Microbial compositions and uses

Abstract

The present invention relates to compositions comprising disintegrated microbial biomass and / or microbial biomass fractions in combination with other ingredients, and methods of producing such compositions. The invention further relates to the use of these composition in commercial products such as food products.

Description

[0001] Microbial Compositions and Uses

[0002] TECHNICAL FIELD

[0003] The present invention relates to compositions comprising disintegrated microbial biomass and / or microbial biomass fractions in combination with other ingredients, and their use in commercial products, including food products.

[0004] BACKGROUND

[0005] Uses of microbial biomass as well as methods to produce fractions or extracts from microbial biomass have been reported in a variety of patents and scientific literature. Examples include US3887431 , which describes methods to fractionate yeast biomass to provide a protein isolate fraction, and US2908661 , which describes a microalgal flour, a method to produce such a flour and its uses.

[0006] In addition to microbial biomass itself, various compositions comprising microbial biomass have also been reported. For example, GB2617170 describes methods and compositions combining fungal microbial biomass with food materials to enhance the functional properties of the resulting food composition. These compositions are mostly combinations of ingredients in which the microbial biomass serves to provide functionality which the other ingredients in the composition do not possess. Functional ingredients are therefore crucial to the development of novel textures or to mimic the attributes of existing products.

[0007] In recent years, the number of ingredients derived from plant sources (e.g., canola, lupin, sunflower, fava bean, mung bean, lentils) and microbial biomass (e.g., yeast, algae, fungi, bacteria) have grown significantly, in particular for applications in the food sector for the preparation of analogues (Kyriakopoulou et al., 2021). Analogues resemble the textural, sensorial and functional properties of an existing product, i.e. mimicking a known product. Some of the most notable examples are cheese and meat analogues in the food sector. However, cheese and meat analogues are difficult to develop and require an extensive list of ingredients, each of which brings an important property or functionality to the analogue. However, there is a shortage of appropriate ingredients that possess the required attributes in order to be suitable for use in meat or cheese analogues. In addition to this, little is understood regarding the synergistic effects various ingredients have in a formulation. While some (albeit limited) knowledge is available based on the functionality of each individual ingredient, the nature of the interactions when blending several ingredients, the potential synergies and the resulting final properties, remain uncertain. Moreover, the growing demand for eco-friendly alternatives to meat and dairy farming, such as meat and cheese analogues among others, is driving a need for more ingredients that can be used in such analogues. In order to do this, synergy between ingredients must be explored. The present invention addresses this need. The inventors have discovered several unique behaviors and synergistic effects when specific microbial compositions are prepared by combining microbial biomass fractions and additional ingredients / additives. The present application describes these compositions and methods for their production.

[0008] SUMMARY OF THE INVENTION

[0009] The present invention relates to compositions comprising disintegrated microbial biomass, and / or fractions of microbial biomass (MBF) and their combination with at least one other material.

[0010] In one aspect of the invention there is provided a composition comprising: a. at least one disintegrated microbial biomass, and / or at least one microbial biomass fraction (MBF) wherein the protein content of the disintegrated microbial biomass or microbial biomass fraction is at least 50%, and b. at least one plant based protein, and / or c. at least one plant based fibre.

[0011] In another aspect of the invention there is provided a composition comprising: a. at least one disintegrated microbial biomass, and / or at least one microbial biomass fraction (MBF) wherein the protein content of the disintegrated microbial biomass or microbial biomass fraction is at least 50%, and b. at least one animal based protein, and / or c. at least one plant based fibre.

[0012] In a preferred embodiment, the disintegrated microbial biomass or microbial biomass fraction has a protein content of more than 60%, preferably between 65-75%. More preferably, the disintegrated microbial biomass or microbial biomass fraction has a protein content of -70%. In another aspect of the invention there is provided a composition comprising a. at least one disintegrated microbial biomass, and / or at least one microbial biomass fraction (MBF) wherein the fibre content of the disintegrated microbial biomass or microbial biomass fraction is at least 20% and b. at least one plant based fibre, and I or c. at least one salt, and / or d. at least one animal based protein.

[0013] In another aspect of the invention there is provided a composition comprising a. at least one disintegrated microbial biomass, and / or at least one microbial biomass fraction (MBF) wherein the fibre content of the disintegrated microbial biomass or microbial biomass fraction is at least 20% and b. at least one microbial based fibre.

[0014] In some embodiments, the at least one disintegrated microbial biomass or microbial biomass fraction has a moisture content of less than 10%.

[0015] In some embodiments, the at least one disintegrated microbial biomass, or microbial biomass fraction has a fibre content of at least 20%.

[0016] In an embodiment, the disintegrated microbial biomass, and / or microbial biomass fraction is subjected to thermal treatment or heat treated at a temperature in the range of approximately 50-200°C, preferably, 50-70°C, 65-75°C or 80-90°C to enhance functional properties, before being incorporated into a composition of the present invention. Preferably, the disintegrated microbial biomass, and / or microbial biomass fraction is dried to form a powder before being heat treated at temperatures of 50-200°C. In some embodiments, heat treatment is carried out for 5-300 minutes, 10-240 minutes, 10-180 minutes, 10-120 minutes, 20-90 minutes, or 15-115 minutes. Alternatively, heat treatment is carried out for up to 24 hours, up to 48 hours, up to 72 hours, up to 96 hours, up to 120 hours or longer.

[0017] In some embodiments, the disintegrated microbial biomass and / or microbial biomass fraction is subjected to thermal treatment or heat treatment at a temperature in the range of approximately 50-200°C, preferably 50-70°C, 65-75°C or 80-90°C with a relative humidity of more than 60%, preferably a relative humidity of more than 70%, more preferably a relative humidity of more than 80%, even more preferably a relative humidity of more than 90%. The relative humidity may be in the range of 50-100%, 60-100%, 70- 100%, 80-100%, 90-100%, 50-90%, 60-90%, 70-90%, 80-90%, 50-80%, 60-80%, 70- 80%, 50-70% or 60-70%. Preferably, the relative humidity is in the range of 60-100% or 70-100%. In some embodiments relative humidity may be 100%.

[0018] In some embodiments the composition is in an emulsion system wherein preferably the emulsion system comprises at least 40% oil.

[0019] In some embodiments, the composition further comprises one or more of a polysaccharide, fat, oil, thickening agent, stabilizing agent, enzyme, animal based protein, plant based protein, plant based fibre or salt.

[0020] In some embodiments the composition further comprises flour.

[0021] In a preferred embodiment, the plant based protein is selected from one or more of potato protein, soya protein, lentil protein, chickpea protein, peanut protein, almond protein, spirulina, quinoa protein, chia seed protein, tofu protein, pea protein, pistachio protein, oat protein or wheat protein.

[0022] In another preferred embodiment, the animal-based protein is selected from one or more of milk protein, egg protein or meat protein. Preferably, when the animal-based protein is meat protein, said meat protein may be selected from one or more of beef protein, pork protein, lamb protein, chicken protein, turkey protein, duck protein, rabbit protein, goat protein, venison protein or pheasant protein, however the skilled person would understand that alternative meat proteins are available and can be used in compositions according to the present invention. Preferably, when the animal-based protein is milk protein, said milk protein may be selected from one or more of casein, whey, lactoglobulin, or lactoferrin, however the skilled person would understand that alternative milk proteins are available and can be used in compositions according to the present invention. Preferably, when the animal-based protein is egg protein, said egg protein may be selected from one or more of ovalbumin, ovotransferrin or ovomucoid, however the skilled person would understand that alternative egg proteins are available and can be used in compositions according to the present invention. Preferably, when the composition comprises at least one animal-based protein, said animal-based protein is milk protein and / or egg protein.

[0023] In another preferred embodiment, the plant based fibre is selected from one or more of psyllium fibre, konjac fibre, oat fibre, lentil fibre, resistant starch, acacia gum, methylcellulose, cellulose, hydroxypropyl methylcellulose, p-glucan, citrus fibre or pectin. Preferably, when the plant based fibre is p-glucan, said p-glucan is derived from oat.

[0024] In an embodiment the composition comprises a ratio of microbial biomass fraction to plant based protein or animal based protein of approximately 25:75 (dry matter).

[0025] In an embodiment the composition comprises a ratio of microbial biomass fraction to plant based protein or animal based protein of 1.7-2.3:1.

[0026] In an embodiment the composition comprises a ratio of microbial biomass fraction to animal based protein or plant based protein of 0.1 -7:1.

[0027] In an embodiment the composition comprises a ratio of microbial biomass fraction to plant based fibre of 1-2:1 (dry matter).

[0028] In an embodiment the composition comprises a ratio of microbial biomass fraction to plant based fibre of 5-6:1 (dry matter).

[0029] In an embodiment the composition comprises a blend of 0.2-0.5:1 , wheat flour to microbial biomass fraction.

[0030] In another aspect of the invention there is provided a composition comprising: a. at least one disintegrated microbial biomass, and / or at least one microbial biomass fraction (MBF) wherein the protein content of the disintegrated microbial biomass or microbial biomass fraction is at least 50% and b. at least one oil or fat.

[0031] In an embodiment, the at least one oil or fat is plant based. In a preferred embodiment the at least one oil or fat is selected from one or more of coconut oil, sunflower oil, olive oil, canola oil, rice oil, vegetable oil, avocado oil, sesame seed oil, flaxseed oil, walnut oil, peanut oil, rice fat, bran fat or coconut fat.

[0032] In some embodiments the composition further comprises one or more of a polysaccharide, thickening agent, stabilizing agent, enzyme or salt.

[0033] In some embodiments the composition comprises a second disintegrated microbial biomass or microbial biomass fraction. Preferably, the second disintegrated microbial biomass, or microbial biomass fraction has a fibre content of at least 20%.

[0034] In another aspect of the invention there is provided a composition comprising a. at least one disintegrated microbial biomass, and / or at least one microbial biomass fraction (MBF) wherein the protein content of the disintegrated microbial biomass or microbial biomass fraction is at least 50%, or at least one disintegrated microbial biomass, and / or at least one microbial biomass fraction (MBF) wherein the fibre content of the disintegrated microbial biomass or microbial biomass fraction is at least 20% and b. at least one salt.

[0035] In a preferred embodiment the salt is CaCh and / or sodium bicarbonate.

[0036] In some embodiments the composition further comprises a thickening agent such as xanthan gum.

[0037] In another aspect of the invention there is provided a composition comprising a. at least one disintegrated microbial biomass, and / or at least one microbial biomass fraction (MBF) wherein the protein content of the disintegrated microbial biomass or microbial biomass fraction is at least 50% and b. at least one enzyme.

[0038] In a preferred embodiment the enzyme is a multi-copper oxidase, a transferase, a peptidase, a transglutaminase or blends of said enzymes. These enzymes may be used with suitable mediators. Suitable mediators refer to molecules that facilitate enzyme activity, such as co-factors. Examples of co-factors include ions such as copper, zinc and iron as well as organic molecules such as vitamins or vitamin derived molecules, including thiamine pyrophosphate and flavin adenine dinucleotide. It is understood that the suitable mediators of an enzyme are determined by the specific enzyme being employed and the reaction intended to be catalysed. The skilled person would be able to select suitable mediators based on their specific needs using routine skill.

