Rubisco GEL product

Abstract

High protein content flowable Rubisco gels and process for preparation thereof are described. The gels are useful for the formulation of high protein consumer products, such as sports gels.

Description

RUBISCO GEL PRODUCTFIELD OF THE DISCLOSURE

[0001] The present disclosure broadly relates to a flowable heat set gel comprising a high concentration of Rubisco protein which has low viscosity and is suitable for use in a flowable food product, and to processes for producing such a flowable heat set gel. The disclosure also relates to food products comprising the flowable heat set gel, and to processes for producing those products.Background to the disclosure

[0002] Ribulose-l,5-bisphosphate carboxylase / oxygenase, known as RuBisCo or Rubisco, is the most abundant protein in the world, being present in all green leaves. Rubisco plays a critical role as the first enzyme involved in fixation of atmospheric carbon dioxide into energy rich molecules during photosynthesis.

[0003] Leaf protein was identified as a food source in the 1940s by British biochemist and virologist Norman Pirie. In the 1950s this protein fraction was characterized as being principally comprised of Rubisco, a white, cytoplasmic protein with a large molecular weight by Wildman et al. In the 1970s, researchers at the USDA Western Regional Research Centre undertook studies to extract Rubisco as a leaf protein concentrate from alfalfa in a process they called "Pro-Xan". Other studies focussed on increasing Rubisco's efficiency for carbon capture / biomass accumulation, crop yield, and seed yield. Rubisco makes up about 30-50% of the soluble protein in plant leaf (Erb TJ, Zarzycki J. Curr Opin Biotechnol. 2018 Feb;49: 100-107).

[0004] In nutritional terms, Rubisco has a complete amino acid profile, and has a digestibility score comparable to beef proteins (PDCAAS score). When isolated in its pure form, it has a neutral flavour and off-white colour.

[0005] There is an ongoing need for alternative protein products available in different forms. Gels having high level of protein (high protein gels) are desirable as they can provide a large serving of protein in a convenient and concentrated form to consumers. High protein gel products are known from animal sources (e.g. Friesland Campinia Whey Protein Gel at 15% protein from whey), however there is an increasing demand for non-animal sourced protein.

[0006] Rubisco as a soluble cytosolic protein is different from other plant proteins typically used as protein sources. These other plant proteins are commonly derived fromseeds, grains, or nuts within which the majority of the proteins are storage proteins which are insoluble and designed to released during germination, include for example legumin, vicilin, convicilin (legumes), glycinin, conglycinin (soy), glutenin, gliadin (wheat), and zein (maize).

[0007] Rubisco protein is known to form structured gels with high gel strength (Martin et al., J. Agric. Food Chem, 2014, 62, 10783-10791) at relatively low Rubisco concentrations. At high Rubisco protein concentrations, the Rubisco gels formed have very high strength, are firm, non-elastic, non-pliable, and brittle. Such hard gels, whilst suitable in some food formats, do not have favourable textural or palatability properties in gels targeted to provide a high level of protein per serve where there is a need for a flowable gel consistency.

[0008] It is an object of the present disclosure to go some way to meeting the abovementioned needs; and / or to at least provide the public with a useful choice.

[0009] Other objects of the disclosure may become apparent from the following description which is given by way of example only.

[0010] Any discussion of documents, acts, materials, devices, articles, or the like which has been included in the present specification is solely for the purpose of providing a context for the present disclosure. It is not to be taken as an admission that any or all of these matters form part of the prior art base or were common general knowledge in the field relevant to the present disclosure as it existed before the priority date.Summary of the disclosure

[0011] In one aspect the invention provides a process for preparing a flowable heat set gel comprising from about 15% to about 28% w / w of Rubisco protein and having a viscosity up to about 18000cP (measured with an ATAGO VISCO 6815 device (A2S spindle, 12rpm, 18°C), the process comprising the steps of: a. Providing an aqueous Rubisco solution comprising from about 7.5% to about 28% non-denatured Rubisco protein, b. Subjecting the aqueous non-denatured Rubisco solution to partial enzymatic hydrolysis in a static or gently agitated environment to provide a partially hydrolysed aqueous Rubisco solution,c. Heating the partially hydrolysed aqueous Rubisco solution to form a heat set gel, d. Subjecting the heat set gel to shear to provide the flowable heat set gel, wherein the heat set gel and / or the flowable heat set gel is subjected to a heating step to deactivate the enzymes and stop enzymatic hydrolysis.

[0012] In one aspect the invention provides a flowable heat set gel when prepared by the process of the invention.

[0013] In one aspect the invention provides a flowable heat set gel comprising from about 15% to about 28 % w / w of Rubisco protein, wherein the flowable heat set gel has a viscosity of up to about 18000cP (measured with an ATAGO VISCO 6815 device, A2S spindle, 12rpm, 18 °C).

[0014] In one aspect the invention provides a food product comprising the flowable heat set gel of the invention, the food product comprising between about 12.5% to about 25 % w / w of Rubisco protein, together with suitable additives selected from any one or more of:(i) sugars and carbohydrates,(ii) fats or oils,(iii) fruit juice, fruit concentrate or puree,(iv) flavours,(v) preservatives,(vi) sweetener,(vii) gums or stabilisers, and(viii) vitamins or other nutritional supplements, wherein the food product has a viscosity of up to about 18000cP (measured with an ATAGO VISCO 6815 device (A2S spindle, 12rpm, 18 °C)).

[0015] In one aspect the invention provides a food product comprising the flowable heat set gel of the invention, the food product comprising between about 12.5% to about 25 % w / w of Rubisco protein, together with suitable additives selected from any one or more of:(i) sugars and carbohydrates,(ii) fats or oils,(iii) fruit juice, fruit concentrate or puree,(iv) flavours,(v) preservatives,(vi) sweetener(vii) gums or stabilisers(viii) vitamins or other nutritional supplements, wherein the food product has a viscosity of between 30 and 1000 cP (measured with an ATAGO VISCO 6815 device, AIS spindle, 200 rpm, 18 °C).

[0016] The flowable heat set gel may have a viscosity between about 1000 and about 18000cP (measured with an ATAGO VISCO 6815 device (A2S spindle, 12rpm, 18 °C).

[0017] The flowable heat set gel may have a a. particle size distribution Dx(10) of from about 1.5 to about 3.5 pm, b. particle size distribution Dx(50) of from about 3.5 to about 15 pm, and / or c. particle size distribution Dx(90) of from about 15 to about 70pm.

[0018] The flowable heat set gel may be combined with additives to provide a formulated gel.

[0019] The flowable heat set gel or the formulated gel may be pasteurised.

[0020] Static or gently agitated in step b. may comprise mixing or agitation of a solution between about 0 rpm to about 30 rpm.

[0021] The enzymes used in the partial enzymatic hydrolysis of step b. may be selected from proteases or transferases which can be thermally deactivated.

[0022] The enzymes used in the partial enzymatic hydrolysis of step b. may be dosed at concentrations of 0.01-1.5% w / w Rubisco protein content.

[0023] The temperature in step b. may be between about 45 °C and about 65 °C.

[0024] The partial enzymatic hydrolysis step b. may occur for a time of between about 5 and about 20 minutes.

[0025] The temperature in step c. may be between about 40 °C to about 75 °C.

[0026] In step d. the shear may be performed for between about 1 to about 20 minutes.

[0027] The formulated gel may comprise between about 70 to 95% w / w of the flowable heat set gel.

[0028] The additives may be selected from any one or more of a. sugars and carbohydrates, b. fats or oils, c. fruit juice, fruit concentrate or puree, d. flavours, e. preservatives, f. sweetener, and g. gums or stabilisers.

[0029] The pasteurised, flowable heat set gel or the pasteurised, formulated gel may undergo a homogenisation step.

[0030] The flowable heat set gel may havea. particle size distribution Dx(10) of from about 1.5 to about 3.5 pm, b. particle size distribution Dx(50) of from about 3.5 to about 15 pm, and / or c. particle size distribution Dx(90) of from about 15 to about 70pm.

[0031] The flowable heat set gel may be pasteurised.

[0032] The food product may have a. particle size distribution Dx(10) of from about 1.0 to about 3.0 pm, b. particle size distribution Dx(50) of from about 3.0 to about 5.0 pm, and / or c. particle size distribution Dx(90) of from about 5.0 to about 15 pm.

[0033] The food product may comprise between about 70 to 95% w / w of the flowable heat set gel.

[0034] The food product may comprise any one or more of:(i) from about 2 to about 10% w / w sugar,(ii) from about 1 to about 6% w / w fats and oils,(iii)from about 1 to about 8% w / w fruit juice, fruit concentrate or puree,(iv)from about 0 to about 10% w / w flavours,(v) from about 0.1 to about 1.5% w / w preservatives,(vi)from about 0 to about 3% w / w sweetener, and(vii) from about 0.1 to about 1% w / w gums or stabilisers.

[0035] The food product may be pasteurised.

[0036] In one aspect, the present disclosure provides a flowable heat set gel comprising from about 15% to about 28 % w / w of Rubisco protein, wherein the heat set gel has a viscosity above about 1000 cP (measured with an ATAGO VISCO 6815 device (A2S spindle, 12rpm, 18 °C).

[0037] In one aspect, the present disclosure provides a flowable heat set gel comprising from about 15% to about 28 % w / w of Rubisco protein, wherein the heat set gel has a viscosity between about 4000 and about 18000cP (measured with an ATAGO VISCO 6815 device (A2S spindle, 12rpm, 18 °C).

[0038] In one aspect, the present disclosure provides a flowable heat set gel comprising from about 15% to about 28 % w / w of Rubisco protein, wherein the flowable heat set gel has a viscosity between about 4000 and about 18000cP (measured with an ATAGO VISCO 6815 device (A2S spindle, 12rpm, 18 °C), and wherein the flowable heat set gel is shelf stable for at least about 90 days at room temperature.

[0039] The present disclosure provides a flowable heat set gel comprising Rubisco protein, wherein the flowable heat set gel has a viscosity up to about 18000cP (measured with an ATAGO VISCO 6815 device (A2S spindle, 12rpm, 18 °C), and wherein the flowable heat set gel is shelf stable for at least about 90 days at room temperature

[0040] The present disclosure provides a flowable heat set gel comprising from about 15% to about 28 % w / w of Rubisco protein, wherein the flowable heat set gel is a pasteurised flowable heat set gel.

[0041] The present disclosure provides a flowable heat set gel comprising one or more of: a. from about 15% to about 28 % w / w of Rubisco protein; b. a viscosity up to about 18000cP (measured with an ATAGO VISCO 6815 device (A2S spindle, 12rpm, 18 °C); c. is shelf stable for at least about 90 days at room temperature; d. is a pasteurised flowable heat set gel.

[0042] The flowable heat set gel may be shelf stable for at least about 6-9 months at room temperature.

[0043] The concentration of Rubisco in the flowable heat set gel may be between about 15% and about 25%w / w, between about 15% and about 23%w / w, between about 20% and about 28%w / w, between about 20% and about 25%w / w, between about 23% and about 28%w / w, or between about 25% and 28%w / w.

[0044] The flowable heat set gel may have a viscosity between about 4000 and about 18000cP, or between about 4000 and about 10000 cP, or between about 4000 and about 7000cP (measured with an ATAGO VISCO 6815 device (A2S spindle, 12rpm, 18 °C).

[0045] The flowable heat set gel may have a viscosity of up to 18000cP (measured with an ATAGO VISCO 6815 device (A2S spindle, 12rpm, 18 °C). In various embodiments, the flowable heat set gel has a viscosity of between 1000 and 18000 cP, for example between 1000 and 160000 cP, between 1000 and 14000 cP, between 1000 and 12000 cP, or between 1000 and 10000 cP, between 1500 and 18000cP, between 1500 and 160000 cP, between 1500 and 14000 cP, between 1500 and 12000 cP, or between 1500 and 10000 cP.

[0046] The flowable heat set gel may have a particle size distribution Dx(10) of from about 1.5 to about 3.5 pm, for example from about 1.5 to about 3.0 pm, about 1.5 to about 2.5 pm

[0047] The flowable heat set gel may have a particle size distribution Dx(50) of from about 3.5 to about 15 pm, for example from about 4.0 to about 15 pm, about 3.5 to about 14 pm, about 4.0 to about 14 pm, about 3.5 to about 12 pm, about 4.0 to about 12 pm, about 3.5 to about 10 pm, or about 4.0 to about 10 pm.

[0048] The flowable heat set gel may have a particle size distribution Dx(90) of from about 15 to about 70pm, for example from about 18 to about 70 pm, about 20 to about 70 pm, about 15 to about 60 pm, about 18 to about 60 pm, about 20 to about 60 pm, about 15 to about 50 pm, about 18 to about 50 pm, or about 20 to about 50 pm.

[0049] The flowable heat set gel may be shelf stable for between about 90 days and 12 months, about 90 days and 180 days, about 90 days and 150 days, or between about 90 days and 120 days, at room temperature.

[0050] The flowable heat set gel may be pasteurised.

[0051] In one aspect, the present disclosure provides a food product (consumer product), the food product comprising between about 70-95% w / w of the flowable heat set gel of the disclosure, together with suitable additives. The food product may comprise between about 70 to 90% w / w of the flowable heat set gel, or about 70 to 85%, or about 70 to 80%, or about 75 to 95%, or about 75 to 90%, or about 80 to 95%, or about 80 to 90% w / w.

[0052] The suitable additives may be selected from any one or more of:(i) sugars and carbohydrates (for example about 2-10% w / w, or 5%),(ii) fats or oils (for example about 1-6% w / w, or 4.5%),(iii) fruit juice, fruit concentrate or puree (for example between about 1-8%, or about 5%),(iv) flavours (for example between about 0-10 %, or about 2%),(v) preservatives (for example between about 0.1-1.5%, or about 0.5%),(vi) sweetener (for example between about 0 -3, or about 1%),(vii) gums or stabilisers (for example between about 0.1-1%, or about 0.2%),(viii) vitamins or other nutritional supplements.

[0053] The additive may be from about 2 to about 10% w / w sugar, for example from about 3 to about 10% w / w, about 4 to about 10% w / w, about 5 to about 10% w / w, about 2 to about 9% w / w, about 3 to about 9% w / w, about 4 to about 9% w / w, about 5 to about 9% w / w, about 2 to about 8% w / w, about 3 to about 8% w / w, about 4 to about 8% w / w, or about 5 to about 8% w / w sugars and carbohydrates.

[0054] The additive may be from about 1 to about 6% w / w fats and oils, for example from about 1 to about 5% w / w, about 1 to about 4.5% w / w, about 2 to about 6% w / w, about 2 to about 5% w / w, about 2 to about 4.5% w / w, about 3 to about 6% w / w, about 3 to about 5% w / w, or about 3 to about 4.5% w / w fats and oils.

[0055] The additive may be from about 1 to about 8% w / w fruit juice, fruit concentrate or puree, for example from about 1 to about 7% w / w, about 1 to about 6% w / w, about 2 to about 8% w / w, about 2 to about 7% w / w, about 2 to about 6% w / w, about 3 to about 8% w / w, about 3 to about 7% w / w, or about 3 to about 6% w / w fruit juice, fruit concentrate or puree.

[0056] The additive may be from about 0 to about 10% w / w flavours, for example about 1 to about 10%, about 2 to about 10%, about 0 to about 9%, about 1 to about 9%, about 2 to about 9%, about 0 to about 8%, about 1 to about 8%, about 2 to about 8%, about 0 to about 7%, about 1 to about 7%, about 2 to about 7%, about 0 to about6%, about 1 to about 6%, about 2 to about 6%, about 0 to about 5%, about 1 to about 5%, or about 2 to about 5% flavours.

[0057] The additive may be from about 0.1 to about 1.5% w / w preservatives, for example about 0.1 to about 1.2% w / w, about 0.1 to about 1.0% w / w, about 0.1 to about 0.8% w / w, about 0.2 to about 1.5% w / w, about 0.2 to about 1.2% w / w, about 0.2 to about 1.0% w / w, about 0.2 to about 0.8% w / w, about 0.3 to about 1.5% w / w, about 0.3 to about 1.2% w / w, about 0.3 to about 1.0% w / w, about 0.3 to about 0.8% w / w, about 0.4 to about 1.5% w / w, about 0.4 to about 1.2% w / w, about 0.4 to about 1.0% w / w, or about 0.4 to about 0.8% w / w preservatives /

[0058] The additive may be from about 0 to about 3% w / w sweetener, for example from about 0.2 to about 3% w / w, about 0.5 to about 3% w / w, about 0 to about 2.5% w / w, about 0.2 to about 2.5% w / w, about 0.5 to about 2.5% w / w, about 0 to about 2% w / w, about 0.2 to about 2% w / w, or about 0.5 to about 2% w / w sweetener.

[0059] The additive may be from about 0.1 to about 1% w / w gums or stabilisers, for example about 0.1 to about 0.8% w / w, about 0.1 to about 0.6% w / w, about 0.1 to about 0.4% w / w, about 0.2 to about 1% w / w, about 0.2 to about 0.8% w / w, about 0.2 to about 0.6% w / w, or about 0.2 to about 0.4% w / w gums or stabilisers.

[0060] The food product may be a formulated consumer gel product, comprising from about 7% to about 25% w / w Rubisco protein, wherein the formulated consumer gel product may have a viscosity between about 1500 and about 4000 cP, or between about 2200 to about 3500 cP (measured with an ATAGO VISCO 6815 device (A2S spindle, 12rpm, 18°C).

[0061] The food product may have a viscosity of up to 18000cP (measured with an ATAGO VISCO 6815 device (A2S spindle, 12rpm, 18 °C).

