GB2516491A

GB2516491A - Edible Fungi

Info

Publication number: GB2516491A
Application number: GB1313234.5A
Authority: GB
Inventors: Timothy John Andrew Finnigan; Muyiwa Akintoye; Reva Mousavi
Original assignee: Marlow Foods Ltd
Current assignee: Marlow Foods Ltd
Priority date: 2013-07-24
Filing date: 2013-07-24
Publication date: 2015-01-28
Anticipated expiration: 2033-07-24
Also published as: GB201413074D0; GB2518726A; GB2516491B; GB2518725A; GB201413076D0; GB201313234D0; GB2518726B; GB2518725B

Abstract

An edible formulation comprises edible fungal particles of a filamentous fungus and at least 2000mg of one divalent or trivalent cation per kilogram of filamentous fungi on a dry basis. The formulation also includes acetate moieties to counter the disagreeable taste of the cations. A preferred formulation includes calcium chloride and calcium acetate. Conventionally egg albumin is used to bind together edible filamentous fungi in meat substitute products. The metal ions are able to increase the elastic modulus of firmness of edible fungus, even in the absence of albumin or hydrocolloids, and provide a meat substitute suitable for vegans.

Description

EDIBLE FUNGI

The invention relates to edible fungi and particularly, although not exclusively, to a process for adjusting the rheology of a mass of filamentous fungus and products resulting therefrom.

It is known, for example from WO 00/15045 (DSM), W096/21362 (Zeneca) and W095/23843 (Zeneca) to use edible filamentous fungi as meat-substitutes, for example in the preparation of burgers and sausages. In such uses, filaments of the fungi are bound together, for example with egg albumin, and are texturised so that the product resembles muscle fibres and therefore has a meat-like appearance and texture. Meat substitutes of the type described have been widely commercially available for many years under the trade mark QUORN In some circumstances, it is desirable to reduce or even eliminate the amount of egg albumin used with edible fungus in the manufacture of meat-substitutes for example on cost grounds or to produce a product suitable for vegans. It may similarly be desirable to reduce the levels of other binding agents or rheology improving agents used. It is one object preferred embodiments of the present invention to address this problem.

On the other hand, the current levels of albumin and/or other rheology-improving agents used in combination with edible fungus may be acceptable but it may nonetheless be desirable for the rheology (e.g. the elastic modulus G' or firmness) of the resulting mass to be increased further, without increasing the levels of albumin and/or other rheology-improving agents. It is one object of a preferred embodiment of the present invention to address this problem.

The invention is based on the discovery that certain metal ions are, even in the absence of albumin, protein and/or other hydrocolloids, able to increase the elastic modulus (G') and firmness of edible fungus. As a result, a foodstuff may be prepared which includes lower levels of albumin, protein and/or other hydrocolloids than previously required; or for the same levels of albumin, protein and/or other hydrocolloids used previously, the elastic modulus and firmness of the foodstuff may be increased.

According to a first aspect of the invention, there is provided an edible formulation comprising edible fungal particles of a filamentous fungus and divalent or trivalent cations.

Said edible formulation suitably includes at least 2000mg, preferably at least 3000mg, more preferably at least 4000mg, especially at least 5000mg of one divalent or trivalent cation per Kg of filamentous fungus on a dry matter. Said edible formulation suitably includes less than 25,000mg, preferably less than 20,000mg, more preferably less than 15,000mg of said one divalent or trivalent cation per Kg of filamentous fungus on a dry matter basis.

Said edible formulation may include intracellular divalent or trivalent cations or extracellular divalent or trivalent cations. Said edible formulation suitably includes the following extracellular levels of said one divalent or trivalent cation: at least 2000mg, preferably at least 3000mg, more preferably at least 4000mg, especially at least 5000mg per Kg of filamentous fungus on a dry matter basis. The maximum extracellular level of said one divalent or trivalent cation is suitably less than 25,000mg, preferably less than 20,000mg, more preferably less than 15,000mg of filamentous fungus on a dry matter basis.