[0039] Most preferably the enzyme is a transglutaminase or a multi-copper oxidase.

[0040] In another aspect of the invention any of the compositions described above may be incorporated into a food product or a cosmetic product, or their precursors.

[0041] In another aspect of the invention there is provided a method of preparing a composition wherein said method comprises combining: a. at least one disintegrated microbial biomass, and / or microbial biomass fraction (MBF), wherein the protein content of the disintegrated microbial biomass or microbial biomass fraction is at least 50%, and b. at least one plant based protein, and / or c. at least one plant based fibre, in an aqueous system or an emulsion system to form a homogenous suspension.

[0042] In another aspect of the invention there is provided a method of preparing a composition wherein said method comprises combining: a. at least one disintegrated microbial biomass, and / or microbial biomass fraction (MBF), wherein the protein content of the disintegrated microbial biomass or microbial biomass fraction is at least 50%, and b. at least one oil or fat, in an aqueous system or an emulsion system to form a homogenous suspension.

[0043] In some embodiments the method further comprises adding one or more of a polysaccharide or salt and / or flour.

[0044] In another aspect of the invention there is provided a method of preparing a composition wherein said method comprises combining: a. at least one disintegrated microbial biomass, and / or at least one microbial biomass fraction (MBF) wherein the fibre content of the disintegrated microbial biomass or microbial biomass fraction is at least 20%, and / or the protein content of the disintegrated microbial biomass or microbial biomass fraction is at least 50%, and b. at least one plant based fibre, and / or c. at least one salt, and / or d. at least one animal based protein in an aqueous system or and emulsion system to form a homogenous suspension.

[0045] In another aspect of the invention there is provided a method of preparing a composition wherein said method comprises combining: a. at least one disintegrated microbial biomass, and / or microbial biomass fraction (MBF), wherein the protein content of the disintegrated microbial biomass or microbial biomass fraction is at least 50%, and b. at least one enzyme in an aqueous or an emulsion system to form a homogenous suspension.

[0046] In another aspect of the invention there is provided a method of preparing a composition wherein said method comprises combining: a. at least one disintegrated microbial biomass, and / or microbial biomass fraction (MBF), wherein the protein content of the disintegrated microbial biomass or microbial biomass fraction is at least 50%, or at least one disintegrated microbial biomass, and / or at least one microbial biomass fraction (MBF) wherein the fibre content of the disintegrated microbial biomass or microbial biomass fraction is at least 20% and b. at least one salt in an aqueous or an emulsion system to form a homogenous suspension.

[0047] In another aspect of the invention there is provided a method of preparing a composition wherein said method comprises combining: a. at least one disintegrated microbial biomass, and / or at least one microbial biomass fraction (MBF) wherein the fibre content of the disintegrated microbial biomass or microbial biomass fraction is at least 20%, and b. at least one animal based protein in an aqueous or an emulsion system to form a homogenous suspension. In some embodiments the method further comprises adding a thickening agent such as xanthan gum.

[0048] In some embodiments the emulsion system comprises at least 40% oil.

[0049] In some embodiments the method further comprises mechanically homogenizing the homogenous suspension. This process applies mechanical shear to a homogenous suspension, in liquid, for example water. In a preferred embodiment, oil is added to the homogenous suspension before mechanical shear is applied in order to produce a homogenous emulsion.

[0050] BRIEF DESCRIPTION OF THE DRAWINGS

[0051] Figure 1- Gelation hardness of gels obtained with blends of MBF (-62% protein) and potato protein.

[0052] Figure 2- Textural attributes of compositions containing MBF and potato protein.

[0053] Figure 3- Textural attributes of gels made with compositions of MBF and konjac fibre.

[0054] Figure 4- Textural attributes of several microbial compositions as fat analogues.

[0055] Figure 5- Melting behaviour of microbial compositions (test 3 and test 4), pork tissue and coconut fat.

[0056] Figure 6- Gelation ability of MBFs (top: MBF with at least 50% protein, 68°C, 1 hour; bottom: MBF with >20% fibre, 82°C, 3 hours).

[0057] Figure 7- Maximum gelation (G’max) of compositions comprising MBF and CaCh.

[0058] Figure 8- Gelation hardness of compositions comprising MBF and transglutaminase (TGM). The protein composition of each MBF was 71.1 % for MBF1 , 70.3% for MBF2 and 64.2% for MBF3. The white bars represent MBF in water at a concentration of 100g / L with transglutaminase (tgm) added at a ratio of 0.35g per gram of MBF and the orange bars represent MBF in water at a concentration of 100g / L without transglutaminase.

[0059] Definitions:

[0060] Microbial biomass and biomass as used in the present description refer to a material produced by growth and / or propagation of microbial cells, or produced as byproduct of fermentation processes. In the context of the present invention a microbial biomass primarily refers to a population of intact microbial cells. Biomass may contain cells and / or intracellular contents as well as extracellular material. Extracellular material includes, but is not limited to, compounds secreted by a cell such as proteins. Microbial biomass as used in the present invention may include microorganisms in one or more of the groups selected from algae, bacteria, fungi and yeast, either grown as single-cells or multi-cellular structures or forming macro-structures such as hyphae.

[0061] Disintegrated microbial biomass or disrupted microbial biomass as used in the present invention refers to a microbial biomass as described above which has been subjected to homogenization. Homogenization can be carried out mechanically, or non-mechanically. Methods of mechanical homogenization include milling and grinding. Meanwhile, methods of non-mechanical homogenization include enzymatic disintegration, chemical disintegration, osmotic shock and thermolysis. It is understood that in the context of the present invention a disintegrated microbial biomass may comprise both intact microbial cells and lysed microbial cells. The terms disintegrated microbial biomass and disrupted microbial biomass can be used interchangeably.

[0062] Microbial biomass fractions are understood in the context of the present invention as extracts, fractions, concentrates, isolates or the like derived from refined components or subcomponents of microbial biomass and / or disintegrated biomass.

[0063] Functionalities is a general term that refers to one or more of several properties or attributes that are necessary or desirable for structuring purposes, in particular in food, feed and / or cosmetic applications. Functionalities may include, but are not limited to, foaming, emulsification, gelation, succulence, firmness, springiness, water holding and oil holding.

[0064] Dry weight (DW) or dry matter as used in the present description means weight determined in the relative absence of water. For example, reference to microbial biomass as comprising a specified percentage of a particular component by dry weight means that the percentage is calculated based on the weight of the biomass after substantially all moisture has been removed.

[0065] Water holding capacity (WHC) as used in the present description refers to the amount of water a sample can hold per unit of weight.

[0066] Oil holding capacity (OHC) as used in the present description refers to the amount of oil a sample can hold per unit of weight.

[0067] Particle size distribution is reported as a volume distribution. Taking D50 as an example it is known in the art that this can also be referred to as Dv50; the terms D50 and Dv50 are used interchangeably herein. The D50 or Dv50 is defined in the art as the maximum particle size, measured by diameter, below which 50% of the sample volume exists, also known as the median particle size (diameter) by volume.

[0068] Relative humidity is a measure of the amount of water vapour in the air compared to the maximum amount of water vapour that the air can hold at the same temperature. Relative humidity is measured as a percentage. When air is holding the maximum amount of water vapour and is at saturation the relative humidity is 100%. Notably, the ability of air to hold water is directly linked to its temperature and warmer air can hold significantly more water than colder air, therefore a specific amount of water in the air will result in a higher relative humidity in cold air compared to warmer air.

[0069] Organoleptic properties refer to characteristics that are perceived by the senses i.e. taste, smell, appearance and feel. Examples of organoleptic properties include but are not limited to, a sweet taste, a sour taste, a bitter taste, a musty taste, a metallic taste, an umami taste, a fruity taste, a floral taste, a cheesy taste, a fruity aroma, a floral aroma, a meaty aroma, a cheesy aroma, an earthy aroma, a crunchy feel, a soft feel, a tough feel, a creamy feel, a chewy feel, a juicy feel, a dark appearance, a light appearance, a foamy appearance, a smooth appearance and a rough appearance. Organoleptic properties can be altered by the specific processing methods applied to a product, the components of a composition and storage conditions, among other factors.

[0070] Methods:

[0071] It should be appreciated that the methods listed below are illustrative only and the skilled person would be able to use alternative methods known in the art to achieve the same objectives.

[0072] Protein content is measured according to standard methods, including Lowry, dumas, BCA, Bradford and Kjeldal, and using a Nitrogen conversion factor of 6.25.

[0073] Fibre content is measured according to standard methods, in particular total dietary fibre, which includes low molecular weight fibre, high molecular weight fibre and resistant starch. Fibre is understood herein as a polysaccharide or polymer which makes part, in general, of the cell assembly (cell wall, organelles) of microbial biomass, seeds, fruits, pulps, skins and other vegetable tissues.

[0074] Moisture content can be determined by the oven method, in which samples are placed in an oven at a temperature typically ranging from 60-100°C for a period of time until there is a constant dry weight. In order to determine a constant dry weight, the sample is weighed at several time intervals. When the weight has stabilized it is then possible to calculate how much of the sample weight was accounted for by moisture. It is understood that the precise temperature and time used for oven drying is dependent on the sample being analysed.

[0075] Gelation hardness can be measured using a texture analyzer TA Plus (lloyd), following a double compression test. In this case, gelation hardness was measured by heating the suspension for 30 minutes at 90°C followed by cooling to room temperature. After the samples are cooled, they were removed from their container and cut into slabs of equal geometry. A double compression test was then applied to each slab and the resulting texture parameters recorded and analyzed.

[0076] Texture profile analysis (TPA) is a common test known in the art to determine the textural properties of food products, cosmetics and pharmaceuticals, in which samples are compressed two times, to get insights into the behavior of samples when they are chewed and to simulate the biting action of the mouth. Textural attributes such as firmness / hardness, springiness, chewiness and cohesiveness are defined according to texture profile analysis. These parameters are calculated from force-time diagrams in a double compression test. Hardness is the peak force measured during the first compression. Springiness is how well a sample springs backs after it has been deformed during the first compression. Cohesiveness is how well a sample will resist a second compression relative to its resistance during the first compression. Chewiness is calculated from the values of hardness, springiness and cohesiveness, and it expresses the energy required to compress (or “chew”) the sample.

[0077] Chewiness = hardness x cohesiveness x springiness

[0078] Melting behaviorwas analyzed by preparing thin slides of sample (approximately 1.1 mm thickness) and subjecting said samples to cooling at approximately - 10°C. After this, the samples were heated up at constant heating rate to 80°C for 30 minutes and constant frequency / shear and the complex shear modulus was recorded. All measurements were conducted in a rheometer (AntonPaar MR102).