[0062] The food product may have a viscosity of between 30 and 450 cP (measured with an ATAGO VISCO 6815 device, AIS spindle, 200 rpm, 18 °C), for example between 40 and 450 cP, between 50 and 450 cP, between 30 and 400 cP, between 40 and 400 cP, between 50 and 400 cP, between 30 and 350 cP, between 40 and 350 cP, between50 and 350 cP, between 30 and 300 cP, between 40 and 300 cP, between 50 and 300 cP, between 30 and 250 cP, between 40 and 250 cP, between 50 and 250 cP, between30 and 200 cP, between 40 and 200 cP, between 50 and 200 cP, between 30 and 150 cP, between 40 and 150 cP, between 50 cP to 150 cP.

[0063] The food product may have a particle size distribution Dx(10) of from about 1.0 to about 3.0 pm, for example from about 1.5 to about 3.0 pm, about 2.0 to about 3.0 pm, about 1.0 to about 2.5 pm, about 1.5 to about 2.5 pm, about 2.0 to about 2.5 pm.

[0064] The food product may have a particle size distribution Dx(50) of from about 3.0 to about 5.0 pm, for example from about 3.5 to about 5.0 pm, about 4.0 to about 5.0 pm, about 3.0 to about 4.5 pm, about 3.5 to about 4.5 pm, or about 4.0 to about 4.5 pm.

[0065] The food product may have a particle size distribution Dx(90) of from about 5.0 to about 15 pm, for example about 5.0 to about 14 pm, about 5.0 to about 12 pm, about 5.0 to about 12 pm, about 5.0 to about 10 pm, about 6.0 to about 15 pm, about 6.0 to about 14 pm, about 6.0 to about 12 pm, about 6.0 to about 12 pm, about 6.0 to about 10 pm, about 8.0 to about 14 pm, about 8.0 to about 12 pm, about 8.0 to about 12 pm, about 8.0 to about 10 pm, about 10 to about 14 pm, about 10 to about 12 pm, or about 10 to about 12 pm.

[0066] The food product may be packaged or contained in suitable container, such as a laminate pouch. The laminate pouch may provide an ultra-high barrier to oxygen and moisture.

[0067] The laminate pouch may contain a single serve amount of the food product.

[0068] The single serve amount may be between 60 g - 150 g food product.

[0069] The food product may comprise between about 7% and about 25% w / wRubisco protein.

[0070] The present disclosure provides a process for preparing a flowable heat set gel comprising from about 7.5% to about 28% w / w of Rubisco protein and having a viscosity up to about 18000cP (measured with an ATAGO VISCO 6815 device (A2S spindle, 12rpm, 18°C).

[0071] The present disclosure provides a process for preparing a flowable heat set gel comprising from about 7.5% to about 28% w / w of Rubisco protein and having a viscosity above about lOOOcP (measured with an ATAGO VISCO 6815 device (A2S spindle, 12rpm, 18°C).

[0072] The present disclosure provides a process for preparing a flowable heat set gel comprising from about 15% to about 28% w / w of Rubisco protein and having a viscosity up to about 18000cP (measured with an ATAGO VISCO 6815 device (A2S spindle, 12rpm, 18°C), the process comprising the steps of: a. Providing an aqueous Rubisco solution comprising non-denatured Rubisco protein; b. Subjecting the aqueous non-denatured Rubisco solution to partial enzymatic hydrolysis to provide a partially hydrolysed aqueous Rubisco solution, c. Heating the partially hydrolysed aqueous Rubisco solution to form a heat set gel, d. Subjecting the heat set gel to shear to provide the flowable heat set gel, wherein the heat set gel and / or the flowable heat set gel is subjected to a heating step to deactivate the enzymes and stop enzymatic hydrolysis.

[0073] The present disclosure provides a process for preparing a flowable heat set gel comprising from about 7.5% to about 28% w / w of Rubisco protein and having a viscosity up to about 18000cP (measured with an ATAGO VISCO 6815 device (A2S spindle, 12rpm, 18°C), the process comprising the steps of: a. Providing an aqueous Rubisco solution comprising from about 7.5% to about 28% non-denatured Rubisco protein; b. Subjecting the aqueous non-denatured Rubisco solution to partial enzymatic hydrolysis in a static or gently agitated environment to provide a partially hydrolysed aqueous Rubisco solution, c. Heating the partially hydrolysed aqueous Rubisco solution to form a heat set gel, d. Subjecting the heat set gel to shear to provide the flowable heat set gel, wherein the heat set gel and / or the flowable heat set gel is subjected to a heating step to deactivate the enzymes and stop enzymatic hydrolysis.

[0074] In step d. the shear may be performed between about 2,000 to about 20,000 rpm, and may be performed for between about 1 to about 20 minutes.

[0075] Step d. may be performed under a vacuum.

[0076] The vacuum may be between about 0.05 bar to about 0.2 bar, for example about 0.1 bar.

[0077] The vacuum may be between about 0.2 bar absolute to about 0.8 bar absolute, for example about 0.3 bar absolute to about 0.7 bar absolute, for example about 0.3 bar.

[0078] The temperature in step c. may be between about 40 °C to about 75 °C, for example about 60 °C to about 75 °, for example about 60 °C. This may be for from about 5 to about 20 minutes, from about 5 to about 15 minutes, or from about 5 to about 10 minutes.

[0079] Step c. may be performed at a pH value close to the native or natural pH of the partially hydrolysed concentrated protein solution. This may be in the pH range of from about pH 5 to about pH 9, for example between pH 5.5 and about 7.5, or at about pH 6.5 to about pH 6.7.

[0080] The temperature to stop enzymatic hydrolysis may be between about 60 °C to about 90 °C, for example about 75 °C to about 80 °, for example about 75 °C.

[0081] The heating step to stop enzymatic hydrolysis may be pasteurisation or ultra heat treatment (UHT).

[0082] Optionally, the process may further comprise the step of a secondary heat treatment, for example pasteurisation or ultra heat treatment (UHT), of the flowable heat set gel.

[0083] The flowable heat set gel may be cooled or frozen and stored.

[0084] The enzymes used in the partial enzymatic hydrolysis of step b. may be selected from proteases or transferases which can be thermally deactivated, and may be dosed at concentrations of 0.01-1.5% w / w Rubisco protein content.

[0085] The temperature in step b. may be between about 45 °C and about 65 °C, for example about 50°C to about 60°C. The enzyme / solution mixture may be gently agitated for about 5 to about 20 minutes.

[0086] The temperature in step b. may be between about 55 °C and about 65 °C, for example to about 60 °C, and the enzyme / solution mixture may be gently agitated for about 20 minutes to about 40 minutes, for example about 25 minutes.

[0087] The aqueous non-denatured Rubisco solution may comprise between about 15% and about 28% Rubisco, for example between about 20% and about 28% Rubisco.

[0088] The present disclosure provides a process for preparing a flowable heat set gel according to the disclosure from an aqueous Rubisco concentrate, the process comprising the steps of: a. Providing an aqueous non-denatured Rubisco concentrate comprising Rubisco protein in solution, b. Subjecting the aqueous Rubisco concentrate to partial enzymatic hydrolysis in a static or gently agitated environment to provide a partially hydrolysed aqueous Rubisco concentrate, c. Heating the partially hydrolysed aqueous Rubisco concentrate to form a heat set gel, d. Subjecting the heat set gel to shear to provide the flowable heat set gel, wherein the heat set gel and / or the flowable heat set gel is subjected to a heating step to deactivate the enzymes.

[0089] Step b. may refer to agitation between about 0 rpm (static) to about 30 rpm, or between about 5 rpm and about 20 rpm.

[0090] The present disclosure provides a process for preparing a flowable heat set gel according to the disclosure from an aqueous Rubisco concentrate, the process comprising the steps of: a. Providing an aqueous non-denatured Rubisco concentrate comprising from about 15% to about 28%, Rubisco protein w / w in solution,b. Subjecting the aqueous Rubisco concentrate to partial enzymatic hydrolysis in a static or gently agitated environment to provide a partially hydrolysed aqueous Rubisco concentrate, c. Heating the partially hydrolysed aqueous Rubisco concentrate to form a heat set gel, d. Subjecting the heat set gel to shear to provide the flowable heat set gel, the shear optionally being performed at about 2,000 to about 20,000 rpm, optionally for about 1 to about 20 minutes. wherein the heat set gel and / or the flowable heat set gel is subjected to a heating step to deactivate the enzymes.

[0091] Step b. may refer to agitation between about 0 rpm (static) to about 30 rpm, or between about 5 rpm and about 20 rpm.

[0092] The aqueous non-denatured Rubisco concentrate in step a. may comprise between about 20% to about 28%, Rubisco protein w / w in solution.

[0093] The temperature in step c. may be between about 40 °C to about 75 °C, for example about 60 °C to about 75 °, for example about 60 °C, and may be for a time from about 5 to about 20 minutes.

[0094] Step c. may be performed at a pH value close to the native or natural pH of the partially hydrolysed concentrated protein solution. This may be in the pH range of from about pH 5 to about pH 9, for example between pH 5.5 and about 7.5, for example at about pH 6.5 to about pH 6.7.

[0095] The temperature to stop enzymatic hydrolysis may be between about 60 °C to about 90 °C, for example about 75 °C to about 80 °, for example about 75 °C.

[0096] The heating step to stop enzymatic hydrolysis may be pasteurisation or ultra heat treatment (UHT).

[0097] Optionally, the process of the disclosure may further comprise the step of a secondary heat treatment, for example pasteurisation or ultra heat treatment (UHT), of the flowable heat set gel.

[0098] The flowable heat set gel from step e. is cooled or frozen and is stored.

[0099] The enzymes used in the partial enzymatic hydrolysis of step b. may be selected from proteases or transferases which can be thermally deactivated, and may be dosed at concentrations of 0.01-1.5% w / w total protein content.

[0100] The temperature in step b. may be between about 55 °C and about 65 °C, for example to about 60 °C, and the enzyme / solution mixture may be gently agitated for about 20 minutes to about 40 minutes, for example about 25 minutes.

[0101] The disclosure provides a process for preparing the food product of the disclosure, the process comprising the steps ofA. combining: a. between about 70%-95% of the flowable heat set gel of the disclosure or when prepared by the process of the disclosure, and b. any one or more of:(i) sugars and carbohydrates (for example about 2-10% w / w, or 5%),(ii) fats or oils (for example about 1-6% w / w, or 4.5%),(iii) fruit juice, fruit concentrate or puree (for example between about 1-8%, or about 5%),(iv) flavours (for example between about 0-10 %, or about 2%),(v) preservatives (for example between about 0.1-1.5%, or about 0.5%),(vi) sweetener (for example between about 0 -3, or about 1%),(vii) gums or stabilisers (for example between about 0.1-1%, or about 0.2%),(viii) vitamins or other nutritional supplements.B. sealing the combination of a. and b. above in a suitable container, for example a laminate pouch, andC. performing a secondary heat treatment, for example a UHT treatment.

[0102] The disclosure provides for a continuous process for preparing a packaged flowable heat set Rubisco gel food product, the process comprising the steps of: a. Providing an aqueous Rubisco concentrate comprising from about 15% or 20% to about 28% non-denatured Rubisco protein w / w in solution, b. Subjecting the aqueous Rubisco concentrate to partial enzymatic hydrolysis in a static or gently agitated environment to provide a partially hydrolysed aqueous Rubisco concentrate, c. Subjecting the partially hydrolysed aqueous Rubisco concentrate to a temperature sufficient to provide a Rubisco heat set gel, d. Subjecting the heat set gel to shear to provide a flowable Rubisco heat set gel having a viscosity between about 4000 and about 18000cP (measured with an ATAGO VISCO 6815 device (A2S spindle, 12rpm, 18 °C), e. Optionally, adding suitable additives to the flowable heat set Rubisco gel, f. Performing a heat treatment suitable to deactivate enzymatic activity (for example pasteurisation or UHT), and g. Packaging the flowable Rubisco heat set gel to provide packaged flowable heat set Rubisco gel food product.

[0103] Step b. may refer to agitation between about 0 rpm (static) to about 30 rpm, or between about 5 rpm and about 20 rpm.

[0104] The temperature in step c. may be between about 58 °C to about 65 °C and may be for a time from about 5 to about 20 minutes.

[0105] Step c. may be performed at a pH value close to the native or natural pH of the partially hydrolysed concentrated protein solution. This may be in the pH range of from about pH 5 to about pH 9, for example between pH 5.5 and about 7.5, or for example at about pH 6.5 to about pH 6.7.

[0106] Steps b. to f. may be performed in a single multi-purpose vessel.

[0107] One or more of steps d. to f. may be performed under vacuum of about 0.05 bar to about 0.2 bar, for example about 0.1 bar.

[0108] One or more of steps d. to f. may be performed under vacuum of about 0.3 bar to about 0.7 bar, for example about 0.3 bar.

[0109] Step d. may be performed under a vacuum.

[0110] Step g. may include hot fill packaging.

[0111] The process may further include the step of cooling the packaged product, for example freezing the packaged product.

[0112] The flowable Rubisco gel food product may have a viscosity between 2000- 4000 cP, and for example between 2200-3500 cP. The product may be a beverage.

[0113] The suitable additives may be selected from any one or more of:(i) sugars and carbohydrates (for example about 2-10% w / w, or 5%),(ii) fats or oils (for example about 1-6% w / w, or 4.5%),(iii) fruit juice, fruit concentrate or puree (for example between about 1-8%, or about 5%),(iv) flavours (for example between about 0-10 %, or about 2%),(v) preservatives (for example between about 0.1-1.5%, or about 0.5%),(vi) sweetener (for example between about 0 -3, or about 1%),(vii) gums or stabilisers (for example between about 0.1-1%, or about 0.2%), and / or(viii) vitamins or other nutritional supplements.

[0114] The enzymes used in the partial enzymatic hydrolysis of step b. may be selected from proteases or transferases which can be thermally deactivated, and may be dosed at concentrations of 0.01-1.5% w / w total protein content.

[0115] The temperature in step b. may be between about 55 °C and about 65 °C, for example to about 60 °C, and the enzyme / solution mixture may be gently agitated for about 20 minutes to about 40 minutes, for example about 25 minutes.

[0116] The disclosure provides a continuous process for preparing a flowable heat set Rubisco gel, the process comprising the steps of: a. Providing an aqueous Rubisco concentrate comprising from about 15% or 20% to about 28% non-denatured Rubisco protein w / w in solution, b. Subjecting the aqueous Rubisco concentrate to partial enzymatic hydrolysis in a static or gently agitated environment to provide a partially hydrolysed aqueous Rubisco concentrate, c. Subjecting the partially hydrolysed aqueous Rubisco concentrate to a temperature sufficient to provide a Rubisco heat set gel, d. Subjecting the Rubisco heat set gel to shear to provide a flowable Rubisco heat set gel having a viscosity between about 4000 and about 18000cP (measured with an ATAGO VISCO 6815 device (A2S spindle, 12rpm, 18 °C), and e. Subjecting the flowable heat set gel to a heating step to deactivate the enzymes wherein the heating is a pasteurisation or UHT step. wherein steps b. to d. are performed in a single multi-purpose vessel.

[0117] Step b. may refer to agitation between about 0 rpm (static) to about 30 rpm, or between about 5 rpm and about 20 rpm.

[0118] The process may further include a step f. packaging the flowable Rubisco heat set gel in shelf stable format.

[0119] Step f. may include hot fill packaging.

[0120] The process may further include a step of cooling the packaged product, for example, freezing the packaged product.

[0121] Suitable additives may be added to the flowable Rubisco heat set gel and may be added before performing the heat treatment in step e.

[0122] The suitable additives may be selected from any one or more of:(i) sugars and carbohydrates (for example about 2-10% w / w, or 5%),(ii) fats or oils (for example about 1-6% w / w, or 4.5%),(iii) fruit juice, fruit concentrate or puree (for example between about 1-8%, or about 5%),(iv) flavours (for example between about 0-10 %, or about 2%),(v) preservatives (for example between about 0.1-1.5%, or about 0.5%),(vi) sweetener (for example between about 0 -3, or about 1%),(vii) gums or stabilisers (for example between about 0.1-1%, or about 0.2%), and / or(viii) vitamins or other nutritional supplements.

[0123] The shear in step d. may be performed between about 2,000 to about 20,000 rpm, and may be for about 1 to about 20 minutes.

[0124] The temperature in step c. may be between about 58 °C to about 65 °C.

[0125] One or more of steps d. to f. may be performed under vacuum of about 0.05 bar to about 0.2 bar, for example about 0.1 bar.

[0126] One or more of steps d. to f. may be performed under vacuum of about 0.3 bar to about 0.7 bar, for example about 0.3 bar.

[0127] Step d. may be performed under vacuum.

[0128] The enzymes used in the partial enzymatic hydrolysis of step b. may be selected from proteases or transferases which can be thermally deactivated, dosed at concentrations of 0.01-1.5% w / w total protein content.

[0129] The temperature in step b. may be between about 55 °C and about 65 °C, for example to about 60 °C, and the enzyme / solution mixture may be gently agitated for about 20 minutes to about 40 minutes, for example about 25 minutes.

[0130] The disclosure provides a flowable heat set Rubisco gel food product produced by process of the disclosure or a flowable Rubisco heat set gel produced by the process of the disclosure.

[0131] The flowable heat set Rubisco gel food product produced by process of the disclosure may comprise between about 7% and about 25% w / w Rubisco protein.

[0132] The flowable Rubisco gel food product may have a viscosity between 2000- 4000 cP, and for example between 2200-3500 cP, and the product may be a drinking product.

[0133] The above embodiments and preferences may relate alone or in any combination of any two or more to any of the above aspects.

[0134] The disclosure may also be said broadly to consist in the parts, elements and features referred to or indicated in the specification of the application, individually or collectively, in any or all combinations of two or more of said parts, elements or features, and where specific integers are mentioned herein which have known equivalents in the art to which the disclosure relates, such known equivalents are deemed to be incorporated herein as if individually set forth.