The total amount of divalent or trivalent metal cations (i.e. including all the different types of divalent and trivalent metal cations) in said edible formulation is suitably at least 11000mg, preferably at least 12500mg, more preferably at least 14000mg, especially at least 15500mg per Kg of filamentous fungus on a dry matter basis. The total amount of divalent or trivalent metal cations may be less than 30,000mg, preferably less than 25,000mg per Kg of filamentous fungus.

Said edible formulation suitably includes the following total (i.e. including all the different types of divalent and trivalent metal cations) extracellular levels of divalent or trivalent metal cations: at least 4000mg, preferably at least 6000mg, more preferably at least 8000mg per Kg of filamentous fungus on a dry matter basis. The total may be less than 30,000mg per Kg of filamentous fungus on a dry matter basis.

Said divalent or trivalent cations are preferably metal cations. Said cations may be selected from Ca2, Mg2 and Fe3.

Preferably, said cations are divalent. Preferably said cations are calcium ions.

In said edible formulation, the ratio of the wt% of calcium ions divided by the wt% of filamentous fungus on a dry matter basis is suitably at least 0.002, preferably at least 0.003, more preferably at least 0.004, especially at least 0.005. Said ratio may be less than 0.025 or less than 0.020.

In said edible formulation, the ratio of the wt% of extracellular calcium ions divided by the wt% of filamentous fungus on a dry matter basis is suitably at least 0.002, preferably at least 0.003, more preferably at least 0.004, especially at least 0.005. Said ratio may be less than 0.025 or less than 0.020.

Said edible formulation may include at least 0.100 wt%, for example at least 0.140 wt% of calcium ions. It may include less than 0.300 wt% of calcium ions.

Said edible formulation may include at least 25 wt% of water. Said formulation may include 75 wt% of water or less.

Said edible formulation may include less than 5 wt% of an edible hydrocolloid (e.g. egg albumin) on a dry matter basis. It may include less than 4 wt% of said edible hydrocolloid. Some formulations may include less than 2 wt%, less than 1 wt% or 0 wt% of edible hydrocolloid (e.g. egg albumin). In some cases, it may include at least 0.5 wt% of egg albumin.

In said edible formulation, the ratio of the wt% of edible hydrocolloid (e.g. egg albumin) divided by the wt% of filamentous fungus (on a dry mailer basis) may be less than 0.5, 0.4, 0.3 or 0.2.

In said edible formulation, the ratio of the wt% of edible hydrocolloid (e.g. egg albumin) on a dry mailer basis to the wt% of calcium ions is suitably less than 100, preferably less than 75, more preferably less than 50. In some cases, the ratio may be 0 (e.g. if there is no edible hydrocolloid (e.g. egg albumin) present).

Said edible formulation may incorporate other ingredients, for example one or more flavouring materials. Said edible formulation may be part of a food product for human consumption, for example selected from mince, a burger, a sausage, or meat-like pieces or strips. In other embodiments, said edible formulation may be a product for use as an ingredient for incorporation with other ingredients to define a food product. In this case, at least 90 wt%, at least 95 wt% or at least 99 wt% of said edible formulation may be made up of filamentous fungus, water and said divalent or trivalent cations.

As described above, said divalent or trivalent cations are preferably calcium cations. However, it is found that if calcium chloride alone is used in the formulation, even at a minimum level to achieve the effect described herein, a disagreeable taste results. Accordingly, it is necessary to take steps to counter the disagreeable taste. It has been found that use of calcium acetate is able to do this. Thus, said edible formulation preferably includes acetate moieties (which may have been initially incorporated into the formulation as calcium acetate).

In said edible formulation, the ratio of the wt% of acetate ions divided by the wt% of filamentous fungus on a dry matter basis is suitably at least 0.005, preferably at least 0.01. Said ratio may be less than 0.03, for example less than 0.02.

Said acetate ions are suitably extra-cellular ions.

Said edible formulation may include at least 0.10 wt%, preferably at least 0.20 wt% of acetate ions. It may include less than 0.50 wt%, for example less than 0.40 wt% of acetate ions.