[0079] Foam capacity was measured using overrun. In the art, “overrun” is a standard measure of foaming capacity i.e. the ability of a substance to incorporate and hold air. Overrun is typically measured as a percentage increase in volume once a liquid is agitated to produce a foam. A higher overrun indicates a greater foaming capacity, i.e. a larger foam can be produced.

[0080] Overrun is typically calculated as: ioo Foam stability was measured according to foam half-time, which is a standard method in the art. Foam half-time is the time it takes for a foam to reduce to half its initial volume. A longer foam half-time indicates that a foam is more resistant to collapse, i.e. the foam is more stable.

[0081] Organoleptic properties were determined by a panel of at least 3-5 members who were trained in identifying different aromas / flavour descriptors. The test was performed by suspending the product of interest in warm water (concentration ~2% DW) and using this suspension as a tasting sample. Each member of the panel provided their descriptors independently.

[0082] DETAILED DESCRIPTION

[0083] The present invention relates to compositions comprising disintegrated microbial biomass, and / or fractions of microbial biomass (MBF) and their combination with at least one other material.

[0084] Described herein are disintegrated microbial biomass, and microbial biomass fractions, and their combination with other materials. Also described are the unique functional properties such compositions possess.

[0085] Compositions of the present invention comprise at least one part of disintegrated microbial biomass, and / or microbial biomass fractions (MBF) and at least one additional component. The additional component may be selected from at least one of proteins, fibres, polysaccharides, salts, oils and / or fats, enzymes, thickening agents and stabilizing agents. However, alternative additional components known to the skilled person may be used. Any number of additional components can be combined with at least one disintegrated biomass, and / or microbial biomass fraction to produce a composition of the present invention.

[0086] A disintegrated microbial biomass, and / or microbial biomass fraction may be subjected to particular conditions such as temperature, humidity and shear to enhance functional properties, before being incorporated into a composition of the present invention. Examples of these functional properties include, gelation capacity, chewiness, foaming capacity, emulsification properties, glazing properties, browning properties, springiness, firmness, mouth feel, succulence, water holding capacity and oil holding capacity.

[0087] The disintegrated microbial biomass, and / or microbial biomass fraction may be subjected to conditions such as temperature, humidity and shear to enhance organoleptic properties, before being incorporated into a composition of the present invention. Examples of these organoleptic properties include: reduced off-flavours, reduced bitterness and astringency, masking of off-notes, reduced fruity / floral smell / taste, enhanced meaty flavour such as an enhanced chicken flavour, and an enhanced roast flavour.

[0088] In an embodiment, the disintegrated microbial biomass, and / or microbial biomass fraction is subjected to temperatures (that is, subjected to thermal treatment) of 50- 200°C, preferably 50-70°C, 65-75°C or 80-90°C to enhance functional properties, before being incorporated into a composition of the present invention. Preferably, the disintegrated microbial biomass, and / or microbial biomass fraction is dried to form a powder before being subjected to temperatures of 50-200°C.

[0089] In a preferred embodiment, when the disintegrated microbial biomass, and / or microbial biomass fraction has a protein content of at least 50% and / or a fibre content of at least 20% and said disintegrated microbial biomass, and / or microbial biomass fraction is subjected to temperatures in the range of approximately 50-70°C or 65-75°C to enhance functional properties before being incorporated into a composition of the present invention. Preferably, the disintegrated microbial biomass, and / or microbial biomass fraction is dried to form a powder before being subjected to temperatures of approximately 50-70°C or 65-75°C.

[0090] In a preferred embodiment, when the disintegrated microbial biomass, and / or microbial biomass fraction has a fibre content of at least 20% said disintegrated microbial biomass, and / or microbial biomass fraction is subjected to temperatures in the range of approximately 80-90°C to enhance functional properties before being incorporated into a composition of the present invention. Preferably, the disintegrated microbial biomass, and / or microbial biomass fraction is dried to form a powder before being subjected to temperatures of approximately 80-90°C. In a preferred embodiment, the thermal treatment is conducted at relative humidity of more than 60%, preferably more than 70%, more preferably more than 80% or more than 90%. In some embodiments, thermal treatment is conducted at a relative humidity of 100%.

[0091] The functional properties described herein are primarily desirable functional properties for use in food manufacturing, although it will be appreciated that such functional properties including gelation, emulsification and / or foaming may also be desirable for other applications, such as cosmetic production.

[0092] The organoleptic properties described herein are primarily desirable sensory properties for use in food manufacturing, food compositions and cosmetic formulations.

[0093] In the context of the present invention, microbial biomass may be selected from unicellular or colonial prokaryotes and eukaryotes and one or more combinations thereof. Preferably, the microbial cells are selected from at least one of heterotrophic microalgae, fungi (unicellular or multicellular), bacteria and yeast. More preferably, the microbial biomass is yeast.

[0094] Non limiting examples of genera from which the microbial biomass may be derived for may be produced are Saccharomyces and Pichia (yeast), Tetraselmis, Chlorella, Arthrospira (algae), Fusarium (fungi), Methylobacterium (bacteria) and Lactobacillus (bacteria). Preferably the microbial biomass is derived from yeast, more preferably from the genus Saccharomyces and / or Pichia. Yeasts which may be used in the present invention include Saccharomyces, such as S. cerevisiae, S. chevalieri, S. boulardii, S. bayanus, S. italicus, S. delbrueckii, S. rosei, S. micro-ellipsodes, S. carlsbergensis, S. bisporus, S. fermentati, S. pastorianis, S. rouxii, or S. uvaruirr, a yeast belonging to the genus Schizo-saccharomyces, such as S. japonicus, S. kambucha, S. octo-sporus, or S. pombe', a yeast belonging to the genus Hansenula, such as H. wingei, H. arni, H. henricii, H. americana, H. canadiensis, H. capsulata, or H. polymorpha a yeast belonging to the genus Candida, such as C. albicans, C. utilis, C. boidinii, C. stellatoidea, C. famata, C. tropicalis, C. glabrata, or C. parapsilosis', a yeast belonging to the genus Pichia, such as P. pastoris, P. kluyveri, P. polymorpha, P. barkeri, P. cactophila, P. rhodanensis, P. cecembensis, P. cephalocereana, P. eremophilia, P. fermentans, or P. kudriavzevir, a yeast belonging to the genus Kluyveromyces, such as K. marxianus and a yeast belonging to the genus Torulopsis, such as T. bovina, or T. glabrata.

[0095] Microbial cells present in microbial biomass contain mostly proteins, carbohydrates, lipids and minerals.

[0096] In the literature there are many examples of what constitutes a microbial biomass fraction (MBF). For example, a microbial biomass fraction can refer to a protein isolate (US3887431), a fibre isolate (US9249235), a protein concentrate (EP3670646), an extract rich in proteins or fibres (US4810509) or fractions containing proteins and / or fibres (NL2026504).

[0097] The methods to produce disintegrated microbial biomass, and / or microbial biomass fractions are well established and known in sufficient detail from technical literature, and as such fall outside of the scope of the present invention.

[0098] Example 1 provides one possible method for producing a protein-rich microbial biomass fraction (MBF) derived from yeast. However, other methods may be employed. It should be understood that compositions of the present invention are not limited by the method used to produce a microbial biomass fraction used in such compositions. Nevertheless, the inventors have found that the method used in Example 1 to produce an MBF results in products with particularly good functional properties. However, other methods of making an MBF can also be used such as those described in US3887431 , US9249235, EP3670646 and US4810509.

[0099] One non-limiting method that may be used to produce microbial biomass fraction suitable for use in compositions of the present invention may comprise the following steps: a) providing a microbial biomass in an aqueous alkaline suspension, at pH 7-11 ; b) mechanically disintegrating the microbial biomass at a temperature below 35°C using a non-denaturing process, such that the disintegrated biomass consists of a population of soluble compounds and suspended fragments characterized by a bimodal distribution and a particle size distribution (psd) at an average of around D50 < 4.5 pm; c) subjecting the disintegrated biomass to a solid-liquid separation process, to separate the disintegrated biomass into an extract rich in small fragments and an extract rich in large fragments, wherein the extract rich in small fragments consists of a population of soluble compounds and suspended fragments characterized by a bimodal distribution and a psd of D50 around <0.5 pm or lower; and the extract rich in large fragments consists of a population of soluble compounds and suspended fragments having a psd of D50>0.5 pm; d) selecting either the extract enriched in small fragments or the extract enriched in large fragments or combining a portion of each of the extract enriched in small fragments and an extract enriched in large fragments.

[0100] The method may optionally further comprise a polishing step to improve the purity of the components and / or a concentration and / or drying step.

[0101] In a preferred embodiment the microbial biomass fraction is polished using technologies and methods known in the art. Such methods include but are not limited to filtration, adsorption, isoelectric precipitation, coagulation and solvent extraction. In a preferred embodiment subjecting the microbial cell extract obtained from disintegration and classification to polishing improves the purity of the main components of said microbial biomass fraction.

[0102] In another embodiment, the microbial biomass fraction may optionally be concentrated and / or dried. In a preferred embodiment the microbial biomass fraction is concentrated and / or dried using technologies and methods known in the art. Such methods include, but are not limited to filtration, evaporation, freeze concentration, pervaporation, sublimation and drying by spray drying, drum drying, or freeze drying.

[0103] The concentration and / or drying step may be carried out in the presence or absence of the optional polishing step. When the concentration and / or drying step is carried out in addition to the optional polishing step, the concentration and / or drying step may be performed before or after the polishing step. In an embodiment, the optional concentration and / or drying step reduces the water content of the microbial biomass fraction. In another embodiment of the aspect of the invention, the method further comprises subjecting the microbial biomass fraction to one or more additional processes, selected from filtration, adsorption, isoelectric precipitation, coagulation and solvent extraction.

[0104] A microbial biomass fraction produced using any method known in the art may be incorporated into a composition of the present invention.

[0105] The inventors have surprisingly found synergistic effects when several ingredients and additives are combined with a disintegrated microbial biomass, and / or a microbial biomass fraction (MBF) which result in surprising functional properties or unique textures.

[0106] Ingredients and additives that can be combined with a disintegrated microbial biomass, and / or microbial biomass fraction (MBF) to produce a composition of the present invention include at least one of a plant based protein (PBP), animal based protein (ABP), plant based fiber (PBF), microbial based fibre, polysaccharide, thickening agent, stabilizing agent, salt, enzyme, oil or fat.

[0107] A disintegrated microbial biomass, and / or microbial biomass fraction can be combined with any number and any combination of a plant based protein (PBP), animal based protein (ABP), plant based fiber (PBF), microbial based fibre, polysaccharide, salt, enzyme, thickening agent, stabilizing agent, oil or fat to produce a composition of the present invention. In some embodiments a disintegrated microbial biomass, and / or microbial biomass fraction can be combined with all of a plant based protein (PBP), animal based protein (ABP), plant based fiber (PBF), microbial based fibre, polysaccharide, salt, enzyme, thickening agent, stabilizing agent, oil and fat to produce a composition of the present invention.

