated by ruminants is exhaled or discharged via the mouth, resulting in a waste of up to 12% of gross caloric intake in the ruminant diet. In addition, producing a single glass of dairy milk from cows consumes up to nine times more land, and significantly more water, than any of the plant- derived milk alternatives.

[0006] Attempts to address these environmental issues with plant-derived milks including soy, almond, or coconut milk, for example, routinely fall short in both flavour and utility. In addition, a major portion of dairy milk's industrial and cultural value originates from its use in derivative goods manufactured from the milk, such as cheese, yoghurt, cream, or butter. While dairy substitute plant-derived milks address some environmental and health problems (and provide sufficient flavour for a minor portion of the consumer population), when exposed to the same processing procedures as dairy milk, they virtually always fail to generate such derivative goods having comparable organoleptic properties, such as taste, colour, odour, and mouthfeel, and they often do not respond in the same way as dairy products to commonly employed cooking procedures, such as they way in which many dairy cheeses respond to being baked, grilled or melted. Current cheese replacements do not match the functionality (including melt behaviour and browning behaviour when cooked or grilled), texture, nutrition, and taste of dairy cheese.

[0007] Furthermore, existing attempts to incorporate plant derived proteins into dairy substitute products including substitute cheeses, curds, yoghurts, and the like have typically failed to be accepted by consumers due to issues including unfavourable discolouration, such as greenish or brownish hues, astringent or bitter flavours or aftertastes, and unfavourable textural properties such as graininess, dryness or insufficient lubricity.

[0008] There is a need then, for alternative dairy substitutes or compositions with desirable flavour and performance characteristics, such as compositions that replicate dairy flavours, whilst minimising foodborne pathogens, and that potentially have a lower environmental impact in production, while retaining the functionality (including, for example, melt behaviour and browning behaviour when cooked or grilled), texture, nutrition, and taste of dairy derived products, and providing a nutritional profile similar to that of, or comparable to, such products manufactured from mammalian-derived milk.

[0009] Another problem that many producers in the field of dairy substitutes have in common is the challenge in scaling at a rapid and cost-effective rate. There is a need then to produce dairy replacement compositions at an industrially applicable scale, having similar structural and functional properties, including in terms of texture, hardness, elasticity, and melt behaviour in the downstream derivative products (such as cheeses) and desirable browning behaviour whencooked or grilled, to those observed in such compositions and downstream derivative products derived from dairy milk.

[0010] It is against this background that the present invention has been developed.

[0011] The preceding discussion of the background art is intended to facilitate an understanding of the present invention only. The discussion is not an acknowledgement or admission that any of the material referred to is or was part of the common general knowledge as at the priority date of the application.SUMMARY OF INVENTION

[0012] The present invention provides substitute dairy food products comprising non-animal derived globular proteins and functionalised plant proteins, as well as processes for the production thereof.

[0013] Functionalisation of plant proteins is achieved by thermo-mechanical pre-treatment, leading to functionalised plant proteins in the form of functional gelled protein particles.

[0014] The addition of these functionalised plant proteins to globular protein based substitute dairy food products surprisingly and unexpectedly leads to enhanced taste, texture, mouthfeel and overall higher product quality, while simultaneously addressing many of the challenges associated with the use of plant proteins in such products, including such challenges as unfavourable discoloration, unfavourable texture (eg; graininess or dryness) and unfavourable flavour profiles (eg; bitterness or astringency).BRIEF DESCRIPTION OF THE DRAWINGS