[0135] It is intended that reference to a range of numbers disclosed herein (for example, 1 to 10) also incorporates reference to all rational numbers within that range (for example, 1, 1.1, 2, 3, 3.9, 4, 5, 6, 6.5, 7, 8, 9, and 10) and also any range of rational numbers within that range (for example, 2 to 8, 1.5 to 5.5, and 3.1 to 4.7) and, therefore, all sub-ranges of all ranges expressly disclosed herein are hereby expressly disclosed. These are only examples of what is specifically intended and all possible combinations of numerical values between the lowest value and the highest value enumerated are to be considered expressly stated in this application in a similar manner.

[0136] In this specification where reference has been made to patent specifications, other external documents, or other sources of information, this is generally for the purpose of providing a context for discussing the features of the disclosure. Unless specifically stated otherwise, reference to such external documents is not to be construed as an admission that such documents, or such sources of information, in any jurisdiction, are prior art, or form part of the common general knowledge in the art.

[0137] To those skilled in the art to which the disclosure relates, many changes in construction and widely differing embodiments and applications of the disclosure will suggest themselves without departing from the scope of the disclosure as defined in the appended claims. The disclosures and the descriptions herein are purely illustrative and are not intended to be in any sense limiting.

[0138] Although the present disclosure is broadly as defined above, those persons skilled in the art will appreciate that the disclosure is not limited thereto and that the disclosure also includes embodiments of which the following description gives examples.Brief description of the figures

[0139] The present disclosure will be described with reference to the accompanying figures, in which:

[0140] Figure 1: Process for the preparation of a flowable heat set gel comprising of a high concentration of Rubisco protein with a low viscosity, as prepared under a nonagitated (static heat set process).

[0141] Figure 2: Process for the preparation of a flowable heat set gel comprising of a high concentration of Rubisco protein with a low viscosity, performed in a concurrent process in a single vessel (concurrent process).

[0142] Figure 3: Concurrent process to prepare formulated flowable high-protein heat set Rubisco gel with packaging.

[0143] Figure 4: Non-concurrent process performed in thermally stable polyethylene bags. The process can be stopped at conclusion of Part A to provide a nonformulated flowable gel.

[0144] Figure 5: Rubisco heat set Gel.

[0145] Figure 6: Granular precipitated protein aggregate formed as a result of excess stirring during gel heat set formation.

[0146] Figure 7: Flowable heat set gel.

[0147] Figure 8: Relationship between viscosity and enzyme concentration for proteolysis indicates a decrease in viscosity with increasing enzyme addition / proteolysis which enables calculation of an enzyme dosing rate to achieve a target viscosity.

[0148] Figure 9: Gelling properties. A change in the viscoelasticity of the heat set gel can be achieved via partial hydrolysis of the concentrated protein feedstock to give a softer gel.

[0149] Figure 10: Aqueous Rubisco (15% TS) can be thermally gelled which causes an increase in the gel elastic component of viscoelasticity (G' storage modulus) as theset gel is formed. Conversely Gelled Rubisco which has been previously gelled then subjected to high shear to form a flowable gel is unable to reform a set gel form when reheated.

[0150] Figure 11 : Particle size distribution of high content Rubisco solution, Partially proteolyzed high protein Rubisco solution, Gelled Rubisco protein mixed by hand mixing (Gelled -HM), intermittent gentle stirring (Gelled -IM) and gentle scraped surface (Gelled -SS).

[0151] Figure 12: Gelling properties of alternative protein materials, showing formation of firm gel for Rubisco, a soft gel for pea, granular particulate form for soy, and whey did not form a gel (not shown). All samples at 15% protein (w / w).

[0152] Figure 13: Formulated heat sheared Rubisco gel of the disclosure provides favourable consumer sensory properties including preferred thickness at 2200-3500 cP viscosity and a non-grainy / non-gritty texture smoothness attribute.

[0153] Figure 14: Formulation of various consumer gel products.

[0154] Figure 15: Process for the preparation of high protein flowable heat set gels and homogenised, formulated high protein content consumer gels.

[0155] Figure 16: Particle size changes from control for homogenised samples where 320-0 is the sheared pasteurised gel with 0% fat content, 320-0-HPH is homogenised pasteurised gel with 0 % fat, 320-1-HPH is homogenised pasteurised gel with 1 % fat, and 320-3-HPH is homogenised pasteurised gel with 3 % fat.

[0156] Figure 17: Relationship between shear and viscosity as measured on the ATAGO VISCO 6815 comparing AIS and A2S spindle for homogenised consumer gel.Definitions

[0157] The term "comprising" as used in this specification and claims means "consisting at least in part of". When interpreting each statement in this specification and claims that includes the term "comprising", features other than that or those prefaced by the term may also be present. Related terms such as "comprise", "comprised" and "comprises" are to be interpreted in the same manner.

[0158] As used herein the term "and / or" means "and" or "or", or both.

[0159] As used herein "(s)" following a noun means the plural and / or singular forms of the noun.

[0160] The term "about" as used herein generally refers to a range of numerical values (e.g. ± 5 to 10% of the recited value) that those skilled in the art would consider equivalent to the recited value. Ranges can be expressed herein as from "about" one particular value, and / or to "about" another particular value. When such a range is expressed, the range is inclusive of the recited values.

[0161] The general chemical and biological terms used, for example, in the formulae herein have their usual meanings.

[0162] The term "total solids" (TS) as used in this specification and claims means the solids remaining after evaporation of a sample to constant weight at 105°C. In a food matrix the total solids content is typically composed of proteins, carbohydrates, fats, secondary metabolites, and minerals (ash).

[0163] The term "protein content" as used herein refers to the weight of protein or polypeptides within a sample which may be expressed as a ratio, percentage, or mass. The protein content can be determined by Kjeldahl, DUMAS or equivalent analytical methods (AOAC 981.10 mod.) for the determination of protein or nitrogen content. Alternatively, protein content can be measured using spectroscopic methods such as near-infrared (NIR) spectroscopy calibrated to protein content values in a real time or near real-time manner.

[0164] The term "Rubisco" or "Rubisco protein" or like terms as used herein refers to the protein ribulose-l,5-bisphosphate carboxylase / oxygenase as extracted from photosynthetic organelles of plant, algae, and microorganisms, using commercial feasible methods known to those in the art. As will be appreciated by someone skilled in the art, an extract from a plant will contain a range of proteins and the use of the word Rubisco does not imply the protein is solely Rubisco protein. The protein extract from plant leaves may also be variously referred to as leaf protein concentrate (LPC) and leaf protein isolate (LPI). In the present disclosure, the Rubisco protein prior to pasteurisation is non-denatured and has a purity of least about 80% to about 90% of the total protein, for example from about 84 to about 86%.

[0165] The term "shelf stable" as used herein refers to a product which retains favourable microbiological, functional and textural attributes stored for an indicated period at an indicated temperature.

[0166] The term "gel" as used herein refers to a colloidal or polymer network created by a cross-linking network within a liquid. The term gel includes "microgels", which refers to a gel comprising colloidal particles in the micron particle range size. Microgels are usually generated by micro-particulation such as high-pressure homogeniser, but also could refer to those generated by high shear mixing or emulsion formulation.

[0167] The term "static or gentle" with reference to "agitation" as used herein refers to mixing or agitation of a solution between about 0 rpm (static) to about 30 rpm, or between about 5 rpm and about 20 rpm, for a time that is sufficient to achieve adequate mixing and dispersion but which does not lead to foaming or disruption of gel cross-linking during thermal gel set up. As will be apparent to a skilled person, the time frames for this mixing or agitation will vary depending on the material and process conditions used in the process with static (Orpm) taking longer than 30rpm agitation to achieve adequate mixing and dispersion of the Rubisco concentrate and the enzyme(s) to achieve partial hydrolysis.

[0168] The term "continuous" or "concurrent" as used herein refers to a process or process step in which more than one technical, process, or conceptual aspects are occurring at the same time, with the benefit of reducing the time required if both aspects were to be performed sequentially.

[0169] The term "pseudoplastic behaviour" as used herein refers to the behaviour of a fluid characterised by experiencing a decrease in viscosity with increasing shear rates (shear-thinning fluid).

[0170] The term "non-denatured" as used herein refers to protein or protein solutions which contain natively folded secondary and tertiary structure. Non-denatured protein (Rubisco) has not been subjected to denaturation conditions and the inventors have found possesses the ability to form a gel structure.

[0171] The term "vacuum" as used herein refers to a partial vacuum applied to a system via a pump capable of generating a vacuum. The partial vacuum may be expressed as value of bar absolute with respect to ambient atmospheric pressure, or equivalently a vacuum bar gauge value expressed as a partial vacuum value below ambient atmosphere, or equivalently, as a negative partial vacuum bar gauge value below ambient atmosphere.Detailed description of the disclosure

[0172] The disclosure in general terms relates to a flowable heat set gel comprising a high concentration of Rubisco protein (about 15% w / w to about 28% w / w) and which has low viscosity suitable for use in a flowable food product, and to processes for producing such a flowable heat set gel. The disclosure also relates to food products including the flowable heat set gel and to processes for producing those products.

[0173] A known use of Rubisco protein in food products has been related to its ability to form strong gels at low concentrations. However, there is also a demand for gels containing high concentrations of Rubisco protein that have a low viscosity.

[0174] The present disclosure provides a flowable heat set gel having a Rubisco protein concentration between about 15 and 28% w / w and a viscosity up to about 18000 cP, for example between about 4000 and about 18000 cP, or for example between about 1000 and about 18000 cP (measured with an ATAGO VISCO 6815 device (A2S spindle, 12rpm, 18 °C)). The flowable heat set gel can be able to be heat treated (a secondary heating step such as pasteurisation or UHT for example). The flowable heat set gel can be shelf stable for at least about 90 days at room temperature. UHT treatment can be used to provide shelf stability for at least about 6-9 months for the flowable heat set gel of the disclosure with desirable characteristics retained.

[0175] The flowable heat set gel of the present disclosure can be used to provide food products, such as high protein gels for use in the sports nutrition or dietary supplementation, or for use in the wider food industry where protein inclusion without high gelation (and hence flowability) is needed or preferred.

[0176] Gels with lower viscosity (e.g. 4000cP to 7000cP, or 1000 to 18000 cP) can be used directly as a flowable food product with no, or limited, additional components. Higher viscosity gels (e.g. 15,000 to 18,000 cP) can be diluted and formulated as food products (e.g. mousse) or stored / transported for later use. Such options are by way of example only and are not intended to be limiting.

[0177] The desired characteristics of a gel product that could be acceptable to consumers in the sports nutrition industry for example could include any one or more of suitable viscosity, mouthfeel, taste, flavour, texture, aroma and appearance. Such characteristics can be desirable in a flowable high protein consumable product for example.

[0178] The processes of the disclosure used to produce the flowable heat set gels of the disclosure can be used to produce flowable heat set gels containing lower levels of Rubisco content (e.g. under 15% to about 7.5% w / w) as well if desired.High protein heat set gel

[0179] There is consumer desire for high protein content gels with protein derived from non-animal sources. Heat set gels prepared at high concentration of Rubisco (> 10%) are known to be firm cross-linked set gel with a non-flowing nature, are nonelastic, non-pliable and have a firm to brittle texture. Such firm gels, generated through thermal treatment of Rubisco protein, are suitable for some food formats at low inclusion rates but are not suitable for use in high protein content flowable consumer gel products as they do not have desirable viscosity, mouthfeel or texture.

[0180] Other commonly available plant proteins do not form firm gels. As seen in Example 6, solutions of Pea protein isolate and Soy protein isolate dispersed into water at 5 to 25% protein content and heated to about 90 °C did not create firm, self-standing gels like those generated by Rubisco solutions (Figure 12). The same is seen for Whey protein isolate. Pea protein formed a soft gel, Soy protein concentrate formed a large granular particulates and Whey protein isolate remained in solution and did not form a gel. Rubisco behaves differently to other proteins, both plant (Pea, Soy) and animal (whey), and has its own characteristics.

[0181] Heating can be used in food preparation as a necessary step for phytosanitary control, such as pasteurisation, UHT and other forms of thermal treatment. This heating can cause formation of undesirable firmness in gels.

[0182] The disclosure solves, or at least ameliorates, the challenge of providing a flowable consumer gel product with a high inclusion rate of Rubisco by decreasing the inherent gel strength of the Rubisco heat set gel. The disclosure describes the partial hydrolysis of Rubisco via enzymatic hydrolysis (for example via protease or transferase enzymes) to give a concentrated Rubisco protein solution, which generates a heat set Rubisco gel which can be subjected to shear to provide a flowable heat set gel with favourable properties, including a flowable viscosity, good mouthfeel and non-grainy or gritty texture, (see Figure 13). The enzymes used to perform the partial hydrolysis are preferably food safe proteases or transferases, dosed at concentrations of 0.01-1% w / w on a protein basis, which can be thermally deactivated.

[0183] In the disclosure, a solution of non-denatured Rubisco (preferably high Rubisco protein content between about 15% and about 28% w / w) is partially digested / hydrolysed with preferably proteolytic enzymes in the presence of mild heating at about 40 to about 65°C; for example about 45 to about 60°C, about 40 to 50°C, or about 50 to 65 °C, for example for a time between about 5 and about 20 minutes or between about 5 and about 15 minutes or between about 5 and about 10 minutes. The partially hydrolysed Rubisco protein solution is then thermally set at above 60 °C, such as 60-70°C or 70-80°C or 80-90°C, to provide a heat set gel that is then subject to shear to produce the flowable heat set gel. Partial enzyme hydrolysis of Rubisco and heating provides a lower strength, firm, gel. The moderation of gel strength is shown in Figure 9, which shows a decrease in gel strength as recorded with rheometric measurements of G' (storage modulus) in the presence partial proteolytic hydrolysis when compared to the non-hydrolysed control, Table 1. High shear mixing and disruption of this partially hydrolysed firm, but lower strength gel provides a flowable heat set gel of the disclosure with reduced viscosity.

[0184] The inventors have found that formation of the heat set gel from the partially hydrolysed Rubisco protein solution needs to be performed under a static or gentle agitation environment to avoid disruption of the protein cross-linking occurring in setting up the heat set gel. Excessive mixing / agitation results in disruption of the gel matrix formation and precipitation of protein having a granular texture. See Figure 5 for a heat set gel formed under static mixing which provides a flowable heat set gel as seen in Figure 7 after high shear mixing, compared to Figure 6 where the heat set gel is formed using excessive mixing / agitation (over about 40 rpm) which gives a granular precipitated protein aggregate after high shear mixing. The static option can be less preferred in a commercial process due to the longer time frames that result to prepare the heat set gel.

[0185] The heat set gel prior to shear to form the flowable heat set gel can be cooled and / or frozen and then stored / transported as desired for later shear treatment to form the flowable heat set gel.Flowable high protein heat set gel

[0186] The heat set Rubisco gel is sheared to produce a flowable Rubisco gel as a homogenous, free-flowing, gel (i.e. a flowable heat set gel) that can contain high levels of Rubisco protein (about 15% to 28% w / w) with a viscosity of up to about 18000cP, for example about 4000 cP to about 18000 cP, or about 1000 and about 18000 cP which exhibits pseudoplastic behaviour. The higher viscosity gels will be less free flowing thanthe lower viscosity gels but are still flowable and therefore can provide one or more advantages to the user.

[0187] The heat set Rubisco gel can be sheared to produce the flowable heat set gel using a high shear mixer, rotor-stator mixer, high pressure homogeniser, ultrasonic homogeniser, microfluidizer, or colloid mill, for example, to disrupt the heat set gel structure to provide a flowable high protein sheared gel (flowable heat set gel). High shear mixing may be performed at about 2,000 to about 20,000 rpm at about 60 °C to about 75 °C, for example 60 °C, to disrupt the heat set gel, with mixing performed for a time sufficient to disrupt the heat set gel into a homogeneous gel (which may be for between about 1 to about 20 minutes, or between about 3 and about 10 minutes), to form the flowable heat set gel of the disclosure.

[0188] The flowable heat set gel may be pasteurised.Process to produce high protein gel products

[0189] The flowable heat set gel formed from the shear step can be formulated into a consumer product by any means that would be known to a skilled person. As used herein, the terms "consumer product", "consumer gel format", "formulated product", "formulated gel product", and "food product" are used interchangeably.

[0190] For example, a product could include one or more of sugars (2-10% w / w, for example 5%), fats / oil (1-6% w / w, for example 4.5%), fruit or vegetable juice, concentrate or puree (1-8%, for example about 5%), flavours (0-10 %, for example about 2%), preservatives (0.1-1.5%, for example about 0.5%), sweetener (for example 0 -3%, for example about 1%), and / or gums / stabilisers (0.1-1%, for example about 0.2%). The flowable high protein heat set gel of the disclosure plus desired additional components can, for example, be blended / mixed for about 5 mins to a homogeneous mixture and then packaged into consumer formats.

[0191] By way of example, a consumer gel format includes individual packages or containers, such as sachets or pouches, into which the flowable high protein heat set gel is packaged. The individual sachets or pouches can provide any desired amount of product such as 50-150 g or 60g - 120g serves for example. The sachets or pouches are preferably ultra-high or high barrier laminate pouches. Such packaging desirably provides a high barrier to oxygen and moisture and is made of material suitable for secondary thermal treatment to facilitate pasteurisation or sterilisation, (e.g. UHT) such as retort pouches, three layer laminated polyester / aluminium foil / polyethylene (LDPE);or metalised polyethylene terephthalate (PET) / LPDE packages, such as RE: MONO laminate pouches (Wastemade™) pouches or equivalent packaging as would be known to a skilled person. As would be clearly apparent to a skilled person, a variety of packaging options could be used to deliver a consumer product including the flowable heat set gel of the present disclosure.