[0108] A plant based protein refers to a protein that is derived from a plant as opposed to an animal or other source. Examples of plant based proteins include but are not limited to potato protein, soya protein, lentil protein, chickpea protein, peanut protein, almond protein, spirulina, quinoa protein, chia seed protein, tofu protein, pea protein, pistachio protein, oat protein or wheat protein. Compositions of the present invention may comprise any number of plant based proteins, for example, a composition of the present invention may comprise, one, two, three, four, five, six, seven, eight, nine, ten or more plant based proteins.

[0109] An animal based protein refers to protein derived from an animal source such as meat, fish, eggs or dairy products. Examples of animal-based proteins include milk protein, egg protein or meat protein such as beef protein, pork protein, chicken protein, turkey protein, duck protein, rabbit protein, goat protein, venison protein or pheasant protein. More specific examples include albumin, casein, collagen, gelatin, lactoglobulin, ovotransferrin, ovoalbumin, ovomucoid, lactoferrin, and whey. The skilled person would understand that alternative animal based proteins are available and can be used in compositions according to the present invention.

[0110] Compositions of the present invention may comprise any number of animal based proteins, for example, a composition of the present invention may comprise one, two, three, four, five, six, seven, eight, nine, ten or more animal based proteins.

[0111] It is understood in the art that animal based proteins can be substituted for plant based proteins and vice-versa. Therefore, any composition according to the present invention or described herein incorporating one or more plant based proteins may be formulated using one or more animal based proteins as an alternative. Likewise, any composition according to the present invention or described herein incorporating one or more animal based proteins may be formulated using one or more plant based proteins as an alternative. The skilled person would be able to decide whether to use plant or animal based proteins according to their specific needs and dietary requirements.

[0112] Examples of plant based fibre include but are not limited to psyllium fibre, konjac fibre, oat fibre, lentil fibre, resistant starch, acacia gum, cellulose, methylcellulose, hydroxypropyl methylcellulose, citrus fibre, p-glucan and pectin.

[0113] Compositions of the present invention may comprise any number of plant based fibres, for example, a composition of the present invention may comprise, one, two, three, four, five, six, seven, eight, nine, ten or more plant based fibres.

[0114] Examples of microbial based fibre include but are not limited to p-glucan and inulin. Compositions of the present invention may comprise any number of microbial based fibres, for example, a composition of the present invention may comprise, one, two, three, four, five, six, seven, eight, nine, ten or more microbial based fibres.

[0115] Examples of oil and fat include but are not limited to coconut oil, sunflower oil, olive oil, canola oil, rice oil, vegetable oil, avocado oil, sesame seed oil, flaxseed oil, walnut oil, peanut oil, rice fat, bran fat and coconut fat.

[0116] It should be understood that any fat or oil suitable for use in food preparations can be used in compositions of the present invention. The skilled person would be able to select an appropriate fat or oil to suit their specific requirements.

[0117] Compositions of the present invention may comprise any number of oils or fats, for example, a composition of the present invention may comprise, one, two, three, four, five, six, seven, eight, nine, ten or more oils or fats.

[0118] Examples of polysaccharides include but are not limited to starches, for example potato starch, tapioca starch, corn starch, wheat starch and pea starch.

[0119] Compositions of the present invention may comprise any number of polysaccharides, for example, a composition of the present invention may comprise, one, two, three, four, five, six, seven, eight, nine, ten or more polysaccharides.

[0120] Examples of salts include but are not limited to calcium chloride, sodium chloride, sodium bicarbonate, magnesium chloride and potassium chloride.

[0121] Compositions of the present invention may comprise any number of salts, for example a composition of the present invention may comprise, one, two, three, four, five, six, seven, eight, nine, ten or more salts.

[0122] Examples of enzymes include but are not limited to multi-copper oxidases, transglutaminase, laccase, proteases, transferases, hydrolases, peptidases and oxidoreductases, and / or suitable mediators. Suitable mediators refer to molecules that facilitate enzyme activity, such as co-factors. Examples of co-factors include ions such as copper, zinc and iron as well as organic molecules such as vitamins or vitamin derived molecules, such as thiamine pyrophosphate and flavin adenine dinucleotide. It is understood that suitable mediators of an enzyme are determined by the specific enzyme being employed.

[0123] Compositions of the present invention may comprise any number of enzymes, for example a composition of the present invention may comprise, one, two, three, four, five, six, seven, eight, nine, ten or more enzymes.

[0124] Compositions of the present invention may comprise any number of suitable mediators, for example a composition of the present invention may comprise, one, two, three, four, five, six, seven, eight, nine, ten or more suitable mediators.

[0125] Examples of thickening agents include but are not limited to xanthan gum, agar-agar, pectin, corn starch and arrowroot.

[0126] Compositions of the present invention may comprise any number of thickening agents, for example a composition of the present invention may comprise, one, two, three, four, five, six, seven, eight, nine, ten or more thickening agents.

[0127] Examples of stabilizing agents include but are not limited to lecithin, and carrageenan.

[0128] Compositions of the present invention may comprise any number of stabilizing agents, for example a composition of the present invention may comprise, one, two, three, four, five, six, seven, eight, nine, ten or more stabilizing agents.

[0129] In an embodiment, the disintegrated microbial biomass, and / or microbial biomass fraction in the composition of the invention comprises 5-100 wt% protein. The disintegrated microbial biomass, and / or microbial biomass fraction may comprise 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%,

[0130] 37%, 38%, 39%, 40%, 41 %, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51 %,

[0131] 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61 %, 62%, 63%, 64%, 65%, 66%,

[0132] 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% protein.

[0133] In a preferred embodiment the disintegrated microbial biomass, and / or microbial biomass fraction in the composition of the invention comprises at least 30 wt% protein. More preferably, the disintegrated microbial biomass, and / or microbial biomass fraction in the composition of the invention comprises at least 50 wt% protein. Even more preferably, the disintegrated microbial biomass, and / or microbial biomass fraction in the composition of the invention comprises more than 50 wt% protein. In some embodiments, the disintegrated microbial biomass, and / or microbial biomass fraction in the composition of the present invention comprises between 50-80 wt% protein, between 55-75 wt%, or preferably between 60-75% protein.

[0134] In some embodiments the disintegrated microbial biomass, and / or microbial biomass fraction has a protein content of approximately 60 wt%.

[0135] In a more preferred embodiment the disintegrated microbial biomass, and / or microbial biomass fraction has a protein content of approximately 70 wt%.

[0136] In an embodiment, the disintegrated microbial biomass, and / or microbial biomass fraction has a fibre content of at least 1-95 wt%, the disintegrated microbial biomass, and / or microbial biomass fraction may comprise, 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%,

[0137] 39%, 40%, 41 %, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51 %, 52%, 53%,

[0138] 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61 %, 62%, 63%, 64%, 65%, 66%, 67%, 68%,

[0139] 69%, 70%, 71 %, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81 %, 82%, 83%,

[0140] 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94% or 95% fibre.

[0141] In an embodiment, the disintegrated microbial biomass, and / or microbial biomass fraction has a fibre content of at least 20 wt%.

[0142] In a preferred embodiment, the disintegrated microbial biomass, and / or microbial biomass fraction has a fibre content of at least 30 wt%. In some embodiments, the disintegrated microbial biomass, and / or microbial biomass fraction has a protein content of at least 50 wt% and a fibre content of less than 20 wt%.

[0143] In some embodiments, the disintegrated microbial biomass, and / or microbial biomass fraction has a fibre content of at least 20 wt% and a protein content of less than 50 wt%.

[0144] In other embodiments, the disintegrated microbial biomass, and / or microbial biomass fraction has a fibre content of at least 20 wt% and a protein content of at least 50 wt%.

[0145] In an embodiment, a composition according to the present invention comprises 0.5%, 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%,

[0146] 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41 %, 42%, 43%, 44%, 45%, 46%, 47%,

[0147] 48%, 49%, 50%, 51 %, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61 %, 62%,

[0148] 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71 %, 72%, 73%, 74%, 75%, 76%, 77%,

[0149] 78%, 79%, 80%, 81 %, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%,

[0150] 93%, 94%, 95%, 96%, 97%, 98% or 99% disintegrated microbial biomass, or microbial biomass fraction.

[0151] Unless stated otherwise, all percentages for components in compositions of the present invention given herein are on a weight-by-weight (w / w) basis. This means that the percentage of a component is based on its weight relative to the total weight of the composition. For example, if a composition comprises 10% microbial biomass fraction, this means that microbial biomass fraction accounts for 10% of the total weight of the composition and other components make up the remaining 90%.

[0152] In some embodiments disintegrated microbial biomass, and / or microbial biomass fraction is included in compositions of the present invention as a powder. In alternative embodiments disintegrated microbial biomass, and / or microbial biomass fraction is included in compositions of the present invention as a liquid.

[0153] In some embodiments a composition according to the present invention comprises disintegrated microbial biomass. In other embodiments, a composition according to present invention comprises microbial biomass fraction. In other embodiments a composition according to the present invention comprises disintegrated microbial biomass and microbial biomass fraction.

[0154] In a preferred embodiment a composition according to the present invention comprises between around 2-20% disintegrated microbial biomass, and / or microbial biomass fraction. In some preferred embodiments a composition according to the present invention comprises around 2, 4, 5, 8, 10 or 15% microbial biomass fraction.

[0155] In an alternative preferred embodiment, a composition according to the present invention comprises around 25% microbial biomass fraction.

[0156] The skilled person would be able to adjust the amount of disintegrated microbial biomass, and / or microbial biomass fraction in a composition of the present invention in order to meet their specific requirements.

[0157] In an embodiment, the composition of the present invention comprises at least one disintegrated microbial biomass, and / or microbial biomass fraction and at least one plant based protein.

[0158] In some embodiments, the composition of the present invention comprises only microbial biomass fraction and plant based protein.

[0159] In some embodiments, the composition of the present invention comprises at least one microbial biomass fraction and at least one animal based protein.

[0160] In some embodiments, the composition of the present invention comprises only microbial biomass fraction and animal based protein.

[0161] In an embodiment, the animal based protein in a composition of the present invention is milk protein.

[0162] In some embodiments, the composition of the present invention comprises at least one disintegrated microbial biomass, and / or microbial biomass fraction, at least one plant based protein and at least one oil or fat. In some embodiments the composition of the present invention comprises at least one disintegrated microbial biomass, and / or microbial biomass fraction, at least one plant based protein, at least one plant based fibre and at least one oil or fat.

[0163] In some embodiments the composition of the present invention comprises at least one disintegrated microbial biomass, and / or microbial biomass fraction, at least one plant based protein and at least one plant based fibre.

[0164] In a preferred embodiment, the composition comprises a ratio of disintegrated microbial biomass or microbial biomass fraction (MBF) to plant based protein of at least 50:50 (dry matter).