[0015] Further features of the present invention are more fully described in the following description of several non-limiting embodiments thereof. This description is included solely for the purposes of exemplifying the present invention. It should not be understood as a restriction on the broad summary, disclosure or description of the invention as set out above. The description will be made with reference to the accompanying drawings in which:Figure 1 is a schematic process flowchart depicting an embodiment of the process for the formation of the functionalised plant protein component of the edible compositions of the present invention, from potato protein (PP) as an exemplary plant protein, at a laboratory scale.Figure 2 is a schematic process flowchart depicting an embodiment of the process for the formation of the functionalised plant protein component of the edible compositions of the present invention, from potato protein (PP) as an exemplary plant protein, at an industrially applicable scale.Figure 3 is a plot of the mean particle size (D[4,3]) of the functionalised plant protein component of the edible compositions of the present invention, produced from potato protein (PP) as an exemplary plant protein, at an industrially applicable scale in accordance with the process depicted in Figure 2 (Pilot plant trials 1 and 2), compared to being produced at a laboratory scale in accordance with the process depicted in Figure 1 (Lab), both before and after homogenisation. The mean particle size (D[4,3]) of the functionalised plant proteins produced is between 2 - 6.3 pm.Figure 4 is a plot of the particle size distribution of the functionalised plant protein component of the edible compositions of the present invention, produced from potato protein (PP) as an exemplary plant protein, at an industrially applicable scale in accordance with the process depicted in Figure 2 (trials 1 and 2; 1sttrial and 2ndtrial), compared to being produced at a laboratory scale in accordance with the process depicted in Figure 1 (STD-Lab), both before and after homogenisation.Figure 5 is a plot of the particle size distribution of the functionalised plant protein component of the edible compositions of the present invention, produced from potato protein (PP) as an exemplary plant protein, at an industrially applicable scale in accordance with the process depicted in Figure 2.Figure 6 is a plot of the the temperature dependent evolution of the complex viscosity p* (mPas) of two embodiments of the functionalised plant protein component (Pre-treated potato protein) of the edible compositions of the present invention, produced from potato protein as an exemplary plant protein, at an industrially applicable (pilot plant) scale in accordance with the process depicted in Figure 2, demonstrating that the functionalised plant proteins are thermally stable up to a temperature of at least 80 °C and will not undergo structural changes (gelation) upon heat-treatment.Figure 7 is a plot of the gelation curves, measured in terms of the complex viscosity p* (mPas) as a function of temperature (°C), for the functionalised plant protein component of the edible compositions of the present invention, produced from potato protein (PP) as an exemplary plant protein, compared to the non-animal derived globular protein component of the edible compositions of the present invention, using kbji protein as an exemplary embodiment of the non-animal derived globular protein component of the edible compositions of the present invention, and compared to a mixture of these two components (at a ratio of functionalised plant protein to non-animal derived globular protein of 40:60) in accordance with an embodiment of the edible composition of the present invention.Figure 8 is a schematic process flowchart, depicting an embodiment (Process A) of the process for the production of an embodiment of the edible composition of the present invention, in the form of a substitute cream cheese comprising the functionalised plant protein component of theedible compositions of the present invention, produced from potato protein as an exemplary plant protein (pre-treated potato protein), and using kdji protein as an exemplary embodiment of the non-animal derived globular protein component of the edible compositions of the present invention, at an industrially applicable scale, wherein a 5% PP functionalised plant protein solution prepared according to the process shown in Figure 2 is added to a substitute cream cheese formulation after pasteurization.Figure 9 is a schematic process flowchart, depicting an embodiment (Process B) of the process for the production of an embodiment of the edible composition of the present invention, in the form of a substitute cream cheese comprising the functionalised plant protein component of the edible compositions of the present invention, produced from potato protein as an exemplary plant protein, and using kdji protein as an exemplary embodiment of the non-animal derived globular protein component of the edible compositions of the present invention, at an industrially applicable scale, wherein the potato protein is added directly with the dry ingredients at the start of the process, and the functionalised plant protein is formed in-situ during the process.Figure 10 is a schematic process flowchart, depicting an embodiment (Process C) of the process for the production of an embodiment of the edible composition of the present invention, in the form of a substitute cream cheese comprising the functionalised plant protein component of the edible compositions of the present invention, produced from potato protein as an exemplary plant protein, and using kdji protein as an exemplary embodiment of the non-animal derived globular protein component of the edible compositions of the present invention, at an industrially applicable scale, wherein the 5% functionalised plant protein solution prepared according to the process shown in Figure 2 is added at the beginning of the process (in contrast to Process A of Figure 8 where it is added after pasteurization of the other ingredients of the edible composition).Figure 11 is a plot of the mean particle size (D[4,3]) of an embodiment of the edible compositions of the present invention (“Hybrid” - right hand side), produced at an industrially applicable scale in accordance with the process depicted in Figure 8 (Process A), from potato protein as an exemplary functionalised plant protein, and amylase (kdji protein) isolated from an Aspergillus sp. culture as an exemplary non-animal derived globular protein, compared to an equivalent edible composition produced with amylase alone (“STD” - left hand side).Figure 12 is a plot of the mean particle size (D[4,3]) of an embodiment of the edible compositions of the present invention [“Hybrid” - left hand side], produced at an industrially applicable scale in accordance with the process depicted in Figure 9 (Process B), from potato protein (PP) as an exemplary functionalised plant protein, and amylase (kdji protein) isolated from an Aspergillus sp. culture as an exemplary non-animal derived globular protein, compared to an equivalent edible composition produced with amylase alone (“STD” - right hand side).Figure 13 is a plot of the mean particle size (D[4,3]) of an embodiment of the edible compositions of the present invention (“Hybrid” - right hand side), produced at an industrially applicable scale in accordance with the process depicted in Figure 10 (Process C), from potato protein (PP) as an exemplary functionalised plant protein, and amylase (kbji protein) isolated from an Aspergillus sp. culture as an exemplary non-animal derived globular protein, compared to an equivalent edible composition produced with amylase alone (“STD” - left hand side). As can be seen from Figures 11 to 13, the edible compositions of the present invention can have similar, smaller or larger particles than the “STD” amylase-only based comparative edible compositions, depending on the process employed (Processes A to C).Figure 14 is a plot of the volume distribution (% Volume Density Vs Particle Size Class pm) of an embodiment of the edible compositions of the present invention (“Hybrid”), produced at an industrially applicable scale in accordance with the process depicted in Figure 8 (Process A), from potato protein (PP) as an exemplary functionalised plant protein, and amylase (kbji protein) isolated from an Aspergillus sp. culture as an exemplary non-animal derived globular protein, compared to an equivalent edible composition produced with amylase alone (“STD”).Figure 15 is a plot of the volume distribution (% Volume Density Vs Particle Size Class pm) of an embodiment of the edible compositions of the present invention [“Hybrid”], produced at an industrially applicable scale in accordance with the process depicted in Figure 9 (Process B), from potato protein (PP) as an exemplary functionalised plant protein, and amylase (kbji protein) isolated from an Aspergillus sp. culture as an exemplary non-animal derived globular protein, compared to an equivalent edible composition produced with amylase alone (“STD”).Figure 16 is a plot of the volume distribution (% Volume Density Vs Particle Size Class pm) of an embodiment of the edible compositions of the present invention (“Hybrid”), produced at an industrially applicable scale in accordance with the process depicted in Figure 10 (Process C), from potato protein (PP) as an exemplary functionalised plant protein, and amylase (koji protein) isolated from an Aspergillus sp. culture as an exemplary non-animal derived globular protein, compared to an equivalent edible composition produced with amylase alone (“STD”).Figure 17 is a plot of the Total Serum Release (TSR%) and Water Holding Capacity (WHC%) of an embodiment of the edible compositions of the present invention (“Hybrid”), produced at an industrially applicable scale in accordance with the process depicted in Figure 8 (Process A), from potato protein (PP) as an exemplary functionalised plant protein, and amylase (kbji protein) isolated from an Aspergillus sp. culture as an exemplary non-animal derived globular protein, compared to an equivalent edible composition produced with amylase alone (“STD”).Figure 18 is a plot of the Total Serum Release (TSR%) and Water Holding Capacity (WHC%) of an embodiment of the edible compositions of the present invention [“Hybrid”], produced at an industrially applicable scale in accordance with the process depicted in Figure 9 (Process B),from potato protein (PP) as an exemplary functionalised plant protein, and amylase (koji protein) isolated from an Aspergillus sp. culture as an exemplary non-animal derived globular protein, compared to an equivalent edible composition produced with amylase alone (“STD”).Figure 19 is a plot of the Total Serum Release (TSR%) and Water Holding Capacity (WHC%) of an embodiment of the edible compositions of the present invention (“Hybrid”), produced at an industrially applicable scale in accordance with the process depicted in Figure 10 (Process C), from potato protein (PP) as an exemplary functionalised plant protein, and amylase (koji protein) isolated from an Aspergillus sp. culture as an exemplary non-animal derived globular protein, compared to an equivalent edible composition produced with amylase alone (“STD”). As can be seen from Figures 17 to 19, the edible compositions of the present invention can have a water holding capacity and total serum release higher, lower or similar to the amylase-only reference standard based edible composition depending on the process used (Process A to Process C).Figure 20 is a plot of the Firmness (Max force [N]) of an embodiment of the edible compositions of the present invention (“Hybrid”), produced at an industrially applicable scale in accordance with the process depicted in Figure 8 (Process A), from potato protein (PP) as an exemplary functionalised plant protein, and amylase (koji protein) isolated from an Aspergillus sp. culture as an exemplary non-animal derived globular protein, compared to an equivalent edible composition produced with amylase alone (“STD”).Figure 21 is a plot of the Firmness (Max force [N]) of an embodiment of the edible compositions of the present invention [“Hybrid”], produced at an industrially applicable scale in accordance with the process depicted in Figure 9 (Process B), from potato protein (PP) as an exemplary functionalised plant protein, and amylase (koji protein) isolated from an Aspergillus sp. culture as an exemplary non-animal derived globular protein, compared to an equivalent edible composition produced with amylase alone (“STD”).Figure 22 is a plot of the Firmness (Max force [N]) of an embodiment of the edible compositions of the present invention (“Hybrid”), produced at an industrially applicable scale in accordance with the process depicted in Figure 10 (Process C), from potato protein (PP) as an exemplary functionalised plant protein, and amylase (koji protein) isolated from an Aspergillus sp. culture as an exemplary non-animal derived globular protein, compared to an equivalent edible composition produced with amylase alone (“STD”). As can be seen from Figures 20 to 22, the edible compositions of the present invention can have a higher or lower firmness compared to the amylase-only reference standard based edible composition depending on the process used (Process A to Process C).Figures 23 are plots of the Viscosity [mPas] of an embodiment of the edible compositions of the present invention (“Hybrid”), produced at an industrially applicable scale in accordance with the process depicted in Figure 10 (Process C), from potato protein (PP) as an exemplaryfunctionalised plant protein, and amylase (koji protein) isolated from an Aspergillus sp. culture as an exemplary non-animal derived globular protein, compared to an equivalent edible composition produced with amylase alone (“STD”), under the conditions; A (8 °C, 7 1 / s); B (25 °C, 7 1 / s); C (8 °C, 500 1 / s); & D (25 °C, 500 1 / s). This exemplary embodiment of the edible composition has a consistently lower viscosity than the reference standard.Figure 24 is a plot of the % structural loss at 8 °C and 25 °C, and 7 and 500 1 / s of an embodiment of the edible compositions of the present invention (“Hybrid”), produced at an industrially applicable scale in accordance with the process depicted in Figure 10 (Process C), from potato protein (PP) as an exemplary functionalised plant protein, and amylase (koji protein) isolated from an Aspergillus sp. culture as an exemplary non-animal derived globular protein, compared to an equivalent edible composition produced with amylase alone (“STD”).Figure 25 is a plot of the Firmness (Max force [N]) of a series of embodiments of the edible compositions of the present invention using koji protein as an exemplary non-animal derived globular protein, and using the following plant proteins both directly, and in pre-prepared functionalised plant protein form; Mung Bean protein, Soy Bean protein and Fava Bean protein; samples with pre-prepared functionalised plant proteins added are labelled “pre-treated” and those where the plant protein is added directly are not. A comparative reference standard of an equivalent edible composition produced with amylase alone labelled “STD” was also analysed. From this analysis it can be seen that in general, the addition of plant proteins to the edible compositions increases their firmness (see sample STD as a comparison). Pre-prepared functionalised plant proteins increase the firmness of the edible compositions to an even higher extent compared to untreated plant proteins.Figure 26 is a plot of sensory evaluation scores of a cohort of 16 panelists ranking three different samples of edible compositions produced at an industrial scale (400 kg) according to the most preferred one, where “Standard” represents a comparative control composition, containing no functionalised plant protein, “Preparation with functionalised potato protein” represents a composition prepared in the same manner as “Standard”, but with the addition of functionalised plant protein, and “Preparation with functionalised potato protein and palm fat” represents a composition prepared identically to “Preparation with functionalised potato protein”, but with palm fat instead of vegetable fat.DETAILED DESCRIPTION OF THE INVENTION