[0192] The high protein content flowable heat set gel of the disclosure having a high protein content and a low viscosity allows its use in the manufacture of multiple foodstuffs, for example but not limited to: athlete high protein gels, athlete food supplements, geriatric dietary supplements, medical food supplement for patients with swallowing difficulties, medical food supplement for patients with high nutrient dietary requirements, medical food supplement for patients with a gastric band, medical food supplement for patients in need of calorific reduction, or dietary supplements for patients with allergies.

[0193] The high protein content flowable heat set gel of the disclosure can also be used to make composite or combined products, for example: addition of vitamins to form a nutritional supplement, nutritional gummies, or capsules; addition of the gel as a soft filling within a product such as a soft filled bar; a gel carrier for condiments, flavouring and sauces; or animal free gelatine replacement, or gel of plant origin; addition to smoothies to increase protein content.

[0194] The Rubisco content of products produced using the flowable heat set gel of the disclosure can be varied as desired by the manufacturer, but may be between about 7% and about 25% Rubisco w / w. The top range of Rubisco in a formulated product is restricted by the top range of the Rubisco content of the flowable heat set gel of the disclosure (about 28%w / w) however the lower range of Rubisco in the products produced can be adjusted as desired. If the flowable heat set gel of the disclosure is the product that is packaged (without formulation additives) then the Rubisco content can be as high as the top range of the flowable heat set gel of the disclosure.

[0195] The Rubisco flowable heat set gel of the disclosure can be included at lower inclusion rates, such as between 7% and 20% w / w with other proteins, preferably plant based such as for example pea or soy protein isolates, where the Rubisco protein provides the flowable gel attribute and the additional proteins provide fortification of the protein content. Such fortification can also be provided by dehydrated Rubisco powder if desired.

[0196] The flowable Rubisco heat set gel of the disclosure may be used as, or as an ingredient in, a pouched sports gel product.

[0197] The sports gel may be formulated by addition of at least one of the following additives to the high protein gel solution: fruit or vegetable juice, concentrate, or puree, sugars, fats, oils, flavours, preservatives, sweeteners, gums, and stabilizers. Such additives can be combined with the high protein flowable heat set gel, and the resulting mixture blended and / or stirred for a suitable time (usually about 5 to about 10 mins) to provide a homogeneous mixture. The viscosity of such a product may be between about 2000 and about 4000 cP, and may be between about 2200 and about 3500 cP, when measured with an ATAGO VISCO 6815 device (A2S spindle, 12rpm, 18 °C).

[0198] The sports gel product is packaged according to any method known to a person skilled in the art. It is also possible to package the flowable heat set gel without additives.

[0199] The packaged sports gel can be produced to a serving size of about 50g - 150 g, or about 60g - 120g, served in packages made of a material suitable for shelf stable storage or for secondary heat treatment (e.g. UHT) to provide an extended shelf life at room temperature. The laminated sachets or pouches referred to previously are preferred option for such packaging.

[0200] A secondary heat treatment may be used to provide a consumer food product with an extended shelf stability without detrimentally affecting the desired characteristics of the flowable heat set gel of the disclosure. The secondary heat treatment can be done by any means known to a skilled person such as by submersing the sealed pouches or sachets (if used) in a water bath at 80-90°C for 5-20 min. The result is a pasteurised product which is stable under refrigerated conditions for up to about 28 days. Alternatively, the sealed pouches are heat treated using known UHT techniques such as, for example, in a retort at about 117-121 °C for about 5-10 min to generate a commercially sterile consumer gel product that is stable in ambient conditions prior to opening for up to at least about 90 days to preferably about 180 days. Alternatively, the product can be subjected to a heat treatment prior to packaging.

[0201] To summarise, a concentrated Rubisco protein solution (15-28% w / w) is mixed with enzyme e.g. Protana, Flavourzyme and Formea (Novozymes), or BS Cone, AKL, PAP, Lypaine (Soufflet Biotechnologies) dosed at between about 0.01%-2% (for example 0.05%-1.5%) with respect to the concentration of the Rubisco protein in theconcentrated protein solution. The concentrated protein solution containing enzyme is sealed in thermally stable polyethylene bags and heated at 45-50° for about 10-90 min at native pH (~6.8) to achieve a partial hydrolysis. The partially hydrolysed protein solution is heated to between about 63-90°C, for example 80-90°C for about 10 min in a static or gently agitated environment. The heat set gel produced is disrupted via shear (by e.g. homogenisation, blender, or high shear mixer) to produce a flowable heat set gel (Figure 4).

[0202] To achieve a consumer formulated gel product, for example at about 15.3% Rubisco protein per serve, the flowable heat set gel of the disclosure (70-95% w / w approximate 15-28% protein content) is combined with one or more of fats / oil (1-6% w / w), sugar (2-10% w / w) fruit juice, fruit concentrate or puree (1-8%), flavours (0- 10%), preservatives (0.1-1.5%), sweetener (0-3%), gums (0.1-1%) then blended / mixed for about 5 mins to a homogeneous mixture. This results in a flowable Rubisco gel product suitable for packing into gel pack formats. The flowable high protein gel product according to the disclosure can be packaged into hermetically sealed thermally stable packaging and retorted / ultra-heat treated (for example about 117°C for 10 mins, or about 121°C for about 5-7 min) to generate a commercially sterile consumer gel product that is stable at ambient conditions prior to opening. Alternatively, the packaged gels can be heat treated at 80-90 °C to generate a refrigerator stable consumer gel product.

[0203] As shown in Figure 4, enzyme is mixed with the non-denatured Rubisco solution starting material (Feed) in thermally stable bags and placed in a water bath. The temperature in the bags is increased to 50°C and held for 30 minutes (Hydrolysis). The temperature is then increased from 50°C to 85°C over a period of about 40 minutes and the temperature is held at 85°C for 15 minutes. In order to achieve 85°C in the centre of the bags, the water bath is set at 95-100°C. As will be apparent there will be a temperature difference between the centre and the outside of the bags, which is a disadvantage of this non-concurrent (or sequential) process using a static (no agitation) gel formation option. This step produces the heat set gel and is a combined heat set, enzyme deactivation and temperature treatment step.

[0204] The heat set gel is then subjected to high shear mixing at 75°C for 15 minutes to produce a flowable heat set gel. The flowable heat set gel is then frozen for storage and / transport.

[0205] To form a formulated product from the frozen, previously flowable, heat set gel, the frozen heat set gel is thawed, desired additives included for the formulated product, and the product is packaged in pouches or as otherwise desired. The packaged product is heat treated at 75°C for 5 minutes and the product is again frozen for storage and / or transport.

[0206] Formulated gel product that provides 20% protein per serve could comprise sheared gel (70-90% w / w) combined with fats / oil (1-6% w / w), fruit juice, fruit concentrate or puree (1-8%), sugars and carbohydrates (2-10% w / w), flavours (0- 10%), preservatives (0.1-1.5%), sweetener (0.0-3%), gums (0.1-1%). The mixture may be blended to a homogeneous mixture for time 5-10mins at pH 6-6.5 and temperature 60-65°C.

[0207] The flowable heat set gel or the formulated gel product can undergo pasteurisation. Pasteurization may occur by heating to 70-75°C with a heated jacketed (85-75°C hot water) for 5-15 mins.

[0208] Pasteurisation can occur before or after formulation. Pasteurisation can occur via any means known to a skilled worker, provided the pasteurisation step reduces / removes the microbial load and optionally the enzyme is deactivated (if not deactivated by other means prior). For example, the pasteurisation can occur for 68°C for 15 seconds.

[0209] The formulated gel may undergo homogenisation at elevated temperature and / or pressure. The temperature may be above 65 °C. The pressure may be above 100 bar.

[0210] Following pasteurisation, the hot pasteurised formulated gel (60-75°C) may be homogenised at high pressure (100-300 bar) to form a micro-particulated emulsion. Homogenisation stabilises the protein-fat mixture for improved quality and shelf-life in a frozen format. The homogenised gel emulsion may be rapidly cooled to via a chilled plate heat exchanger and further chilled in a holding tank to about 4°C prior to packaging.

[0211] High pressure homogenisation uses a pump to push liquid materials thought a narrow valve, forcing the material against an impact ring generating high pressure and multiple sheer planes. This leads to the disruption and reorganisation of large particles / oil bodies into smaller uniform particles. In food applications this is can be used to manipulate particle size and produce nanoemulsions, resulting in changes to mouthfeel and flavour characteristics.

[0212] The pasteurised formulated gel can be transferred to a homogenisation unit to undergo a multi-stage homogenisation process with the first stage set to 150 bar and the second stage homogenisation set to 50 bar, to achieve reduced particle size distributions with Dx(90) less than 10 pm, and Dx(50) less than 5 pm, which have improved mouthfeel and flavour characteristics.Table A: Particle sizes before and after homogenisation

[0213] As previously discussed, Rubisco protein naturally has a low denaturation temperature and forms hard, brittle gels when heated (Martin et al., J. Agric. Food Chem, 2014, 62, 10783-10791). EP3160254 discloses pasteurisation on a Rubisco green juice fraction and state the pasteurisation makes it easier to separate the green juice e.g. by centrifugation into a chlorophyll concentrate and a clear juice. EP316254 also states "Proteins isolated after precipitation by heat combined with low pH are partly denatured and are only valuable as animal feed and thus not for purposes where the function is needed". Therefore, a skilled worker would expect that the conditions required for pasteurisation would result in degradation / denaturation of the Rubisco protein which would not form a high protein flowable gel. Surprisingly, in the current invention, the conditions required for pasteurisation and homogenisation do not result in degradation of the Rubisco protein or the production of "off" flavours. This may be partly achieved by minimising the time the temperature of the gel products is elevated whiledelivering the effects of pasteurisation, and / or due to the partial hydrolysis of the Rubisco protein prior to gelation and pasteurisation.

[0214] Without being bound by theory, the partial hydrolysis of Rubisco protein provides for, in combination with static or gentle agitation, the formation of a weak gel when compared to non-hydrolysed Rubisco during the gel-set step. The inventors have found that the no or gentle agitation gel set up step is required prior to pasteurisation to ensure the flowable heat-set gel is able to be pasteurised without forming gritty precipitates or blocking equipment.

[0215] In accordance with the current disclosure, the process steps are such as to reduce particle size, prior to pasteurisation.

[0216] A test panel of ten tasters strongly favoured the homogenised pasteurised gel over than non-homogenised gel, indicating favourable consumer attributes including any one or more of smoother and creamier texture, less chalky mouthfeel and less particulate, grainy, sandy texture.

[0217] As previously discussed, high pressure homogenisation can be used to manipulate the mouthfeel and flavour characteristics by disrupting large particles and oil bodies. It has been found that homogenisation enables production of a flowable heat set gel product that can be formulated with a lower oil content while maintaining acceptable creaminess and mouthfeel, a visually glossy consistency and a smoothing of flavour notes. Foods and food products with reduced oil or fat content are a desirable consumer food option (Example 16).

[0218] A test panel of ten tasters favoured a lower fat content homogenised pasteurised gel, indicating that lower fat levels or oil inclusion rates in a homogenised gel could provide a similar textural mouth feel to a higher fat content or oil inclusion rate in a non-homogenised gel.

[0219] High pressure homogenisation can provide benefits including any one or more of:• particle size reduction and uniformity (see Figure 16, and Table A above)• reduction of oil inclusion rate (see Example 16) visually glossier and more vibrant productincrease in creaminess and favourable mouthfeel• removal of grainy and chalky textures• reduction in astringency• smoothing of flavour notes.

[0220] The homogenisation at elevated temperature can occur under conditions that also pasteurise the formulated gel or flowable heat set gel.

[0221] The chilled, homogenised formulated gel or chilled, homogenised flowable heat set gel may then be packaged into thermally stable packaging and blast frozen to < -12°C. Fast chilling may prevent the emulsion from splitting which would result in a product with undesirable textures.Concurrent process for production of high protein flowable heat set gel and gel product

[0222] Alternatively in a continuous or concurrent process, the high-protein content aqueous Rubisco concentrate starting material, comprising about 15% or about 20% to about 28% non-denatured Rubisco protein w / w in solution, as discussed previously is heated to between about 40 to about 65°C, for example between about 45 °C and about 50 °C, in a static or gently agitated environment in a single multi-purpose vessel, such as in a jacketed, variable-shear mixing vessel. This process can use lower concentration Rubisco solutions as the starting material if desired as well, however the Rubisco content of the flowable heat set gel produced will be lower as well.

[0223] Enzyme may be added in a range from about 0.01 to about 1.5% with respect to the total protein content of the solution for the partial hydrolysis of the Rubisco protein. The temperature may then be increased to between about 55 °C and about 65 °C, for example to about 60 °C, and the enzyme / solution mixture gently agitated via a scaped surface mixing blade for about 20 mins to about 40 mins, for example about 25 mins at 8.5 rpm. The partial enzyme hydrolysis and gelation of the Rubisco solution to form the heat set gel is performed concurrently.

[0224] As shown in Examples 13 and 14, and Table 14, when carried out at scale the enzyme / solution mixture may be gently agitated at about 50 to 55°C for about 10 to 15 minutes, followed by a slight increase in temperature to about 55 to 60°C for 10 to 40 minutes to set the gel.

[0225] High shear mixing is then performed at about 2,000 to about 20,000 rpm at between about 55 °C and about 65 °C, for example at about 60 °C, (for example the temperature may be held at the same temperature as during the gentle heat set gelation step), to disrupt the heat set gel, with mixing performed for about 1 to 20 minutes, for example about 3 to about 10 minutes, to form the flowable heat set gel with a flowable viscosity of the disclosure.

[0226] The enzymes used for partial hydrolysis can be any one or more of the enzymes discussed previously. The inventors have also found that partial protein hydrolysis can be performed during the gentle agitation and shear process with subsequent deactivation in a secondary heat treatment step (for example pasteurisation or UHT) which is completed to extend shelf life as has been discussed previously.

[0227] As shown in Figure 3, the enzyme hydrolysis occurs during heat setting under gentle agitation and shear of the heat set gel (whilst the enzymes are still active) over a combined time period of about 30 to about 60 minutes (45 minutes as shown in Fig 3). This time period can be adjusted as required such as for example between about 30 minutes and about 60 minutes.

[0228] The hydrolysis is stopped at the partial hydrolysis stage with the secondary heat treatment (e.g. increase temperature to 75-80°C for about 15 minutes or alternatives as described previously). The secondary heat treatment (e.g. pasteurisation or UHT) therefore performs a dual function of enzyme deactivation and also extension of shelf life.

[0229] The formulation additives (if added) to form the final gel product can also be added prior to or as the temperature is increased and the secondary heat treatment may then also perform a third role of microbiological control. The process can be completed in a single multi-purpose vessel allowing for production and process efficiencies.

[0230] Following the secondary heat treatment, the formulated gel, at about 75°C, is transferred to a homogenisation unit to undergo a multi-stage homogenisation process. The first stage of the multi-stage homogenisation process may be set to 150 bar. The second stage of the multi-stage homogenisation process may be set to 50 bar. This can achieve reduced particle size distributions with Dx(90) < 10 micrometers. The homogenised flowable gel upon exiting the homogeniser is immediately cooled to about 4°C via a chilled plate heat exchanger and maintained chilled in a holding tank.

[0231] The formulated flowable heat set gel product can then be packaged as part of the process, for example using suitable pouches or similar containers capable of a hot fill as would be known in the art. Allowing the formulated flowable heat set gel product to cool prior to packaging is also an option. The packaged product can be cooled to a suitable temperature (e.g. freezing) for storage and / or transport. It is also possible for the process not to include additives and the product produced to be the flowable heat set gel. In such a case, the flowable heat set gel could be packaged and stored for inclusion in products or used directly as desired.

[0232] This truncation and concurrent-ness of process to produce a product that can be packaged for consumer use increases the efficiency and speed of the process providing cost and production advantages.

[0233] It can also be preferable that one or more of the process steps between increasing the temperature for enzyme hydrolysis with gentle agitation and the end of the secondary heat treatment are performed under a vacuum of about 0.05 bar to about 0.2 bar, for example about 0.1 bar. The vacuum may be applied over the shear steps. This is very preferable as it serves to reduce foaming and flashes off volatile components which can contribute to undesirable flavours and aromas in the final product. Reduction in foaming improves the processability of the flowable heat set gel into formulated products. The ability to use a single multi-purpose vessel facilitates the use of a vacuum over the course of the process resulting a reduction of such undesirable outcomes. The secondary heat treatment may also be performed under a vacuum of 0.2 to 0.7 bar, such as 0.3 bar.

[0234] For formulated flowable heat set gel product, vacuum may only be applied prior to addition of the ingredients to minimise the loss of desirable flavour and fragrance compounds.

[0235] Gentle agitation can be performed as discussed previously such as, by way of example, using a scraped surface mixer which facilitates heat transfer into the enzyme / solution mixture. As discussed previously, a sufficiently slow mixing speed resulting in gentle agitation is required to prevent disruption of the protein cross linking which results in precipitation of protein. Agitation is performed is between about 0 rpm (static) to about 30 rpm, for example between about 5 rpm and about 20 rpm, or about 8 rpm to about 10 rpm to achieve gel formation.

[0236] Formulation additives are also as discussed previously such as one or more of sugars (2-10% w / w, for example 5%), fats / oil (1-6% w / w, for example 4.5%), fruit or vegetable juice, concentrate or puree (1-8%, for example about 5%), flavours (0-10 %, for example about 2%), preservatives (0.1-1.5%, for example about 0.5%), sweetener (for example 0 -3%, for example about 1%), and / or gums / stabilisers (0.1- 1%, for example about 0.2%).

[0237] The formulated product, including the flowable heat set gel of the disclosure, for consumption can have a viscosity suitable for the product produced. This may be between about 2000- about 4000 cP, or between 2200-3500 cP, for a drinking product, through to dessert-type mousses (10K-18K cP) when measured with an ATAGO VISCO 6815 device (A2S spindle, 12rpm, 18 °C).