In a first preferred embodiment, said edible formulation may include: -at least 2000mg, for example at least 5000mg, of calcium ions per Kg of filamentous fungus on a dry matter basis; -at least 25 wt% of water.

In a second preferred embodiment, said edible formulation may include: -5000mg of calcium ions per Kg of filamentous fungus on a dry matter basis; -less than 25,000mg of calcium ions per Kg of filamentous fungus on a dry matter basis; -at least 25 wt% of water; -less than 5 wt% of egg albumin.

In said first and second preferred embodiments, preferably in said edible formulation, the ratio of the wt% of acetate ions divided by the wt% of filamentous fungus on a dry matter basis is at least 0.005 and is less than 0.03.

Said edible formulation preferably includes at least 0.10 wt% and less than 0.40 wt% of acetate ions.

As described, fungal particles suitably comprise a filamentous fungus. Said filamentous fungus preferably comprises fungal mycelia and suitably at least 80 wt%, preferably at least 90 wt%, more preferably at least 95 wt% and, especially, at least 99 wt% of the fungal particles in said formulation comprise fungal mycelia. Some filamentous fungi may include both fungal mycelia and fruiting bodies. Said fungal particles preferably comprise a filamentous fungus of a type which does not produce fruiting bodies. Where, however, a filamentous fungus of a type which produces fruiting bodies is used, the fungal particles in said composition suitably include at least wt%, preferably at least 90 wt%, more preferably at least 95 wt% of fungal mycelia.

Preferably, said fungal particles comprise substantially only fungal mycelia -that is, said fungal particles in said composition preferably do not include any fruiting bodies.

Preferred fungi for said fungal particles have a cell wall which includes chitin and/or chitosan. Preferred fungi have a cell wall which includes polymeric glucosamine. Preferred fungi have a cell wall which includes 131-3 and 1-6 glucans.

Said fungal particles may include fungal cells of the order Mucorales as described in WO 00115045 (DSM).

Said fungal particles preferably comprise fungus selected from fungi imperfecti.

Preferably, said fungal particles comprise, and preferably consist essentially of, cells of Fusarium species, especially of Fusarium venenatum A3/5 (formerly classified as Fusarium graminearum) (IMI 145425; ATCC PTA-2684 deposited with the American Type Culture Collection, 10801 University Boulevard, Manassas, VA.) as described for example in W096/21 361 (Zeneca) and W095/23843 (Zeneca).

Preferably, said fungal particles are non-viable. Preferably, said fungal particles have been treated to lower the level of RNA which they contain. Thus, the level of RNA in the fungal particles used is preferably less than the level in an identical fungus when in a viable state.

Fungal particles in said formulation may comprise filaments having lengths of less than 1000 pm, preferably less than 800 pm. Said filaments may have a length greater than 100 pm, preferably greater than 200 pm. Preferably, fewer than 5 wt%, preferably substantially no, fungal particles in said formulation have lengths of greater than 5000pm; and preferably fewer than 5 wt %, preferably substantially no, fungal particles have lengths of greater than 2500 pm. Preferably, values for the number average of the lengths of said fungal particles in said formulation are also as stated above.

Fungal particles in said formulation may comprise filaments having diameters of less than 20 pm, preferably less than 10 pm, more preferably 5 pm or less. Said filaments may have diameters greater than 1 pm. preferably greater than 2 pm.

Preferably, values for the number average of said diameters of said fungal particles in said formulation are also as stated above.

Fungal particles in said formulation may comprise filaments having an aspect ratio (length/diameter) of less than 1000, preferably less than 750, more preferably less than 500. especially of 250 or less. The aspect ratio may be greater than 10.

preferably greater than 40, more preferably greater than 70. Preferably, values for the average aspect ratio of said fungal particles (i.e. the average of the lengths of the particles divided by the average of the diameters of the fungal particles) in said formulation are also as stated above.

According to a second aspect ot the invention, there is provided a method of making an edible formulation, the method comprising: (i) selecting an aqueous formulation comprising edible fungal particles of a filamentous fungus; (ii) contacting said aqueous formulation with divalent or trivalent cations.