[0016] The present invention enables the use of non-animal derived proteins like plant proteins and globular proteins such as enzymes and / or protein mixtures isolated from microbial fermentations, to produce animal-free “substitute” dairy food products (e.g. cheeses, etc) withsuperior functional and / or organoleptic properties, compared to existing substitute dairy food products.

[0017] When formulating such substitute food products, the simple combination of a plant protein and a globular protein often does not lead to desirable product attributes. The key bottleneck in the application of plant proteins in foods is that they usually have an astringent, dry and rough mouthfeel or texture, thus limiting their utilization in higher quantities. Functionalizing plant proteins enhances their performance in food products such that they significantly improve the products' sensorial, organoleptic and / or techno-functional properties. The present inventors have surprisingly found that changes in the globular protein gel network caused by the incorporation of such functionalised plant protein particles, significantly improves the overall quality of the resultant substitute dairy food products.

[0018] Thermo-mechanical pre-treatment (ie; “functionalisation”) of plant proteins results in small physically cross-linked gel-bead like particles with the ability to swell and provide a lubrication effect during oral processing. This enables the incorporation of plant proteins into substitute dairy foods (e.g. koji protein based cheeses). The creaminess is enhanced while the sandy, floury mouthfeel compared to a globular protein only-based product is significantly reduced. The resultant “hybrid” substitute dairy food has the potential to reduce the fat content in the formulation without negative impact on the mouthfeel thus offering a healthier product. Furthermore, the yellowness of the product is reduced, resulting in an overall whiter product leading to increased consumer acceptance. For example, compared to the globular protein (amylase) only-based cream cheese, the hybrid version has more dairy-notes, an overall more complex flavor profile and the acidity is significantly reduced.Functionalised plant proteins

[0019] The functionalisation of plant proteins in accordance with the present invention is achieved via a thermo-mechanical pre-treatment process, involving adjusting a solution of plant protein to a neutral or near neutral pH, heating the solution to a suitable elevated temperature for a suitable period of time, cooling the solution to form a gel, and breaking down the gel via a mechanical means, to form a functionalised plant protein.

[0020] Throughout this specification, the term “functionalised plant protein” or “functionalized plant protein”, and grammatical equivalents thereof, shall be understood to refer to any plant derived protein source or any protein or mixture of proteins isolated from a plant, which has been subjected to the thermo-mechanical treatment process in accordance with the present invention (Figures 1 and 2). Functionalised plant proteins prepared in accordance with the thermo-mechanical treatment process of the present invention may be in the form of a suspension of plant protein particles, and / or may be in the form of a plant protein microgel.

[0021] In accordance with the present invention, functionalised plant proteins may be preprepared by thermo-mechanical treatment of plant proteins, prior to their incorporation into edible dairy substitute food products. Alternatively, functionalised plant proteins may be prepared in-situ during the formation of edible dairy substitute food products.Edible compositions

[0022] In a first aspect, the disclosure herein provides an edible composition comprising; i. a non-animal derived globular protein, wherein the non-animal derived globular protein is at least partially denatured; and ii. at least one functionalised plant protein.

[0023] In a preferred embodiment of the first aspect, the disclosure herein provides an edible composition comprising; i. a non-animal derived globular protein, wherein the non-animal derived globular protein comprises one or more non-animal derived enzymes selected from the group consisting of; a lactase, an amylase and a cellulase; and wherein the non-animal derived globular protein is at least partially denatured; and ii. at least one functionalised plant protein.

[0024] Throughout this specification, the term “globular protein”, and grammatical equivalents thereof, shall be understood to refer to any protein (or mixture of proteins) having a native globular or spheroprotein tertiary structure. Thus, the term “globular protein” includes for example, and without limitation, globins and enzymes, and mixtures thereof. The term “non- animal derived globular protein” shall be understood to refer to any globular protein (including mixtures of different species of globular proteins) derived from a source other than an animal, Thus, the term “non-animal derived globular protein” includes for example, and without limitation, any globular protein (or mixture of globular proteins) derived from a microbe, or a plant, or any globular protein (or mixture of globular proteins) that is recombinantly produced, or is synthetically produced.

[0025] In some embodiments, the non-animal derived globular protein comprises an isolate of a fungal culture.

[0026] Throughout this specification, the term “isolate”, and grammatical equivalents thereof, shall be understood to refer to things that have been isolated from the word or phrase to which the term applies. For example, and without limitation, “an isolate of a fungal culture” may refer to an extract or fermentate of the fungal culture, or anything that has been isolated from the fungalculture, including complex mixtures isolated from the fungal culture. Thus, where the specification defines the non-animal derived globular protein as comprising “an isolate of a fungal culture”, the isolate of the fungal culture may include more than one species of globular protein.

[0027] In some embodiments, the non-animal derived globular protein comprises an isolate of an Aspergillus sp. culture.

[0028] In some embodiments, the non-animal derived globular protein comprises an isolate of a koji culture.

[0029] In some embodiments, the non-animal derived globular protein comprises a non-animal derived enzyme.

[0030] In some embodiments, the non-animal derived globular protein comprises one or more enzymes selected from the group consisting of; a lactase, a glucosidase, a cellulase, an amylase, an invertase, a protease, xylanase, glutenase, phytase, lipase, gelatinise, glucose oxidase, transglutaminase, pectinase, beta amylase, pullulanase, naringinase, limoninase, aminopeptidase, laccase, tyrosinase, cutinase, superoxide dismutase, endoglycosidase, glycocyl transferase, glucose isomerase, amidase, lignin peroxidase, invertase, and a kinase.

