Plant-based food gel

Abstract

Disclosed herein is an alternative foodstuff, which may be formulated as a plant-based cheese analogue. Also disclosed herein are methods for forming the foodstuff articles, and uses of the foodstuff.

Description

PLANT-BASED FOOD GELCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] The present application claims priority from Australian Provisional Patent Application No 2024904216 filed on 19 December 2024, the contents of which are incorporated herein by reference in their entirety.TECHNICAL FIELD

[0002] The present disclosure provides alternative foodstuff, including in the form of a plant-based cheese analogue. Also disclosed herein are methods for forming the foodstuff articles.BACKGROUND

[0003] The development of plant-based food items, such as cheese analogs, has gained traction lately as consumers are seeking plant-based alternatives to animal-sourced food due to environmental, ethical, and health concerns. At present, several issues have limited the consumer perception and this includes the acceptance of plant-based cheese analogs. This includes poor sensory attributes (such as taste, aroma, meltability, and mouthfeel), and nutritional content. In particular, the meltability of plant-based cheese has been one of the greatest hurdles in mimicking dairy cheese.

[0004] There is a needed to identify alternative approaches to develop and obtain new plant-based foodstuff, that could be used in place of animal-sourced products.

[0005] Any discussion of documents, acts, materials, devices, articles or the like which has been included in the present specification 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 of each of the appended claims.SUMMARY

[0006] In a first aspect, disclosed herein is a plant-based foodstuff comprising:(i) at least one gelling agent, optionally in an amount of between about 0.1% (w / w) to about 2.0% (w / w), about 0.1% (w / w) to about 1.5% (w / w), about 0.1% (w / w) to about 1.0% (w / w), about 0.2% (w / w) to about 0.8% (w / w) or about 0.2% (w / w) to about 0.6% (w / w);(ii) at least one starch, optionally in an amount of between about 4% (w / w) to about 20% (w / w);(iii) at least one plant-based oil; and(iv) optionally at least one source of protein, wherein:(a) in use, the plant-based foodstuff transitions from the form of a first gel network to a second gel network following the application of heat; or(b) in use, the plant-based foodstuff is in the form of a first gel network at a first pre-determined temperature and a second gel network at a second predetermined temperature, and wherein the storage (G1) and loss (G") moduli measured in a rheometer of the first gel network present solid-like properties at room temperature and transition into a softer gel network with lower storage and loss moduli upon heating and during the formation of the second gel network.

[0007] In a second aspect disclosed herein is a plant-based foodstuff comprising:(i) at least one gelling agent in an amount of between about 0.1% (w / w) to about1.0% (w / w), about 0.2% (w / w) to about 0.8% (w / w) or about 0.2% (w / w) to about 0.6% (w / w);(ii) at least one starch in an amount of between about 4% (w / w) to about 20% (w / w);(iii) at least one plant-based oil; and(iv) optionally at least one source of protein, wherein, in use, the plant-based foodstuff transitions from the form of a first gel network to a second gel network following the application of heat.

[0008] In a third aspect, disclosed herein is a plant-based foodstuff comprising:(i) at least one gelling agent;(ii) at least one starch;(iii) at least one plant-based oil; and(iv) optionally at least one source of protein, wherein, in use, the plant-based foodstuff is in the form of a first gel network at a first pre-determined temperature and a second gel network at a second pre-determined temperature, and wherein the storage (G1) and loss (G") moduli measured in a rheometer of the first gel network present solid-like properties at room temperature and transition into a softer gel network with lower storage and loss moduli upon heating and during the formation of the second gel network.

[0009] In a fourth aspect, disclosed herein is a method of producing a plant-based foodstuff, the method comprising:(i) mixing a solution of at least one source of protein and at least one plant-based oil to form an emulsion at a pre-determined temperature;(ii) adding at least one gelling agent to the emulsion; and(iii) adding at least one starch to the emulsion, wherein when steps (i) to (iii) are performed in order an emulsion gel is formed, and wherein the emulsion gel is (iv) optionally cooled to form the plant-based foodstuff.

[0010] In a fifth aspect, disclosed herein is a plant-based foodstuff produced according to the method of the fourth aspect.

[0011] In a sixth aspect, disclosed herein is a plant-based foodstuff according to the first, second or third aspect, when produced according to the method of the third aspect.

[0012] In a seventh aspect, disclosed herein is the use of a plant-based foodstuff, wherein the plant-based foodstuff is defined according to the first, second or third aspect, as a plant-based cheese analogue, a fat analogue and / or a fat component in plant-based meat.

[0013] In an eighth aspect, disclosed herein is the use of a plant-based foodstuff, wherein the plant-based foodstuff is defined according to the first, second or third aspect,when produced according to the method of the fourth aspect, as a plant-based cheese analogue, a fat analogue and / or a fat component in plant-based meat.

[0014] Other aspects and embodiments relating to the present disclosure are described herein.

[0015] It will be appreciated that each example, aspect and embodiment of the present disclosure described herein is to be applied mutatis mutandis to each and every other example, aspect or embodiment unless specifically stated otherwise.

[0016] The present disclosure is not to be limited in scope by the specific examples described herein, which are intended for the purpose of exemplification only. Functionally-equivalent substituents, compositions, methods and processes are clearly within the scope of the disclosure as described herein.BRIEF DESCRIPTION OF DRAWINGS

[0017] Whilst it will be appreciated that a variety of embodiments disclosed herein may be utilised, described herein are a number of examples with reference to the following drawings:

[0018] Figure 1 - Average Synchrotron FTIR spectra and laboratory -based FTIR spectra. A-B. The average spectra of GRF emulsion gel samples were randomly selected from 15 carbohydrate, protein, and lipid-rich regions. C. Laboratory -based FTIR spectra for heated and unheated emulsion gel and constituent polysaccharides.

[0019] Figure 2 - Photograph of KC-GRF emulsion gel and schematic representation of changes in internal structure during heating. A. Dairy mozzarella. B. 8% GRF emulsion gel. C. Schematic representation of the changes in internal structure at different phases. Phase 1 : Chilled, rigid gel product to be applied on food. Phase 2: Thermal reversion of KC network when heated to intermediate temperature. Phase 3: Gelatinization of the starch network at high temperature. Phase 4: Soft gel after partial cooling for consumption.

[0020] Figure 3 - Confocal Laser Scanning Microscopy (CLSM) images of PPLKC- GRF emulsion gel. A. CLSM images before cooking (Phase 1) of PPI-KC-GRF emulsion gel at various KC, GRF and oil content. B. CLSM images after cooking (Phase 4) (3 min, 90 °C) of PPI-KC-GRF emulsion gel at various KC, GRF and oil content. Red: lipid; Blue: Starch; Green: PPI, KC, starch. 8% GRF, 0.6% KC, and 15% oil are identical samples.

[0021] Figure 4 - Split-channel confocal laser scanning microscopy (CLSM) images of 8% GRF emulsion gel. Channels are split to images excited at 488 nm (Nile red, lipid) 633 nm (Nile blue A, protein and KC), and 861 nm (FITC, starch).

[0022] Figure 5 - Synchrotron-FTIR chemical images depicting the location of lipid, carbohydrates, and protein within KC-GRF emulsion gel before and after heating at 8% and 12% GRF. The scale bar in the image represents 10pm.

[0023] Figure 6 - Frequency sweep and temperature sweep of KC-GRF emulsion gel. A-C. Frequency sweep at 25 °C of KC-GRF emulsion gel at various KC, GRF or oil content. D-F. Temperature sweep of KC-GRF emulsion gel at various KC, GRF or oil content.

[0024] Figure 7 - 0% GRF and 4% GRF sample after heating and partial cooling. No gel was seen for the 0% GRF sample after cooling to 50°C after heating at 90°C for 15 minutes and incubation at 50°C for 30 minutes.

[0025] Figure 8 - Frequency sweep and amplitude sweep of KC-GRF emulsion gel at 50°C. A-C. Frequency sweep of KC-GRF emulsion gel at various KC, GRF or oil content. D-F. Amplitude sweep of KC-GRF emulsion gel at various KC, GRF or oil content.

[0026] Figure 9. Freeze-thaw stability of PPI-KC-GRF emulsion gel as indicated by percentage weight (%) change of emulsion gels at varying GRF content (A); at varying KC content (B); at varying Oil content (C), and dairy cheeses and PBCC (D) after each cycle up to 5 freeze-thaw cycles.

[0027] Figure 10 - Meltability of PPI-KC-GRF emulsion gel, dairy cheese, and commercial plant-based cheese analogs. Al -3. Percentage increase in sample area afterheating of PPI-KC-GRF emulsion gel at varying GRF, KC, and oil content. Bl -3. Circular representation of increased area after heating of emulsion gel and commercial dairy cheeses (90 °C, 7 min). D. Photograph of shredded dairy mozzarella (DI), PPI-KC-GRF emulsion gel (D2), PBMC1 (D3) and PBMC2 (D4) before heating over a pizza base. E. Photograph of shredded dairy mozzarella (El), PPI-KC-GRF emulsion gel (E2), PBMC1 (E3), and PBMC2 (E4) after heating over a pizza base (180 °C, 10 min).

[0028] Figure 11 - Image of plant-based cheddar cheese (PBCC) before (Al) and after heating (A2) and circular representation of meltability (B). Color legend for circular representation: Pink: PBCC, Grey: Initial size, Orange, Dairy mozzarella, Black: Dairy Cheddar. PBCC recorded slight shrinkage after a modified Schreiber test at 90°C for 7 minutes.

[0029] Figure 12 - Temperature sweep of PPI-KC-GRF emulsion gel (Phase 2 - Phase 3). A. Varying GRF content. B. Varying KC content. C. Varying oil content. D. Mozzarella and PBMC1.DESCRIPTION OF EMBODIMENTSGeneral Terms

[0030] In the following description, reference is made to the accompanying drawings which form a part hereof, and which is shown, by way of illustration, several embodiments. It is understood that other embodiments may be utilised and structural changes may be made without departing from the scope of the present disclosure.

[0031] With regards to the definitions provided herein, unless stated otherwise, or implicit from context, the defined terms and phrases include the provided meanings. Unless explicitly stated otherwise, or apparent from context, the terms and phrases below do not exclude the meaning that the term or phrase has acquired by a person skilled in the relevant art. The definitions are provided to aid in describing particular embodiments, and are not intended to limit the claimed invention, because the scope of the invention is limited only by the claims. Furthermore, unless otherwise required by context, singular terms shall include pluralities and plural terms shall include the singular.

[0032] All publications discussed and / or referenced herein are incorporated herein in their entirety.

[0033] Throughout this disclosure, unless specifically stated otherwise or the context requires otherwise, reference to a single step, composition of matter, group of steps or group of compositions of matter shall be taken to encompass one and a plurality (i.e., one or more) of those steps, compositions of matter, groups of steps or groups of compositions of matter. Thus, as used herein, the singular forms “a”, “an” and “the” include plural aspects unless the context clearly dictates otherwise. For example, reference to “a” includes a single as well as two or more; reference to “an” includes a single as well as two or more; reference to “the” includes a single as well as two or more and so forth.