[0238] The concurrent process as illustrated with reference to Figure 2 and Figure 3, provides one or more of the following beneficial aspects:(i) enzyme hydrolysis and gel heat setting under gentle agitation can be performed simultaneously which is favourable from a time perspective,(ii) enzyme dosing can be adjusted during the process(iii) the enzyme / protein solution mixture can be immediately heated to about 75 °C to about 80 °C (secondary heat treatment) following high shear mixing to deactivate the enzymes,(iv) the rate and distribution of heat transfer into the solution is improved with the use of mild agitation which minimises the time to reach a suitable temperature for optimum enzyme hydrolysis, and minimises the time to reach a suitable temperature for enzyme deactivation which provides greater process optimisation and control of viscosity,(v) the introduction of formulation additives to the sheared flowable heat set gel prior to secondary heat treatment favourably enables the simultaneous operations of deactivating the enzymes to halt proteolysis and a heat treatment step for microbiological control of the formulated product, which is beneficial as it obviates the need for subsequent heat treatment and minimises the generation of undesirably flavours or aromas from repeated heat cycling.(vi) minimising the time and temperature for processing is beneficial to limit the formation of undesirable flavours and aromas,(vii) the use of larger vessels provides for larger process volumes which is important for commercial production efficiencies,(viii) performing the process in a single multi-purpose vessel has a number of beneficial aspects: supporting productivity of throughput by minimising the transfer of materials between equipment, minimising exposure to the environment and limiting inadvertent introduction of microorganisms, enabling the use of a single terminal heat treatment which provides to simultaneously deactivate the enzymes and to minimise microorganisms levels enabling a final consumer aseptic product pack-out, and(ix) performing high pressure homogenisation of the sheared flowable heat set gel after the secondary heat treatment enables particle size reduction to provide sensory improvements.High protein gel product

[0239] The flowable heat set Rubisco gel may have a viscosity of up to about 18000cP, for example about 4000 cP to about 18,000 cP, or a viscosity of about 4000 cP to about 10000 cP and can be mixed with additives to provide a consumer gel with favourable texture, flavour and aroma properties and a viscosity of about 1500 cP to about 4000 cP, or a viscosity of about 2200 to about 3500 cP, measured with an ATAGO VISCO 6815 device (A2S spindle, 12rpm 18 °C). The consumer gel may comprise between about 7% and about 25% Rubisco protein w / w as previously discussed. Higher viscosity products can also be produced (e.g. mousse at about 18000cP) as discussed previously.

[0240] The flowable heat set gel can be formulated according to consumer preferences such as for both sweet or savoury profiles through the inclusion of ingredients which perform sweetness, functional, taste, structure or texture modifying roles to achieve a final formulated consumer gel product.

[0241] A consumer gel format can be presented as a sachet or sealed pouch containing a composition comprising:i. the flowable heat set gel (about 70- about 85% w / w, or about 70 to about 90% w / w) combined with additives suitable for consumption that can be selected in a non-exclusive manner from any one of more of: ii. sugars (2-10% w / w, for example 5%): including simple carbohydrates such as glucose, sucrose, fructose, galactose, lactose, maltose, maltodextrin, honey, maple syrup, coconut sugar, agave nectar, lucuma powder; complex carbohydrates such as corn starch, potato starch, tapioca starch, dextran, cyclodextrin, fructo-oligosaccharides, galacto-oligosaccharides; and digestion resistant sugars such as maltodextrin, microcrystalline cellulose, carboxymethylcellulose for texturing properties; polyols and isomalts, polydextrose, glycerol etc), iii. fats / oil (1-6% w / w, for example 4.5%): including vegetable oils, polyunsaturated oils, mono-unsaturated oils, such as coconut oil, canola oil (rapeseed oil), soybean oil, corn oil, rice bran oil, sunflower, palm oil, avocado oil, almond oil, cottonseed oil, medium chain triglyceride (MCT) oil,

[0242] fruit concentrate (such as fruit juice), fruit puree, or vegetable concentrate or vegetable puree (1-8%, for example about 5%): including raspberry, strawberry, blueberry, passionfruit, mango, cocoa, banana, apple, blackcurrant, pear, lemon, yuzu, orange, kumara, pumpkin, capsicum, potato, carrot, mushroom, tomato, or similar options as will be known to a skilled person i. flavours (0-10 %, for example about 2%), natural and synthetic flavours, extracts, and concentrates including flavours, extracts, flavour suppressors, flavour maskers such as ClearIQ, Smoothenol (Burnfix), herbs and spices such as paprika, cumin, cardamon, pineapple, cinnamon, chilli, parmesan, soy sauce, vanilla, chocolate, yoghurt, cream, mint, menthol, Biscoff, coffee, umami, kokumi, salt, monosodium glutamate, ii. sweeteners (0 -3, for example about 1%), including aspartame, saccharin, sucralose, xylitol, erythritol, sorbitol, acesulfame potassium, neotame, stevia, thaumatin, monk fruit extract, brazzein, sweetease, iii. preservatives (0.1-1.5%, for example about 0.5%), including Potassium sorbate, sorbic acid, Ascorbic palmitate, ascorbic acid, vitamin C, Tocopherols (vitamin E), butylated Hydroxyanisole (BHA), butylated Hydroxytoluene (BHT), EDTA, citric acid, rosemary extract,iv. gums (0.1-1%, for example about 0.2%) including xanthan gum, guar gum, agar, carrageenan, gum arabic, locust bean gum, konjac gum, gelatin for gelling and thickening properties, v. Salt, pH, and acidity modifiers: including citric acid, tartaric acid, lactic acid, sodium chloride.Process to produce flowable heat set gelAqueous Rubisco starting material

[0243] The high protein content gel of the present disclosure is prepared from an aqueous Rubisco solution starting material that is a concentrate comprising between about 15% or 20% to about 28 %w / w, for example at least about 21% protein w / w, of non-denatured Rubisco protein. The aqueous Rubisco solutions can be produced by any Rubisco isolation process using methods disclosed in prior art, including by way of nonlimiting examples: heat treatment, isolation and concentration via salt precipitation and solubilisation (Libouga et al, 1996), acid precipitation (Nissen, at al 2021), ion or anion exchange chromatography (Martin et al, 2014), size exclusion chromatography (SEC) (Martin et al 2014), tangential filtration, cross flow filtration or membrane concentration (Martin et al 2019; Nieuwland et al, 2021, Plantible US2022 / 0259259A1), spray drying (Kohler, et al 1982) or freeze drying (Lamsal et al 2005, Plantible US2022 / 0259259A1) with reconstitution of a Rubisco protein powder (Tan et al, 2023, Plantible US2022 / 0259259A1, Martin et al 2014). Methods which lead to irreversible aggregation or precipitation of the protein solution of Rubisco are not favoured such as, for example, acid precipitation (Nissen, at al 2021) or heat precipitation (Kinsella, et al 1976).

[0244] The concentrated protein solution may be diluted with water to a desired total solids level which will produce the required protein level in the final product. For example, a 27% w / w total solids concentrated protein solution could be diluted to 25% w / w TS to target a 20% protein per serve consumer product.

[0245] In an exemplary embodiment the aqueous Rubisco solution starting material is prepared by the following process: Biomass comprising non-denatured Rubisco protein is harvested and juiced / macerated using a screw press to produce a green juice. The resulting green juice is heated for about 60-120 seconds at about 57-60°C, then cooled via a heat exchanger to less than about 12°C within about 30 seconds of the heating cycle to coagulate the green protein fraction. The cooled juice is clarified in a clarifier to "de-green" the juice. The resulting clarified juice undergoes membrane concentrationand purification in a purifier, followed by further membrane concentration if necessary to provide aqueous Rubisco concentrate starting material for the process.

[0246] The biomass can be selected from any suitable source of Rubisco such as plant or the plant parts from the family Fabaceae, such as alfalfa, kudzu or forage peas; from the family Gramineae (also known as Poaceae), such as oats; from the family Lemnoideae, such as duckweed; or spinach; from the family Brassicaceae, such as kale; or roadside crops, cocksfoot, Italian ryegrass, Raphno® (kale-radish hybrid), clover, Persian clover, rapeseed leaves, carrots, radishes, Jerusalem artichoke; from the family Amaranthaceae, such as beets, spinach, fodder beet; triticale, white clover, barley, tobacco, chicory, sugar cane, or fava bean leaves. The biomass is preferably alfalfa and is preferably freshly harvested to maximise the Rubisco content of the biomass. Biomass comprising Rubisco protein may be harvested according to known methods.

[0247] The aqueous Rubisco solution starting material may also be prepared via dissolution or dispersion of a non-denatured Rubisco powder into an aqueous solution using any means known to a skilled person.Partial hydrolysis

[0248] A process to produce a high protein content flowable heat set gel of the disclosure is shown in Figure 1.

[0249] As shown in Figure 1, the aqueous high protein content Rubisco solution (Rubisco concentrate) used as the starting material is subjected to an enzyme hydrolysis step to partially hydrolyse the Rubisco protein. As previously discussed, the inventors have found that partially hydrolysing the Rubisco protein with an enzyme assists with the formation of a high protein content gel that has a suitable viscosity for use in a flowable high protein consumable product for example.

[0250] Partial enzymatic hydrolysis (or partial hydrolysis) refers to the enzyme mediated hydrolysis of the Rubisco protein in the Rubisco solution into smaller proteins and peptides, sufficient to enable a reduction in the viscosity of the flowable heat set gel produced to the preferred viscosity. This is determined in practice by adjusting the enzyme: protein ratio in response to the viscosity in the flowable heat set gel produced by the process to achieve the target viscosity.

[0251] Partial enzyme hydrolysis of Rubisco provides a lower strength gel. The moderation of gel strength is shown in Figure 9, which shows a decrease in gel strength as recorded with rheometric measurements of G' (storage modulus) in the presencepartial proteolytic hydrolysis when compared to the non-hydrolysed control, Table 1.High shear mixing and disruption of this partially hydrolysed firm gel provides a flowable gel with reduced viscosity.

[0252] The preferred enzymes for the partial hydrolysis are proteases or transferases. Proteases, also called hydrolases or peptidases are class Enzyme Commission Number EC 3.4.X.X, including thiol protease, serine proteases, aspartate proteases, and metalloproteases, which cleave peptide bonds. Transferases are class Enzyme Commission Number EC 2.6. l.X and act to transfer a functional group from one substance to another. In various embodiments, the enzyme may be a mixture of one or more enzymes. Exemplary enzymes are Protana, Flavourzyme and / or Formea Prime, all available from Novozymes, or Prolyve brand enzymes from Soufflet Biotechnologies. Preferably, enzymes are used which can be thermally deactivated by elevated temperatures commensurate with typical food processing conditions to halt hydrolysis as desired. Such temperatures can be provided by pasteurisation or UHT procedures and can therefore not only deactivate enzymatic activity but also extend the shelf life of the flowable heat set gel or products containing the flowable heat set gel.

[0253] Prior to enzyme addition, the aqueous Rubisco solution may be agitated and pre-heated at 30-70 RPM in a heated jacket vessel (62°C hot water) until the temperature of the solution reaches 50°C for 15-30mins at pH 7.5-8.5. This pre-heating step allows the enzyme to be added at conditions for at least 80% relative enzyme activity.

[0254] The pH may be adjusted as necessary to provide an optimal pH according to the enzyme used. The pH may be adjusted using food-grade buffering agents known to those skilled in the art such as hydrochloric acid, phosphoric acid, citric acid / citrate, acetic acid / acetate, sodium phosphate, sodium carbonate, sodium hydroxide, potassium hydroxide, and other pH buffering salts or agents to achieve the desired pH of the protein solution.

[0255] The enzyme is added directly to the aqueous Rubisco solution in a concentration range from 0.01%-1.5% (for example 0.05%-0.2%) with respect to the total Rubisco protein content of the concentrate, and mixed to disperse into solution manually. Alternatively, the solution can be mixed in an agitation tank to disperse the concentrate and enzyme. The mixed concentrated protein solution is heated to a temperature between about 45 and 60°C, for example between about 45 and about 50°C. The heated concentrated protein solution is kept at this temperature for betweenabout 15 and about 90 min. The heating step can be done using any suitable means known to a skilled person such as using a jacketed vessel (e.g. tank) under mild, gentle, agitation sufficient to mix the enzyme and concentrated protein solution (for example between about 8-60 RPM, or between about 8 to about 25 RPM, for example 8.5 RPM)). Alternatively, for example, the heating step can be done in sealed, thermally-stable, polyethylene bags (for example 1 kg portions) which are immersed in a 45-50°C water bath.

[0256] The enzyme may be added at a concentration of 0.05 to 1% of the feed's total solid content. The ratio of protein to total solids is generally 75-85%.

[0257] The aqueous Rubisco solution with added enzyme may be agitated for 5-20 minutes to allow hydrolysis to occur.

[0258] Optionally a 0.3 to 0.8 bar vacuum can be maintained during the pre-heat and hydrolysis stages to remove air bubbles for downstream texture and shelf-life improvements.

[0259] The partial hydrolysis step leads to a marginal increase or decrease in viscosity of the concentrated protein solution, typically from about 20-25 cP measured in the Rubisco concentrate feed increasing to about 35-50 cP at the conclusion of hydrolysis, or conversely decreasing from 54 cP in the Rubisco feed concentration to 24.5 cP as measured with a ATAGO VISCO 6815 device (A2S spindle, 12rpm, 18 °C).Table 1 : Enzyme mediated decrease in viscosity as measured by G' and viscosity

[0260] Particle size distribution analysis performed on a Mastersizer 3000+(Malvern Panalytical) indicates that enzymatic hydrolysis causes a small increase in the Dx(50) particle size distribution, increasing from 2.15 to 5.44, but a decrease in the large aggregate particles (>100 um) which are disrupted, such that Dx(90) decreased from 109 to 22, (Figure 11, Table 2). Without being bound by theory, it is assumed that a proportion of the Rubisco molecules are associating with each other under the elevated temperature at which the hydrolysis is performed (45-50 °C), whilst simultaneously the enzyme mediated hydrolysis is disrupting both these small increases in particle size and disrupting large particulate aggregates which may have formed during the concentration of the feed stream to make the concentrated gel feed. Collectively under optimal conditions these competing hydrolytic and aggregating processes may lead to a small increase in particle size, and a marginal increase in viscosity in the hydrolysed pre-gel concentrated protein solution. By increasing the concentration of enzymes or changing the identity of the enzymes the particle size distribution can be modified.Table 2: Partide size distribution of high content Rubisco solution subjected to proteolysis, gelation and shearThe extent of partial protein hydrolysis can be measured by a variety of methods, including as examples: free amine quantitation using ninhydrin colorimetric assay, size exclusion chromatography, HPLC peptide analysis, amino acid functionalisation and quantitation, pH titration, and other known methods. The degree of functional hydrolysis may also be determined via empirical correlation between protein content, enzyme dosing, and time for hydrolysis, and the resultant measured viscosity of the shear high protein heat set gel.

[0261] Subjecting the partially hydrolysed gel feed to thermal gel setting conditions (75-95 °C) under static conditions generates a heat set gel (Heat set gel in Figure 1).While Figure 1 refers to static conditions, gentle agitation can also be used, as has been described previously. The heat set gel does not have a measurable viscosity, and particles size cannot be measured by Mastersizer instrumentation until after it is disrupted. This heat set gel is blended via shear mixing to disrupt the macromolecular structure to create a smooth, flowable heat set gel (Flowable gel in Figure 1). The particle sizes within this flowable heat set gel are on average larger than in the hydrolysed gel feed for both Dx(50) and Dx(90). In the concurrent process, the average particle size is similarly observed to increase.

[0262] Enzyme dosing rates were determined on a weight / weight basis of enzyme weight or enzyme solution added to a protein weight to achieve a target enzyme: protein ratio (w / w, %) which upon proteolytic hydrolysis delivers a desired viscosity value of 4000-18000 cP, or 1000-18000 cP, as measured following gel disruption prior to addition of formulation ingredients.

[0263] The quantity of enzyme added to provide a targeted % ratio of enzyme: protein weight can be added as a powder or solution, with the quantity added being the amount to achieve a targeted enzyme activity specification. For enzyme solutions, the quantity of enzyme solution added is corrected for the dilution factor of enzyme solution. As will be appreciated by those skilled in the art, optimisation of enzyme quantity added may also include measurement and correction of the enzyme batch proteolytic activity.

[0264] As will be understood by those skilled in the art, in order to achieve a desired level of hydrolysis and resultant viscosity of the aqueous Rubisco concentrate, the ratio of enzyme addition to protein content (%), the proteolytic activity units of the enzyme, the time period for which the proteolysis is performed, the pH, and the temperature at which the proteolysis is performed all and variously may be optimised and controlled to deliver the desired viscosity.Gel formation

[0265] The partially hydrolysed protein solution is heated in a static or gently agitated environment (0-30 rpm, for example 8.5 rpm) at a temperature of between about 60 to about 90 °C, for example between about 80 or 85 to about 90 °C for about 5 to about 20 mins. This step in the sequential process provides at least three outcomes: forming a cross-linked and set Rubisco gel, deactivating the protease enzymes, and thermally treating the solution as a form of phytosanitary control. This heating step can be performed using any suitable means known to a skilled person such as using ajacketed vessel (e.g. tank), a multi-functional jacketed, variable-shear mixing vessel, alternatively, for example, the heating step can be done in sealed, thermally-stable, polyethylene bags (1 kg portions) which are immersed in a 85-90°C water bath.

[0266] Concentrated protein solution (25% TS) or partially hydrolysed protein solution may be heated to 60-65°C using a jacketed vessel (75°C hot water) for 15 to 40mins with low shear agitation / scraping close to the surface of the heated vessel (10- 40 RPM).

[0267] The heating of the Rubisco protein / enzyme solution generates a firm heat set-gel, Figure 5.