[0031] In some embodiments, the non-animal derived globular protein is denatured by hydrolysis, or by heat treatment, or by mechanical shearing, or any combination thereof.

[0032] In some embodiments, the non-animal derived globular protein is completely denatured.

[0033] In some embodiments, the functionalised plant protein comprises one or more plant proteins derived from plant parts selected from the group consisting of; fruits, tubers, roots, seeds, legumes, pulses, cereals, and nuts.

[0034] In some embodiments, the functionalised plant protein comprises one or more plant proteins derived from plants selected from the group consisting of; solanaceous plants, and fabaceous plants.

[0035] In some embodiments, the functionalised plant protein comprises potato protein, or mung bean protein, or fava bean protein, or soy protein, or sunflower protein, or rapeseed / Canola protein, or pea protein, or hemp protein, or gluten or a combination thereof.

[0036] In some embodiments, the edible composition further comprises one or more lipids, or one or more fats, or one or more sweetening agents, or one or more carbohydrates, or one or more polysaccharides, or one or more sulphated polysaccharides, or one or more emulsifiers, or one or more lecithins, or one or more glycerophospholipids, or any combination thereof.

[0037] In some embodiments, the edible composition is a non-animal derived substitute dairy product.

[0038] In some embodiments, the edible composition is a non-animal derived substitute food product in the form of a curd, milk, butter, cheese, yoghurt, ripened yoghurt, custard, cream cheese, soft cheese, medium-hard cheese, hard cheese, pasta filata cheese, or ripened cheese.Processes for the production of edible compositions

[0039] In a second aspect, the disclosure herein provides a process for preparing the edible compositions of the first aspect, wherein the process comprises; i. functionalising at least one plant protein via a thermo-mechanical functionalisation treatment; and ii. at least partially denaturing a non-animal derived globular protein.

[0040] In some embodiments, the thermo-mechanical functionalisation treatment comprises the steps of; a. subjecting a solution or suspension of a plant protein to a heat treatment at an elevated temperature; b. cooling the heated solution or suspension of a plant protein from step a. to form a gel; and c. subjecting the gel from step b. to a mechanical shearing process.

[0041] In some embodiments, the elevated temperature of step a. is a temperature falling within the range of 65 °C to 98 C.

[0042] In some embodiments, step b. comprises cooling the heated solution or suspension of a plant protein from step a. to a temperature falling within the range of 1 °C to 60 °C.

[0043] In some embodiments, the mechanical shearing process comprises treatment with a mechanical blender, or a homogenizer.

[0044] In some embodiments, step ii. of at least partially denaturing the non-animal derived globular protein comprises thermal denaturation, and / or mechanical denaturation, and / or chemical denaturation, and / or denaturation by fermentation.

[0045] In some embodiments, step i. is performed separately from step ii., and then the functionalised plant protein is added to the non-animal derived globular protein, prior to step ii. of at least partially denaturing the non-animal derived globular protein.

[0046] In some embodiments, step i. is performed concomitantly or simultaneously with step ii.General

[0047] Those skilled in the art will appreciate that the invention described herein is susceptible to variations and modifications other than those specifically described. The invention includes all such variations and modifications. The invention also includes all of the process steps, features, formulations, compositions and compounds referred to or indicated in the specification, individually or collectively and any and all combinations or any two or more of the steps or features.

[0048] Each document, reference, patent application or patent cited in this text is expressly incorporated herein in their entirety by reference, which means that each cited document, reference, patent application or patent should be read and considered by the reader as part of this text. That the document, reference, patent application or patent cited in this text is not repeated in this text is merely for reasons of conciseness.

[0049] Any manufacturer’s instructions, descriptions, product specifications, and product sheets for any products mentioned herein or in any document incorporated by reference herein, are hereby incorporated herein by reference, and may be employed in the practice of the invention.

[0050] The present invention is not to be limited in scope by any of the specific embodiments described herein. These embodiments are intended for the purpose of exemplification only. Functionally equivalent products, formulations and methods are clearly within the scope of the invention as described herein.

[0051] The invention described herein may include one or more range of values (eg. Particle size, concentration, viscosity etc). A range of values will be understood to include all values within the range, including the values defining the range, and values adjacent to the range which lead to the same or substantially the same outcome as the values immediately adjacent to that value which defines the boundary to the range. Accordingly, unless indicated to the contrary, the numerical parameters set forth in the specification and claims are approximations that may vary depending upon the desired properties sought to be obtained by the present invention. Hence “about 80 %” means “about 80 %” and also “80 %”. At the very least, each numerical parameter should be construed in light of the number of significant digits and ordinary rounding approaches.

[0052] Throughout this specification, unless the context requires otherwise, the word “comprise” or variations such as “comprises” or “comprising”, will be understood to imply the inclusion of a stated integer or group of integers but not the exclusion of any other integer or group of integers. It is also noted that in this disclosure and particularly in the claims and / or paragraphs, terms such as “comprises”, “comprised”, “comprising” and the like can have the meaning attributed to it in U.S. Patent law; e.g., they can mean “includes”, “included”, “including”, and the like; and that terms such as “consisting essentially of’ and “consists essentially of’ have the meaning ascribed to them in U.S. Patent law, e.g., they allow for elements not explicitly recited, but exclude elements that are found in the prior art or that affect a basic or novel characteristic ofthe invention.

[0053] Other definitions for selected terms used herein may be found within the detailed description of the invention and apply throughout. Unless otherwise defined, all other scientific and technical terms used herein have the same meaning as commonly understood to one of ordinary skill in the art to which the invention belongs.

[0054] The following examples serve to more fully describe the manner of using the abovedescribed invention, as well as to set forth the best modes contemplated for carrying out various aspects of the invention. It is understood that these methods in no way serve to limit the true scope of this invention, but rather are presented for illustrative purposes.EXAMPLES

[0055] Further features of the present invention are more fully described in the following non-limiting Examples. This description is included solely for the purposes of exemplifying the present invention. It should not be understood as a restriction on the broad description of the invention as set out above.Example 1 - Preparation of functionalised plant proteins (Laboratory scale)

[0056] MaterialsPotato protein isolate, Solanic300, Avebe, Netherlands (90% protein content).Sodium hydroxide, food grade, general lab supplier.

[0057] MethodTo prepare an exemplary functionalized plant protein solution at laboratory scale a potato protein isolate was hydrated in tap water at a concentration of 15% w / w in a beaker and stirred until dissolved. The pH was adjusted to 7 using 1 M NaOH. Alternatively, a predefined amount of 1 M NaOH can be added to the tap waterbefore stirring in the plant protein to achieve a pH of 7. The solution was hydrated at room temperature for 2 h. Then, the beaker was put into a pre-heated water bath set to 90 °C for 30 min. Alternatively, the solution can be transferred into plastic foils which are tightly knotted on both ends and placed into the pre-heated water bath under the same conditions. After 30 min the beaker (or plastic foil) was placed into an ice bath to rapidly cool down the gels and stored at 4 °C overnight. Next, the gels were diluted with tap water at a ratio 1 :2 to achieve a final protein concentration of 5% w / w. The diluted gels were broken down using a household hand mixer at normal speed for 1 min. Finally, the diluted solution was homogenized using a high pressure homogenizer (PandaPlus 1000, GEA) at 250 / 50 bar (1st / 2nd stage) for three cycles. The process is presented in Figure 1 .Example 2 - Preparation of functionalised plant proteins (Pilot plant scale)

[0058] MaterialsPotato protein isolate, Solanic300, Avebe, Netherlands (90% protein content).Sodium hydroxide, food grade, general lab supplier.