[0165] In an embodiment, the plant based protein in a composition of the present invention is potato protein.

[0166] In an embodiment, when the plant based protein is potato protein, said potato protein comprises ~90 wt% protein.

[0167] In an embodiment, a composition of the present invention comprises at least 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21 %, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31 %, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41 %, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50% or more plant based protein, preferably said plant based protein is potato protein. Preferably a composition of the present invention comprises 1-10% plant based protein, more preferably 4% plant based protein wherein preferably said plant based protein is potato protein.

[0168] In an embodiment, a composition of the present invention comprises at least 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21 %, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31 %, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41 %, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50% or more animal based protein, wherein preferably said animal based protein is milk protein. In an embodiment, a composition of the present invention comprises around 0.1-10g of disintegrated microbial biomass or microbial biomass fraction (MBF) per gram of animal based protein, wherein preferably said animal based protein is milk protein. In some embodiments, a composition of the present invention comprises around 0.5-5g, 1-10g, 0.1 -1g, 0.1 -0.5g, 0.5-2g, 2-6g, 6-10g of disintegrated microbial biomass or microbial biomass fraction (MBF) per gram of animal based protein, wherein preferably said animal based protein is milk protein.

[0169] In an embodiment a composition according to the present invention comprises 4% plant based protein, 8% microbial biomass fraction (MBF), 40% oil, with the remainder being water. Preferably the microbial biomass fraction and plant based protein are added as powders. Alternatively, the microbial biomass fraction and plant based protein are added as liquids.

[0170] In an embodiment a composition according to the present invention comprises a blend of microbial biomass fraction and potato protein at a ratio of 25:75 on a dry matter basis. Such compositions exhibit superior gelation hardness compared to alternative blend ratios, including those with a higher protein content. This is shown in Example 2.

[0171] The texturizing ability of ingredients is crucial for their development as analogues that can mimic the sensory attributes of meat, cheese, egg, sauces, cakes and other commercial products. Particularly important properties are hardness, chewiness, cohesiveness and springiness; these attributes are desired for many reasons including the provision of bite and succulence.

[0172] The inventors have found that a composition comprising a microbial biomass fraction with a protein content of at least 50 wt%, preferably at least 70 wt% and potato protein in an oil-water suspension results in a texture with high firmness, springiness and chewiness compared to a microbial biomass fraction alone or potato protein alone , this is shown in Example 3 where sunflower oil was used in the oil-water suspension, however, similar results are observed using alternative oils such as rapeseed oil and coconut oil, or indeed other plant based oils. In an embodiment, a microbial biomass fraction with a protein content of at least 50% yields high firmness and chewiness when in an emulsion system comprising an oil-water suspension and the amount of oil in the oil-water suspension is set between 30-60 v%.

[0173] In a preferred embodiment the disintegrated microbial biomass, and / or microbial biomass fraction is in an emulsion system wherein said emulsion system is an oil-water suspension with approximately 35-55 v% oil.

[0174] In another preferred embodiment a composition of the present invention is in an emulsion system wherein said emulsion system is an oil-water suspension comprising approximately 40 v% oil.

[0175] In an embodiment, the ratio of disintegrated microbial biomass or microbial biomass fraction to plant based protein is 1.7-2.3:1 when prepared in an emulsion system comprising an oil-water suspension comprising at least 40 v% oil.

[0176] In an embodiment, the composition of the present invention comprises at least one disintegrated microbial biomass, and / or microbial biomass fraction and at least one plant based fibre.

[0177] In some embodiments, the composition of the present invention comprises only microbial biomass fraction and plant based fibre.

[0178] In an embodiment, the composition of the present invention comprises at least one disintegrated microbial biomass, and / or microbial biomass fraction at least one plant based fibre and at least one oil or fat.

[0179] Compositions of the present invention comprising at least one microbial biomass fraction in combination with at least one plant based fibre were found to exhibit unique textures when blended at various ratios. For example, a microbial biomass fraction with a protein content of at least 50% blended with psyllium fibre results in a texture with good firmness, springiness and chewiness, resembling the texture of meat.

[0180] In an embodiment, a composition of the present invention comprises at least O.1%, 0.2%, 0.3%, 0.4% 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11 %, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21 %, 22%, 23%, 24%,

[0181] 25%, 26%, 27%, 28%, 29%, 30%, 31 %, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%,

[0182] 40%, 41 %, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51 %, 52%, 53%, 54%,

[0183] 55%, 56%, 57%, 58%, 59%, 60%, 61 %, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%,

[0184] 70%, 71%, 72%, 73%, 74%, 75% or more plant based fibre, preferably said plant based fibre is p-glucan (preferably oat p-glucan), psyllium fibre or konjac fibre.

[0185] In a preferred embodiment a composition of the present invention comprises 0.5-5% plant based fibre, more preferably between 1.5-3% plant based fibre, even more preferably 1.5% or 3% plant based fibre, wherein preferably said plant based fibre is konjac fibre.

[0186] In a preferred embodiment a composition of the present invention comprises 8% microbial biomass fraction with a protein content of at least 50%, 1.5 to 3% plant based fibre and the remainder being water, wherein preferably said plant based fibre is konjac fibre.

[0187] In another preferred embodiment a composition of the present invention comprises 8% microbial biomass fraction with a protein content of at least 50%, 1.5 to 3% plant based fibre, wherein preferably said plant based fibre is konjac fibre and 20-60% oil, preferably 40% oil, wherein preferably, the oil is selected from at least one of sunflower oil, rapeseed oil, coconut oil, or any other oil suitable for use in food preparations; and the remainder is water.

[0188] In another embodiment, a composition according to the present invention comprises at least 1% microbial biomass fraction, at least 0.5% w / w oat p-glucan, the remainder being made up of water and oil. The inventors have found that such a composition provides a texture with unique mechanical properties such as firmness, springiness, cohesiveness, elasticity, good bite and juiciness that resemble meat products.

[0189] In a preferred embodiment, a composition according to the present invention comprises 0.5-20% w / w microbial biomass fraction, 0.1-15% w / w p-glucan (preferably oat p- glucan), 10-60% w / w oil wherein preferably, the oil is selected from at least one of sunflower oil, rapeseed oil, coconut oil, or any other oil suitable for use in food preparations; and the remainder is water. In another embodiment, a composition of the present invention comprises around 5% microbial biomass fraction with a protein content of at least 50%, 2% psyllium fibre, 50% oil wherein preferably, the oil is selected from at least one of sunflower oil, rapeseed oil, coconut oil, or any other oil suitable for use in food preparations; and the remainder is water.

[0190] In another embodiment, a composition of the present invention comprises around 4% microbial biomass fraction with a protein content of at least 50%, 3% citrus fibre, 50% oil; wherein preferably, the oil is selected from at least one of sunflower oil, rapeseed oil, coconut oil, or any other oil suitable for use in food preparations; and the remainder is water.

[0191] In another embodiment, a composition of the present invention comprises around 2% microbial biomass fraction with a protein content of at least 50%, 1% methylcellulose, and the remainder is water.

[0192] In another embodiment, a composition of the present invention comprises around 2% microbial biomass fraction with a protein content of at least 50%, 2% pectin, 20% oil, wherein preferably, the oil is selected from at least one of sunflower oil, rapeseed oil, coconut oil, or any other oil suitable for use in food preparations; and the remainder is water.

[0193] In some embodiments, a composition of the present invention comprises at least one disintegrated microbial biomass and / or microbial biomass fraction with a protein content of at least 50% and a thickening agent wherein the thickening agent accounts for around 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.1 %, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1 %, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, 3%, 3.1%, 3.2%, 3.3%, 3.4%, 3.5%, 3.6%, 3.7%, 3.8%, 3.9%, 4%, 4.1%, 4.2%, 4.3%, 4.4%, 4.5%, 4.6%, 4.7%, 4.8%, 4.9%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, 10%, 10.5%, 11 %, 11.5%, 12%, 12.5%, 13%, 13.5%, 14%, 14.5%, 15%, 15.5%, 16%, 16.5%, 17%, 17.5%, 18%, 18.5%, 19%, 19.5% or 20% of the composition. In some preferred embodiments the thickening agent is xanthan gum and accounts for around 0.1 % of the composition. In some embodiments a composition of the present invention comprises at least one disintegrated microbial biomass and / or microbial biomass fraction and at least one thickening agent, wherein the thickening agent accounts for around 0.01-20%, 0.01-5%, 0.02-1 % or 0.05-0.5% of the composition; preferably the thickening agent is xanthan gum and accounts for 0.05-0.5% of the composition.

[0194] In some embodiments, a composition of the present invention comprises at least one disintegrated microbial biomass and / or microbial biomass fraction with a protein content of at least 50% and at least one salt wherein the salt accounts for around 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.1%, %, 0.11%, 0.12%, 0.13%, 0.14%, 0.15%, 0.16%, 0.17%, 0.18%, 0.19%, 0.2%,%, 0.21%, 0.22%, 0.23%, 0.24%,

[0195] 0.25%, 0.26%, 0.27%, 0.28%, 0.29%, 0.3%, %, 0.31%, 0.32%, 0.33%, 0.34%, 0.35%,

[0196] 0.36%, 0.37%, 0.38%, 0.39%, 0.4%, %, 0.41%, 0.42%, 0.43%, 0.44%, 0.45%, 0.46%,

[0197] 0.47%, 0.48%, 0.49%, 0.5%, %, 0.51 %, 0.52%, 0.53%, 0.54%, 0.55%, 0.56%, 0.57%,

[0198] 0.58%, 0.59%, 0.6%, 0.61%, 0.62%, 0.63%, 0.64%, 0.65%, 0.66%, 0.67%, 0.68%, 0.69%, 0.7%, 0.71 %, 0.72%, 0.73%, 0.74%, 0.75%, 0.76%, 0.77%, 0.78%, 0.79%, 0.8%, %, 0.81%, 0.82%, 0.83%, 0.84%, 0.85%, 0.86%, 0.87%, 0.88%, 0.89%, 0.9%%, 0.91%, 0.92%, 0.93%, 0.94%, 0.95%, 0.96%, 0.97%, 0.98%, 0.99%, 1%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, 3%, 3.1%, 3.2%, 3.3%, 3.4%, 3.5%, 3.6%, 3.7%, 3.8%, 3.9%, 4%, 4.1%, 4.2%, 4.3%, 4.4%, 4.5%, 4.6%, 4.7%, 4.8%, 4.9% or 5% of the composition. In some preferred embodiments salt is sodium bicarbonate and accounts for around 0.12% of the composition.

[0199] In some embodiments a composition of the present invention comprises at least one disintegrated microbial biomass and / or microbial biomass fraction and at least one salt, wherein the at least one salt accounts for around 0.01-20%, 0.01-5%, 0.02-1 % or 0.05- 0.5% of the composition; preferably the salt is sodium bicarbonate and accounts for around 0.05-0.5% of the composition.