[0034] Those skilled in the art will appreciate that the disclosure herein is susceptible to variations and modifications other than those specifically described. It is to be understood that the disclosure includes all such variations and modifications. The disclosure also includes all of the examples, steps, features, methods, processes, and compositions, referred to or indicated in this specification, individually or collectively, and any and all combinations or any two or more of said steps or features.

[0035] The term “and / or”, e.g., “X and / or Y” shall be understood to mean either “X and Y” or “X or Y” and shall be taken to provide explicit support for both meanings or for either meaning.

[0036] Unless otherwise indicated, the terms “first,” “second,” etc. are used herein merely as labels, and are not intended to impose ordinal, positional, or hierarchical requirements on the items to which these terms refer. Moreover, reference to a “second” item does not require or preclude the existence of lower-numbered item (e.g., a “first” item) and / or a higher-numbered item (e.g., a “third” item).

[0037] As used herein, the phrase “at least one of’, when used with a list of items, means different combinations of one or more of the listed items may be used and only one of the items in the list may be needed. The item may be a particular object, thing, or category. In other words, “at least one of’ means any combination of items or number ofitems may be used from the list, but not all of the items in the list may be required. For example, “at least one of item A, item B, and item C” may mean item A; item A and item B; item B; item A, item B, and item C; or item B and item C. In some cases, “at least one of item A, item B, and item C” may mean, for example and without limitation, two of item A, one of item B, and ten of item C; four of item B and seven of item C; or some other suitable combination.

[0038] As used herein, the term “about”, unless stated to the contrary, typically refers to a range of up to + / - 10% of the designated value, and includes smaller ranges therein, for example + / - 5% or + / - 1% of the designated value.

[0039] It is to be appreciated that certain features that are, for clarity, described herein in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, various features that are, for brevity, described in the context of a single embodiment, may also be provided separately or in any sub-combination.

[0040] Throughout the present specification, various aspects and components of the invention can be presented in a range format. The range format is included for convenience and should not be interpreted as an inflexible limitation on the scope of the invention. Accordingly, the description of a range should be considered to have specifically disclosed all the possible sub-ranges as well as individual numerical values within that range, unless specifically indicated. For example, description of a range such as from 1 to 5 should be considered to have specifically disclosed sub-ranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 5, from 3 to 5 etc., as well as individual and partial numbers within the recited range, for example, 1, 2, 3, 4, 4.5, 4.75, and 5, unless where integers are required or implicit from context. This applies regardless of the breadth of the disclosed range. Where specific values are required, these will be indicated in the specification.

[0041] Throughout this specification the word "comprise", or variations such as "comprises" or "comprising", will be understood to imply the inclusion of a stated element, integer or step, or group of elements, integers or steps, but not the exclusion of any other element, integer or step, or group of elements, integers or steps.

[0042] Throughout this specification, the term "consisting essentially of' is intended to exclude elements which would materially affect the properties of the claimed composition, method or process.

[0043] The terms "comprising", "comprise" and "comprises" herein are intended to be optionally substitutable with the terms "consisting essentially of', "consist essentially of', "consists essentially of, "consisting of, "consist of' and "consists of, respectively, in every instance.

[0044] Herein “weight %” may be abbreviated to as “wt%” or “wt.%”. The weight % may be w / w or w / v, unless specifically indicated or clear from context.Specific Terms

[0045] Herein, the terms “storage modulus or moduli” or “elastic modulus or moduli” or “G” or “E'” refers to the amount of energy needed to distort a material, for example a first and / or a second gel network.

[0046] Herein, the terms “loss modulus or moduli” or “viscous modulus or moduli” or “G"’ or “E"” refers to the amount of energy lost when a material, for example a first and / or a second gel network, returns to its original shape after being distorted or deformed.

[0047] Herein a “solid” refers to a substance that displays structural rigidity, and resistance to a force applied to at least one surface. Unlike a liquid, a solid object does not flow to take on the shape of its container, nor does it expand to fill the entire available volume like a gas. In other words, a first and / or a second gel network as referred to herein, may be considered having a solid phase when the storage modulus is higher than the loss modulus.

[0048] Herein a “semi-solid” refers to a state of matter that displays one or more properties of a solid and a liquid. For example, the semi-solid may not hold its shape, but does not flow like a liquid. In other words, a first and / or a second gel network asreferred to herein, may be considered having a semi-solid phase when the storage modulus and loss modulus are about equal.

[0049] Herein “liquid” refers to a phase of matter that conforms to the shape of its container but retains a nearly constant volume independent of pressure. In other words, a first and / or a second gel network as referred to herein, may be considered having a liquid phase when the storage modulus is less than the loss modulus.Foodstuff

[0050] Disclosed herein is a plant-based foodstuff comprising:(i) at least one gelling agent in an amount of between about 0.1% (w / w) to about 2.0% (w / w), about 0.1% (w / w) to about 1.5% (w / w), about 0.1% (w / w) to about 1.0% (w / w), about 0.2% (w / w) to about 0.8% (w / w) or about 0.2% (w / w) to about 0.6% (w / w);(ii) at least one starch in an amount of between about 4% (w / w) to about 20% (w / w);(iii) at least one plant-based oil; and wherein, in use, the plant-based foodstuff transitions from the form of a first gel network to a second gel network following the application of heat.

[0051] Also disclosed herein is a plant-based foodstuff comprising:(i) at least one gelling agent;(ii) at least one starch;(iii) at least one plant-based oil; and(iv) optionally at least one source of protein, wherein, in use, the plant-based foodstuff is in the form of a first gel network at a first pre-determined temperature and a second gel network at a second pre-determined temperature, and wherein the storage (G1) and loss (G") moduli, optionally measured in a rheometer of the first gel network present solid-like properties at room temperature and transition into a softer gel network with lower storage and loss moduli upon heating and during the formation of the second gel network.

[0052] Also disclosed herein is a plant-based foodstuff comprising:(i) at least one gelling agent;(ii) at least one starch;(iii) at least one plant-based oil; and(iv) optionally at least one source of protein, wherein, in use, the plant-based foodstuff is in the form of a first gel network at a first pre-determined temperature and a second gel network at a second pre-determined temperature, and wherein the storage (G1) and loss (G") moduli, optionally measured in a measured in a rheometer, of the second gel is lower than the first gel.

[0053] Herein the G' and G" can be measured by any appropriate method known in the art, for example by using ASTM D7175 - 15, as approved 1 July 2015, or ISO 3219- 1 :2021. Alternatively, a rheometer with a parallel plate geometry can be used, to conduct a temperature sweep between 25°C and 90°C. The sample may be heated at a rate of 5°C on a Peltier heater. The G' and G" of the gel may be measured at a constant 0.5% strain and 1.0 rad / s.

[0054] In one or more embodiments the plant-based foodstuff is in the form of a solidphase.

[0055] In one or more embodiments the first gel network above the first pre-determined temperature and below the second pre-determined temperature is in the form of a liquidphase and / or the second gel network at the second pre-determined temperature is in the form of a semi-solid phase.

[0056] In one or more embodiments, the storage (G1) and loss (G") moduli for the first gel network at the first pre-determined temperature is between about 900 Pa to 42000 Pa (G1) and 200 to 7000 (G"). The G' and / or G" may be measured at a temperature (°C) of about, or at least about: 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95 or 100. The G' and / or G" may be measured at the first pre-determined temperature as defined herein. The G' and / or G" may be measured at a temperature (°C) of less than about: 100, 95, 90, 85, 80, 75, 70, 65, 60, 55, 50, 45, 40, 35, 30, 25, 20, or 15. The temperature may be in a range provided by any two of these upper and / or lower values. In one embodiment the temperature may be 25 °C, about 25 °C, 90°C or about 90 °C.

[0057] In one or more embodiments, the G' for the first gel network at the first predetermined temperature as defined herein, is about or at least about (in Pa): 900, 1000, 2000, 3000, 4000, 5000, 6000, 7000, 8000, 9000, 10000, 11000, 12000, 13000, 14000, 15000, 16000, 17000, 18000, 19000, 20000, 21000, 22000, 23000, 24000, 25000, 26000, 27000, 28000, 29000, 30000, 31000, 32000, 33000, 34000, 35000, 36000, 37000, 38000, 39000, 40000, 41000, or 42000. In one or more embodiments, the G' for the first gel network at the first pre-determined temperature as defined herein, is less than about (in Pa): 42000, 41000, 40000, 39000, 38000, 37000, 36000, 35000, 34000, 33000, 32000, 31000, 30000, 29000, 28000, 27000, 26000, 25000, 24000, 23000, 22000, 21000, 20000, 19000, 18000, 17000, 16000, 15000, 14000, 13000, 12000, 11000, 10000, 9000, 8000, 7000, 6000, 5000, 4000, 3000, 2000, 1000, or 900. In one or more embodiments, the G" for the first gel network at the first pre-determined temperature as defined herein may be in a range provided by any two of these upper and / or lower values

[0058] In one or more embodiments, the G" for the first gel network at the first predetermined temperature as defined herein, is about or at least about (in Pa): 200, 400, 600, 800, 1000, 1200, 1400, 1600, 1800, 2000, 2200, 2400, 2600, 2800, 3000, 3200, 3400, 3600, 3800, 4000, 4200, 4400, 4600, 4800, 5000, 5200, 5400, 5600, 5800, 6000, 6200, 6400, 6600, 6800, or 7000. In one or more embodiments, the G" for the first gel network at the first pre-determined temperature as defined herein, is less than about (in Pa): 7000, 6800, 6600, 6400, 6200, 6000, 5800, 5600, 5400, 5200, 5000, 4800, 4600, 4400, 4200, 4000, 3800, 3600, 3400, 3200, 3000, 2800, 2600, 2400, 2200, 2000, 1800, 1600, 1400, 1200, 1000, 800, 600, 400, or 200. In one or more embodiments, the G" for the first gel network at the first pre-determined temperature as defined herein may be in a range provided by any two of these upper and / or lower values.

[0059] In one or more embodiments, the G' for the second gel network at the second pre-determined temperature as defined herein, is about or at least about (in Pa): 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500, 525, 550, 575, 600, 625, 650, 675, 700, 725, 750, 775, 800, 825, 850, 875, or 900. In one or more embodiments, the G' for the second gel network at the second pre-determined temperature as defined herein, is less than about (in Pa): 900, 875, 850, 825, 800, 775, 750, 725, 700, 675, 650, 625, 600,575, 550, 525, 500, 475, 450, 425, 400, 375, 350, 325, 300, 275, 250, 225, or 200. In one or more embodiments, the G" for the second gel network at the second pre-determined temperature as defined herein may be in a range provided by any two of these upper and / or lower values.

[0060] In one or more embodiments, the G" for the second gel network at the second pre-determined temperature as defined herein, is about or at least about (in Pa): 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, or 500. In one or more embodiments, the G" for the second gel network at the second predetermined temperature as defined herein, is less than about (in Pa): 500, 475, 450, 425, 400, 375, 350, 325, 300, 275, 250, 225, 200, 175, 150, 125, 100, 75, or 50. In one or more embodiments, the G" for the second gel network at the second pre-determined temperature as defined herein may be in a range provided by any two of these upper and / or lower values.