Contact may be effected so as to produce an edible formulation with cations as described in the first aspect andlor at levels described in the first aspect. Thus, the edible formulation prepared may be as described in the first aspect.

The method may comprise contacting said aqueous formulation with calcium chloride. Said aqueous formulation may be contacted with an aqueous solution of calcium chloride. The ratio of the pads by weight (herein "pbw") of edible fungal particles on a dry matter basis to the pbw of calcium chloride on a dry matter basis contacted in the method may be at least 50, preferably at least 100, more preferably at least 150. Said ratio may be less than 400, preferably less than 300, more preferably less than 250, especially less than 200.

The method may comprise contacting said aqueous formulation with calcium acetate. The ratio of the pbw of edible fungal particles on a dry matter basis to the pbw of calcium acetate on a dry matter basis may be at least 10, suitably at least 25, preferably at least 40, especially at least 50. Said ratio may be less than 300, suitably less than 200, preferably less than 100, especially less than 80.

Said method may include contacting said aqueous formulation with both calcium chloride and calcium acetate, either independently or concurrently. The ratio of the pbw of calcium chloride to the pbw of calcium acetate contacted with said edible fungal particles may be at least 0.1, preferably at least 0.2, more preferably at least 0.3. Said ratio may be less than 1, preferably less than 0.75, more preferably less than 0.5, especially less than 0.4. Preferably, said ratio is in the range 0.30 to 0.40.

In the method, said aqueous formulation and cations are preferably intimately mixed to produce said edible formulation.

Said edible formulation may include less than 1 wt% of cations contacted with particles in step (U). It may include at least 0.2 wt% or at least 0.4 wt% of cations which suitably comprise calcium chloride and calcium acetate as described.

Said edible formulation suitably includes at least 20 wt% water; it may include wt% or less of water.

Said method may include contacting said edible fungal particles and cations with one or more flavours.

Said method may include contacting said edible fungal particles and cations with an edible hydrocolloid, for example egg albumin.

According to a second aspect of the invention, there is provided the use of divalent or trivalent cations (especially the use of calcium ions) for increasing the G' modulus of a filamentous fungus.

Preferably, calcium ions are used, suitably to provide an edible formulation as described in the first aspect. Suitably, calcium ions are used to make an edible formulation in accordance with the second aspect.

According to a third aspect of the invention, there is provided a method of increasing the G modulus of a filamentous fungus, the method comprising: (i) selecting a filamentous fungus; (ii) selecting a compound comprising divalent or trivalent cations; (iii) contacting the filamentous fungus with said compound.

Preferably, the method comprises selecting a formulation comprising calcium ions and contacting the filamentous fungus with said ions. The method preferably produces an edible formulation in accordance with the first aspect.

Any feature of any aspect of any invention or embodiment described herein may be combined with any other invention described herein mutatis mutandis.

Specific embodiments of the invention will now be described, by way of example, with reference to the accompanying figures, in which: Figure 1 is a graph of elastic modulus G' v. calcium concentration for frozen and non-frozen mycoprotein paste; Figure 2 is a graph of elastic modulus G' v calcium concentration for non-frozen, steamed and frozen mycoprotein paste; Figure 3 is a comparison of the elastic modulus C' of formulations of mycoprotein, mycoprotein plus calcium and mycoprotein plus sodium chloride; Figure 4 is a comparison of the elastic modulus C' of a standard egg albumin-containing formulation and a formulation containing calcium and a lower level of egg albumin; Figure 5 is a schematic representation of steps in the production of mycoprotein-containing products for human consumption.