[0200] In an embodiment a composition of the present invention comprises around 0.1-10% microbial biomass fraction and 0.05-0.5% thickening agent (preferably xanthan gum), with the remainder of the composition being water and the microbial biomass fraction preferably has a protein content of at least 50%. In an embodiment a composition of the present invention comprises around 0.1-10% microbial biomass fraction and 0.05-0.5% salt (preferably sodium bicarbonate), with the remainder of the composition being water and the microbial biomass fraction preferably has a protein content of at least 50%.

[0201] In an embodiment a composition of the present invention comprises around 0.1-10% microbial biomass fraction, 0.05-0.5% thickening agent (preferably xanthan gum) and 0.5-0.05% salt (preferably sodium bicarbonate), with the remainder of the composition being water, and the microbial biomass fraction preferably has a protein content of at least 50%.

[0202] In a specific embodiment a composition of the present invention comprises around 2% microbial biomass fraction with a protein content of at least 50%, 0.1 % xanthan gum, 0.12% sodium bicarbonate and the remainder is water.

[0203] In another specific embodiment a composition of the present invention comprises microbial biomass fraction with a protein content of at least 50% and xanthan gum wherein the xanthan gum accounts for around, 1-20%, 1-10% or 3-5% (preferably 3.5- 4.5%) of the weight of the microbial biomass fraction wherein the microbial biomass fraction and xanthan gum mixture is combined with sodium bicarbonate at a ratio of 1 :0.02-0.1 , preferably 1 :0.05 (wt).

[0204] In another embodiment a composition of the present invention comprises microbial biomass fraction with a protein content of at least 50% and sodium bicarbonate, wherein preferably the ratio of microbial biomass fraction to sodium bicarbonate is 1 :0.02-0.1 , preferably 1 :0.05 (wt).

[0205] In an embodiment, a composition according to the present invention comprises a ratio of disintegrated microbial biomass or microbial biomass fraction to plant based fibre of 1- 2:1 , preferably 1.6:1 (dry matter).

[0206] In an alternative embodiment a composition according to the present invention comprises a ratio of disintegrated microbial biomass or microbial biomass fraction to plant based fibre of 5-6:1 , preferably 5.3:1 , blended in an emulsion with an oil content of at least 30%, preferably 40%.

[0207] In yet another embodiment, the ratio of disintegrated microbial biomass or microbial biomass fraction to plant based fibre is 6-7:1 , preferably 6.6:1 , blended in an emulsion with an oil content of at least 30%, preferably 40%.

[0208] A blend of at least one microbial biomass fraction with a protein content of at least 60% and konjac fibre when combined in a water-oil suspension comprising at least 40% oil, yields a texture with high hardness, springiness and chewiness. In contrast, textures obtained with a microbial biomass fraction or konjac fibre alone and / or in different oilwater ratios were weaker and therefore unsuitable for use in food compositions.

[0209] A composition according to the present invention may comprise at least one, at least two, at least three, at least four, at least five, or more disintegrated microbial biomasses, and / or microbial biomass fractions (MBFs).

[0210] In a preferred embodiment, a composition of the present invention comprises one or two disintegrated microbial biomasses or microbial biomass fractions.

[0211] In some embodiments a composition according to the present invention comprises a mixture of at least one oil or fat, at least one polysaccharide, a first microbial biomass fraction with a protein content of at least 50% and a second microbial biomass fraction with a fibre content of at least 20% wherein the ratio of the oil / fat and polysaccharide mixture to the first microbial biomass fraction to the second microbial biomass fraction is at least 1 :0.5:0.5, preferably 1 :0.5:5.

[0212] In an alternative embodiment, there is provided a composition according to the present invention comprising a first microbial biomass fraction with a protein content of at least 50% and a second microbial biomass fraction with a fibre content of at least 20% are in an aqueous suspension in the range of 50gL-150g / L wherein the ratio of the first microbial biomass fraction to the second microbial biomass fraction is in the range of 1.5:1 to 1 :1.5. Compositions of microbial biomass fractions can also be obtained by subjecting disintegrated microbial biomass and / or microbial biomass fractions to specific combinations of time and temperature. High temperatures have a detrimental effect on the functional properties of proteins, mostly due to denaturation, radicalisation, polymerisation and other chemical reactions. However, the inventors have surprisingly found that when exposing a microbial biomass fraction with a protein content of at least 50%, preferably at least 60%, more preferably around 70%, to a specific temperaturetime combination of preferably around 68°C for approximately 1 hour, when moisture is not higher than 10%, the functional properties, such as gelation performance of the resulting composition are drastically enhanced. In another embodiment, a composition exhibiting enhanced functional properties can also be obtained by exposing a microbial biomass fraction with a fibre content >20% to a specific combination of temperature and time (example 6).

[0213] A disrupted microbial biomass, and / or microbial biomass fraction used in a composition of the present invention may comprise a moisture content of approximately 0.1%, 0.5%, 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%,

[0214] 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41 %, 42%, 43%, 44%, 45%, 46%, 47%,

[0215] 48%, 49%, 50%, 51 %, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61 %, 62%,

[0216] 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71 %, 72%, 73%, 74%, 75%, 76%, 77%,

[0217] 78%, 79%, 80%, 81 %, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90%.

[0218] Preferably a disrupted microbial biomass and / or microbial biomass fraction used in a composition of the present invention has a moisture content of at most between 10-15%.

[0219] In another preferred embodiment a disintegrated microbial biomass and / or microbial biomass fraction used in a composition of the present invention has a moisture content of less than 10%.

[0220] In one embodiment, the at least one microbial biomass fraction has a protein content of at least 50%, and a moisture content of at most 15% and said microbial biomass fraction is preferably heated to 60-90°C for 160-200 minutes, more preferably 60-75°C for 45-90 minutes, most preferably approximately 68°C for approximately 1 hour This is shown in example 5 and figure 6. In a preferred embodiment, the at least one microbial biomass fraction has a fibre content of at least 20% and a moisture content of at most 10% and said microbial biomass fraction is heated to a temperature of 75-85°C for 170-190 minutes. This is shown in example 5 and figure 6.

[0221] The inventors have found that the texture properties of an analogue can be improved when a microbial biomass fraction with a fibre content of at least 20% is blended with konjac fibre in a water-oil suspension comprising at least 40% oil, this is shown in Example 3.

[0222] The inventors have surprisingly found that foam stability of the disintegrated biomass or microbial biomass fraction can be increased by subjecting the disintegrated biomass and / or microbial biomass fraction with a protein content of at least 50% and a moisture content of < 15% (in powder form) to thermal treatment at temperatures in the range of 90-200°C, preferably 90-180°C, as shown in example 8.

[0223] Surprisingly the inventors have found that the functional properties of the microbial biomass fractions can be fine-tuned by further controlling the relative humidity during the thermal treatment. In particular, interfacial properties and viscoelastic properties can be drastically improved. Moreover, the inventors found that controlling the ranges of temperature and relative humidity can be used to alter the organoleptic characteristics of the microbial biomass fractions, as shown in example 9. Examples of relative humidities that are appropriate for thermal treatment of the microbial biomass or microbial biomass fraction are 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, or any intermediate value. Preferably the relative humidity is more than 60%, more preferably the relative humidity is more than 80%.

[0224] In an embodiment, the composition of the present invention comprises at least one disintegrated microbial biomass and / or microbial biomass fraction and at least one polysaccharide.

[0225] In some embodiments, the composition of the present invention comprises only microbial biomass fraction and polysaccharides, optionally in water. Sources of polysaccharides that may be used in compositions of the present invention include flour. Examples of flour suitable for use in compositions of the present invention include but are not limited to wheat flour, almond flour, rye flour, oat flour, corn flour, rice flour and chickpea flour.

[0226] In an embodiment, the composition of the present invention comprises at least one disintegrated microbial biomass and / or microbial biomass fraction and at least one oil or fat. Preferably said oil or fat is plant based.

[0227] In some embodiments, the composition of the present invention comprises only microbial biomass fraction and oil or fat.

[0228] In an embodiment, the composition of the present invention comprises at least one disintegrated microbial biomass and / or microbial biomass fraction and at least one flour and at least one oil or fat.

[0229] In some embodiments, the composition of the present invention comprises only microbial biomass fraction, flour and oil or fat.

[0230] The inventors have found compositions that mimic the viscoelastic and melting properties of animal fats. For example, a microbial biomass fraction with a protein content of at least 50% when prepared in combination with a wheat flour delivers a texture that is suitable for use in the preparation of animal fat analogues, as shown in Example 5. In addition to this, the inventors have found that if appropriate plant-based lipid / fat is selected the melting properties of animal-fat tissue can also be mimicked, shown in Example 4.

[0231] In an embodiment, a composition of the present invention comprises at least 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21 %, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31 %, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41 %, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50% or more flour, preferably said flour is wheat flour.

[0232] In a preferred embodiment, a composition of the present invention comprises 10-20%, more preferably 15% flour, preferably said flour is wheat flour. In another preferred embodiment a composition of the present invention comprises 15% flour, preferably wheat flour, around 5-8% microbial biomass fraction, 40% oil and the remainder is water.

[0233] In an embodiment, a composition of the present invention comprises at least one microbial biomass fraction with a protein content of at least 50% and flour at a ratio of 0.2:0.5, preferably 0.34, preferably the flour is wheat flour. Wherein said composition is preferably prepared in an emulsion comprising a water-oil suspension comprising 30- 50% oil, preferably 40% oil, wherein said oil is preferably selected from at least one of sunflower oil, rapeseed oil, coconut oil, or any other oil suitable for use in food preparations.

[0234] In an embodiment, a composition of the present invention comprises at least 0.5%, 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%,

[0235] 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41 %, 42%, 43%, 44%, 45%, 46%, 47%, 48%,

[0236] 49%, 50%, 51 %, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61 %, 62%, 63%,

[0237] 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71 %, 72%, 73%, 74%, 75%, 76%, 77%, 78%,

[0238] 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% or 90% oil or fat, preferably said oil or fat is a plant based oil or fat, more preferably said oil or fat is sunflower oil, rapeseed oil or coconut fat. It should be understood that any oil suitable for use in food preparations can be used in any composition of the present invention.

[0239] In a preferred embodiment a composition of the present invention comprises 50% coconut fat, 10% microbial biomass fraction with the remainder being water.

[0240] In an embodiment, disintegrated microbial biomass and / or microbial biomass fractions comprising proteins and / or fibres can be made into microbial compositions by combining said disintegrated microbial biomass and / or microbial biomass fraction with salts. The inventors have found that such combinations have unexpected functional properties. For example, blending a microbial biomass fraction with a fibre content of at least 20% with Ca, in particular CaCh results in a composition with enhanced gelation properties, as shown in Example 6. In an embodiment, a composition of the present invention comprises at least one microbial biomass fraction with a protein content of at least 50% and / or a fibre content of at least 20% and between 5-5000 ppm CaCh. Preferably, said composition comprises 20-1000 ppm CaCh, more preferably in the range of 200-800 ppm CaCh.