[0061] In one or more embodiments the at least one gelling agent is selected from: pectin, sodium alginate, guar, konjac, xanthan gum, cellulose, glucose, methylcellulose, K-carrageenan, i-carrageenan, high-acyl gellan gum, curdlan gum, agar, konjac gum, locust bean gum, tara gum, cassia gum, and mixtures thereof.

[0062] In one or more embodiments the at least one gelling agent is selected from: K- carrageenan, i-carrageenan, high-acyl gellan gum, curdlan gum, agar, konjac gum, and mixtures thereof.

[0063] In one or more embodiments the at least one gelling agent is selected from: K- carrageenan, i-carrageenan, high-acyl gellan gum, and mixtures thereof.

[0064] In one or more embodiments the at least one gelling agent is present in an amount of about, or at least about (% w / w): 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0. 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9 or 2.0. In one or more embodiments the at least one gelling agent is present in an amount of less than about (% w / w): 2.0, 1.9, 1.8, 1.7, 1.6, 1.5, 1.4, 1.3, 1.2, 1.1, 1.0, 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2 or 0.1. In one or more embodiments the at least one gelling agent is present in an amount in a range or any two of the above values, for example in a range of about 0.1% (w / w) to about 2.0% (w / w),about 0.1% (w / w) to about 1.5% (w / w), about 0.1% (w / w) to about 1.0% (w / w), about 0.2% (w / w) to about 0.8% (w / w), about 0.2% (w / w) to about 0.4% (w / w) or about 0.2% (w / w) to about 0.6% (w / w).

[0065] In one or more embodiments the at least one starch is amylopectin starch, optionally a high amylopectin starch. For example, the at least one starch may be one or more of: waxy potato starch, waxy corn starch, arrowroot starch, sweet potato starch, rice starch, waxy rice starch, waxy wheat starch, waxy barley starch, waxy sorghum starch, waxy millet starch, tapioca starch, taro starch, and mixtures thereof. For example, the at least one starch may be one or more of: waxy potato starch, arrowroot starch, sweet potato starch, rice starch, tapioca starch, and mixtures thereof. For example, the at least one starch may be one or more of: waxy potato starch, arrowroot starch, sweet potato starch, rice starch, and mixtures thereof.

[0066] In one or more embodiments the at least one starch is glutinous rice flour.

[0067] In one or more embodiments the at least one starch is present in an amount of about, or at least about (% w / w):l, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 or 25. In one or more embodiments the at least one starch is present in an amount of less than about (% w / w): 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2 or 1. In one or more embodiments the at least one starch is present in an amount in a range or any two of the above values, for example in a range of about 8% (w / w) to about 12% (w / w), or about 4% (w / w) to about 20% (w / w).

[0068] In one or more embodiments the first pre-determined temperature is about or at least about (in °C): 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, or 80. In one or more embodiments the first pre-determined temperature is less than about (in °C): 80, 75, 70, 65, 60, 55, 50, 45, 40, 35, 30, or 25. In one or more embodiments the first pre-determined temperature is in a range or any two of the above values, for example in a range of about 45 °C to about 75 °C, or about 50 °C to about 65 °C.

[0069] In one or more embodiments the second pre-determined temperature is about or at least about (in °C): 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95 or 100. In one or more embodiments the second pre-determined temperature is less than about (in °C):100, 95, 90, 85, 80, 75, 70, 65, 60, 55, 50, 45, 40, or 35. In one or more embodiments the second pre-determined temperature is in a range or any two of the above values, for example in a range of about 75 °C to about 95 °C, or about 80 °C to about 90 °C.

[0070] In one or more embodiments comprises at least one source of protein. The source of protein may be selected from: a pea protein isolate, soy protein isolate, potato protein isolate, rice protein isolate, fava bean protein isolate, chickpea protein isolate, peanut protein isolate, hemp protein isolate, almond protein isolate, sunflower seed protein isolate, mung bean protein isolate, and mixtures thereof.

[0071] In one or more embodiments the at least source of protein is present in an amount of about, or at least about (% w / w):l, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or15. In one or more embodiments the at least one source of protein is present in an amount of less than about (% w / w): 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2 or 1. In one or more embodiments the at least one source of protein is present in an amount in a range or any two of the above values, for example in a range of about 3% (w / w) to about 10% (w / w).

[0072] In one or more embodiments the plant-based oil is selected from: canola oil, rapeseed oil, olive oil, avocado oil, walnut oil, hemp oil, almond oil, avocado seed oil, corn oil, cottonseed oil, flax seed oil, grapeseed oil, peanut oil, sesame oil, sunflower oil, safflower oil, rice bran oil, coconut oil (for example medium chain triglyceride coconut oil), soybean oil, palm oil, and mixtures thereof.

[0073] In one or more embodiments, the at least one plant-based oil is present in an amount of about, or at least about (% w / w): 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15,16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, or 40. In one or more embodiments the at least one plant-based oil is present in an amount of less than about (% w / w): 40, 39, 38, 37, 36, 35, 34, 33, 32, 31, 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2 or 1.. In one or more embodiments the at least one plant-based oil is present in an amount in a range or any two of the above values, for example in a range of about 5% (w / w) to about 30% (w / w).

[0074] In one or more embodiments, the plant-based foodstuff may further comprise one or more optional additives. Examples of additives include, but are not limited to one or more of: flavours, seasonings, colours, antioxidants, stabilisers and mixtures thereof. For example, the plant-based foodstuff may comprise at least one of lactic acid, citric acid, sodium chloride, calcium chloride or beta-carotene. For example, the plant-based foodstuff may comprise at least one of lactic acid, citric acid or beta-carotene.

[0075] In one or more embodiments the plant-based foodstuff is a plant-based cheese analogue (or a fat analogue).

[0076] In one or more embodiments the plant-based foodstuff is as a fat analogue which may be used as a fat component in plant-based meat.Methods

[0077] One or more advantages of the present disclosure are provided by the novel method steps, as described here, for producing a plant-based foodstuff. The inventors have surprisingly found that by first forming a stable emulsion from at least one source of protein and at least one plant-based oil, the internal structure of the plant-based foodstuff may be controlled. It is contemplated that the formation of the stable emulsion may advantageously provide at least one of the following: (i) improved sensory attributes such as taste, aroma, meltability, and / or mouthfeel, and (ii) improved nutritional content.

[0078] Disclosed herein is a method of producing a plant-based foodstuff, the method comprising:(i) mixing a solution of at least one source of protein and at least one plant-based oil to form an emulsion at a pre-determined temperature;(ii) adding at least one gelling agent to the emulsion; and(iii) adding at least one starch to the emulsion, wherein when steps (i) to (iii) are performed in order an emulsion gel is formed, wherein the emulsion gel is (iv) optionally cooled to form the plant-based foodstuff.

[0079] It will be appreciated that formation of the emulsion in step (i), as described here, may advantageously allow flexibility for forming two interchanging gel matrices, i.e. (a) from addition of at least one gelling agent and (b) from addition of at least onestarch. In other words, when steps (i) to (iii) are performed in order an emulsion gel is formed. One or more advantages of the present disclosure are provided by performing method steps (i) to (iii) in order, and may advantageously provide at least one of the following: (i) a stable emulsion to form the emulsion gel in the water phase, (ii) even distribution of the at least one gelling agent in the water phase, and (iii) formation of a plant-based foodstuff, i.e. a cold-set gel, on addition of the at least one starch during optional cooling step (iv).

[0080] In one or more embodiments mixing step (i) further comprises:(a) optionally rehydrating the at least one source of protein in water to form a rehydrated protein solution,(b) pre-heating the rehydrated protein solution at a pre-determined temperature A, and / or(c) optionally homogenizing the rehydrated protein solution, prior to addition of the at least one plant-based oil.

[0081] In one or more embodiments the pre-determined temperature A is about or at least about (in °C): 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100. In one or more embodiments the pre-determined temperature A is less than about (in °C): 100, 99, 98, 97, 96, 95, 94, 93, 92, 91, 90, 89, 88, 87, 86, 85, 84, 83, 82, 81, 80, 79, 78, 77, 76, 75, 74, 73, 72, 71, or 70. In one or more embodiments the second pre-determined temperature is in a range or any two of the above values, for example in a range of about 70 °C to about 99 °C, or about 80 °C to about 99 °C.

[0082] In one or more embodiments mixing step (i) further comprises (d) optionally cooling and / or storing the emulsion.

[0083] In one or more embodiments step (ii) further comprises:(a) heating the emulsion to a pre-determined temperature B; and (b) optionally stirring the emulsion for a first period of time.

[0084] In one or more embodiments the pre-determined temperature B is about or at least about (in °C): 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67,68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100. In one or more embodiments the pre-determined temperature B is less than about (in °C): 100, 99, 98, 97, 96, 95, 94, 93, 92, 91, 90, 89, 88, 87, 86, 85, 84, 83, 82, 81, 80, 79, 78, 77, 76, 75, 74, 73, 72, 71, 70, 69, 68, 67, 66, 65, 64, 63, 62, 61, 60, 59, 58, 57, 56, 55, 54, 53, 52, 51, or 50. In one or more embodiments the second pre-determined temperature is in a range or any two of the above values, for example in a range of about 50 °C to about 99 °C, about 70 °C to about 99 °C , about 70 °C to about 99 °C, or about 80 °C to about 99 °C.

[0085] In one or more embodiments, the first period of time is (in minutes), about or at least about: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, or 40. In one or more embodiments, the first period of time mate be in a range (in minutes), of less than about: 40, 39, 38, 37, 36, 35, 34, 33, 32, 31, 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1. In one or more embodiments the first period of time is in a range or any two of the above values, for example in a range of about 1 to about 40 minutes or about 5 to about 30 minutes.

[0086] In one embodiment, the first period of time is a sufficient time for the emulsion to be uniform or substantially uniform.

[0087] In one or more embodiments step (iii) further comprises:(a) heating the emulsion to a pre-determined temperature C; and(b) optionally stirring the emulsion for a second period of time.

[0088] In one or more embodiments the pre-determined temperature C is about or at least about (in °C): 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67,68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90,91, 92, 93, 94, 95, 96, 97, 98, 99, or 100. In one or more embodiments the pre-determined temperature C is less than about (in °C): 100, 99, 98, 97, 96, 95, 94, 93, 92, 91, 90, 89,88, 87, 86, 85, 84, 83, 82, 81, 80, 79, 78, 77, 76, 75, 74, 73, 72, 71, 70, 69, 68, 67, 66,65, 64, 63, 62, 61, 60, 59, 58, 57, 56, 55, 54, 53, 52, 51, or 50. In one or more embodiments the second pre-determined temperature is in a range or any two of theabove values, for example in a range of about 50 °C to about 99 °C, about 70 °C to about 99 °C , about 70 °C to about 99 °C, or about 80 °C to about 99 °C.