[0059] MethodTo prepare the functionalized plant protein solution at an industrially applicable pilot plant scale a potato protein isolate, tap water, and a pre-defined amount of 1 M NaOH to achieve a pH of 7 were hydrated in a batch cooker for 2 min. If necessary, an antifoaming agent can be added to prevent foam formation. Then the mixture was heated to 90 °C under vacuum to avoid foam formation using direct steam. Once the temperature of the mix reached 90 °C, the mixture was stirred for another 1 min before adding cooling water to achieve a protein content of 5% w / w and a temperature < 60 °C. The cooled down mix was stirred once again for 1 min and finally homogenized using a high pressure homogenizer (TwinPanda 400, GEA) at 250 / 50 bar (1st / 2nd stage) for one cycle. The process is presented in Figure 2.Example 3 - Characterisation of particle size of functionalised plant proteins

[0060] The particle sizes of functionalized plant proteins before and after homogenization, produced in accordance with examples 1 and 2 (potato protein), were measured using a Mastersizer 3000 (Malvern Panalytical) using a refractive index of 1 .47, an absorption index of 0.005, and water as a dispersant (refractive index 1 .33).

[0061] Figure 3 shows the mean particle size (d[4,3]) of the functionalised plant proteins before and after homogenization. The data show that after homogenization the particle size of the functionalised plant protein is significantly reduced. At laboratory scale, the particle size is slightly lower compared to the pilot plant process, however, they are in a similar range. Due to the reduction of the particle size of the functionalised plant protein, the otherwise astringent mouthfeel of such protein particles is reduced and a ball bearing effect on the tongue is created leading to increased lubricity.

[0062] Figure 4 shows the volume-based particle size distribution of the functionalized plant protein before and after homogenization. It can be seen that after homogenization not only the particle size is reduced but also the distribution is narrower pointing towards a more stable and homogeneous mix. Figure 5 shows the volume-based particle size distribution of the functionalized plant protein produced at pilot plant scale.Example 4 - Gelation properties of functionalised plant proteins

[0063] Analysis of the thermal gelation properties of functionalised plant proteins are carried out by small deformation oscillatory tests. One commonly applied test procedure to study the rheological properties of protein gels are oscillation tests using a rheometer. The basic principle of these small deformation oscillation tests can be described using concentric cylinder geometry.

[0064] In a rheometer, applying oscillation tests the viscoelastic behavior of a sample can be described using either the complex viscosity (mPa s) or the storage modulus, G' (given in Pa). The complex viscosity q* is a measure of the total resistance to flow as a function of angular frequency and is given by the quotient of the maximum stress amplitude and maximum strain rate amplitude. The G' value is a measure of the deformation energy stored in the material during the shear process. G' represents the elastic behavior of the sample. A high gel elasticity corresponds to a high elastic component (high G' and q*). The temperature at which q* increases reflect thermally induced changes that lead to the formation of a gel.

[0065] Figure 6 shows the temperature dependent evolution of q* of two functionalised plant proteins (using potato protein isolate, prepared according to Figure 2, and Example 2). No change is observed until 80 °C and only a small increase is observed upon further heating to 90 °C implying that the protein does not undergo further changes during processing therefore being able to act as an inert filler material in the at least partially denatured globular protein matrix of a substitute dairy food product in accordance with the present invention.Example 5 - Investigation of hybrid gel structures

[0066] MaterialsKoji protein powder (Breatec), protein content 70.45%.Potato protein, Solanic 300 (Avebe)

[0067] A heat-induced gel was produced from koji protein powder in water. The addition of functionalized plant protein altered the gelation and the gel properties. The gelation and gel properties were measured by rheological methods using small deformation oscillation tests.

[0068] MethodThe plant protein (potato protein) was briefly hydrated in tap water containing NaOH to reach a final pH of 6-7 then heated to 80°C, and cooled, followed by homogenization at 250 / 50 bar to provide a functionalised plant protein. The final protein concentration was 3.9% (sample: PP).Koji powder (as an exemplary globular protein) was added to water at a rate of 5% protein weight / volume (5 grams protein in 100 ml total volume). The pH was adjusted to pH 4, providing a globular protein suspension (sample: koji protein).10.8 ml of the 5% koji protein suspension and 7.2 ml of the functionalised 3.9% potato protein suspension were mixed, to provide a sample of functionalised plant protein and globular protein in a ratio of 40:60 (sample: Mix).All three samples were heated at a rate of 5 K / min to a final temperature of 85°C to denature the protein(s) and induce coagulation and simultaneously measure the process of coagulation inside the rheometer, followed by cooling the gels thus formed to room temperature at a rate of 5 K / min.

[0069] Analysis of rheological properties (gel properties) within rheometerOne commonly applied test procedure to study the rheological properties of protein gels are oscillation tests using a rheometer. The basic principle of these small deformation oscillation tests can be described using concentric cylinder geometry.In a rheometer, applying oscillation tests the viscoelastic behavior of a sample can be described using either the complex viscosity (mPa s) or the storage modulus, G' (given in Pa). The complex viscosity q* is a measure of the total resistance to flow as a function of angular frequency and is given by the quotient of the maximum stress amplitude and maximum strain rate amplitude. The G' value is a measure of the deformation energystored in the material during the shear process. G' represents the elastic behavior of the sample. A high gel elasticity corresponds to a high elastic component (high G' and q*) .The data (Figure 7) show the temperature dependent evolution of q* of the koji protein preparation (globular protein), potato protein preparation (functionalised plant protein) and a mixed system comprising both, upon heating until 85 °C. The temperature at which q* increases reflects thermally induced changes that lead to the formation of a gel. No increase was detected for the functionalised plant protein sample (PP). Compared to pure globular protein (koji protein), the increase in q* starts at a higher temperature and reaches a lower final value if functionalised plant protein (PP) is mixed with globular protein (koji protein).Example 6 - Preparation of edible compositions

[0070] Materials5.14% koji protein, Breatec2.61% plant protein, e.g. potato protein isolate, Solanic 300, Avebe2.39% nutriose FM10, Roquette0.6% carrageenan LC-05, CP Kelco29.88% Redusat S40G, Fuji Oil0.3% LecicoSun FM580, Lecico / Avril Group1.6% 1 M NaOH, Spolchemie0.6% salt0.6% sugar0.1 % Vega FreshQ 101 , Christian Hansen0.1 % Vega Premium, Christian Hansen tap water.