[0241] In an embodiment, a composition of the present invention comprises at least one microbial biomass fraction with a protein content of at least 50% and Ca at a ratio of 250:1.

[0242] In another embodiment, a composition of the present invention comprises at least one microbial biomass fraction with a fibre content of at least 20% and Ca at a ratio of 250: 1.

[0243] In a further embodiment, the texturizing ability of disintegrated microbial cell biomass and / or microbial biomass fractions can be significantly improved with a composition is made comprising at least one disintegrated microbial cell biomass and / or microbial biomass fraction and at least one enzyme belonging to the group multicopper oxidases or transglutaminases.

[0244] Multicopper oxidases are a diverse group of enzymes that couple the oxidation of a variety of substrates to the reduction of dioxygen and include notable members such as tyrosinases, laccases, ascorbate oxidase, Fet3p, CueO and ceruloplasmin. Multicopper oxidases are widely distributed in nature and are involved in many different processes, including metal homeostasis, ascorbate metabolism, and phenolic substrate oxidation.

[0245] Transglutaminases are enzymes that catalyse the formation of a covalent bond between glutamine and lysine residues of proteins and peptides.

[0246] Preferably, when the composition comprises at least one microbial biomass and at least one multicopper oxidase, preferably the multicopper oxidase is a tyrosinase or a laccase.

[0247] In a preferred embodiment, when the composition comprises at least one enzyme, the enzyme is transglutaminase.

[0248] In some embodiments a composition of the present invention comprises at least one disintegrated microbial biomass and / or microbial biomass fraction and at least one enzyme (preferably transglutaminase) wherein the disintegrated microbial biomass and / or microbial biomass fraction is suspended in water to be at a concentration of between around 5-500 g / L, 10-400g / L, 20-300 g / L, 30-200 g / L, or 40-150g / L.

[0249] In some embodiments a composition of the present invention comprises at least one disintegrated microbial biomass and / or microbial biomass fraction and at least one enzyme (preferably transglutaminase) wherein the disintegrated microbial biomass and / or microbial biomass fraction is suspended in water to be at a concentration of around 5, 10, 15, 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, 160, 165, 170, 175, 180, 185, 190,

[0250] 195, 200, 205, 210, 215, 220, 225, 230, 235, 240, 245, 250, 255, 260, 265, 270, 275,

[0251] 280, 285, 290, 295, 300, 305, 310, 315, 320, 325, 330, 335, 340, 345, 350, 355, 360,

[0252] 365, 370, 375, 380, 385, 390, 395, 400, 405, 410, 415, 420, 425, 430, 435, 440, 445,

[0253] 450, 455, 460, 465, 470, 475, 480, 485, 490, 495 or 500 g / L. In a preferred embodiment the disintegrated microbial biomass and / or microbial biomass fraction is suspended in water to be at a concentration of around 100 g / L.

[0254] In some embodiments a composition of the present invention comprises at least one disintegrated microbial biomass and / or microbial biomass fraction and at least one enzyme wherein the disintegrated microbial biomass and / or microbial biomass fraction is suspended in water and the enzyme is added at a concentration of between around 0.01-1g, 0.05-0.7, 0.1-0.5, or 0.2-0.5g per gram of disintegrated microbial biomass and / or microbial biomass fraction, preferably the enzyme is transglutaminase.

[0255] In some embodiments a composition of the present invention comprises at least one disintegrated microbial biomass and / or microbial biomass fraction and at least one enzyme wherein the disintegrated microbial biomass and / or microbial biomass fraction is suspended in water and the at least one enzyme is added at a concentration of around 0.01 , 0.015, 0.02, 0.025, 0.03, 0.035, 0.04, 0.045, 0.05, 0.055, 0.06, 0.065, 0.07, 0.075, 0.08, 0.085, 0.09, 0.1 , 0.15, 0.2, 0.25, 0.3, 0.35, 0.4, 0.45, 0.5, 0.55, 0.6, 0.65, 0.7, 0.75, 0.8, 0.85, 0.9, 0.95 or 1g per gram of disintegrated microbial biomass and / or microbial biomass fraction. In a preferred embodiment the at least one enzyme is added at a concentration between around 0.2-0.5g per gram of disintegrated microbial biomass and / or microbial biomass fraction and the enzyme is preferably transglutaminase. More preferably the at least one enzyme is added at a concentration of around 0.3 or 0.35g per gram of disintegrated microbial biomass and / or microbial biomass fraction. Most preferably the at least one enzyme is added at a concentration of around 0.35g per gram of disintegrated microbial biomass and / or microbial biomass fraction.

[0256] In some embodiments, compositions of the present invention comprising at least one disintegrated microbial biomass and / or microbial biomass fraction and at least one enzyme (preferably transglutaminase) are subjected to incubation at between around 20-60, 25-50, or 30-40°C, preferably around 35°C for a period of 10-180, 20-120, or SO- 75 minutes, preferably around 60 minutes.

[0257] In embodiments of the invention where the composition comprises at least one disintegrated microbial biomass and / or microbial biomass fraction and at least one enzyme the skilled person would be able to adjust the concentration of the enzyme added and the duration and temperature of incubation depending on the specific enzyme being used and their specific requirements.

[0258] In a specific embodiment a composition of the present invention comprises at least one microbial biomass fraction with a protein content of at least 50% suspended in water at a concentration of around 100 g / L and has been incubated with around 0.35g of transglutaminase per gram of microbial biomass fraction at around 35°C for around 60 minutes.

[0259] Compositions according to the present invention are suitable for use in food analogues such as meat analogues, dairy analogues and egg analogues.

[0260] Examples of meat include but are not limited to chicken, beef, lamb, pork, venison, or veal analogues.

[0261] Examples of dairy analogues include but are not limited to cheese, yogurt, butter, or cream analogues.

[0262] In another aspect of the invention there is provided a method of making a composition, comprising at least one disintegrated microbial biomass, and / or fractions of microbial biomass (MBF) in combination with at least one other material. Said other material may be selected from at least one of a plant based protein (PBP), animal based protein (ABP), plant based fiber (PBF), microbial based fibre, polysaccharide, salt, oil or fat.

[0263] In one embodiment a composition of the present invention is made in an aqueous system. In this embodiment at least one disintegrated microbial biomass, and / or microbial biomass fraction and at least one other material selected from one or more of a plant based protein (PBP), animal based protein (ABP), plant based fiber (PBF), microbial based fibre, polysaccharide, salt, oil or fat is dispersed in water and optionally mixed to obtain a homogenous suspension. Said suspension may then optionally be subjected to mechanical homogenization, preferably at room temperature. Preferably, the at least one disintegrated microbial biomass, and / or microbial biomass fraction and at least one other material are blended to ensure equal distribution of components before being dispersed in water.

[0264] In a preferred embodiment a composition of the present invention is made in an emulsion system. Emulsions are a class of disperse systems consisting of two immiscible liquids. In this embodiment, at least one disintegrated microbial biomass, and / or microbial biomass fraction and at least one other material selected from one or more of a plant based protein (PBP), plant based fiber (PBF), microbial based fibre, polysaccharide, enzyme, thickening agent, stabilizing agent, salt, oil or fat is dispersed in an emulsion and optionally mixed to obtain a homogenous emulsion. Said emulsion may then optionally be subjected to mechanical homogenization, preferably at room temperature, Preferably, the at least one disintegrated microbial biomass, and / or microbial biomass fraction and at least one other material are blended to ensure equal distribution of components before being dispersed in the emulsion.

[0265] Preferably, the emulsion system used to make a composition of the present invention is an oil-water suspension wherein the amount of oil in the oil-water suspension is set between 20-60 v%. Preferably said emulsion system comprises at least 40 v% oil or between 55-60 v% oil, most preferably the emulsion system comprises approximately 40% oil.

[0266] In another aspect of the invention there is provided a method of processing a composition comprising at least one disintegrated microbial biomass, and / or microbial biomass fraction (MBF) in combination with at least one other material wherein said other material may be selected from at least one of a plant based protein (PBP), animal based protein (ABP), plant based fiber (PBF), microbial based fibre, polysaccharide, salt, enzyme, thickening agent, stabilizing agent, oil or fat; however, any other suitable ingredient or additive known in the art may also be used.

[0267] In some embodiments, processing a composition of the present invention enhances the functional properties of said composition. Functional properties that may be enhanced as a result of such processing include but are not limited to gelation capacity, chewiness, foaming capacity emulsification properties, glazing properties, browning properties, firmness, springiness, succulence, water holding capacity and oil holding capacity.

[0268] In an embodiment there is provided a method of processing a composition according to the present invention using at least one of hydration, blending, thermal processing, sieving, granulation, cutting / chopping, whipping, cooling, frying, dewatering, pressing, kneading, homogenizing or mixing.

[0269] Thermal processing may be carried out at a temperature of 60-300°C for 1-300 minutes. The time and temperature at which thermal processing is carried out would be adjusted by the skilled person according to the specific composition being processed.

[0270] In some embodiments there is provided a method of processing a composition according to the present invention using at least one of hydration, blending and thermal processing at a temperature of at least ~90°C for at least 10 minutes to form a matrix that resembles dairy cheese.

[0271] In a preferred embodiment, a composition according to the present invention is processed using all of hydration, blending and thermal processing at a temperature of at least ~90°C for at least 10 minutes to form a matrix that resembles dairy cheese.

[0272] In some embodiments, there is provided a method of processing a composition according to the present invention by heating. The skilled person would be able to adjust the time and temperature that the composition is heated to in order to meet their specific needs and taking into account the amount of the composition to be heated. In some embodiments the composition may be heated to any temperature between 40 - 300°C. Heating may take place for 1-300 minutes. Preferably, the composition is heated to any temperature between 60-200°C for 20-90 minutes. Heating may take place at relative humidities of 30-100%, 40-100%, 50-100%, 60-100%, 70-100%, 80-100% or 90-100% preferably relative humidity is above 60%, more preferably above 70%, even more preferably above 80%.

[0273] In an embodiment, there is provided a method of processing a composition according to the present invention by heating said composition to 60-90°C for 40-80 minutes. Preferably, a composition according to the present invention is heated to ~68°C for 50- 70 minutes. It is understood that the skilled person would be able to adjust heating temperatures and times in order to meet their specific needs.

[0274] In some embodiments, controlling the temperature, relative humidity and duration of processing of a disintegrated microbial biomass, microbial biomass fraction or composition of the present invention improves the organoleptic properties of said microbial biomass, a microbial biomass fraction or composition. Non-limiting examples of organoleptic attributes that can be improved are bitterness, astringency, mustiness, fruity / flora flavours / aromas, reduced metallic flavours and masking, reduction or elimination of other off-flavours.

[0275] In another aspect of the invention there is provided a method of incorporating the composition of the present invention into a food product or cosmetic.

[0276] Also within the scope of the invention are food products and cosmetics comprising the composition of the present invention.