[0089] In one or more embodiments, the second period of time is (in minutes), about or at least about: 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, or 160. In one or more embodiments, the second period of time mate be in a range (in minutes), of less than about: 160, 155, 150, 145, 140, 135, 130, 125, 120, 115, 110, 105, 100, 95, 90, 85, 80, 75, 70, 65, 60, 55, 50, 45, 40, 35, 30, 25, or 20. In one or more embodiments the first period of time is in a range or any two of the above values, for example in a range of about 20 to about 160 minutes or about 30 to about 150 minutes.

[0090] In one embodiment, the second period of time is a sufficient time to ensure the complete rehydration of the gelling agent.

[0091] In one or more embodiments for the methods as defined herein, the at least one gelling agent is as defined herein. For example, the at least one gelling agent is selected from: pectin, sodium alginate, guar, konjac, xanthan gum, cellulose, glucose, methylcellulose, K-carrageenan, i-carrageenan, high-acyl gellan gum, curdlan gum, agar, konjac gum, locust bean gum, tara gum, cassia gum, and mixtures thereof. In one or more embodiments of the methods as defined herein the at least one gelling agent is present in an amount of about, or at least about (% w / w): 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9 or 2.0; or present in an amount of less than about (% w / w): 2.0, 1.9, 1.8, 1.7, 1.6, 1.5, 1.4, 1.3, 1.2, 1.1, 1.0, 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2 or 0.1. In one or more embodiments the at least one gelling agent is present in an amount in a range or any two of the above values, for example in a range of about 0.1% (w / w) to about 2.0% (w / w), about 0.1% (w / w) to about 1.5% (w / w), about 0.1% (w / w) to about 1.0% (w / w), about 0.2% (w / w) to about 0.8% (w / w), about 0.2% (w / w) to about 0.4% (w / w), or about 0.2% (w / w) to about 0.6% (w / w).

[0092] In one or more embodiments for the methods as defined herein, the at least one starch is a amylopectin starch, optionally a high amylopectin starch. For example, the at least one starch may be one or more of: waxy potato starch, waxy com starch, arrowrootstarch, sweet potato starch, rice starch, waxy rice starch, waxy wheat starch, waxy barley starch, waxy sorghum starch, waxy millet starch, tapioca starch, taro starch, and mixtures thereof. For example, the at least one starch may be one or more of: waxy potato starch, arrowroot starch, sweet potato starch, rice starch, and mixtures thereof. In one or more embodiments of the methods as defined herein, the at least one starch is glutinous rice flour. In one or more embodiments of the methods as defined herein the at least one starch is present in an amount of about, or at least about (% w / w):l, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 or 25; or present in less than about (% w / w): 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2 or 1. In one or more embodiments the at least one starch is present in an amount in a range or any two of the above values, for example in a range of about 8% (w / w) to about 12% (w / w), or about 4% (w / w) to about 20% (w / w).

[0093] In one or more embodiments of the methods as defined herein, the source of protein may be selected from: a pea protein isolate, soy protein isolate, potato protein isolate, rice protein isolate, fava bean protein isolate, chickpea protein isolate, peanut protein isolate, hemp protein isolate, almond protein isolate, sunflower seed protein isolate, mung bean protein isolate, and mixtures thereof.

[0094] In one or more embodiments of the methods as defined herein, the at least source of protein is present in an amount of about, or at least about (% w / w): l, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or 15; or present in less than about (% w / w): 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2 or 1; or present in an amount in a range or any two of the above values, for example in a range of about 3% (w / w) to about 10% (w / w).

[0095] In one or more embodiments of the methods as defined herein the plant-based oil is selected from: canola oil, rapeseed oil, olive oil, avocado oil, walnut oil, hemp oil, almond oil, avocado seed oil, corn oil, cottonseed oil, flax seed oil, grapeseed oil, peanut oil, sesame oil, sunflower oil, safflower oil, rice bran oil, coconut oil, medium chain triglyceride coconut oil, soybean oil, palm oil, cacao butter, and mixtures thereof. In one or more embodiments of the methods as defined herein the plant-based oil is selected from: canola oil, olive oil, avocado oil, walnut oil, hemp oil, almond oil, avocado seed oil, corn oil, cottonseed oil, flax seed oil, grapeseed oil, peanut oil, sesame oil, sunfloweroil, safflower oil, rice bran oil, coconut oil (for example medium chain triglyceride coconut oil), soybean oil, palm oil and mixtures thereof. In one or more embodiments of the methods as defined herein, the at least one plant-based oil is present in an amount of about, or at least about (% w / w): 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, or 40; or is present in an amount of less than about (% w / w): 40, 39, 38, 37, 36, 35, 34, 33, 32, 31, 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2 or 1; or in a range or any two of the above values, for example in a range of about 5% (w / w) to about 30% (w / w).

[0096] Also disclosed herein is a plant-based foodstuff produced according to any method as described herein.Uses

[0097] Disclosed herein is the use of a plant-based foodstuff, as defined herein as a plant-based cheese analogue.

[0098] Disclosed herein is the use of a plant-based foodstuff, as defined herein as a plant-based cheese analogue, wherein the plant-based foodstuff is produced according to a method as described herein.

[0099] Disclosed herein is the use of a plant-based foodstuff, as defined herein as a fat analogue.

[0100] Disclosed herein is the use of a plant-based foodstuff, as defined herein as a fat component in plant-based meat.EXAMPLE EMBODIMENTS

[0101] The present disclosure may be described by one or more of the following example embodiments:1. A plant-based foodstuff comprising:(i) at least one gelling agent in an amount of between about 0.1% (w / w) to about 1.0% (w / w), about 0.2% (w / w) to about 0.8% (w / w) or about 0.2% (w / w) to about 0.6% (w / w);(ii) at least one starch in an amount of between about 4% (w / w) to about 20% (w / w);(iii) at least one plant-based oil; and(iv) optionally at least one source of protein, wherein, in use, the plant-based foodstuff transitions from the form of a first gel network to a second gel network following the application of heat.2. A plant-based foodstuff comprising:(i) at least one gelling agent;(ii) at least one starch;(iii) at least one plant-based oil; and(iv) optionally at least one source of protein, wherein, in use, the plant-based foodstuff is in the form of a first gel network at a first pre-determined temperature and a second gel network at a second pre-determined temperature, and wherein the storage (G1) and loss (G") moduli measured in a rheometer of the first gel network present solid-like properties at room temperature and transition into a softer gel network with lower storage and loss moduli upon heating and during the formation of the second gel network.3. The plant-based foodstuff according to example embodiment 1 or example embodiment 2, wherein the foodstuff is in the form of a solid-phase.4. The plant-based foodstuff according to any one of the preceding example embodiments, wherein the first gel network above the first pre-determined temperature and below the second pre-determined temperature is in the form of a liquid-phase and the second gel network at the second pre-determined temperature is in the form of a semisolid phase.5. The plant-based foodstuff according to the any one of the preceding example embodiments, wherein the storage (G1) and loss (G") moduli for the first gel network at the first pre-determined temperature is between about 900 Pa to 42000 Pa (G1) and 200 to 7000 (G") and the storage (G1) and loss (G") moduli for the second gel network at the second pre-determined temperature is between about 200 to 900 Pa (G1) and 50 to 500 Pa (G).6. The plant-based foodstuff according to any one of the preceding example embodiments, wherein the at least one gelling agent is selected from: K-carrageenan, i- carrageenan, high-acyl gellan gum, curdlan gum, agar, konjac gum, and mixtures thereof.7. The plant-based foodstuff according to any one of the preceding example embodiments, wherein the at least one gelling agent is selected from: K-carrageenan, i- carrageenan, high-acyl gellan gum, and mixtures thereof.8. The plant-based foodstuff according to any one of the preceding example embodiments, wherein the at least one gelling agent is present in an amount of between about 0.2% (w / w) to about 0.8% (w / w) or about 0.2% (w / w) to about 0.4% (w / w).9. The plant-based food stuff according to any one of the preceding example embodiments, wherein the at least one starch is a amylopectin starch, optionally a high amylopectin starch.10. The plant-based food stuff according to any one of the preceding example embodiments, wherein the at least one starch is glutinous rice flour.11. The plant-based foodstuff according to any one of the preceding example embodiments, wherein the at least one starch is present in an amount of between about 8%(w / w) to about 12% (w / w).12. The plant-based foodstuff according to any one of the preceding example embodiments, wherein the first pre-determined temperature is in a range of between about 45 °C to about 75 °C.13. The plant-based foodstuff according to any one of the preceding example embodiments, wherein the first pre-determined temperature is in a range between about 50 °C to about 65 °C.14. The plant-based foodstuff according to any one of the preceding example embodiments, wherein the second pre-determined temperature is in a range of between about 75 °C to about 95 °C.15. The plant-based foodstuff according to any one of the preceding example embodiments, wherein the second pre-determined temperature is in a range between about 80 °C to about 90 °C.16. The plant-based foodstuff according to any one of the preceding example embodiments, further comprising a source of protein.17. The plant-based foodstuff according to example embodiment 16, wherein the source of protein is selected from: a pea protein isolate, soy protein isolate, potato protein isolate, rice protein isolate, fava bean protein isolate, chickpea protein isolate, peanut protein isolate, hemp protein isolate, almond protein isolate, sunflower seed protein isolate, and mixtures thereof.18. The plant-based foodstuff according to any one of the preceding example embodiments, wherein the at least one source of protein is present in an amount of between about 3% (w / w) to about 10% (w / w).19. The plant-based foodstuff according to any one of the preceding example embodiments, wherein the plant-based oil is selected from: canola oil, olive oil, avocadooil, walnut oil, hemp oil, almond oil, avocado seed oil, corn oil, cottonseed oil, flax seed oil, grapeseed oil, peanut oil, sesame oil, sunflower oil, safflower oil, rice bran oil, , coconut oil (for example medium chain triglyceride coconut oil), soybean oil, palm oil and mixtures thereof.20. The plant-based foodstuff according to any one of the preceding example embodiments, wherein the at least one plant-based oil is present in an amount of between about 5% (w / w) to about 30% (w / w).21. The plant-based foodstuff according to any one of the preceding example embodiments, further comprising one or more optional additives, optionally one or more flavours, seasonings, colours, antioxidants, stabilisers and mixtures thereof.22. The plant-based foodstuff according to any one of the preceding example embodiments, wherein the plant-based foodstuff is a plant-based cheese analogue or a fat analogue.23. A method of producing a plant-based foodstuff, the method comprising:(i) mixing a solution of at least one source of protein and at least one plant-based oil to form an emulsion at a pre-determined temperature;(ii) adding at least one gelling agent to the emulsion; and(iii) adding at least one starch to the emulsion, wherein when steps (i) to (iii) are performed in order an emulsion gel is formed, wherein the emulsion gel is (iv) optionally cooled to form the plant-based foodstuff.24. The method according to example embodiment 23, wherein mixing step (i) further comprises:(a) optionally rehydrating the at least one source of protein in water to form a rehydrated protein solution,(b) pre-heating the rehydrated protein solution at a pre-determined temperature A, optionally in a range about 70 °C to about 99 °C, or in a range of about 80 °C to about 99 °C, and / or(c) optionally homogenizing the rehydrated protein solution, prior to addition of the at least one plant-based oil.25. The method according to example embodiment 23 or example embodiment 24, wherein mixing step (i) further comprises (d) optionally cooling and / or storing the emulsion.26. The method according to any one of example embodiments 23 to 25, wherein step (ii) further comprises:(a) heating the emulsion to a pre-determined temperature B, optionally in a range about 50 °C to about 99 °C, about 60 °C to about 99 °C, about 70 °C to about 99 °C, or in a range of about 80 °C to about 99 °C; and(b) optionally stirring the emulsion for a first period of time, and optionally for about 1 to about 40 minutes or about 5 to about 30 minutes.27. The method according to any one of example embodiments 23 to 26, wherein step (iii) further comprises:(a) heating the emulsion to a pre-determined temperature C, optionally in a range about 50 °C to about 99 °C, about 60 °C to about 99 °C, about 70 °C to about 99 °C, or in a range of about 80 °C to about 99 °C; and(b) optionally stirring the emulsion for a second period of time, and optionally for about 20 to about 160 minutes or about 30 to about 150 minutes.28. The method according to any one of example embodiments 23 to 27, wherein the at least one gelling agent is as defined in example embodiment 6 or example embodiment 7.29. The method according to any one of example embodiments 23 to 28, wherein the at least one gelling agent is present in an amount of about 0.2% (w / w) to about 0.8% (w / w) or about 0.2% (w / w) to about 0.6% (w / w).30. The method according to any one of example embodiments 23 to 29, wherein the at least one starch is as defined in example embodiment 9 or example embodiment 10.31. The method according to any one of example embodiments 23 to 30, wherein the at least one starch is present in an amount of about 4% (w / w) to about 20% (w / w).32. The method according to any one of example embodiments 23 to 31, wherein the at least one source of protein is as defined in example embodiment 17.33. The method according to any one of example embodiments 23 to 32, wherein the at least one source of protein is present in an amount of about 3% (w / w) to about 10% (w / w).34. The method according to any one of example embodiments 23 to 33, wherein the at least one plant-based oil is as defined in example embodiment 19.35. The method according to any one of example embodiments 23 to 34, wherein the at least one plant-based oil is present in an amount of about 5% (w / w) to about 30% (w / w).36. A plant-based foodstuff produced according to the method of any one of example embodiments 23 to 35.37. A plant-based foodstuff according to any one of example embodiments 1 to 22, when produced according to the method of any one of example embodiments 23 to 35.38. Use of a plant-based foodstuff, wherein the plant-based foodstuff is defined according to any one of example embodiments 1 to 22, as a plant-based cheese analogue, a fat analogue and / or a fat component in plant-based meat.39. Use of a plant-based foodstuff, wherein the plant-based foodstuff is defined according to any one of example embodiments 1 to 22, when produced according to the method of any one of example embodiments 23 to 35, as a plant-based cheese analogue, a fat analogue and / or a fat component in plant-based meat.EXAMPLES