[0268] The heating applied to the Rubisco protein solution to generate a heat setgel (Set Gel in Figure 1) is conducted at a temperature of greater than 60 °C, which is the denaturation temperature of Rubisco protein in solution. The temperature may be between about 70 °C and 90 °C.

[0269] The inventors have found that it is important that there is only a static environment or gentle agitation such that there is no or limited mixing / shear applied to the partially hydrolysed concentrated protein solution in this heating step. The environment should therefore be a static or gently agitated (for example at about 8.5 rpm) mixing. Shear in this heating step has been found to disrupt the formation of the 3D network cross-link gel structure which is set up upon heating at temperatures of above about 60 °C. The formation of gel structure prevents the protein from precipitating when denaturing, where such precipitation results in grittiness (solid particles) rather than a desired smooth fluid. The gentle agitation performed is between about 0 rpm (static) to about 30 rpm, for example between about 5 rpm and about 20 rpm, or about 8 rpm to about 10 rpm to achieve the desired gel formation.

[0270] Excess stirring or agitation of the Rubisco protein solution causes protein precipitation and formation of granular aggregates at temperature greater than about 60 °C, which have an undesirable texture, an example of which is shown in Figure 6.

[0271] Heat set gel formation may be performed at a pH value close to the native or natural pH of the partially hydrolysed concentrated protein solution, specifically in the pH range of about pH 5 to about pH 9, for example a pH range between pH 5.5 and about 7.5, or at about pH 6.5 to about pH 6.7.

[0272] The form, flavour, and microstructure of the Rubisco heat set gel formed by the heating step are suitably stable at this stage that the process can be optionally halted by way of freezing the heat set Rubisco gel (Set Gel in Figure 1). The frozen heat set gel can be transported and thawed for later use to form the flowable heat set gel of the disclosure for example.

[0273] The inventors have found that the thermal gelation of Rubisco is dependent on the presence of non-denatured Rubisco protein in solution to enable the formation of the 3D network of linkages within the gel structure. Denatured Rubisco, such as that prepared by thermal precipitation or which has formed a gel and been disrupted via high shear (as per the present disclosure), is not able to form additional gels.Applying shear to form the flowable heat set gel (Flowable gel Figure 1 )

[0274] The heat set gel from Step (c) is then shear mixed using, for example: a homogeniser, a blender, or a high-shear mixer, to form a homogenous, flowable, heat set gel of the present disclosure comprising between about 15% to about 28 % w / w of Rubisco protein. The shear may be applied at about 2,000 rpm to about 20,000 rpm for about 1 to about 20 minutes. The flowable heat set gel may be homogenous, glossy, runny, with a viscosity of between about 4000 cP to about 18000 cP, for example 4000- lOOOOcP, or 1000-18000cP, or between about 4000 and about 7000cP, (measured with an ATAGO VISCO 6815 device (A2S spindle, 12rpm, 18°C)) and exhibits pseudoplastic behaviour, (see: Figure 7). Optionally, the heat set Rubisco gel may be cooled to about 4 to 50°C prior to shear mixing.

[0275] The flowable heat set gel may have a viscosity of up to 18000cP (measured with an ATAGO VISCO 6815 device (A2S spindle, 12rpm, 18 °C), the flowable heat set gel has a viscosity of between 1000 and 18000 cP, for example between 1000 and 160000 cP, between 1000 and 14000 cP, between 1000 and 12000 cP, or between 1000 and 10000 cP, between 1500 and 18000cP, between 1500 and 160000 cP, between 1500 and 14000 cP, between 1500 and 12000 cP, or between 1500 and 10000 cP.

[0276] A skilled worker will appreciate that viscosity measurements are dependent on the measurement technique used, including the spindle diameter and spindle speed. At a lower speed the shear rate is less, hence for pseudoplastic fluids the apparent viscosity will be higher than measurements at faster spindle speeds (higher shear rates). A skilled worker will be able to convert between parameters such as spindle diameter and spindle speed to calculate the shear rate. A skilled worker will understand the need to change spindle types depending on the viscosity of the material in order to minimiseinaccuracies during measurement due to over-torquing the instrument. Conversion between different spindle speeds and spindle types for the ATAGO VISCO 6815 are demonstrated in Example 15 and Figure 17, where the same sample is measured at different spindle speeds (5, 12, 20, 30, 50, 60, 100, 150, 200, 250 rpm) on both the AIS and A2S spindle. "Al" and "A2" refer to the specific spindle provided with an ATAGO VISCO 6815 and "S" refers to the beaker in which the samples are measured. For clarity, AIS and A2S are used in this specification to denotate the specific spindle and beaker used during analysis.

[0277] As such, viscosities of the flowable heat set gel are presented as measurements on an A2S spindle at 12rpm, 18 °C and viscosities of the formulated gel are presented as measurement on an AIS spindle at 200 rpm, 18 °C. This is because the consumer formulated gel is less viscous than the flowable heat set gel.

[0278] Differences in viscosity are often considered significant when they span more than an order of magnitude. Smaller differences, such as from 100 cP to 500 cP, are minimally significant compared with a change from 100 cP to 1000 cP, and are also dependent on the sensitivity of the process or application.

[0279] The viscosity of the flowable het set gel, or high protein consumer gel product may vary between "batches" due to the feedstock, the enzyme used, enzyme digestion time, total batch time, heat set temperature, and / or the additives in the consumer gel.

[0280] The inventors found that high shear mixing of the heat set gel provided a flowable heat set gel which possesses a desirable non-rigid, semi-solid state, with the ability to flow. Flowability of the gel refers to the ability of the gel to deform on application of shear / gravity, such that it will, for example, pour out of a bowl.Surprisingly the disclosure provides a flowable gel despite a high Rubisco protein content which would previously have been expected to result in a rigid gel that would not have flowable characteristics.

[0281] Surprisingly, the inventors have found that a dispersant is not required during the shear step to obtain these desired characteristics (e.g. homogeneous, flowy gel with a viscosity of about 4000-18000 cP). Avoiding dispersant provides process and cost efficiencies to the user and removes the need for additional components in the flowable heat set gel.

[0282] The flowable heat set gel (Flowable gel, Figure 1) is suitably stable at this stage that it can be stored including by way of freezing. The frozen heat set gel can be transported and thawed for later use. If the heat set gel is frozen and later thawed, the shear mixing step from above is repeated to re-form the flowable heat set gel of the present disclosure.

[0283] If desired, the flowable heat set gel of the disclosure can be dehydrated, by spray drying, freeze-drying or similar techniques as are known, to achieve a dehydrated powder. The flowable heat set gel may be diluted sufficiently (if needed) to achieve a viscosity / fluidity to allow transit through a spray dryer nozzle, thus enabling spray drying.

[0284] The dehydrated protein powder prepared from the flowable heat set gel can be formulated with other ingredients and additives to generate a high protein content food product without significant loss of beneficial characteristics (e.g. viscosity / taste / texture). The dehydrated powder has similar composition attributes as the flowable heat set gel, except that it has been dried to about 4-6% residual moisture.

[0285] Dehydrated Rubisco protein powder, including that formed from the flowable heat set gel of the present disclosure, can be added to the flowable heat set gel of the current disclosure should an increase or fortification in the protein content of the flowable heat set gel be required or otherwise desired. If dehydrated flowable heat set gel powder is used this will achieve a fortification without a commensurate increase in viscosity. This protein fortification may be from about 0% to about to about 60% on a protein w / w basis as shown in Example 3.Specific embodiments of the disclosure

[0286] The process of the disclosure can therefore be seen to comprise a process for preparing a flowable heat set gel comprising from about 7.5% to about 28% w / w, for example from about 15% or 20% to 28%w / w, of Rubisco protein and having a viscosity up to about 18000cP (measured with an ATAGO VISCO 6815 device (A2S spindle, 12rpm, 18°C), the process comprising the steps of: a. Providing an aqueous Rubisco solution comprising from about 7.5% to about 28% non-denatured Rubisco protein. The content of the aqueous non-denatured Rubisco may be from about 15% or 20% to 28% Rubisco protein as this will provide a flowable heat set gel having a higher, and related, Rubisco content.b. Subjecting the aqueous non-denatured Rubisco solution to partial enzymatic hydrolysis in a static or gently agitated environment to provide a partially hydrolysed aqueous Rubisco solution. The enzyme(s) may be selected from proteases and transferases that can be thermally deactivated. The enzymes are dosed at concentrations of from about 0.01 to about 1.5% w / w total Rubisco protein content and the enzyme: Rubisco ratio is adjusted based on the desired viscosity in the flowable heat set gel. The enzyme and the aqueous Rubisco solution are mixed and partial enzymatic hydrolysis is carried out at a temperature that may be between about 55 °C and about 65 °C, for example to about 60 °C, and the enzyme / solution mixture is gently agitated for about 20 minutes to about 40 minutes, for example about 25 minutes. Static or gentle agitation is between about 0 rpm (static) to about 30 rpm to avoid disruption of the gel formation. c. Heating the partially hydrolysed aqueous Rubisco solution to form a heat set gel. The temperature may be between about 40 °C to about 90 °C, for example about 60 °C to about 85°C, for example about 75°C or 60 °C. for about 5 to about 20 minutes. This step can optionally be at a temperature and for a time sufficient to also deactivate the enzymes and provide a temperature treatment (e.g. pasteurisation or UHT). Heat set gel formation may be be performed at a pH value close to the native or natural pH of the partially hydrolysed protein solution This may be in the pH range from about pH 5 to about pH 9, for example between pH 5.5 and about 7.5, or at about pH 6.5 to about pH 6.7. d. Subjecting the heat set gel to shear to provide the flowable heat set gel. Shear may be between about 2,000 to about 20,000 rpm, and may be about 1 to about 20 minutes in duration. e. Subjecting the flowable heat set gel to a heating step to deactivate the enzymes and stop enzymatic hydrolysis if not already completed in step c. Optionally, this step could also achieve pasteurisation or UHT.

[0287] Any one or more of the process steps b. c. and d. may be under vacuum and performed in a single multi-purpose vessel. Step d. at least may be under vacuum to minimise foaming. The vacuum may be between about 0.05 bar to about 0.2 bar, for example about 0.1 bar.

[0288] Any one or more of the process steps b. c. and d. may be under vacuum and performed in a single multi-purpose vessel. Step d. at least, may be under vacuum to minimise foaming. The vacuum may be between about 0.3 bar to about 0.7 bar, for example about 0.4 bar.

[0289] The Rubisco content of the flowable heat set gel may be from about 7.5% to about 28% w / w, between 15% and 25% w / w, or between 20% and 25% w / w. The viscosity may be from 4000cP to about 18,000cP, between 4000cP and 10,000cP, or between 4000cP and 7000cP.

[0290] The flowable heat set gel can be formulated into products as desired by the manufacturer and the consumer using additives that would be known to a skilled person. Such products can have a Rubisco protein content as desired but may be from about 15% to about 25% Rubisco protein. The products can have a viscosity commensurate with the product which can be, for example, between about 2000- about 4000 cP, and for example between 2200-3500 cP, for a drinking product, and between about 10,000 and about 18,000 cP for dessert-type mousses, when measured with an ATAGO VISCO 6815 device (A2S spindle, 12rpm, 18 °C).

[0291] If the Rubisco protein content of the flowable heat set gel is below about 15% w / w, dehydrated Rubisco protein powder can be added to the flowable heat set gel to increase the Rubisco protein content up to about 20%w / w.

[0292] The process of the disclosure can also be seen to comprise a process for preparing a flowable heat set gel comprising from about 15% to 28%w / w, of Rubisco protein and having a viscosity from about 4000cP to 18000cP (measured with an ATAGO VISCO 6815 device (A2S spindle, 12rpm, 18°C), the process comprising the steps of: a. Providing an aqueous Rubisco solution comprising from about 15% to about 28% non-denatured Rubisco protein, b. Subjecting the aqueous non-denatured Rubisco solution to partial enzymatic hydrolysis using protease or transferase enzymes that can be thermally deactivated, at a dose concentration of from about 0.01 to about 1.5% w / w total Rubisco protein content, in a static or gently agitated environment to provide a partially hydrolysed aqueous Rubisco solution by mixing the enzyme and the aqueous Rubisco solution, and gently agitating the mixture at between 1 rpm and 30 rpm for about 20 minutes to about 40 minutes at a temperature between about 55 °C and about 65 °C,c. Heating the partially hydrolysed aqueous Rubisco solution from about 70°C to about 90°C at a pH from about 5.5 and about 7.5 to form a heat set gel, d. Subjecting the heat set gel to shear of from about 2,000 to about 20,000 rpm for about 1 to about 20 minutes, under a vacuum of from about 0.05 bar to about 0.2 bar, to provide the flowable heat set gel, e. Adjusting the dose concentration of enzyme in step b. if needed based on the desired viscosity of the flowable heat set gel of step d.

[0293] The process of the disclosure can also be seen to comprise a process for preparing a flowable heat set gel comprising from about 15% to 28%w / w, of Rubisco protein and having a viscosity from about 4000cP to 18000cP (measured with an ATAGO VISCO 6815 device (A2S spindle, 12rpm, 18°C), the process comprising the steps of: a. Providing an aqueous Rubisco solution comprising from about 15% to about 28% non-denatured Rubisco protein, b. Subjecting the aqueous non-denatured Rubisco solution to partial enzymatic hydrolysis using protease or transferase enzymes that can be thermally deactivated, at a dose concentration of from about 0.01 to about 1.5% w / w total Rubisco protein content, in a static or gently agitated environment to provide a partially hydrolysed aqueous Rubisco solution by mixing the enzyme and the aqueous Rubisco solution, and gently agitating the mixture at between 1 rpm and 30 rpm for about 20 minutes to about 40 minutes at a temperature between about 55 °C and about 65 °C, c. Heating the partially hydrolysed aqueous Rubisco solution from about 70°C to about 90°C at a pH from about 5.5 and about 7.5 to form a heat set gel, d. Subjecting the heat set gel to shear of from about 2,000 to about 20,000 rpm for about 1 to about 20 minutes, under a vacuum of from about 0.05 bar to about 0.2 bar, to provide the flowable heat set gel, e. Adjusting the dose concentration of enzyme in step b. if needed based on the desired viscosity of the flowable heat set gel of step d.

[0294] The process of the disclosure can also be seen to comprise a process for preparing a formulated product comprising from about 12.5% to 25%w / w, of Rubisco protein and having a viscosity from about 2000cP to 18000cP (measured with an ATAGO VISCO 6815 device (A2S spindle, 12rpm, 18°C), the process comprising the steps of: a. Providing an aqueous Rubisco solution comprising from about 15% to about 28% non-denatured Rubisco protein, b. Subjecting the aqueous non-denatured Rubisco solution to partial enzymatic hydrolysis using protease or transferase enzymes that can be thermally deactivated, at a dose concentration of from about 0.01 to about 1.5% w / w total Rubisco protein content, in a static or gently agitated environment to provide a partially hydrolysed aqueous Rubisco solution by mixing the enzyme and the aqueous Rubisco solution, and gently agitating the mixture at between 1 rpm and 30 rpm, c. Heating the partially hydrolysed aqueous Rubisco solution from about 55°C to about 65°C at a pH from about 5.5 and about 7.5 to form a heat set gel, d. Subjecting the heat set gel to shear of from about 2,000 to about 20,000 rpm for about 1 to about 20 minutes, at a temperature of from about 70°C to about 85°C under a vacuum of from about 0.05 bar to about 0.2 bar, to provide a flowable heat set gel having a viscosity of between about 4000cP and about 18,000cP, e. Adding formulation components to the flowable heat set gel to create the formulated product, f. Optionally heat treating the formulated product to between 75°C and 85°C for about 5 to about 20 minutes prior to or after packaging the formulated product, wherein steps b. and c. are concurrent and are at a temperature of between about 55 °C and about 65 °C and for a time of from about 35 minutes to about 55 minutes, and wherein the dose concentration of enzyme in step b. (and thus the enzyme: protein ratio) is adjusted if needed based on the viscosity of the flowable heat set gel of step d. and / or of the formulated product of step e. and wherein steps b., c., and d. are performed in a single multipurpose vessel.

[0295] The disclosure can also be seen to comprise a flowable heat set gel comprising from about 15% to about 25 % w / w of Rubisco protein, wherein the heat set gel has a viscosity between about 4000 and about 7000cP (measured with an ATAGO VISCO 6815 device (A2S spindle, 12rpm, 18 °C) wherein the flowable heat set gel has been pasteurised or UHT.

[0296] The disclosure can also be seen to comprise a food product (consumer product), the food product comprising between about 12.5% to about 25 % w / w of Rubisco protein, wherein the heat set gel has a viscosity between about 4000 and about 18000cP (measured with an ATAGO VISCO 6815 device (A2S spindle, 12rpm, 18 °C), together with suitable additives optionally selected from any one or more of:(i) sugars and carbohydrates (for example about 2-10% w / w, or 5%),(ii) fats or oils (for example about 1-6% w / w, or 4.5%),(iii) fruit juice, fruit concentrate or puree (for example between about 0-8%, or about 5%),(iv) flavours (for example between about 1-10 %, or about 2%),(v) preservatives (for example between about 0.1-1.5%, or about 0.5%),(vi) sweetener (for example between about 0 -3, or about 1%),(vii) gums or stabilisers (for example between about 0.1-1%, or about 0.2%),(viii) vitamins or other nutritional supplements, wherein the food product has a viscosity of between about 2000cP and about 18,000cP.

[0297] The food product may be pasteurised or UHT. The food product may have a shelf life of about 6 to about 9 months.