The following materials are referred to hereinafter: Mycoprotein paste -Mycoprotein paste-refers to a visco-elastic material comprising a mass of edible filamentous fungus derived from Fusarium venenatum A3/5 (formerly classified as Fusarium graminearum Schwabe) (IMI 145425; ATCC PTA-2684 deposited with the American type Culture Collection, 12301 Parklawn Drive, Rockville Md. 20852) and treated to reduce its RNA content to less than 2% by weight by heat treatment. Further details on the material are provided in W096/21 362 and W095/23843. The material may be obtained from Marlow Foods Limited of Stokesley, U.K. It comprises about 23-25 wt % solids (the balance being water) made up of non-viable RNA reduced fungal hyphae of approximately 400-750 pm length, 3-pm in diameter and a branching frequency of 2-3 tips per hyphal length.

Calcium chloride solution -refers to a 36 wt% solution of calcium chloride.

Free range albumin -refers to anhydrous commercially-available egg albumin typically including 97-98 wt% protein on a dry basis.

In a study by Applicant into the firmness of a mixture of mycoprotein paste and egg albumin during mixing, it was noted that addition of calcium chloride produces a rise in firmness and, unexpectedly, this rise in firmness could be achieved even in the absence of egg albumin. Thus, it appeared the calcium chloride was interacting with the mycoprotein paste to increase its firmness or strength. This observation was further investigated, as described below.

The following general procedures and/or preparatory processes were used.

Example I

Mycoprotein paste was mixed with 0.3 wt%, 0.6% and 0.9 wt% of 36 wt% aqueous solutions of calcium chloride to make pastes including different concentrations of calcium chloride. A solution of calcium chloride was added and the water level adjusted to ensure all materials included the same level of water for comparison purposes. The prepared samples were then divided into separate parts.

One part was transferred to a freezer at -21°C.and held there for at least a week before being defrosted prior to rheological measurements. These samples are referred to as "frozen paste'. Another sample referred to as non-frozen", was prepared as described but was not frozen. A third sample was steamed at 100°C for 25 minutes.

Example 2

Solutions of egg albumin protein (16°k and 12% wlw) were prepared using deionised waler. The egg albumin protein powder was mixed with water and optional calcium chloride using a high shear kitchen blender until all the powders were dissolved. The pH of the solutions was then adjusted to pH=6 using either concentrated NaOH or sulphuric acid. The egg albumin solutions were then poured into 17 mm diameter cellulose casings that were then tied up at both sides. The gelation of the albumin sam pIes was induced by heating the casings at 100°C in a steam oven for 20 minutes. After steaming, the gels were cooled down in a chiller to 5°C and held overnight at this temperature before any textural measurement.

Example 3 -Rheology measurements To measure rheology, a Bohlin CVO 50 Rheorneter (Bohlin Instruments, UK) with parallel plate geometry (diameter 20mm) suitable for working in high shear rates at two gap spacings of iSOOpm and 2000pm and 25°C was used. Samples were carefully placed on the centre of the rheometer stage, using a spatula. The rotating element was then gently lowered to a distance of 1550pm or 2050pm, excess sample trimmed off and the rotating element lowered a further SOprn to produce gap sizes of lSOOpm or 2000pm, respectively. Samples were then left for 2 minutes before measurement to reach a temperature of 25°C and to reduce any stress generated during sample loading. Gap size for fresh paste and raw material was carried out at a gap distance of l500pm, while the measurement for steamed and frozen products was performed at 2000pm.

In this study dynamic (oscillation) experiments were carried out to measure the response of the material to stress.

Example 4

A series of rheological measurements was carried out to establish how material microstructure or the interaction between cells might increase by inclusion of calcium ions in a mycoprotein mixture. Samples were prepared as described in Example 1 and tested as described generally in Example 3. Results are provided in Figure 1.

It is clear from Figure 1 that for both products the elastic modulus 0' is increased by inclusion of calcium ions in the mycoprotein paste. The elastic modulus for non-frozen material was increased from l2000Pa for the standard material (with no calcium) to around 25000Pa for the material containing 0.9% calcium; and the modulus for frozen material was increased from around 35000Pa to over 80000Pa. it is also clear that the materials 0' increases at a quicker rate for frozen material compared to the non-Frozen material.