[0102] The present disclosure may be described by one or more of the following examples.Materials and MethodsMaterials

[0103] Pea protein isolate (85% protein) was purchased from Bioflex Nutrition Pty Ltd. (Grove, TAS, Australia), kappa-carrageenan (KC) was purchased from The Melbourne Food Depot (Melbourne, VIC, Australia). GRF from Erawan Marketing Co., Ltd. (Bangkok, Thailand) and canola oil were purchased from a local supermarket. Commercial plant-based mozzarella / pizza cheese shreds from Bio (PBMC1) and Made with Plants (PBMC2), plant-based cheddar cheese from Bio (PBCC), dairy mozzarella, and dairy cheddar were purchased from a local supermarket. All other reagents used were analytical grade.Emulsion gel preparation

[0104] An emulsion of (pea protein isolate) PPI, water, and oil was first prepared. PPI was rehydrated and dispersed in MiliQ water at room temperature (21±1 °C). The PPI dispersion was then heated in an 85 °C water bath for 30 minutes before the addition of oil. The oil and PPI dispersion was then pre-mixed using an overhead mixer at 800 rpm for 3 minutes. The mixture was then homogenized using a high-shear homogenizer at 12,000 rpm for 3 minutes (IKA Ultra Turrax T25, Staufen, Germany). The emulsion wasallowed to cool to room temperature after homogenization and refrigerated pending further use.Table 1. Final Composition of KC-GRF samples. All percentages depicted are w / w.Sample PPI (%) Oil (%) KC (%) GRF (%) Water (%)0% GRF 5 15 0.6 0 79.44% GRF 5 15 0.6 4 75.48% GRF 5 15 0.6 8 71.412% GRF 5 15 0.6 12 67.40.2% KC 5 15 0.2 8 71.80.4% KC 5 15 0.4 8 71.60.8% KC 5 15 0.8 8 71.25% Oil 5 5 0.6 8 81.410% Oil 5 10 0.6 8 76.420% Oil 5 20 0.6 8 66.4

[0105] The PPI / canola oil emulsion were measured by weight with MilliQ water added to samples of varying GRF and KC content according to their final composition. The mixture were then heated to 80 °C and held for 10 min while stirring at 800 rpm with a magnetic stirrer. KC was added to the mixture and was allowed to fully rehydrate and disperse at 80 °C for 2 hours. The mixture was removed from the heater stirrer after 2 hours and allowed to cool to 65 °C before GRF was added to the mixture and stirred using an overhead mixer at 800 rpm for 5 minutes. The mixture was then poured into a square mold and allowed to cool and set before refrigeration. The final composition of PPI-KC-GRF emulsion gel samples are presented in Table 1.Rheological measurements

[0106] Rheological measurements of the PPI-KC-GRF emulsion gels were conducted using a strain-controlled rheometer (TA Instruments ARES-G2, New Castle, USA) with the Advanced Peltier System (APS) for temperature control. A 40 mm diameter steel cross-hatched geometry was used (upper and lower). To create thin slices of samples, 4 g of emulsion gel mixture was cast into a square plastic mold and rotated to ensure an even spread. The samples were then placed over a level surface to set and refrigerated overnight before testing. A 38.5 mm diameter ring cutter was then used to extract acircular cutting of the sample. The sample discs were gently transferred to the lower geometry and compressed to a 1 mm gap. Silicone oil was applied to the perimeter of the sample and the geometries were placed under a solvent trap to minimize evaporation.Frequency Sweep

[0107] A frequency sweep was conducted twice, initially on the sample at 25 °C and after heating-cooling at 50 °C. The viscoelasticity of the emulsion gel was evaluated between a frequency range of 0.1 to 100 rad / s. The storage modulus (G1) and loss modulus (G") were measured for both tests at different temperatures.Temperature Sweep

[0108] A temperature sweep of the emulsion gel was measured to study the gel-sol -gel transition during heating. Immediately following the frequency sweep at 25 °C, the sample was heated to 90 °C at a rate of 5°C / min. G’ and G’ ’ were measured at a constant 0.5% strain and a frequency of 1.0 rad / s. The sample was held at 90 °C for 1 minute before being cooled to 50 °C at a rate of 5 °C / min for frequency and amplitude sweeps.Amplitude sweep

[0109] An amplitude sweep of each emulsion gel sample was conducted after a frequency sweep at 50 °C. The samples were subjected to an oscillation strain between 0.01 to 1000% strain at a frequency of 1 rad / s. The G’ and G” were recorded to identify the change in gel strength.Confocal laser scanning microscopy (CLSM)

[0110] A confocal laser scanning microscope (CLSM, Zeiss LSM 800, Jena, Germany) was employed to observe the microstructure of the plant-based cheese. Nile blue A and Nile red were added to the warm emulsion / polysaccharide mixture to stain the PPI and oil, respectively. Fluorescein isothiocyanate (FITC) was added to the mixture as a fluorescence stain agent for starch due to a high affinity for gelatinized starch. A small portion of the stained mixture was placed onto a microscope slide and covered with a coverslip. Images were captured with an AxioCam 506 camera using a 10x objective lens by exciting at 488 nm for Nile Red, 633 nm for Nile Blue A, and 561 nm for FITC.Synchrotron macro ATR-FTIR microspectroscopy

[0111] The synchrotron-FTIR experiment was performed on Infrared Microspectroscopy (IRM) beamline (Australian Synchrotron, part of ANSTO, Victoria, Australia), using a Bruker Vertex 80v spectrometer coupled with a Hyperion 3000 FTIR microscope and a liquid nitrogen-cooled narrow-band mercury cadmium telluride (MCT) detector (Bruker Optik GmbH, Ettlingen, Germany). The synchrotron-FTIR spectra were obtained within a spectral range of 3800-700 cm'1using 4 cm'1spectral resolution. Three characteristic bands, corresponding to v(C=O) of esters from lipid triglycerides (1740 cm-1), amide I (1670 cm-1), and polysaccharides (1055-984 cm-1), were integrated to create chemical images that revealed the distribution of lipids, proteins, and polysaccharides specific for canola oil, pea protein, KC and GRF, based on the laboratory-based FTIR-spectra of the raw ingredients (Figure 1). Average spectra were derived from 15 individual spectra of lipid-rich, protein-rich, and polysaccharide-rich areas.Freeze-thaw stability

[0112] The freeze-thaw stability of the emulsion gel is important for frozen food applications. The freeze-thaw stability of the emulsion gels was measured using a procedure modified by Cui et al. (2022). Briefly, 12 g of warm emulsion gel mixture was poured into a 50 ml centrifuge tube and was left to set at room temperature and refrigerated overnight before freezing. Each emulsion gel-loaded tube was frozen in a - 20 °C freezer for 22 hours, after which the samples were thawed in a 30 °C water bath for 2 hours. Moisture was removed from the top of the gel using a sheet of Kim wipe. The difference in weight before and after the removal of moisture was recorded. Triplicates of each formulation were tested over five consecutive cycles. The water syneresis was calculated using equation (1).Where Wnis the weight of the sample at the n111cycle and Wois the initial weight of the sample prior to the first cycle.Texture analysis

[0113] Texture analysis was conducted on the emulsion gels before and after heating. Texture Profile Analysis (TP A) was used to measure the mechanical properties of the emulsion gel before heating. Emulsion gel samples were twice compressed using a P / 50 uniaxial probe. All samples were cut into a 1.8 cm x 1.8 cm x 1.8 cm cube for testing. The samples were tested using the following parameters: 75% strain, Imm / s test speed, 5 seconds wait time, and 5 mm / s post-test speed.

[0114] The gel strength of the heated samples was measured using an adapted procedure for gelatin bloom strength (Chandra & Shamasundar, 2014). Briefly, 8 g of samples were loaded in small glass vials and heated in a 90 °C water bath for 15 minutes to ensure the complete gelatinization of starch. The samples were then placed in a 50 °C water bath for 30 minutes for the temperature to equilibrate before testing. A P / 10 probe was used to penetrate the heated gels at a depth of 4 mm. Hence, the gel strength is the maximum force required to achieve the depth.Meltability

[0115] Emulsion gels were cut into discs of 29 mm diameter and 7 mm height. The sample was then placed over an aluminum baking pan and heated in a forced air oven at 90 °C for 7 minutes. An image of the sample before and after heating was captured using a DSLR camera (D5600, Nikon, Tokyo, Japan). ImageJ was used to measure the change in area before and after heating. All samples were allowed to warm to room temperature for 1 hour before heating.