[0071] Process AIn accordance with the process steps outlined in Figure 8, all dry ingredients except globular protein (e.g. koji protein powder) and functionalised plant protein (e.g. potato protein) are mixed in a batch cooker with tap water for 2 min at room temperature. Then, direct steam is injected until the temperature has reached 75-80 °C to pasteurize the mixture. The cold functionalized plant protein solution (4-5°C) produced as described above (Examples 1 and 2) is added to the batch cooker to cool down the mass and mixed for 2 min. Next, the globular protein is added and mixed in for 12 min under vacuum to prevent foam formation. Thereafter, the pre-melted fat and lecithin are added to the cooker, mixed in for 2 min at high shear under vacuum to create a pre-mix. Thepre-mix is homogenized at 150 / 50 bar (1st / 2nd stage) using a high-pressure homogenizer (TwinPanda 400, GEA). After addition of starter cultures (Vega FreshQ, Vega Premium), the mix is incubated (fermented) at 35 °C for 16 h. Next, the mass is transferred to the batch cooker once again and heated to 80 °C using direct steam under continuous stirring. Then, the heated mass is cooled down e.g. in buckets. Once the temperature of the product has reached < 60 °C it is homogenized at 150 bar using a high pressure homogenizer (TwinPanda 400, GEA). The final product is filled into airtight buckets and stored at 4-5 °C.

[0072] Process BIn accordance with the process steps outlined in Figure 9, all dry ingredients except globular protein (e.g. koji protein powder), but including dry plant protein (e.g. potato protein powder - in this process, the plant proteins are functionalised in situ) are mixed in a batch cooker with tap water for 2 min at room temperature. Then, direct steam is injected until the temperature has reached 75-80 °C to pasteurize the mixture. Cold tap water is added to the batch cooker to cool down the mass and mixed for 2 min. Next, the globular protein is added and mixed in for 12 min under vacuum to prevent foam formation. Thereafter, the pre-melted fat and lecithin are added to the cooker, mixed in for 2 min at high shear under vacuum to create a pre-mix. The pre-mix is homogenized at 150 / 50 bar (1st / 2nd stage) using a high pressure homogenizer (TwinPanda 400, GEA). After addition of starter cultures, the mix is incubated (fermented) at 35 °C for 16 h. Next, the mass is transferred to the batch cooker once again and heated to 80 °C using direct steam under continuous stirring. Then, the heated mass is cooled down e.g. in buckets. Once the temperature of the mass has reached < 60 °C it is homogenized at 150 bar using a high pressure homogenizer (TwinPanda 400, GEA). The final product is filled into buckets and stored at 4-5 °C.

[0073] Process CIn accordance with the process steps outlined in Figure 10, all dry ingredients, including globular protein powder (e.g. koji protein powder), as well as tap water and the functionalized plant protein solution produced as described above (Examples 1 and 2) are mixed in a batch cooker for 10 min at room temperature. Then, the pre-melted fat and lecithin are added to the cooker, mixed in for 2 min at high shear under vacuum to create a pre-mix. The pre-mix is homogenized at 150 / 50 bar (1st / 2nd stage) using a high pressure homogenizer (TwinPanda 400, GEA). After addition of starter cultures, the mix is incubated (fermented) at 35 °C for 16 h. Next, the mass is transferred to the batch cooker once again and heated to 80 °C using direct steam under continuous stirring.Then, the heated mass is cooled down e.g. in buckets. Once the temperature of the mass has reached < 60 °C it is homogenized at 150 bar using a high pressure homogenizer (TwinPanda 400, GEA). The final product is filled into buckets and stored at 4-5 °C.

[0074] Process DIn a further alternative process, tap water, 1 M NaOH and antifoaming agent are mixed in a batch cooker at room temperature for 1 min. Then all dry ingredients except globular protein powder (e.g. koji protein powder), but including dry plant protein, are added and mixed for another 2 min at room temperature. Next, the solid fat and lecithin are added to the batch cooker and mixed in for 1 min. Then, direct steam is injected until the temperature has reached 75-80 °C to pasteurize the mixture while melting the fat. After adding cold tap water to cool down the mass and mixing for 1 min the globular protein is added and mixed in for 12 min. The mix is homogenized at 250 / 50 bar (1st / 2nd stage) using a high-pressure homogenizer (TwinPanda 400, GEA). After addition of starter cultures, the mix is incubated (fermented) at 35 °C for 16 h. Next, the mass is transferred to the batch cooker once again and heated to 80 °C using direct steam under continuous stirring. Then, the heated mass is cooled down e.g. in buckets. Once the temperature of the mass has reached < 60 °C it is homogenized at 150 bar using a high pressure homogenizer (TwinPanda 400, GEA). The final product is filled into buckets and stored at 4-5 °C.

[0075] Analysis of particle size of the edible compositionsThe particle size of the edible compositions were measured using a Mastersizer 3000 (Malvern Panalytical) with a refractive index of 1 .45, an absorption index of 0.01 , and water as a dispersant. The volume-based particle size distribution was analyzed using the Mie-theory. All measurements were carried out in triplicate. Values presented in the figures represent the average.Figures 11-13 illustrate the mean particle size (d[4,3]) of cream cheese style substitute dairy compositions made with and without functionalized potato protein (thereafter referred to as “Hybrid” and “STD” respectively. The ratio of functionalised potato protein to koji protein in the cream cheeses is 0 / 100 and 40 / 60, respectively).The data show that the effect of thermo-mechanically pre-treated (i.e. functionalized) potato protein addition to the koji protein-based cream cheese composition on the mean particle size is dependent on the process. According to Figures 12 and 13, when the cream cheese is prepared according to process B and C, the particle size of the hybridcream cheese is larger compared to the STD. However, when the cream cheese is produced according to process A, the resulting mean particle size of the hybrid cream cheese composition is smaller. This shows that the process chosen influences the final product properties.Figures 14-16 illustrate the volume-based particle size distributions of the cream cheeses with and without functionalised plant protein produced using the different processes A, B and C. The data show that the process can affect the particle size differently. While process C leads to trimodal particle size distributions for both hybrid and non-hybrid (STD) cream cheese variants, process A resulted in a bimodal distribution for both, and process B led to a bimodal distribution in case of the non-hybrid (STD) cream cheese and a trimodal distribution in case of the hybrid cream cheese. Furthermore, in process A the particle size distributions of the koji protein based cream cheese and the one containing functionalized potato protein are very similar, while in process B and C the size distribution of the ones containing potato protein are slightly shifted towards larger particle sizes reflecting a change in the microstructure of the cheese, i.e. especially protein-fat interactions.

[0076] Analysis of Water Holding Capacity and Total Serum Release of the edible compositionsThe water holding capacity / total serum release both play an important role not only in the sensory perception of food products but also in the quality. In cream cheeses the water holding capacity (WHC) usually is correlated to the mouthfeel and a high total serum release (TSR) is often times accompanied by a lower consumer acceptance.The WHC and TSR were measured by filling aliquots of the cheese sample including any expelled “whey” into three centrifuge tubes weighing 35 + / - each. The tubes are centrifuged at the same time at a relative centrifugal force (RFC) of 3000 x g and a holding time of 10 min and a temperature of 10 °C for the first step. Then, the RCF is increased to 15,000 x g, and 21 ,000 x g for the two following steps with the same holding time each, respectively. After each step, the serum is poured out and weighed by pushing the cream cheese away with a flat spatula to create a channel between the cheese mass and the wall of the tube.The TSR is calculated by adding up the amount removed after each step, thus expressed as a percentage of the total sample weight:Total Serum Release (TSR)% = (( g serum per step) / (g initial weight - g tube))x 100The WHC is calculated as follows:WHC %= ((W(0) - W) / W(0)) x 100, whereW(0): g of initial water content (moisture content)W: g of serum released after centrifugation after step 1 and 2Figures 17-19 depict the WHC and TSR of edible cream cheese style substitute dairy compositions with and without functionalized plant protein produced using different processes. The data show that the impact of functionalized plant protein addition to the koji protein based cream cheese composition is process dependent. While in process C almost no change in WHC nor TSR is observed, process A led to an increase in TSR and a reduction in WHC in the hybrid cheese compared to the non-hybrid (STD) and vice versa in process B. The differences in the WHC and TSR can be explained by the different ability of the globular protein to bind water upon addition of the functionalized plant protein and a change in the protein-water interactions.