[0298] The following non-limiting examples are provided to illustrate the present disclosure and in no way limit the scope thereof.Examples

[0299] It is not the intention to limit the scope of the disclosure to the mentioned examples only. As would be appreciated by a skilled person in the art, many variations are possible without departing from the scope of the disclosure.Example 1 - High protein flowable heat set gels and Formulated high protein content consumer gels using a scalable process

[0300] 8 kg of concentrated Rubisco protein (non-denatured) solution (21.0% solids, 75% protein purity w / w) was added into a Limitech-LAIMKPS21-16 insulated mixing vessel. The product was slowly agitated (24.3 RPM) while hot water at 55 °C was used in the surrounding vessel jacket to heat the solution to the target temperature of 50°C over 35 mins. Prolyve BS2 enzyme solution (12.42 ml of 10% solution of powder in water, Soufflet Biotechnologies) was added and dispersed into solution with mixing. The water jacket temperature was increased to 90 °C to bring the protein solution to 60 °C and to set the gel, with agitation (15.7 RPM). The heat set gel took 30 mins to set up. The gel was blended / sheared using the high shear mixer (2300 RPM) for 5 mins into a smooth, thin, flowing gel (flowable heat set gel). This product can be heat treated, cooled and aseptically bagged, as below, without formulation inclusion.

[0301] The formulation composition (Table 3 below) was added and blended into the flowable heat set gel. The product was brought to 75-80°C and held for 10 mins using steam injection into the vessel jacket with mixing and agitation (21.3 RPM). The formulated gel was cooled to 40 °C using cold water circulated in the jacket, then removed via the exit port, aseptically bagged into polyethylene food safe bladders and cooled in an ice bath, prior to blast freezing at -30 °C. The formulated gel product was smooth, thin, glossy, with an acceptable smell and a pleasant taste. The viscosity of the formulated consumer gel formed was 2250 cP measured with a ATAGO VISCO 6815 device (A2S spindle, 12rpm).Table 3 Formulation composition proportions added to 8 kg Rubisco protein solution batch

[0302] Formulated gels were stored at 4 °C and monitored for microbial content for 2 weeks, see Table 4 below. Negligible microbial growth was observed over 14 days for the formulated high protein consumer gel when compared to the non-heat-treated concentrated Rubisco protein solution.Table 4: Microbiological content of heat-treated formulated gelsExample 2 - Formulated high protein content consumer gels using static gel preparation

[0303] Part A: Concentrated Rubisco protein (non-denatured) solution 94 kg, 23.4% solids, 80% protein w / w was weighed into thermally stable polyethylene bags (1 kg portions) and dosed with 1.9% Prolyve BS Cone (10% solution of enzyme powder in water) and 0.4% Prolyve ALK 3000 (10% solution of enzyme powder in water) on v / w protein basis and mixed to disperse into the mixture. The bags were sealed and enzymatic hydrolysis performed at 45-50 °C for 50 mins by immersion in a water bath, followed by transfer to 95 °C water bath and gel heat-set for 1 hour. The heat set gels were removed from the bags, combined in 30 kg batches and blended / sheared with a Waring Commercial Blender (650W) for 3 minutes. Viscosity of the starting protein solution was 12.41 cP; flowable heat set gel post blending was 5900 cP, measured with a ATAGO VISCO 6815 device (A2S spindle, 12rpm). The flowable heat set gel can be used for formulation or could be frozen in a blast freezer at -30 °C for subsequent formulation, Figure 4.

[0304] Part B: The formulation composition (Table 5 below) was added and blended into the flowable heat set gel matrix. The mixture was heated to 75 °C in a heating vessel for 10 mins to provide the 8.3 kg flavoured gel which was aseptically dispensed into individual 100 g consumer pouches and sealed hermetically with a twist plastic cap. The viscosity of the formulated consumer gel formed was 2498 cP with a ATAGO VISCO 6815 device (A2S spindle, 12rpm).Table 5: Formulation composition proportions added to 6.7 kg protein gel batchExample 3 - Ranges of protein content into formulated gels

[0305] The inclusion of Rubisco protein powder to the shear protein gels (flowable heat set gel) can be used to increase the protein composition (% w / w) in the gel, without unfavourably affecting taste or texture attributes to achieve a formulated protein content of about 10% to about 25% protein content w / w.

[0306] By way of example, a gel with increased protein content can be prepared by addition of dehydrated gel powder during formulation, where the dehydrated powder has a protein content of about 74% to about 85 % protein content w / w on total solids is added to the flowable heat set gel which has a protein content of about 15 % to about 21% protein content w / w. The resultant protein gel has a protein content that is an average of the protein individual components, Table 6.

[0307] The increased protein content gel can be formulated to provide consumer gel with higher protein content.

[0308] For example, 703 g of flowable heat set protein gel with 15.6% protein content w / w is hand mixed with a spatula with 134 g of dehydrated Rubisco powder containing 74.4% protein content w / w and then homogenised with a DAIHAN Scientific homogenizer at power 3 for 1-2 min to achieved complete dispersion and to generate the modified gel material with a composite protein content of 25% protein w / w.Formulating additives are combined at the appropriate quantities, as described in Table 6, and homogenised for mixing to achieve a formulated gel content of 19.4% protein content w / w, which had an acceptable flowable consistency and a high protein content.

[0309] As will be understood, the addition of additives to the high protein flowable heat set Rubisco gel for provision of textures, aroma, or texture properties will decrease the protein content proportionate to the volume of additions, for example from about 20% to about 28% protein w / w to about 15% to about 23.4% w / w to a maximum of about 25% after formulation addition, Table 6, depending on the amount of additives used. As such, it is desirable to have a high protein content in the flowable heat set Rubisco gel, which is greater than 20% protein w / w to enable the provision of a formulated consumer product with greater than 15% protein w / w. This can of course be adjusted as desired as would be known to a skilled person.Table 6: Increased protein content gel ingredients (g)Example 4 - Formulated high protein content gels using a concurrent process

[0310] Following the general process of Figure 3, 52 kg of concentrated protein solution (21.0% solids, 75% protein purity w / w) was heated to 45 °C in an agitated vessel (25 rpm) using a 55 °C heated jacket. Once the product reached 45 °C 1.5% BS2 Liquid (124.5 ml of 10% solution of enzyme powder in water) and 0.4% ALK liquid (33.2 ml of 10% solution of enzyme powder in water) enzyme on a protein basis was added and allowed to hydrolyse for 35 minutes. The hydrolysed material was transferred into an Almix vessel (TetraPak) and heated gently to 60 °C with intermittent steam applied to the jacket (temp 85 °C increased to 99 °C) and the vessel gently agitated (7.5-8RPM). The Rubisco protein solution material took 15 mins to start to set, and a further 10 mins to fully set to a heat set gel. Once set the heat set gel was blended (shear mixed) for 5 mins (2500 rpm) to form a smooth and flowing gel (flowable heat set gel). Gums and preservatives were added (Table 7) and blended in, and the mixture heated in a secondary heating step to 75-80 °C for 10 mins with shear and agitation. During this heating step a vacuum of 0.1 bar was applied to minimise foaming. After lOmins the product was cooled with 10 °C water for 30 mins. The product was bagged off and frozen. The final product was flowable, lush, glossy.Table 7: Scaled gel formation

[0311] The disclosure therefore provides opportunity as per Example 1 and per Example 2 to provide a non-formulated gel (i.e. no formation additives) which may be refrigerated or frozen. This non-formulated high protein flowable heat set gel can be subsequently formulated according to consumer or manufacturer requirements. As per the present Example, the high protein flowable heat set gel may also contain only gelation and gum agents to facilitate formulation.Example 5 - Enzyme variations (% inclusion)

[0312] Concentrated Rubisco protein solution (500 mL, 22.0% solids, 78% protein purity, pH 6.8) was mixed with varying addition rates of Lypaine enzyme (Soufflet Biotechnologies) (Table 8), sealed in thermally stable polyethylene bags and heated at 50-55 °C for 30 min to enable partial proteolytic hydrolysis. The hydrolysed protein solution is subsequently heated to 80-90 °C in a water bath for 40 min to generate a firm non-flowing heat set gel. The heat set gel was blended with a high shear mixer (Living8<.Co 200W stick blender, 12,000 rpm) for 3 minutes until a free flowing texture is achieved as a flowable heat set gel. The viscosity was measured with an ATAGO VISCO 6815 device (A2S spindle, 12rpm, 18°C).

[0313] The measured viscosity of the flowable heat set gel is inversely proportional to enzyme dosing rate (Table 8, Figure 8). Concentrated Rubisco protein solution was measured to have a pre-hydrolysis viscosity of about 19.39 - 23.86 cP which increases to about 34.1 - 42.71cP on heating. After heat set gel formation and high shear was applied to form the flowable heat set gel, it was observed that higher enzyme dosing rates (%) lead to a decrease in measured viscosity in the flowable heat set gel. In the absence of enzyme (Lyp 0.0%) the viscosity reading was outside the measurement range of the instrument (HHH, > 25,000 cP). This method and approach enables calculation of an optimal enzyme dosing rate to achieve a desired viscosity.

[0314] The target viscosity for the sheared gel is about 4000 cP to about 18000 cP. It can be seen from Table 9 that the Lyp concentration to achieve the targeted viscosity would be between 0.5% and 0.75%. The Lyp concentration in the process would be adjusted accordingly to achieve the targeted viscosity. The targeted Lyp concentration would change according to the protein content in the concentrate used in the process.Table 8: Enzyme concentration dosing relationship to sheared high protein gelExample 6 - High protein content Rubisco gel powder compared to protein isolates

[0315] High protein content Rubisco solutions can form firm gels upon thermal treatment, Figure 12. This differs from other common protein sources which do not have this ability.

[0316] Concentrated protein solutions of pea protein isolate (Roquette, NUTRALYS® S), soy protein concentrate (ADM Arcon® SM) and whey protein isolate (Scorpion Whey, NZ) were dispersed in water without pH adjustment to generate solution with 5 to 25% protein content (300 mL). Sample were sealed in thermally stable polyethylene bags and heated to 90 °C in a water bath for 40 minutes. The Pea, Soy and Whey protein solutions did not create strong, self-standing gel like Rubisco, Figure 12. Pea protein formed a soft gel, soy protein concentrate formed a large granular particulate form, whey protein isolate did not form a gel. This shows that Rubisco behaves differently to other proteins, both plant (Pea, Soy) and animal (whey), and has its own characteristics. This data is consistent with De Berardinis et al (De Berardinis, L.; Plazzotta, S.; Manzocco, L. Optimising Soy and Pea Protein Gelation to Obtain Hydrogels Intended as Precursors of Food-Grade Dried Porous Materials. Gels 2023, 9, 62. https: / / doi.org / 10.3390 / gels9010062), and Martin et al. J. Agric. Food Chem. 2014, 62, 10783-10791, dx.doi.org / 10.1021 / jf502905g.Example 7 -Agitated or mixed gel formation in high protein content Rubisco solutions

[0317] Concentrated Rubisco protein solution (~23% TS, 77% protein content, 0.3L) was mixed with enzyme composition Protana (Novozymes) and Flavourzyme (Novozymes) dosed at 0.05%-1.5% enzymes (w / w protein basis) until fully dispersed and heated at 45 °C for 45 mins to perform partial enzyme hydrolysis. The resultant hydrolysed protein solution was added to a Thermomix 6 (Vorwerk) mixing bowl and slowly heated to 90 °C on slowest mix setting (slow = 40 rpm). Under these low shear / mixing conditions, the protein solution precipitated to granular protein particles with phase separation illustrating that even low shear (40 rpm) was detrimental to the formation of a stable gel due to excessive disruption of the microgel structure, Figure 6. Gentle or no agitation (as described previously) is needed to avoid disruption of the gel structure.Example 8 - Viscosity

[0318] The disclosure provides a high protein content flowable Rubisco heat set gel which has preferrable textural attributes for consumers. Preferably, the viscosity is between 4000-18000 cP for the flowable heat set gel. Preferably, for a consumer product, the viscosity will be between 2000-4000 cP, and more preferably between 2200-3500 cP, for a formulated high protein content flowable Rubisco heat set gel product for consumer consumption as a drink, as measured on an ATAGO VISCO 6815 device (A2S spindle, 12rpm, 19 °C).

[0319] Following the method of Example 7, flowable heat-set gels were prepared with different viscosities, and formulated with additives such as discussed in Example 4 to prepare formulated products. These formulated products were assessed for viscosity and textural attributes. The viscosity (cP) was measured with an ATAGO VISCO 6815 device (A2S spindle, 12rpm, 18°C) and the values were compared to gel fluidity textural attributes to enable different consumer formats, including drinking viscosity (2000-4000 cP) through to dessert-type mousses (10K-18K cP), Table 10.

[0320] The flowable heat set gel products with viscosity 2200-3500 cP had a favourable mouthfeel which was smooth, velvety, non-granular or grainy, Figure 13.Table 10: Correlation of viscosity measurement to textural attribute*Values recorded as HHH indicated a viscosity above the measurement range of the instrument (> 25,000 cP for A2S spindle)

[0321] The addition of additives to the high protein flowable heat set Rubisco gel for formulation, will affect the viscosity through proportional dilution effects decreasing theviscosity from about 4000 - 18000 cP in the precursor unformulated flowable gel to about 2000 - 4000 cP, more preferably a viscosity of about 2200 to about 3500 cP in the formulated consumer gel, (Table 10).

[0322] Adjustment of viscosity could also be achieved through the combination of different gels with different viscosity attributes (Table 11).Table 11: Viscosity measurements of different gels and gel combinationsExample 9 - Formulation

[0323] The consumer gel can be formulated for both sweet or savoury profiles through the inclusion of ingredients which perform sweetness, functional, taste, structure or texture modifying roles to achieve a final formulated consumer gel product, Figure 14. The protein, fat and carbohydrate composition can be formulated to provide varying nutritional attributes.

[0324] To achieve a consumer formulated gel at 10 - 20% protein per serve, flowable heat set gel (70-85% w / w) made by the process of Example 1 or Example 2 is combined with sugars (4%), fats / oil (4.5-5% w / w), fruit concentrate or puree (1-8%), flavours (0.6%), sweeteners (0.5%), preservatives (0.1%), and gums (0.2%). Gums are hydrated in the water prior to inclusion, first by hand mixing with a spatula, then added to the rest of the ingredients. The formulated mix is blended with a homogenizer (HG- 15A Homogeniser, DAIHAN Scientific) at power 30 for 1-2 min to achieve a homogeneous mixture suitable for packing into gel pack formats. The blended viscosityof the consumer gels produced in this example below was measured to be about 2000 cP to about 3100 cP with a ATAGO VISCO 6815 device (A2S spindle, 12rpm, 18°C).

[0325] Example Unflavoured gel base. On a per 100 g basis combine the following ingredients at room temperature. 86.58 g of Rubisco sheared gel is mixed with 3.3 g of water into which 4.8 g of sucrose, 0.038 g of Guar gum, 0.010 g of Xanthan gum and 1 g of maltodextrin had previously been dispersed and allowed hydrate. 3.6 g of MCT oil is added and mixed to combine, followed by the addition of 0.4 of natural flavour, 0.009 g of thaumatin, and 0.1 g of flavour clarifier. The mixture is stirred followed by the addition of 0.086 g of potassium sorbate, 0.09 g of ascorbate palmitate and 0.017 g of tocopherols. The mixture is heated to 75 °C in a Thermomix for 10 mins to provide the unflavoured gel base. As will be appreciated, flavour ingredients can be formulated into the gel base by substituting a commensurate volume of water to generate different flavour properties.

[0326] Example Berry Flavoured gel at 50 kg. To a stirred tank introduce 37.750 kg of Rubisco sheared gel followed by 0.45 kg of blueberry concentrate and 1.25 kg of raspberry concentrate and mix at 50 rpm. Separately, add 4.15 kg of cold water to 2.41 kg of sucrose, 53 g of Guar gum, 3.5 g of Xanthan gum and 500 g of maltodextrin, blend under dispersed and allowed to sit to hydrate. Add the sugar and gum solution to the tank and mix until combined. Add 1.8 kg of MCT oil and mix until combined. Separately, combine 100 g of vanilla flavour, 226.5 g of berry flavour, 225 g of natural flavour, 4.4 g of thaumatin, 25 g of flavour clarifier with 1 kg of water, and add to tank and mix until combined. Add 43 g of potassium sorbate, 4.5 g of ascorbate palmitate, and 8.5 g of tocopherols. Heat the tank to 75 °C and hold for 10 mins to prepare the final formulated consumer gel, which can then be dispensed into packaging.

[0327] Example Chocolate flavoured gel: On a per 100 g basis combine the following ingredients at room temperature. 60.2 g of Rubisco sheared gel and 8.53 g of spray dried gel is mixed with 2.0 g of cocoa powder, 0.4 g of chocolate flavour, 0.10 g of cocoa enhancer and 19.7 g of water into which 4.1 g of sucrose, 0.026 g of Guar gum, 0.007 g of Xanthan gum and 0.1 g of maltodextrin had previously been dispersed and allowed hydrate. 3.86 g of MCT oil is added and mixed to combine followed by the addition of 0.54 of natural flavour, and 0.1 g of flavour clarifier. The mixture is stirred followed by the addition of 0.098 g of potassium sorbate, 0.01 g of ascorbate palmitate and 0.020 g of tocopherols. The mixture is heated to 75 °C in a Thermomix for 10 mins to provide a chocolate flavoured gel.

[0328] Consumer sensory preference panels were conducted assessing formulated gel attributes for: thickness, smoothness, sweetness, flavour, flavour strength, aroma strength, aftertaste and dryness. Average liking was recorded on a 0-100 scale where 0 = extremely disliking to 100 = extremely liking, and average ratios calculated between the experienced attribute score and the ideal attribute score, and depicted on a radial spider diagram, Figure 13.Example 10 - High protein content Rubisco gel powder

[0329] The high protein flowable heat set gels such as produced in the processes, for example, in Example 1, Example 2 or Example 5 can be spray dried to provide a powder.

[0330] 5 kg of flowable heat set gel was diluted 1 : 1 with water and spray dried on aS-300 Buchi Mini Spray Dryer S-300 220C (190 °C inlet, 75-80 °C outlet, 800 ml / h spray gas, 25 m3drier gas) to provide 0.69 kg of a dehydrated powder.