[0116] For visual comparison, 12 g of shredded plant-based mozzarella analog, dairy mozzarella, and 8% GRF (yellow food coloring added) were added over a 5 cm x 10 cm commercial pizza base each and heated at 180 °C for 10 minutes. Images before and after heating were captured before and after heating using a DSLR camera (D5600, Nikon, Tokyo, Japan).Extensibility

[0117] The extensibility of the emulsion gels and reference cheese samples were measured through an adapted method by Akarca et al. (2023) of lateral hand extensionof heated emulsion gels. 10 g of shredded emulsion gels and cheese were loaded into a 50 ml centrifuge tube and heated at 90 °C for 15 minutes. The sample was then removed from the centrifuge tube and slowly pulled laterally from either end of the sample until the gel disconnected. The maximum length was measured using an electronic calipers.Statistical analyses

[0118] Triplicates were conducted for the above tests, with results reported as average and their standard deviations. A one-way ANOVA and Duncan’s test were carried out using SPSS (version 26.0, IBM Corp., Armonk, NY) to determine statistical differences at P<0.05. All figures were prepared using OriginPro (version 2023, OriginLab Corporation, US).Results and discussionAppearance and mechanism of KC-GRF cheese analog

[0119] After the cold-set gelation of KC in the emulsion-poly saccharide mixture, the gel presents as a free-standing soft material that is yellowish-white. A 1.5 xl.5xl.5 cm3sample was photographed and can be seen in Figure 2B with a cube of dairy mozzarella cheese for comparison (Figure 2A). Dairy cheese appeared predominantly yellow in contrast to the PPI-KC-GRF cheese analog.

[0120] A schematic of possible mechanisms of changes in the PPI-KC-GRF emulsion gel during cooking is presented in Figure 2C. Starting from a rigid gel (Phase 1), as seen in Figure 2B, the gel transitions into a “melted” state due to the helix-coil transition of KC at elevated temperatures (Phase 2). At this phase, the gel structure was significantly weakened,, and the mixture was in a flowable state where spread could occur. Upon further heating, the gelatinization of starch occurs and forms a second network (Phase 3). The starch gel remains soft upon cooling to eating temperatures (Phase 4). These observations and their implications on the structural formation of the emulsion gel were discussed in the following sections through microstructural, rheological, and textural changes.Microstructure - Confocal laser scanning microscopy

[0121] The CLSM micrographs of the PPI-KC-GRF cheese analog with various GRF, KC, and oil content can be seen in Figure 3. The micrographs show the samples in Phase 1 (Figure 2A) and Phase 2 (Figure 3). PPI-KC and oil content were seen to have a greater impact on oil droplet size than GRF, which seems to be largely consistent between 0- 12%. As KC was added during heating, an increased amount of KC may have increased the stability of the droplet by increasing viscosity, thereby providing more repellent force between the droplets. An increase in oil content had minimal visible size differences up to 15% oil, while oil leaking and coalescence of droplets were observed for 20% oil.

[0122] Phase separation between the dispersed phase (lipid) and the continuous phase (water) was observable at 0.8% KC (Figure 3A). A high concentration of KC appeared to have led to the depletion flocculation of oil droplets in the gel matrix due to the increase in osmotic pressure induced by the hydrophilicity of KC and incompatibility between protein and KC. Nonetheless, the degree of flocculation was observed to a lesser extent when GRF content was increased. This may be due to the emulsion-holding ability of native rice starch and protein with oil.

[0123] The micrographs support the mechanism outlined in Figure 2C, as can be seen from the FITC labeled starch (blue). Figure 3A and Figure 4 show starch present as granules dispersed throughout the gel matrix in Phase 1 (Figure 2C). Gelatinization of starch after heating at 90 °C was seen in Figure 3B, where a phase change of starch from granular to swollen was observed. The morphology of the gelatinized starch seen in Figure 3B and Figure 4 is consistent with those observed for high amylopectin starch under CLSM. Thus, the results indicated that heating had induced a morphological change in the emulsion gel. Moreover, heating appeared to have increased the oil droplet size. Nonetheless, the absence of obvious oil leakage for all samples supports the emulsion gel's stability after heating.Microstructure - Synchrotron macro ATR-FTIR microspectroscopy

[0124] Synchrotron macro ATR-FTIR microspectroscopy was used to examine the differences between 8% GRF and 12% GRF samples to further elucidate the impact of heating n both morphology and chemical structures in these plant-based cheeses. Figure5 shows the chemical-images of lipids (1740 cm-1), polysaccharides (1055-984 cm-1), and proteins (1670 cm-1) of 8% GRF and 12% GRF samples before and after heating. The images reveal the spatial distribution and relative concentration of these biomolecular components within the emulsion gel, as indicated by their localization and absorption intensity. In particular, protein molecules were co-localized with lipid droplets at their interface between the dispersed phase and the continuous aqueous phase. An increase in polysaccharide-rich regions was observed when the GRF content increased from 8% to 12% before heating.

[0125] The chemical-images for polysaccharides were found to correlate with CLSM observations (Figure 3A and 3B), where polysaccharide-concentrated regions were evident in both 8% GRF and 12% GRF samples prior to heating. After heating, these localised regions disappeared, and the polysaccharide signals appeared to distribute more uniformly across the images, as shown in Figure 5 (B2 and B4). This change aligns with Figure 3 A and 3B, showing that starch granules initially in Phase 1 granular state (Figure 3A) underwent swelling upon heating (Figure 3B). The absorption intensity of the polysaccharide signals was higher for 12% GRF sample than 8% GRF sample after heating indicating that the observed signal primarily originates from starch and that increased starch concentration led to enhanced swelling and gelatinization. These findings are supported by the chemical image of polysaccharide-rich areas (Figure 1), which shows a reduction in intensity at peaks around 1047 cm'1indicative of a decreased crystallinity. A similar trend was observed in the crystallinity / amorphous ratio (1047 cm' V 1022 cm'1), where heating reduced this ratio for both 8% GRF and 12% GRF.Rheology - Frequency sweep at 25 °C

[0126] Frequency sweep at 25 °C reveals the gel strength of the KC network at Phase 1, where its mechanical strength is essential to the shelf stability of plant-based cheese before consumption, as well as its resistance to stress during processing (i.e. cooking). As seen in Figure 6 A-C, all samples displayed solid-like rheology where G ’ > G ” across the frequency range measured (0.1-100 rads / s). A weak gel was observed for all samples as G ’ and G ’ ’ were frequency dependent.

[0127] An increase in GRF, KC, and oil content all increased the G ’ of the gel. but with different roles in the emulsion gels from each ingredient. The greatest impact on G’ was observed from altering the total KC content from 0.2%-0.8% (w / w) , to further confirm that KC was the principal gel matrix at Phase 1. The effect of increased GRF content on G’ may have originated from the reduction in porosity of the KC network, which has increased the density of the emulsion gel. On the other hand, the lipid phase appeared to be an active filler in Phase 1, where an increase in G’ was observed from increasing oil content. Like the observation from Figure 3A, where increasing GRF content did not induce flocculation of oil droplets, the existence of native rice starch granules could have assisted in the maintenance of emulsion stability even as oil content increased. This could be due to the existence of surface protein on these granules (6.7% (w / w) protein), providing hydrophobic sites for lipid stabilization while interacting with the continuous KC network through electrostatic interactions.Rheology - Temperature sweep

[0128] The temperature sweep provides insights into the rheological properties of the emulsion gel during cooking during Phases 2 and 3. Figure 6 D-F and 12 show the temperature sweep of emulsion gels at varying GRF, KC, and oil content between 25-90 °C. Samples displayed thermal reversibility of the KC network during heating. At lower temperatures (up to 50 °C), the gel strength of the samples reflects that of the frequency sweep at 25 °C. A weakening of the emulsion gel between 50 °C- 75 °C, presented as a significant reduction in the difference between G' and G" was seen in all samples. In which, the G' of 8% GRF samples reduced from being 5.00x (25.19 °C) to 1.45x (72.11 °C) of G". The initial reduction in G' and G" indicates the helix-coil transition of KC and the melting of a solid-like emulsion gel, which could be a proxy for the melting behavior of cheese (Phase 2). The gelatinization of starch and later a starch-based gel was observable after the temperature reached 70 °C, where G' increases after reaching a minimum. The recorded temperature appeared largely in line with the pasting temperature of GRF in literature. In contrast, no subsequent increase in G' was recorded for 0% GRF samples even at the conclusion of heating at 90 °C (Figure 6D). The trendingof G' towards G" due to increasing temperature was also similar to that seen in dairy mozzarella up to 50°C (Figure 12D).

[0129] The profile of the temperature sweep varies as the emulsion gel’s composition changes with smaller reductions in G' observed for samples with increasing GRF, KC, and oil content. Particularly, increases in KC content appeared to have the greatest effect. The concave profile that stemmed from the aforementioned “melting” of KC and gelatinization of GRF was minimized for 0.8% KC samples (Figure 6E). This may be due to the increased energy and time required to complete the helix-coil transition of KC, which has overlapped with the onset of gelatinization of starch. The delay is consistent where an increase in KC content increases the gel melting enthalpy. On the other hand, the onset of thermal reversibility for KC appeared to be affected by the addition of starch, where samples containing starch saw a reduction in G' earlier than 0% GRF samples (Figure 6D).Rheology - Frequency and amplitude sweep at 50 °C

[0130] The frequency sweep of emulsion gels measured at 50 °C can be seen in Figure 8 A-C and 13 A for dairy mozzarella and PBMC1. The temperature was selected to simulate the typical eating temperature for cheese products that would be perceived as hot. At 50 °C, the gel strength of the emulsion gel is positively affected by GRF, KC (up to 0.6% KC), and oil (up to 15% oil). The greatest change was observed for GRF content, where its addition from 0% to 8-12% saw a one magnitude increase in G' (Figure 8A). Altering the KC content increases G'up to 0.6% KC, with samples at 0.8% KC showing a reduced gel strength comparable to 0.4% KC samples (Figure 8B). A similar trend was observed for altering oil content, where a maximum G' was achieved at 15% oil. This may be due to the coalescence of oil droplets and leaking (Figure 8B), which undermined the structure of the gel. 8% GRF and 10% oil achieved comparable G' to dairy mozzarella at 50 °C. Nonetheless, the tan 5 of dairy mozzarella remains higher than 8% GRF or 10% oil samples (Figure 8C), indicating stronger elasticity over mozzarella.