[0077] Analysis of firmness (texture) of the edible compositionsThe impact on the product firmness was evaluated using a texture analyzer (TA.XT. Plus C, Stable Micro Systems). The cream cheese samples (150g in round cups, cold) were penetrated to 75% of their initial height using a cream probe. The force needed to penetrate each sample was recorded.The firmness of the cream cheese samples is shown in Figures 20-22. The hybrid cream cheeses (Hybrid) can have a higher or lower firmness than the globular protein-only based cheeses (STD) depending on the process used, pointing towards a change in the protein network and the overall microstructure upon addition of the functionalized plant protein.

[0078] Analysis of viscosity of the edible compositionsThe effect on the edible composition product viscosity was measured using the MCR 102e rheometer (Anton Paar) and a parallel plate geometry PP25 / S at 8 and 25 °C.For the viscosity measurement, a small sample amount of the edible composition is placed between the plate geometry. The measuring gap is 1 mm and excess sample is removed before starting the measurement. The viscosity is analysed by an increasing ascending shear ramp rate (0...500 1 / s), a holding section at 500 1 / s and a decreasing descending shear rate ramp (500...0 1 / s). The viscosity in mPas is derived from the curves at a certain shear rate. The loss in structure can be calculated as the percentage of viscosity before and after shearing. The measurements are performed at 8°C and 25°C to see the impact of temperature shift.Figures 23A-23D show the viscosity, and Figure 24 shows the percentual structure loss of the cream cheeses at 8 and 25 °C at 7 and 500 1 / s. In process C the viscosity of the hybrid cream cheese containing functionalised plant protein is lower than that of the globular protein only-based cheese. The percentual structure loss is temperature dependent. These results may be explained by microstructural differences upon addition of functionalized plant protein.

[0079] L*a*b* colour analysis of the edible compositionsThe colour of the cream cheese was measured using the CR-10 Plus colorimeter (Konica Minolta) using a D65 illuminant, an 8° illumination angle and a measurement area of 8mm. All measurements were carried out in triplicate.Colour can be described using different color spaces. The CIELAB, also referred to as L*a*b* coordinates, is one of them and widely used across many industries.L* stands for the lightness and represents the perceived lightness of the colour. It ranges from 0 to 100 where 0 is black and 100 is white. a* stands for the green-red balance and ranges from approx. -128 to 127 where negative values indicate green and positive values indicate red. b* stands for the blue-yellow balance and ranges from -128 to 127 where negative values indicate blue and positive values indicate yellow.The absolute difference between two colours is referred to as DeltaE and describes the absolute difference between the L*, a* and b* coordinates between two samples.Table 1 illustrates the L*a* and b* coordinates as well as DeltaE of the globular protein based cream cheese (STD) and the hybrid cream cheese containing functionalised plant protein.Table 1. L*a*b* and DeltaE of cream cheese with and without functionalized potato protein produced using different processes:The colour of the hybrid cheeses is generally less yellow than the comparative standard (STD) protein-based cream cheese containing no functionalised plant protein (lower b* value). Lightness (L*) and red-green balance (a*) are similar between both cheese variants. The total difference between standard and hybrid cream cheese is greater than 1 , which is a noticeable difference to the human eye.

[0080] Sensory evaluation of the edible compositionsA descriptive sensory analysis performed by a trained sensory panel consisting of seven panellists revealed the following product attributes listed in Table 2.Table 2. Descriptive test of cream cheeses: globular protein only-based cream cheese hybrid cream cheese containing functionalized plant protein (40 / 60)harsh acidity, aftertaste fresh, mild, slight sourness, buttery, milkyThe hybrid edible cream cheese compositions in accordance with the present invention were demonstrably creamier, and exhibited significantly more positive organoleptic properties that the equivalent composition prepared without functionalised plant protein.Example 7 - Preparation and firmness (texture) analysis of further edible compositions

[0081] MaterialsGlobular protein: Koji protein powder (GP 121.31 Breatec), protein content 70.45%.Plant proteins: Potato protein (Avebe)Mung bean protein (Bioway)Fava bean protein (Top Health Ingredients) Soy protein (ADM).Edible compositions: 6.13% protein30% vegetable fat0.6% salt0.6% sugar2.4% maltodextrin0.6% carrageenan 0.3% sunflower lecithin Tap water.

[0082] Method & ResultsPlant protein preparations were added to a globular (koji) protein-based cream cheese formulation and the impact on the product firmness were evaluated using a texture analyzer. The cheese sample was penetrated to 75% of its initial height using a cream probe. The force needed to penetrate the sample was recorded.Plant proteins were either added directly, without pre-treatment or prepared using the same procedure as described above in Examples 1 and 2, as functionalised plant proteins. The globular protein I plant protein ratio was 60 / 40. The impact of plant protein addition to the koji-protein based cream cheese is shown in Figure 25.In general, the addition of plant proteins to globular protein-based cream cheese increases its firmness (see sample STD as a comparison). Addition of functionalised plant proteins increases the firmness of the cream cheese to an even higher extent compared to untreated plant proteins.Example 8 - Preparation of edible compositions at an industrial scale

[0083] Preparation of functionalised plant protein on industrial scale (400 kg)On an industrial scale, functionalised potato protein (as an exemplary plant protein) was produced by first preparing a 1 M NaOH solution for pH control, second the addition of an antifoaming agent thereto, at a concentration of 0.2% and third, the addition of the potato protein thereto, at a concentration of 22.22%, with mixing, at ambient temperature. After hydration of the dry ingredients, functionalisation was performed by heating the mixture with direct steam to 85°C for 2 min, smoothening it, followed by a homogenisation step (300 bar, 1 -stage). The product was hot filled into plastic bags (15 kg each) and cooled to 2-4 °C. The functionalised plant protein remains stable for several months and can be used batchwise for further purposes.

[0084] Preparation of standard (comparative example) edible composition on industrial scale (400 kg)Firstly, all dry ingredients (salt 0.65%, sugar 0.65%, stabilising agent 0.65% and fibre 2.6%) were mixed together with water, vegetable fat (32.3%) and emulsifier (0.3%). The mixture was then pasteurised at 74°C with direct steam (addition of water with steam), cooled below 50°C (addition of water), koji protein (9.2%) was added and hydrated for at least 8 min. After the homogenisation step (2 stages, 150 / 50 bar), starter cultures were added at a temperature below 42°C. The product was fermented in a tank for 16 h at 35°C, pasteurised, smoothened and filled into packages.

[0085] Preparation of edible composition with functionalised plant protein and vegetable fat on an industrial scale (400 kg)Firstly, all dry ingredients (salt 0.65%, sugar 0.65%, stabilising agent 0.65% and fibre 2.6%) were mixed together with water, vegetable fat (32.3%) and emulsifier (0.3%). The mixture was then pasteurised at 74°C with direct steam (addition of water with steam), cooled below 50°C (addition of water), functionalised potato protein (14.6%) and koji protein (5.6%) were added and hydrated for at least 8 min. After the homogenisation step (2 stages, 150 / 50 bar), starter cultures were added at a temperature below 42°C. The product was fermented in a tank for 16 h at 35°C, pasteurised, smoothened and filled into packages.