[0331] Alternatively, 33 kg of flowable heat set gel was loaded at 20-25% solids on NIRO FSD 4.0 (200 °C inlet, 75-90 °C outlet, 30 kg / h product feed rate, 750 kg / h drier gas) to provide 7.6 kg of a dehydrated powder.

[0332] The dehydrated gel powder produced can be used as an additive as described in Example 3 to increase the relative protein content of the formulated consumer product.

[0333] An aqueous solution of 15.0 % w / w of spray-dried Rubisco flowable heat set gel powder was prepared by dissolving non-denatured Rubisco powder in water or 20 mM HEPES buffer pH 7.0, with stirring for 1 h to ensure complete solubilisation. The aqueous protein dispersions were sealed in thermally stable polyethylene bags and heated at 80 °C for 30 min in a water bath and then cooled in cold water followed by storage at 4 °C overnight. The Rubisco gel protein, derived from powder dispersion stayed in solution after the heat-cool cycle and did not form a gel indicating that the functional gelation properties of Rubisco are expended after the first gelling process followed by spray drying.Example 11 -Thermal treatment and packaging

[0334] High protein content gel products formed from the flowable heat set gels of the disclosure can be packaged into single serve pouches or trays which offerconvenience to the consumer, which may be presented as frozen, chilled, refrigerated, or shelf stable packaged formats.

[0335] Consumer formulated gels (as from Example 1) including the flowable heat set gel of the present disclosure were packaged into single serve ultra-high barrier laminate pouches (60 g - 120 g serves), where the pouches are made of material suitable for the secondary heat treatment such as three layer laminated polyethylene (LDPE) / metalised polyethylene terephthalate (PET) / LPDE packages, such as RE: MONO laminate pouches (Wastemade™). The formulated product is aseptically filled into the pouches using a sterile 50 ml syringe, and the pouches closed hermetically with a twist plastic cap and stored under refrigeration (4 °C) or frozen (-18 °C).

[0336] Consumer formulated gels including the flowable heat set gel of the present disclosure were packed into hermetically-sealed thermally stable pouches and heat treated in a 75 °C water bath for 5 min (60 g samples) or 11 min (120 g samples). Samples are cooled down rapidly in a blast freezer (-32 °C) until internal temperature reaches 10 °C (typically 10-15 min for 60 g pouches) and then transferred to a 4 °C fridge (refrigerated samples) or blast frozen and transferred to a -20 °C freezer (frozen samples).

[0337] Consumer formulated gels including the flowable heat set gel of the present disclosure were packed into hermetically-sealed thermally stable pouches and ultra-heat treated (UHT) in a retort (Steriflow, Microflow 911R) under the following protocol:1. Heating from 4 °C to 90 °C, 1 bar, 10 min.2. Heating from 90 °C to 117 °C, 2 bar, 10 min.3. Holding at 117 °C, 2 bar, 10 min.4. Cooling from 117 °C to 90 °C, 2 bar, 4 min.5. Cooling from 90 °C to 50 °C, 1.5 bar, 4 min.6. Cooling from 50 °C to 35 °C, 0.6 bar, 5 min.7. Cooling from 35 °C to 30 °C, 0 bar, 10 min.

[0338] Following UHT treatment the formulated consumer gels are held are ambient temperature (21 °C) for storage. Testing indicated the absence of microbiological organisms (Table 12) and the retention of flavour and aroma, and no observed syneresis or textural changes.

[0339] Table 12: Microbial stability assessment following UHT treatment for samples held at -18 °C

[0340] Formulated (Example 1) and non-formulated (Example 5) high protein gels including the flowable heat set gel of the present disclosure can be aseptically packaged as hot solution directly from the vessel, shear mixer, or homogeniser into larger volume sterile thermally-stable polyethylene bags.Example 12 - Effect of enzyme hydrolysis on gel strength

[0341] Two 500 mL Rubisco protein solutions at 25% TS was prepared. One solution was set aside as a control, and one was hydrolysed with 0.1% Flavourzyme at 60 °C for 5 minutes. Both samples were run on an Anton Paar MCR 302 rheometer over a temperature ramp of 25 - 95 °C, constant shear strain of 0.001 s-1, interval of 0.5 °C / s, using a parallel plate PP50 geometry. Partial enzyme proteolytic hydrolysis of Rubisco provides a lower strength gel as shown in Figure 9, with a decrease in measured gel strength G' from 65557 Pa for the control and 30772 Pa for the hydrolysed gel (see Table 1).To determine the repeatability of gel formation, two 500 mL Rubisco protein solutions at 15% TS were prepared. One sample was hydrolysed with 0.1% Flavourzyme at 60 °C for 5 minutes and then cooled on ice. The second sample was hydrolysed with 0.1% Flavourzyme at 60 °C for 5 minutes and then transferred to a 90 °C water bath for 30 mins to form the heat hydrolysed gel, following by high shear mixing for form the flowable heat set gel which was cooled in an ice bath. Both samples were run on a MCR 302 rheometer as described in the above experiment. As shown in Figure 10, the aqueous Rubisco which had not been subjected to thermal gelation was able to form heat set gel as evidenced by the increase in viscoelasticity, however the sample which had already been gelled and then sheared to a flowable gel was unable to form a new secondary gel, demonstrating that the gelation process requires non-denatured Rubisco protein.Example 13 - High protein flowable heat set gels and homogenised, formulated high protein content consumer gels using a scalable process

[0342] 99.5 kg of concentrated Rubisco protein (non-denatured) solution (26.4% solids, 79% protein purity w / w, viscosity 66.75 cP (ATAGO VISCO 6815 device, AIS spindle, 200 rpm, 18 °C) was preheated to 45 °C and pumped into a Scanima SRB200 high shear mixer. The product was slowly agitated (42 RPM) while hot water at 62 °C was used in the surrounding vessel jacket to heat the solution to the target temperature of 50°C over 5 mins. Prolyve BS CONC enzyme solution (170.6 ml of 10% solution of powder in water, Soufflet Biotechnologies) was added and dispersed into solution with mixing. The surrounding vessel temperature was left at 62 °C for 15 mins and the product heated to 55 °C within this time. The water jacket temperature was increased to 75 °C to bring the protein solution to 60 °C and to set the gel, with gentle agitation. During this time, optionally, a partial vacuum of 0.4 to 0.7 bar atmosphere was applied to the vessel to remove gases and assist with foam control. The heat set gel took 13 mins to set up. The gel was blended / sheared using the high shear mixer for 10 mins into a smooth, thin, flowing gel (flowable heat set gel, viscosity 2032 cP (ATAGO VISCO 6815 device, A2S spindle, 12 rpm, 18 °C)). This product can be heat treated, cooled and aseptically bagged, as below, without formulation inclusion.

[0343] The formulation composition (Table 13 below) was added and blended into the flowable heat set gel. The product was brought to 72°C and held for 11 mins using 82 °C hot water into the surrounding water jacket with mixing and agitation (42 RPM). The formulated gel product was smooth, thin, glossy, with an acceptable smell and a pleasant taste.

[0344] The formulated consumer gel is pumped into a Bertoli BH31008 2 stage homogeniser. The 1ststage pressure was 180 bar and the 2ndstage pressure was 20 bar. The viscosity of the homogenised formulated consumer gel was 95.98 cP measured with a ATAGO VISCO 6815 device (AIS spindle, 200 rpm, 18 °C). It is chilled to <5 °C through a plate heat exchanger and pumped into a filling hopper. 107 g of formulated gel is filled into a 100 mL consumer pouch using with a Hunter Filling Systems Series 300 Volumetric Filler. The consumer pouches are placed in a blast freezer at -30 °C and frozen.Table 13: Formulation composition proportions added to 99.5 kg Rubisco protein solution batchExample 14 - Effect of homogenisation on viscosity

[0345] Gel feed (28% total solids, 84.5% crude protein / dry matter) was divided into batches as outlined in Table 14 below. Each batch followed the parameters for hydrolysis, heat set, shear, formulation, pasteurisation, and homogenisation temperatures in Table 14. A Limitech P4 mixer was used for the hydrolysis, heat set, shear, formulation, and pasteurisation steps. A GEA Lab Homogeniser Panther 3006 was used for the homogenisation step. Viscosity measured on ATAGO VISCO 6815 device, AIS spindle, 200 rpm.

[0346] The viscosity of the pasteurised consumer gel and homogenised consumer gel demonstrate the homogenisation significantly reduces the viscosity of the sample. Sensory testing in a panel of 10 tasters showed the homogenised gel was strongly preferred compared to the non-homogenised consumer gel, having a creamier mouthfeel with less astringency and chalkiness.Table 14: Batch detailsTable 15: Ingredients added per batch in gExample 15 - High protein smoothie and nutritional beverage

[0347] A frozen portion of pasteurised, homogenised unflavoured consumer gel was combined in a blender (Ninja® Personal Blender) with frozen, chilled or fresh fruit, vegetables and juices and blended until smooth, to provide a smoothie or nutritional beverage with increased protein content in a convenient serve.Table 16: Ingredients per serve for plant protein supplemented smoothieTable 17: Nutritional informationExample 15 - Flowable heat set gel preparations and formulated consumer gel preparations

[0348] Flowable heat set gels were prepared at the conditions outlined in Table 18 on the Limitech F4 mixer except for trial T383 which was a Scanima SRB200, and were homogenised on the GEA Lab Homogeniser Panther 3006 at 200 bar 1st stage pressure, 0 bar second stage pressure except for T282 which was homogenised on the Bertoli from example 13 at 180 bar. The flowable heat set gels were formulated and pasteurised at 75 °C for 11 minutes. The viscosities of the flowable heat set gel were measured using the A2S spindle at 12 rpm and on the AIS spindle at 12 rpm, 100 rpm, and 200 rpm and are provided in Table 19.Table 18: Preparation conditions for six batches of flowable heat set gel and formulated gelTable 19: Viscosity data for the flowable heat set gels (ATAGO VISCO 6815, 18 °C)

[0349] As expected due to shear thinning / pseudoplastic behaviour, the flowable heat set gel ranged in viscosity at the different speed ranges (shear rate); when measured on the A2S spindle the highest measured viscosity was 17504 cP, at 12 rpm and lowest was 1735 cP, at 12 rpm. Some of the flowable heat set gels had viscosities too high to be measured accurately at other speeds (HHH result).Table 20: Viscosity data for the formulated consumer geis (ATAGO VISCO 6815 AIS. 200 rpm, 18 °C)

[0350] The corresponding viscosity data for the formulated consumer demonstrates the viscosity reduction after ingredient addition and homogenisation, and range between 34.89 cP to 432.2 cP when measured at 200 rpm (Table 19).Example 16 - Reduced oil or fat content formulated consumer gel preparations

[0351] Heat set gel (25.3% total solids, 85% crude protein / dry matter) prepared using the method of Example 13 was divided into equal portions, combined with binders and variable amounts of oil and homogenised on a bench top homogeniser (HG-15A Homogeniser, DAIHAN Scientific) at power 30 for 1-2 min to achieve a homogeneous mixture. The resultant sheared product was homogenised on a GEA Lab Homogenizer PandaPLUS 2000 with a 150 bar first stage and a 50 bar second stage. The remaining ingredients in Table 21 were added and homogenised on the bench top homogeniser (HG-15A Homogeniser). The product was packed into 100 mL pouches and pasteurised, cooled and frozen to provide formulated consumer gel preparations with 5%, 3%, 1% and 0% oil content.Table 21 : Ingredients added per batch in gOil Inclusion rate

[0352] The texture and mouthfeel of the homogenised consumer gel with different oil content was evaluated by a sensory panel of 10 tasters which showed that the high pressure homogenised gel with a reduced oil content (3% and 1%) provided an increase in creaminess and mouthfeel, a visually glossy consistency, a smoothing of flavour notes and minimal chalkiness which was similar or superior in texture to a high oil content (5%) in a non-high pressure homogenised gel.

Claims

1. CLAIMS1. A process for preparing a flowable heat set gel comprising from about 15% to about 28% w / w of Rubisco protein and having a viscosity up to about 18000cP (measured with an ATAGO VISCO 6815 device, A2S spindle, 12rpm, 18°C), the process comprising the steps of: a. Providing an aqueous Rubisco solution comprising from about 7.5% to about 28% non-denatured Rubisco protein, b. Subjecting the aqueous non-denatured Rubisco solution to partial enzymatic hydrolysis in a static or gently agitated environment to provide a partially hydrolysed aqueous Rubisco solution, c. Heating the partially hydrolysed aqueous Rubisco solution to form a heat set gel, d. Subjecting the heat set gel to shear to provide the flowable heat set gel, wherein the heat set gel and / or the flowable heat set gel is subjected to a heating step to deactivate the enzymes and stop enzymatic hydrolysis.

2. The process of claim 1, wherein the flowable heat set gel has a viscosity between about 1000 and about 18000cP (measured with an ATAGO VISCO 6815 device, A2S spindle, 12rpm, 18 °C).

3. The process of claim 1 or claim 2, wherein the flowable heat set gel has a a. particle size distribution Dx(10) of from about 1.5 to about 3.5 pm, b. particle size distribution Dx(50) of from about 3.5 to about 15 pm, and / or c. particle size distribution Dx(90) of from about 15 to about 70pm.

4. The process of any one of claims 1 to 3, wherein the flowable heat set gel is combined with additives to provide a formulated gel.

5. The process of any one of claims 1 to 4, wherein the flowable heat set gel or the formulated gel is pasteurised.

6. The process of any one of claims 1 to 5, wherein static or gently agitated in step b. comprises mixing or agitation of a solution between about 0 rpm to about 30 rpm.

7. The process of any one of claims 1 to 6, wherein the enzymes used in the partial enzymatic hydrolysis of step b. are selected from proteases or transferases which can be thermally deactivated.

8. The process of any one of claims 1 to 7, wherein the enzymes used in the partial enzymatic hydrolysis of step b. are dosed at concentrations of 0.01-1.5% w / w Rubisco protein content.

9. The process of any one of claims 1 to 8, wherein the temperature in step b. is between about 45 °C and about 65 °C.

10. The process of any one of claims 1 to 9, wherein the partial enzymatic hydrolysis step b. occurs for a time of between about 5 and about 20 minutes.

11. The process of any one of claims 1 to 10, wherein the temperature in step c. is between about 40 °C to about 75 °C.

12. The process of any one of claims 1 to 11, wherein in step d. the shear is performed for between about 1 to about 20 minutes.

13. The process of any one of claims 4 to 12, wherein the formulated gel comprises between about 70 to 95% w / w of the flowable heat set gel.

14. The process of any one of claims 4 to 13, wherein the additives are selected from any one or more of a. sugars and carbohydrates, b. fats or oils, c. fruit juice, fruit concentrate or puree, d. flavours, e. preservatives, f. sweetener, andg. gums or stabilisers.

15. The process of claim 5, wherein the pasteurised, flowable heat set gel or the pasteurised, formulated gel undergoes a homogenisation step.

16. A flowable heat set gel when prepared by the process of any one of claims 1 to 15.

17. A flowable heat set gel comprising from about 15% to about 28 % w / w of Rubisco protein, wherein the flowable heat set gel has a viscosity of up to about 18000cP (measured with an ATAGO VISCO 6815 device, A2S spindle, 12rpm, 18 °C).

18. The flowable heat set gel of claim 17, wherein the flowable heat set gel has a. particle size distribution Dx(10) of from about 1.5 to about 3.5 pm, b. particle size distribution Dx(50) of from about 3.5 to about 15 pm, and / or c. particle size distribution Dx(90) of from about 15 to about 70pm.

19. The flowable heat set gel of any one of claims 17 or claim 18 that is pasteurised.

20. A food product comprising the flowable heat set gel of any one of claims 16 to 19, the food product comprising between about 12.5% to about 25 % w / w of Rubisco protein, together with suitable additives selected from any one or more of:(i) sugars and carbohydrates,(ii) fats or oils,(iii) fruit juice, fruit concentrate or puree,(iv) flavours,(v) preservatives,(vi) sweetener,(vii) gums or stabilisers,, and(viii) vitamins or other nutritional supplements,wherein the food product has a viscosity of up to about 18000cP (measured with an ATAGO VISCO 6815 device (A2S spindle, 12rpm, 18 °C)).

21. A food product comprising the flowable heat set gel of any one of claims 16 to 19, the food product comprising between about 12.5% to about 25 % w / w of Rubisco protein, together with suitable additives selected from any one or more of:(i) sugars and carbohydrates,(ii) fats or oils,(iii) fruit juice, fruit concentrate or puree,(iv) flavours,(v) preservatives,(vi) sweetener(vii) gums or stabilisers(viii) vitamins or other nutritional supplements, wherein the food product has a viscosity of between 30 and 1000 cP (measured with an ATAGO VISCO 6815 device, AIS spindle, 200 rpm, 18 °C).

22. The food product of claim 20 or claim 21, wherein the food product has a a. particle size distribution Dx(10) of from about 1.0 to about 3.0 pm, b. particle size distribution Dx(50) of from about 3.0 to about 5.0 pm, and / or c. particle size distribution Dx(90) of from about 5.0 to about 15 pm.

23. The food product of any one of claims 20 to 22, the food product comprising between about 70 to 95% w / w of the flowable heat set gel.

24. The food product of any one of claims 20 to 23 comprising any one or more of:(i) from about 2 to about 10% w / w sugar,(ii) from about 1 to about 6% w / w fats and oils,(iii) from about 1 to about 8% w / w fruit juice, fruit concentrate or puree,(iv) from about 0 to about 10% w / w flavours,(v) from about 0.1 to about 1.5% w / w preservatives,(vi) from about 0 to about 3% w / w sweetener, and(vii) from about 0.1 to about 1% w / w gums or stabilisers.

25. The food product of any one of claims 20 to 24 that is pasteurised.

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