Example 5

To assess he effect of calcium on real product texturization, three levels of calcium as per those used in Example 1 were also applied to mixtures containing egg albumin. The materials were then steamed, frozen and stored in freezer in the same way as for commercially available mycoprotein products sold under the Trade Mark QUORN. Figure 2 compares the rheology of the material after mixing (raw material), after steaming (steamed product) and after freezing and frozen storage (frozen material). Results show the elastic modulus of the products (a rigidity indicator of the materials) increased for all raw, steamed and frozen products as the level of calcium increased from 0.3 wt% to 0.9 wt%. However, the extent of increase in 0 in all materials was not the same. The enhancement of 0' for frozen material was much higher than for steamed material which was higher than for raw material. As Figure 2 shows, the G' value for frozen material was increased from around 150000 Pa for the material with no calcium to over l90000Pa for the material with 0.9% calcium, whereas the value for steamed material varied from around 52000 Pa to around 70000 Pa for the same comparison. The G' for raw material also increased, albeit at a lower rate (12000 Pa to 20000 Pa). The results clearly demonstrate the effect of calcium inclusion in mycoprotein product texturization.

Example 6 -Assessment of effect of monovalent salt By a process similar to that described in Example 5, the elastic modulus of raw, steamed and frozen formulations of mycoprotein alone (referred to as Standard (SI)), mycoprotein plus calcium and mycoprotein plus sodium chloride were assessed.

Results are provided in Figure 3 which shows that the effect of calcium on elastic modulus (i.e. mycoprotein texturization) was significantly higher than the effect of sodium (i.e. NaCI) for raw, steamed and frozen formulations. The elastic modulus (G') for frozen was increased from 152000 Pa for Standard Frozen product up to about 190000 Pa for the calcium-containing formulation, whereas the sodium-containing formulation had a G' only a little above the Standard. The change in G' between the frozen Standard and calcium-containing formulations was very pronounced.

Example 7 -Effect of reduced levels of egg albumin As discussed in the introduction of this specification, it may be desirable to reduce the level of egg albumin in mycoprotein-containing formulations. By a process similar to that detailed in Example 5; the elastic modulus of raw, steamed and frozen formulations of a standard egg albumin-containing formulation (referred to as "Pieces with Existing Egg(E)" and the same formulation but containing 75 wt% of the amount of egg albumin were compared. Results are provided in Figure 4 from which it will be noted that the reduced egg calcium-containing samples in the second series of results have similar elastic modulus compared to the first series of results. Thus, the level of egg albumin can be reduced and the same elastic modulus achieved.

The aforementioned examples used calcium chloride exclusively as the calcium source and this was found to affect rheology as described. However, calcium chloride is found to have a bitter taste at some levels at which it may be incorporated into mycoprotein-containing formulations, particularly formulations with reduced (or zero) egg albumin. Investigations were, therefore, undertaken to establish if it was possible to mask the bitter taste and thereby allow the calcium chloride to be used at desired levels, while still affecting mycoprotein rheology.

It was found that a calcium blend comprising calcium chloride and calcium acetate could advantageously be used. Calcium acetate itself has a sweet, relatively unpleasant taste. However, when combined with calcium chloride, it produces a mixture with an acceptable, slightly salty taste. Use of a calcium chloride/calcium acetate mixture with mycoprotein is found to produce the rheological effects described above, along with an acceptable slightly salty/savoury taste.

Furthermore, advantageously, the calcium acetate and calcium chloride have high water solubility so can readily be incorporated into mycoprotein-containing formulations.

It is found that a combination comprising 0.15 parts by weight (pbw) of calcium ch!oride on a solids basis and 0.40 pbw of calcium acetate on a solids basis provides an acceptable taste and also delivers the rheological properties described. Such a combination may be introduced into mycoprotein paste by use of the 36 wt% calcium chloride solution described together with solid calcium acetate.