[0131] Figures 8 D-F show the amplitude sweep of all emulsion gel, dairy mozzarella, andPBMCl samples measured at 50 °C. The transition from the linear viscoelastic region(LVR) to non-linearity was visible for all samples as both G' and G" declined when the strain was increased from 0.01% to 1000%. Like the frequency sweep in Figure 8 A-C, the gel strength at the LVR increases as GRF content increases, indicating increased gel strength. Starch was observed to be the dominant network at 50 °C within the LVR, as KC and oil content appeared to have limited effects on G'. However, no discernible differences were observed in critical strain (yc) and the crossover point where G -G" for all samples, which indicates a complete loss of viscoelasticity.

[0132] The changes in microscale deformations as indicated in the rheological properties of emulsion gels, will be further investigated under macroscale deformations under texture profile analysis in the following sections.Freeze-thaw stability

[0133] Figure 9 show the freeze-thaw stability of the emulsion gels at Phase 1, dairy cheeses, and PBCC over five consecutive cycles. Overall, a high degree of freeze-thaw stability was observed for all samples except 0% GRF and 0.2% KC, which had a higher syneresis rate than other samples. The freeze-thaw stability of the emulsion gels was also similar to that of the dairy cheeses, and PBCC up until the 4thcycle, while parity was only achieved with cheddar and PBCC at the conclusion of the 5thcycle. Samples with varying GRF (4%-12%) and KC (0.4%-0.8%) content did not see significant differences in freeze-thaw stability over five cycles, with the final weight being 93.22%-92.27% and 93.76-88.74% for GRF and KC, respectively. The enhancement of freeze-thaw stability with increasing KC was in line with the rheological measurements (Figure 6B) and literature, where freeze-thaw stability increases as KC concentration increases due to enhanced gel strength. Poor freeze-thaw stability was recorded for pure KC gels, which further confirmed that GRF is essential in producing stable emulsion gels. GRF may have interacted with its surrounding KC network, thereby increasing the network's strength.Texture analysis

[0134] TPA was used to analyze the texture of the emulsion gel at Phase 1 and to benchmark against commercial plant-based cheese analog and dairy cheese (mozzarella and cheddar). Table 2 shows the respective TPA parameters measured for all samples.Increasing GRF, KC, and oil content, all had a statistically significant positive effect on hardness and a generally insignificant effect on springiness and cohesiveness. Hardness increased by 1.83 times and 4.60 times between the lowest and highest of the respective GRF and KC content tested. Only varying KC content was also seen to increase the springiness of the emulsion gel with statistical significance. All PPI-KC-GRF emulsion gels are significantly softer than dairy cheese, and commercial plant-based cheddar analog. The increase in hardness as GRF, KC, and oil content resonates with the data obtained from the frequency sweep at 25 °C (Figures 6 A-C).

[0135] Since the emulsion gel is significantly softer after heating, a gel strength measurement at 50 °C was conducted by penetration to measure the firmness of the emulsion gel for eating (Phase 4). The gel strength at 50 °C reflects similar relationships as the frequency and amplitude sweep at 50 °C (Figure 8). Increasing GRF content increases the gel strength. Notably, samples with 0% GRF could not form a solid-like gel network at 50 °C (Figure 7). This further supports that GRF is the dominant network at this temperature. Increasing KC content appeared to increase the gel strength up to 0.6% KC. At 0.8% KC, a significant weakening of gel strength was observed, possibly due to the phase separation observed in Figure 3B. Oil content appeared to have an insignificant impact on the gel strength at 50 °C. Again, dairy cheese was significantly firmer than PPI-KC-GRF emulsion gels, which are themselves also firmer than commercial plantbased cheese analogs.

[0136] Overall, texture analysis conducted at the two temperatures measured reflects their mechanical properties at various phases. The TPA measurements at 25 °C reflect the potential processing and handling characteristics of the emulsion gel. As cheese could be sliced or shredded to fit its role in the final food product, it is important for an analog to maintain structural integrity during these processes. The TPA results show that the mechanical characteristics of the gel are a product of GRF, KC, and oil content. Thus, these characteristics are tunable for flexible applications of the emulsion gel. Texture measurement at 50 °C is more reflective of the texture at eating temperatures. Improvements were seen over commercial plant-based cheese analogs using coconut oil. This was achieved even as canola oil was used as the lipid phase, which would notpartially crystallize upon cooling. This shows that the gel matrix of PPI-KC-GRF emulsion gels is superior to that seen in commercial plant-based cheese. Given the trend of increasing gel strength seen in increasing GRF content further optimization of GRF content in emulsion gels may bring the gel strength better inline with dairy cheese.Meltability

[0137] The modified Schreiber test was conducted to measure the melting characteristics of PPI-KC-GRF emulsion gels, dairy cheese, and plant-based cheddar analogs. Furthermore, qualitative observations were made for melted PPI-KC-GRF emulsion gels, shredded plant-based mozzarella analog, and dairy mozzarella cheese heated over a pizza base at 180 °C for 10 min. The test was chosen to provide a more suitable comparison to pre-shredded plant-based mozzarella analogs.

[0138] A plot and corresponding circular representation showing the change in the area after heating can be seen in Figure 10 A-C. The meltability of the emulsion gel appeared to be inversely affected by increasing GRF and KC content, with oil content presenting insignificant differences. The GRF content of the emulsion gel appeared to have the highest impact, reducing meltability from +190.47% (0% GRF) to +15.16% (12% GRF). On the other hand, increasing KC content led to a more modest reduction, from +70.70% (0.2% KC) to +12.39% (0.8% KC). Although all samples underperformed dairy cheese (Mozzarella: +271.36%, Cheddar: +284.67%), they performed remarkably better than the plant-based cheddar analog, which slightly shrank (-3.89%) after heating (Figure 11). The results are reminiscent of the temperature sweep shown in. Figure 6D, where a smaller decrease in the moduli corresponded to lower meltability. The combination of temperature-sweep and modified Schreiber test was able to provide a holistic observation of melting in PPI-KC-GRF emulsion gels.

[0139] Figure 10 D-E shows a comparison between dairy mozzarella, plant-based mozzarella analogs, and 8% GRF (dyed yellow). After high-temperature heating, dairy mozzarella samples and 8% GRF were seen to congeal into a continuous mass after heating, with little of the original shredded strands seen. Contrastingly, congealing of cheese strands was not seen to the same degree in both plant-based mozzarella cheese samples. Individual strands were still visible after heating with some softening andcongealing. This comparison shows that PPI-KC-GRF emulsion gels were more visually comparable to dairy mozzarella than plant-based mozzarella cheese analog. This may be due to the greater loss of gel structure from the helix-coil transition of KC, as shown in this study.Extensibility

[0140] The extensibility of dairy cheese, plant-based mozzarella cheese analog, and PPI-KC-GRF are recorded in Table 2. The manual stretchability measurement serves as a preliminary analysis of the emulsion gel’s susceptibility to lateral extension typically seen in cooked food. Like the textural analysis conducted at 50 °C, no gel network was formed at 0% GRF. An increase in GRF content saw an increase in the extensibility of the emulsion gel, with 12% GRF samples 92% more stretchy than 4% GRF. Since the GRF network is the dominant network at 50 °C, the textural quality is attributed to the high amylopectin content of glutinous rice starch. An increase in oil content saw an increase in extensibility up to 15% oil. The trend was reversed at 20% Oil, which saw a significant decrease in extensibility and aligns with previous rheological and microstructural observations. Overall, the extensibility was able to match that of plantbased mozzarella analogs but fell short of the extensibility of dairy mozzarella.Table 2. Texture Profile Analysis, gel strength and stretchiness of KC-GRF cheese and dairy cheeseMoz az4arell ± 5519907..9328 « 54 ±0.02 00.557 / ± ±00.00351±794468057±121549390227 / 44.2222±±3311.4411Cheddar47 ±77355..404100.1188 ±±00.002? 00.1199±±00.0000±262365358 ±6132755018% GRF, 0.6% KC and 15% Oil are identical samples.2PBMC 1 and PBMC 2 are shredded products.3Different superscript letters in the same column of the table indicate statistical significance in ANOVA (p < 0.05).4ANOVA was not conducted for TPA results (hardness, springiness, cohesiveness, and chewiness) between dairy cheeses, commercial plant-based cheese, and emulsion gels due to large differences in measured means. Instead, ANOVA was conducted individually for emulsion gels and reference samples.Discussion

[0141] This study presents a novel approach to creating cheese-like characteristics using plant-based materials. The lack of casein in plant-based cheese analog is the primary challenge in creating desirable thermal and mechanical characteristics of plant-based cheese. The proposed PPI-KC-GRF formulation successfully created two gel networks that individually exist at varying thermal and temporal phases. This was achieved by heating and exploiting the gap between the helix-coil transition temperature and the gelatinization temperature of starch. This technique allowed for the tailoring of important gel characteristics at different phases in processing and cooking.

[0142] A firm KC gel network was responsible for the structural integrity of the emulsion gel prior to cooking (Phase 1). This allows for the handling and processing of the emulsion gel into various shapes. Processes such as shredding are important in some cheese applications. As seen in Figure 10D2, the PPI-KC-GRF emulsion gel was able to maintain structural integrity after shredding. The formulation’s suitability as a food ingredient was then reinforced by the excellent freeze-thaw stability observed. The stability enables the emulsion gel to be used on frozen foods, such as pizzas and ready- to-eat meals, with extended shelf-life. Nonetheless, the current formulation is softer compared to PBCC and dairy cheeses, this is likely a result of the lack of solid fat content in the emulsion gel as compared to the coconut oil and milk fat seen in PBCC and dairy cheeses. The heating of the emulsion gel beyond 50 °C first melts and subsequently gels again. This was seen by the convergence and subsequent divergence of G' and G" as temperature increases, as seen in Figure 6 D-F. A soft, starch-structured emulsion gel for consumption was formed after partial cooling to 50 °C. Improvements over commercial plant-based cheese analogs were observed throughout the study. Sensory characteristics such as textual, meltability, and extensibility were also found to be tunable and may be further customized for various cheese types.

[0143] The proposed formulation was able to achieve melting with a significant reduction in G'in a temperature range that is closer to dairy mozzarella. The post-cooking rheology of the emulsion gel also better resembles mozzarella than commercial plantbased alternatives. In literature, the melting of mozzarella cheese initiates at 40 °C and completely melts at 60°C, which the current findings, where G1and G" cross between 48- 50 °C (Figure 12) are consistent. This is in contrast to other studies, where a gradual decline of G' was observed during heating at temperatures up to 100°C. This was made possible by the thermal gap between the reversal of KC and starch gelatinization wherewithin the gap, little gel structure exists which is distinct from other formulations. Although all studies, including the current one, do not show the complete loss of elasticity during heating, the initiation of melting at a temperature range more in line with dairy cheese may improve the consumer perception of plant-based cheese.

[0144] The melting characteristics observed in this study also stem from the ability to create stable emulsions from pea protein, which could accommodate the processing requirements of the two polysaccharides added. This differs from commercial solutions by negating the use of starch as the stabilizer of oil. Pre-heating of pea protein suspension at high concentrations prior to high shear emulsification may have increased the stability of the emulsion through pre-aggregation. This enabled full rehydration and solubilization of KC during extended heating at 80 °C, and no significant phase separation was observed during subsequent cooling and blending of GRF at 65 °C. The existence of protein within the gel matrix would also improve the nutritional profile of the emulsion gel, as current commercial plant-based cheese contains little to no protein.