[0086] Preparation of edible composition with functionalised plant protein and palm fat on an industrial scale (400 kg)Firstly, all dry ingredients (salt 0.65%, sugar 0.65%, stabilising agent 0.65% and fibre 2.6%) were mixed together with water, palm fat (32.3%) and emulsifier (0.3%). The mixture was then pasteurised at 74°C with direct steam (addition of water with steam), cooled below 50°C (addition of water), functionalised potato protein (14.6%) and koji protein (5.6%) were added and hydrated for at least 8 min. After the homogenisation step (2 stages, 150 / 50 bar), starter cultures were added at a temperature below 42°C. The product was fermented in a tank for 16 h at 35°C, pasteurised, smoothened and filled into packages.

[0087] Sensory evaluation of the edible compositions produced on an industrial scale (400 kg)Tested products were the three types produced above on an industrial scale, viz. standard recipe (comparative example, containing no functionalised plant protein) preparation with functionalised potato protein and preparation with functionalised potato protein and palm fat. Three samples were evaluated by panelists (N = 16) using blind codes. The panelists were asked to rank the samples according to the most preferred one. With a confidence level of 90%, as shown in Figure 26, the samples containing thefunctionalised potato protein scored higher (preparation with functionalised potato protein: 31% and preparation with functionalised potato protein and palm fat: 56%) than the standard without functionalised potato protein (13%). The only difference between the standard recipe and the preparation with functionalised potato protein is that in the latter one part of the koji protein is substituted by functionalised potato protein. Overall, panelists mentioned a more balanced flavor profile, reduced sourness, and better texture / increased creaminess for both samples containing the functionalised potato protein compared to the standard.

[0088] Sensory evaluation of edible composition with functionalised plant protein and vegetable fat produced on an industrial scale (400 kg), compared to a commercially available diary substitute cream cheese productSince the edible composition produced with functionalised potato protein and palm fat on an industrial scale scored so highly as the preferred edible composition in the above described sensory evaluation, we decided to conduct a comparative sensory evaluation of this composition compared to a market leading commercially available cream cheese composition. The commercially available product chosen for comparison was Stockeld™ cream cheese, which lists its ingredients as follows:Ingredients: legume milk (water, lentil protein, chickpea protein, live cultures (incl. Bifidobactetrium lactis (HN019TM), Lactobacillus acidophilus (NCFM®)), coconut oil, starch blend (potato starch, modified food starch), canola oil, less than 2% of potato protein, salt, calcium phosphate, Xanthan gum, sugar, Rowansberry extract (to preserve freshness).Samples were evaluated by panellists (N = 17) using blind codes. The panellists were asked to rank the samples on a 9-point hedonic scale according to their overall liking, flavour liking and texture liking. The resulting average for each attribute is given in Table 3. The letter behind the number indicates whether the difference was significant (different letter) or not (same letter). The samples were also ranked according to the most preferred sample. The percentage of panelists who preferred one sample over another is displayed in Table 3. Overall, the preparation with functionalised potato protein and palm fat scored higher than the Stockeld™ cream cheese. The details as to why panellists preferred one sample over the other are listed in the comments section of Table 3.Table 3. Results of sensory evaluation compared to commercially available product

Claims

CLAIMS1 . An edible composition comprising; i. a non-animal derived globular protein, wherein the non-animal derived globular protein comprises one or more non-animal derived enzymes selected from the group consisting of; a lactase, an amylase and a cellulase; and wherein the non- animal derived globular protein is at least partially denatured; and ii. at least one functionalised plant protein.

2. The edible composition of claim 1 , wherein the non-animal derived globular protein comprises an isolate of a fungal culture.

3. The edible composition of claim 1 or claim 2, wherein the non-animal derived globular protein comprises an isolate of an Aspergillus sp. culture.

4. The edible composition of any one of claims 1 to 3, wherein the non-animal derived globular protein comprises an isolate of a kbji culture.

5. The edible composition of one of claims 1 to 4, wherein the non-animal derived globular protein is denatured by hydrolysis, or by heat treatment, or by mechanical shearing, or any combination thereof.

6. The edible composition of any one of claims 1 to 5, wherein the non-animal derived globular protein is completely denatured.

7. The edible composition of any one of claims 1 to 6, wherein the functionalised plant protein comprises one or more plant proteins derived from plant parts selected from the group consisting of; fruits, tubers, roots, seeds, legumes, pulses, cereals and nuts.

8. The edible composition of any one of claims 1 to 7, wherein the functionalised plant protein comprises one or more plant proteins derived from plants selected from the group consisting of; solanaceous plants, and fabaceous plants.

9. The edible composition of any one of claims 1 to 8, wherein the functionalised plant protein comprises potato protein, or mung bean protein, or fava bean protein, or soy protein, or sunflower protein, or rapeseed / Canola protein, or pea protein, or hemp protein, or gluten or a combination thereof.

10. The edible composition of any one of claims 1 to 9, wherein the edible composition further comprises one or more lipids, or one or more ats, or one or more sweetening agents, or one or more carbohydrates, or one or more polysaccharides, or one or more sulphated polysaccharides, or one or more emulsifiers, or one or more lecithins, or one or more glycerophospholipids, or any combination thereof.11 . The edible composition of any one of claims 1 to 10, wherein the edible composition is a non-animal derived substitute dairy product.

12. The edible composition of any one of claims 1 to 11 , wherein the edible composition is a non-animal derived substitute food product in the form of a curd, milk, butter, cheese, yoghurt, ripened yoghurt, custard, cream cheese, soft cheese, medium-hard cheese, hard cheese, pasta filata cheese, or ripened cheese.

13. A process for preparing the edible composition of any one of claims 1 to 12, wherein the process comprises the steps of; i. functionalising at least one plant protein via a thermo-mechanical functionalisation treatment to provide a functionalised plant protein; and ii. at least partially denaturing a non-animal derived globular protein.

14. The process of claim 13, wherein the thermo-mechanical functionalisation treatment comprises the steps of; a. subjecting a solution or suspension of a plant protein to a heat treatment at an elevated temperature; b. cooling the heated solution or suspension of a plant protein from step a. to form a gel; and c. subjecting the gel from step b. to a mechanical shearing process.

15. The process of claim 14, wherein the elevated temperature of step a. is a temperature falling within the range of 65 °C to 98 °C.

16. The process of claim 14 or claim 15, wherein step b. comprises cooling the heated solution or suspension of a plant protein from step a. to a temperature falling within the range of 1 °C to 60 °C.

17. The process of any one of claims 14 to 16, wherein the mechanical shearing process comprises treatment with a mechanical blender, or a homogenizer.

18. The process of any one of claims 13 to 17, wherein step ii. of at least partially denaturing the non-animal derived globular protein comprises thermal denaturation, and / or mechanical denaturation, and / or chemical denaturation, and / or denaturation by fermentation.

19. The process of any one of claims 13 to 18, wherein step i. is performed separately from step ii., and then the functionalised plant protein is added to the non-animal derivedglobular protein, prior to step ii. of at least partially denaturing the non-animal derived globular protein.

20. The process of any one of claims 13 to 19, wherein step i. is performed concomitantly or simultaneously with step ii.

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