Figure 5 provides an overview of process steps taken to prepare mycoprotein-containing foods. in the process mycoprotein (the fungus product of a fermentation process after RNA reduction and dewaterng by centrifugation to approximately 25 wt% solids content) is mixed with other ingredients it is desired to incorporate to produce a substantialiy homogenous mass of a rnycoprotein-containing foodstuff (for example mince, burgers, pieces/strips). The homogenous mass is put through a former and then a steamer (e.g. over 87°C for 25-30 minutes). The steamed product is then chilled (e.g. -Sto -10°C for about 20 minutes) which improves the texture of the product by making it slightly firmer. There follows an ophonal size reduction process followed by a second texturization step involving freezing. Thereafter, products are weighed and packaged prior to the final texturization step at -18°C in a cold store for at least 7 days. Thereafter product can be delivered to retail outlets for sale to customers.

The calcium chloride/calcium acetate mixture may be used to reduce the amount of egg albumin (or other texture producing materials such a proteins) needed in mycoproteincontaining formulations and may allow a vegan product to be formulated which does not contain any egg albumin or other animal proteins.

In Lable 1, Examples A and B provide details of formulations used to make mince; Examples C and D provide details of formulations used to make burgers; and Examples E and F provide details of formulations used to make pieces/strips.

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The egg albumin level in mycoprotein-containing products (mince, burgers and pieces/strips) ranged from 2 to 4 wt%. This can be reduced by about 25% by use of a calcium blend of calcium chloride and calcium acetate. The mycoprotein paste level in products such as sausages may be about 30 wt% whereas it is over 90 wt% for mince and pieces. It is found that the egg albumin reduction effect was better in products with higher mycoprotein paste levels. Selected flavours may be added to the products at a level of 1.8 to 4 wt%. In addition, other additives such as whey protein, oil, diced onion, textured wheat protein, fat and oils may be added at various levels depending on the product.

In the products referred to, the only ingredient which was reduced was the egg albumin and a blend of calcium acetate and calcium chloride was added to compensate for a part of this reduction. The level of water was adjusted accordingly.

Products containing the calcium chloride/calcium acetate mixture were assessed for taste, texture and changes of the product during its declared shelf-life by a trained panel of individuals and compared to current products. They were found to be commercially acceptable.

The invention is not restricted to the details of the foregoing embodiment(s). The invention extends to any novel one, or any novel combination, of the features disclosed in this specification (including any accompanying claims, abstract and drawings), or to any novel one, or any novel combination, of the steps of any method or process so disclosed.