[0145] The present disclosure supports the case of using double-polysaccharide systems to target the thermal properties of dairy cheese. The focus on thermal changes, both visually and in rheology, and texture could provide a solid base for future development could take place. Areas such as taste, aroma, and potential nutritional fortifications could further enhance the appeal of a double-polysaccharide cheese analog.Conclusions

[0146] Contemporary commercial plant-based cheese is limited by its inability to balance cheese-like characteristics and nutritional content, calling for a better solution. The present disclosure presents a new template for tailoring gel characteristics in plantbased cheese. Through the use of two polysaccharides, KC and GRF, an emulsion gel tailored for cheese application could be formed. The texture of the gel prior to and after heating could also resemble what is required of the emulsion gel at each respective stage. Meltability was achieved through gradual heating of the emulsion gel, which sequentially triggered the thermal reversibility of KC and gelatinization of GRF. Examples of the plant-based foodstuff disclosed herein show improvement in meltability, texture, and / or excellent freeze-thaw stability that enables its use as a plant-based cheese analog. Furtheroptimization at low KC content and increased GRF content may improve the meltability of the formulation while maintaining a rigid initial gel. Optimization of GRF content may also improve strength of gel and stretchability of the emulsion gel after heating which is essential for use in place of mozzarella cheese. Encapsulation of flavoring compounds and carotenoids for coloring may also be explored to improve the nutritional profile of the formulation and assess its overall sensory acceptability. Since the sample was created with only five ingredients, including water, and without the need for additional emulsifiers, it may assist in addressing consumer demand for clean-label foods. This study provides a foundation for future innovations in plant-based cheese, bridging the gap between sustainable ingredients, enhanced functionalities, and ultimately greater consumer acceptance.

[0147] It will be appreciated by persons skilled in the art that numerous variations and / or modifications may be made to the above-described embodiments, without departing from the broad general scope of the present disclosure. The present embodiments are, therefore, to be considered in all respects as illustrative and not restrictive.

Claims

CLAIMS:

1. A plant-based foodstuff comprising:(i) at least one gelling agent, optionally in an amount of between 0.1% (w / w) to about 2.0% (w / w), 0.1% (w / w) to about 1.5% (w / w), about 0.1% (w / w) to about 1.0% (w / w), about 0.2% (w / w) to about 0.8% (w / w) or about 0.2% (w / w) to about 0.6% (w / w);(ii) at least one starch, optionally in an amount of between about 4% (w / w) to about 20% (w / w);(iii) at least one plant-based oil; and(iv) optionally at least one source of protein, wherein:(a) in use, the plant-based foodstuff transitions from the form of a first gel network to a second gel network following the application of heat; or(b) in use, the plant-based foodstuff is in the form of a first gel network at a first pre-determined temperature and a second gel network at a second pre-determined temperature, and wherein the storage (G1) and loss (G") moduli measured in a rheometer of the first gel network present solid-like properties at room temperature and transition into a softer gel network with lower storage and loss moduli upon heating and during the formation of the second gel network.

2. The plant-based foodstuff according to claim 1, wherein the foodstuff is in the form of a solid-phase.

3. The plant-based foodstuff according to any one of the preceding claims, wherein the first gel network above the first pre-determined temperature and below the second pre-determined temperature is in the form of a liquid-phase and the second gel network at the second pre-determined temperature is in the form of a semi-solid phase.

4. The plant-based foodstuff according to the any one of the preceding claims, wherein the storage (G1) and loss (G") moduli for the first gel network at the first predetermined temperature is between about 900 Pa to 42000 Pa (G1) and 200 to 7000 (G")and the storage (G1) and loss (G") moduli for the second gel network at the second predetermined temperature is between about 200 to 900 Pa (G1) and 50 to 500 Pa (G1).

5. The plant-based foodstuff according to any one of the preceding claims, wherein: the at least one gelling agent is selected from: K-carrageenan, i-carrageenan, high-acyl gellan gum, curdlan gum, agar, konjac gum, and mixtures thereof; and / or the at least one gelling agent is selected from: K-carrageenan, i-carrageenan, high-acyl gellan gum, and mixtures thereof; and / or the at least one gelling agent is present in an amount of between about 0.2% (w / w) to about 0.8% (w / w), about 0.2% (w / w) to about 0.6% (w / w), or about 0.2% (w / w) to about 0.4% (w / w).

6. The plant-based food stuff according to any one of the preceding claims, wherein: the at least one starch is a amylopectin starch, optionally a high amylopectin starch; and / or the at least one starch is glutinous rice flour; and / or the at least one starch is present in an amount of between about 4% (w / w) to about 20% (w / w), or between about 8%(w / w) to about 12% (w / w).

7. The plant-based foodstuff according to any one of the preceding claims, wherein: the first pre-determined temperature is in a range of between about 45 °C to about 75 °C; and / or the first pre-determined temperature is in a range between about 50 °C to about 65 °C.

8. The plant-based foodstuff according to any one of the preceding claims, wherein: the second pre-determined temperature is in a range of between about 75 °C to about 95 °C; and / or the second pre-determined temperature is in a range between about 80 °C to about 90 °C.

9. The plant-based foodstuff according to any one of the preceding claims, further comprising a source of protein, wherein: optionally the source of protein is selected from: a pea protein isolate, soy protein isolate, potato protein isolate, rice protein isolate, fava bean protein isolate, chickpea protein isolate, peanut protein isolate, hemp protein isolate, almond protein isolate, sunflower seed protein isolate, mung bean protein isolate, and mixtures thereof; and / or optionally the at least one source of protein is present in an amount of between about 3% (w / w) to about 10% (w / w).

10. The plant-based foodstuff according to any one of the preceding claims, wherein: the at least one plant-based oil is selected from: canola oil, olive oil, avocado oil, walnut oil, hemp oil, almond oil, avocado seed oil, corn oil, cottonseed oil, flax seed oil, grapeseed oil, peanut oil, sesame oil, sunflower oil, safflower oil, rice bran oil, coconut oil, a medium chain triglyceride coconut oil, soybean oil, palm oil, cacao butter, and mixtures thereof; and / or the at least one plant-based oil is present in an amount of between about 5% (w / w) to about 30% (w / w).

11. The plant-based foodstuff according to any one of the preceding claims, further comprising one or more optional additives, optionally one or more flavours, seasonings, colours, antioxidants, stabilisers and mixtures thereof.

12. The plant-based foodstuff according to any one of the preceding claims, wherein the plant-based foodstuff is a plant-based cheese analogue or a fat analogue.

13. A method of producing a plant-based foodstuff, the method comprising:(i) mixing a solution of at least one source of protein and at least one plant-based oil to form an emulsion at a pre-determined temperature;(ii) adding at least one gelling agent to the emulsion; and(iii) adding at least one starch to the emulsion, wherein when steps (i) to (iii) are performed in order an emulsion gel is formed, wherein the emulsion gel is (iv) optionally cooled to form the plant-based foodstuff.

14. The method according to claim 13, wherein mixing step (i) further comprises:(a) optionally rehydrating the at least one source of protein in water to form a rehydrated protein solution,(b) pre-heating the rehydrated protein solution at a pre-determined temperature A, optionally in a range about 70 °C to about 99 °C, or in a range of about 80 °C to about 99 °C, and / or(c) optionally homogenizing the rehydrated protein solution, prior to addition of the at least one plant-based oil.

15. The method according to claim 13 or claim 14, wherein mixing step (i) further comprises (d) optionally cooling and / or storing the emulsion.

16. The method according to any one of claims 13 to 25, wherein step (ii) further comprises:(a) heating the emulsion to a pre-determined temperature B, optionally in a range about 50 °C to about 99 °C, about 60 °C to about 99 °C, about 70 °C to about 99 °C, or in a range of about 80 °C to about 99 °C; and(b) optionally stirring the emulsion for a first period of time, and optionally for about 1 to about 40 minutes or about 5 to about 30 minutes.

17. The method according to any one of claims 13 to 16, wherein step (iii) further comprises:(a) heating the emulsion to a pre-determined temperature C, optionally in a range about 50 °C to about 99 °C, about 60 °C to about 99 °C, about 70 °C to about 99 °C, or in a range of about 80 °C to about 99 °C; and(b) optionally stirring the emulsion for a second period of time, and optionally for about 20 to about 160 minutes or about 30 to about 150 minutes.

18. The method according to any one of claims 13 to 17, wherein: the at least one gelling agent is selected from: K-carrageenan, i-carrageenan, high-acyl gellan gum, curdlan gum, agar, konjac gum, and mixtures thereof; and / or the at least one gelling agent is selected from: K-carrageenan, i-carrageenan, high-acyl gellan gum, and mixtures thereof; and / or the at least one gelling agent is present in an amount of between about 0.2% (w / w) to about 0.8% (w / w), about 0.2% (w / w) to about 0.6% (w / w), or about 0.2% (w / w) to about 0.4% (w / w).

19. The method according to any one of claims 13 to 18, wherein: the at least one starch is a amylopectin starch, optionally a high amylopectin starch; and / or the at least one starch is glutinous rice flour; and / or the at least one starch is present in an amount of between about 4% (w / w) to about 20% (w / w), or between about 8%(w / w) to about 12% (w / w).

20. The method according to any one of claims 13 to 19, wherein: the source of protein is selected from: a pea protein isolate, soy protein isolate, potato protein isolate, rice protein isolate, fava bean protein isolate, chickpea protein isolate, peanut protein isolate, hemp protein isolate, almond protein isolate, sunflower seed protein isolate, mung bean protein isolate, and mixtures thereof; and / or the at least one source of protein is present in an amount of between about 3% (w / w) to about 10% (w / w).

21. The method according to any one of claims 13 to 20, wherein:the at least one plant-based oil is selected from: canola oil, rapeseed oil, olive oil, avocado oil, walnut oil, hemp oil, almond oil, avocado seed oil, corn oil, cottonseed oil, flax seed oil, grapeseed oil, peanut oil, sesame oil, sunflower oil, safflower oil, rice bran oil, coconut oil, a medium chain triglyceride coconut oil, soybean oil, palm oil, cacao butter, and mixtures thereof; and / or the at least one plant-based oil is present in an amount of between about 5% (w / w) to about 30% (w / w).

22. A plant-based foodstuff: produced according to the method of any one of claims 13 to 21; and / or according to any one of claims 1 to 12, when produced according to the method of any one of claims 13 to 21.

23. Use of a plant-based foodstuff, wherein the plant-based foodstuff is defined according to any one of claims 1 to 12, as a plant-based cheese analogue, a fat analogue and / or a fat component in plant-based meat.

24. Use of a plant-based foodstuff, wherein the plant-based foodstuff is defined according to any one of claims 1 to 12, when produced according to the method of any one of claims 13 to 21, as a plant-based cheese analogue, a fat analogue and / or a fat component in plant-based meat.

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