Claims

Claims 1. An edible formulation comprising edible fungal particles of a filamentous fungus and divalent or trivalent cations.
2. A formulation according to claim 1. which includes at least 2000mg of one divalent or trivalent cation per Kg of filamentous fungus on a dry matter basis.
3. A formulation according to claim 1 or claim 2, which includes less than 25,000mg of said one divalent or trivalent cation per Kg of filamentous fungus on a dry matter basis.
4. A formulation according to any preceding claim, wherein said cations are selected from Ca2, Mg2 and Fe3.
5. A formulation according to any preceding claim, wherein said cations are calcium ions.
6. A formulation according to any preceding claim, wherein the ratio of the wt% of calcium ions divided by the wt% of filamentous fungus on a dry matter basis is at least 0.002 and is less than 0.025.
7. A formulation according to any preceding claim, which includes at least 0.100 wt% calcium ions and less than 0.300 wt% calcium ions.
8. A formulation according to any preceding claim, which includes at least 25 wt% of water.
9. A formulation according to any preceding claim, which includes less than 5 wt% of an edible hydrocolloid, for example egg albumin.
10. A formulation according to any preceding claim, which includes acetate moieties.
11. A formulation according to any preceding claim, wherein the ratio of the wt% of acetate ions divided by the wt% of filamentous fungus on a dry matter basis is at least 0.005.
12. A formulation according to any preceding claim, which includes at least 0.10 wt% of acetate ions and less than 0.50 wt% of acetate ions.14. A formulation according to any preceding claim, wherein said filamentous fungus comprises fungal mycelia, wherein as least 80 wt% of the fungal particles in said formulation comprise fungal mycelia.15. A method of making an edible formulation, the method comprising: i) selecting a aqueous formulation comprising edible fungal particles of a filamentous fungus: U) contacting said aqueous formulation with divalent or trivalent cations.16. A method according to claim 15, wherein said formulation is in accordance with any of claims ito 14..17. A method according to claim 15 or claim 16, which comprises contacting said aqueous formulation with calcium chloride.18. A method according to any of claims 15 to 17, wherein the ratio of the parts by weight ot edible fungal particles on a dry matter basis to the parts by weight of calcium chloride on a dry mafter basis contacted in the method is at least 50 and is less than 400.19. A method according to any of claims 15 to 18, which comprises contacting said aqueous formulation with calcium acetate.20. A method according to any of claims 15 to 19, wherein the ratio of the pbw of edible fungal particles on a dry matter basis to the pbw of calcium acetate on a dry matter basis is at least 10 and is less than 300, 21. A method according to any of claims 15 to 20, which includes contacting said aqueous formulation with both calcium chloride and calcium acetate.22. A method according to any of claims 15 to 21, wherein the ratio of the pbw of calcium chloride to the pbw of calcium acetate contacted with said edible fungal particles is at least 0.1 and is less than 1.23. The use of divalent or trivalent cations (especially the use of calcium ions) for increasing the G' modulus of a filamentous fungus.Amendments to the Claims have been filed as follows Claims 1. An edible formulation in the form of a meat substitute comprising edible fungal particles of a filamentous fungus and divalent or trivalent cations, wherein said formulation includes acetate moieties and wherein said formulation includes at least 2000mg of one divalent or trivalent cation per Kg of filamentous fungus on a dry matter basis.2. A formulation according to claim 1, which includes less than 25,000mg of said one divalent or trivalent cation per Kg of filamentous fungus on a dry matter basis.3. A formulation according to claim 1 or claim 2. wherein said cations are selected from Ca2, Mg2 and Fe3t ct-4. A formulation according to any preceding claim, wherein said cations are calcium ions.5. A formulation according to any preceding claim, wherein the ratio of the wt% of calcium ions divided by the wt% of filanientous fungus on a dry matter basis is at least 0.002 and is less than 0.025.6. A formulation according to any preceding claim, which includes at least 0.100 wt% calcium ions and less than 0.300 wt% calcium ions.7. A formulation according to any preceding claim, which includes at least 25 wt% of water.8. A formulation according to any preceding claim, which includes less than 5 wt% of an edible hydrocolloid.9. A formulation according to any preceding claim, wherein the ratio of the wt% of acetate ions divided by the wt% of filamentous fungus on a dry matter basis is at least 0.005.10. A formulation according to any preceding claim, which includes at least 0.10 wt% of acetate ions and less than 0.50 wt% of acetate ions.11. A formulation according to any preceding claim, wherein said filamentous fungus comprises fungal mycelia, wherein as least 80 wt% of the fungal particles in said formulation comprise fungal mycelia.12. A method of making an edible formulation in the form of a meat substitute according to any preceding claim, the method comprising: i) selecting a aqueous formulation comprising edible fungal particles of a filamentous fungus; ii) contacting said aqueous formulation with divalent or trivalent cations.
13. A method according to claim 12, which comprises contacting said aqueous formulation with calcium chloride.
14. A method according to claim 12 or claim 13, wherein the ratio of the parts by O weight of edible fungal particles on a dry matter basis to the parts by weight of calcium chloride on a dry matter basis contacted in the method is at least 50 and is less than r 400.%1
15. A method according to any of claims 12 to 14, which comprises contacting said aqueous formulation with calcium acetate.
16. A method according to any of claims 12 to 15, wherein the ratio of the pbw of edible fungal particles on a dry matter basis to the pbw of calcium acetate on a dry mailer basis is at least 10 and is loss than 300,
17. A method according to any of claims 12 to 16, which includes contacting said aqueous formulation with both calcium chloride and calcium acetate.
18. A method according to any of claims 12 to 17, wherein the ratio of the pbw of calcium chloride to the pbw of calcium acetate contacted with said edible fungal particles is at least 0.1 and is less than 1.