[0277] Although primarily described herein with reference to preparations obtained from yeast cells, the invention is not limited to the same. Various other microorganisms can be used. In embodiments, the microbe may be selected from fungi, including yeast (preferably Saccharomyces sp, more preferably brewer’s or baker’s yeast, or Pichia sp); plants, in particular microalgae (including Tetraselmis sp or Chlorella sp, for example C. vulgaris)', and cyanobacteria (including Arthrospira sp, preferably A. platensis). The microbe may also be selected from bacteria, for example Methylobacterium or lactic acid bacteria.

[0278] Other variations to the disclosed embodiments can be understood and effected by those skilled in the art in practicing the claimed invention, from a study of the drawings, the disclosure, and the appended claims. In the claims, the word "comprising" does not exclude other elements or steps, and the indefinite article "a" or "an" does not exclude a plurality. The mere fact that certain measures are recited in mutually different dependent claims does not indicate that a combination of these measured cannot be used to advantage.

[0279] While the foregoing disclosure provides a general description of the subject matter encompassed within the scope of the present invention, including methods, as well as the best mode thereof, of making and using this invention, the following examples are provided to further enable those skilled in the art to practice this invention and to provide a complete written description thereof. However, those skilled in the art will appreciate that the specifics of these examples should not be read as limiting on the invention, the scope of which should be apprehended from the claims and equivalents thereof appended to this disclosure. Various further aspects and embodiments of the present invention will be apparent to those skilled in the art in view of the present disclosure.

[0280] Examples

[0281] Example 1 : Method for producing a microbial biomass fraction

[0282] This example provides a method for producing a microbial biomass fraction (MBF) with a protein content of at least 50%.

[0283] A microbial biomass fraction was produced according to the method described in PCT / EP2021 / 075137 (WO2022 / 058287). In summary, said microbial biomass fraction was produced by i) providing an aqueous suspension comprising yeast cells at -100 g / L and adjusting this suspension to -pH 9 with NaOH; ii) subjecting said suspension to mechanical cell disintegration at a temperature of <25°C, to obtain an aqueous suspension comprising disintegrated microbial cells; and iii) separating the suspension using centrifugation to provide a population of suspended cell fragments in an aqueous mixture with soluble compounds.

[0284] This suspension was subjected to mild centrifugation to yield a fraction enriched in small cell fragments (FESF) with an average d50 < 0.5 pm and a fraction enriched in large cell fragments (FELF) with an average d50 > 0.5 pm. Both or either fractions are optionally polished with methods known in the technical field, and / or are concentrated and / or dehydrated using methods known in the technical field. The FESF was subjected to polishing by means of filtration and drying, meanwhile the FELF was subjected to drying only. This process results in two MBFs with the following composition.

[0285] Table 1 : %DW of proteins, carbohydrates, lipids ash and fibre in two microbial biomass fractions produced with the method described in Example 1. It should be noted that total fibre is accounted for by carbohydrates in each microbial biomass fraction.

[0286] Example 2: Compositions containing potato protein

[0287] An MBF (MBF1) with a protein content of 72.3 wt% and a moisture content of 5.3 wt% produced according to the method described in Example 1 , was blended with potato protein obtained from a commercial brand (Solanic® 200; Avebe, Veendam, The Netherlands) as a dry powder. The powders are blended thoroughly to make sure that the content of both proteins is evenly distributed. Furthermore, the resulting blended powder is dispersed in water until a homogenous suspension, with a solids content of 10% is obtained. The suspension is then subjected to a gelation method whereby the suspension was heated up to 90°C followed by cooling to 10°C to allow gel formation and the hardness of the resulting gels was measured. Gel hardness for various ratios of MBF to potato protein is shown in Figure 1. It is evident that there is a unique composition of a blend in the range of 25:75 (MBF : Potato Protein), in which the maximum gelation hardness is obtained. Gelation hardness is a crucial parameter.

[0288] In addition to hardness, the overall textural attributes can be further improved when an MBF (MBF1) with a protein content of 72% wt is combined with potato protein in an emulsion system to form oleogels. These structures are important in structuring food analogues, with particular interest to mimic the behaviour of muscle tissue, fat tissue or composites. Several tests were conducted in which blends of MBF1 and potato protein were added to an oil-water system using sunflower oil, followed by mechanical homogenization (Thermomix®) at room at room temperature followed by gelation at 90°C for 30 minutes and cooling to ~10°C. The resulting gel was then cut into slabs of constant geometry and subjected to texture profile analysis. The most illustrative results are shown in Figure 2; a structure with excellent properties can be derived when a composition comprising MBF with a protein content of 72.3% (MBF1) and potato protein in an emulsion containing around 40% oil.

[0289] Example 3: Compositions comprising fibres

[0290] Compositions comprising MBF with a protein content of at least 50% (MBF1) (produced as described in example 1 , and milled konjac fibre from a commercial source (Amorphophallus konjac; Pit & Pit, Hoogstraten, Belgium) were prepared by blending the ingredients in either an aqueous suspension or a water-oil system using sunflower oil while vigorously mixing (Thermomix®) to allow a complete dispersion and formation of a homogenous suspension / emulsion. After this, samples were heated up to 90°C followed by cooling to 10°C to allow gel formation. The resulting texture is then cut into slabs of equal geometry and then assessed using texture profile analysis (TPA). The results are shown in Figure 3; a texture with superior attributes was achieved with the unique combination of the MBF and konjac fibre in an emulsion where the oil content is -40%. The improvement in textural attributes was particularly strong regarding hardness and chewiness.

[0291] Other fibres in unique combinations with MBFs yield textures with clear attributes which could not have been obtained with the ingredients alone. An overview of illustrative examples is given in Table 2.

[0292] Table 2: Textural attributes of compositions of fibres and MBFs (MBF1).

[0293] Example 4: Animal fat analogue

[0294] This example illustrates the textural attributes of several microbial compositions (produced using the method described in Example 1) that resemble the textural behaviour of animal fat tissues (Figure 4). In all cases, the MBF with a protein content of at least 50% (MBF1) and fibres were added as powder and were thoroughly mixed at room temperature (Thermomix ®) in the corresponding water-oil system until a homogenous, smooth creamy emulsion was obtained. The emulsion was then heated to 90°C for 30 minutes to induce gelation and then cooled down to ~10°C to set the gels. The resulting textures were then analysed using texture profile analysis (TPA) and rheology to assess their melting behaviour. These results are presented in Figure 4 and Figure 5.

[0295] The textural attributes of the resulting analogues are greatly improved when the microbial composition includes the MBF in combination with wheat flour in a water-oil system containing 40% sunflower oil. Hardness and chewiness are significantly improved, while springiness and cohesiveness are maintained or marginally reduced.

[0296] In addition to texture, the melting properties of fats are crucial to mimic the sensory characteristics of animal tissues. Figure 5 shows how compositions made with the MBF in combination with coconut oil has a melting profile that resembles that of pork fat tissue. On the contrary, only coconut fat shows complete melting. The inventors have found several recipes in which, by selecting the right type of oil and fibre, both the melting and the structural properties of animal fat tissue can be mimicked. In this case it important to select an oil with a similar melting profile to animal fat such as coconut oil; the inventors have also found that using wheat fibre in such a composition provides appropriate structure and rigidity to mimic animal fat tissue. Example 5: Thermal MBF Compositions

[0297] This example shows the functional properties of microbial compositions prepared by exposing an MBF (prepared according to Example 1) to a specific combination of temperature and time. The MBF with a protein content of at least 50% (MBF1) and the MBF with a fibre content >20% (MBF2), both of which have a moisture content no larger than 10% were exposed to convective conditions at a specific temperature-time composition after which the MBF was collected, and its functional properties were measured. In this example an MBF with a protein content of -72% (MBF1) was subjected to around 68°C for approximately 1 hour and an MBF with a fibre content of more than 20% (MBF2) was subjected to around 82°C for approximately 3 hours. The results are shown in Figure 6; in all cases a significant enhancement of the gelation properties is achieved.

[0298] Example 6: Compositions comprising salts

[0299] This example illustrates compositions comprising MBF and Ca+, such compositions resulted in superior rheological properties. In this example, an MBF with a protein content of at least 50% (MBF1) was blended with CaCh to give an overall MBF content of approximately 15% DW, and the gelation behaviour in terms of storage modulus G’ was measured. MBF dispersed in tap water was also measured as a control. Figure 7 clearly shows that combining MBF with CaCh was able to enhance the gelation properties of the composition. Specifically, compositions with 20-1000 ppm Ca exhibited superior gelation properties, with particularly high gelation being observed for compositions comprising 600 ppm Ca.

[0300] Example 7: compositions comprising enzymes

[0301] This example illustrates compositions comprising MBF and enzymes. Multiple MBFs with a protein content of at least 50% (MBF1) were suspended in water to reach a concentration of 100g / L. This suspension was incubated at 35°C for 1 hour with 0.35g of transglutaminase per gram of MBF. After the incubation period, the gelation properties of the suspension were measured using texture profile analysis. Figure 8 shows that in all cases where transglutaminase was present, there was a significant improvement in gelation hardness.

[0302] Example 8: Foaming capacity and stability of thermally treated microbial biomass fractions. A microbial biomass fraction MBFs with a protein content of at least 50% (MBF1) was prepared according to the method of Example 1. Said MBF had a moisture content <15% and was subjected to thermal treatment under specific combinations of time and temperature. After thermal treatment the resulting fractions were evaluated for their foaming properties. The results are shown in Table 3, as measured by foam half-time. In particular, the data in Table 3 shows that the duration of thermal treatment has a substantial impact on foam stability. For example, when a microbial biomass fraction as described above was heated to 150°C for 60 minutes this resulted in a very stable foam with a half-time of 304 minutes. However, shortening the duration of treatment to 18 minutes drastically reduced foam stability, with a half-time of only 15 minutes being achieved. Thermal treatment for too long has a negative effect on foam stability. For example, when treatment at 150°C was extended to 102 minutes, the foam half-time was 191 minutes, substantially less than what was achieved with thermal treatment for 60 minutes. Overall, this suggests that the duration of heat treatment should be optimised to maximise foam stability.

[0303] In contrast, foam capacity, as measured by the overrun, remained relatively with little change in overrun in response to changes in the thermal treatment parameters.

[0304] Table 3: Effect of thermal treatment on foaming capacity and stability on MBF1 .

[0305] Example 9: Organoleptic properties after thermal treatment Microbial biomass fractions (MBF1 and MBF2) were produced as described in Example 1. A microbial biomass fraction comprising at least 50% protein (MBF1) and a microbial biomass fraction comprising at least 20% fibre (MBF2), having a moisture content of less than 15%, were subjected to various conditions for thermal treatment, as indicated in Table 4. Note that MBF1 was produced in different production batches, the batch of production is indicated in Table 4.

[0306] The temperature, duration of treatment and relative humidity at which thermal treatment was carried out was varied between samples. After thermal treatment, the microbial biomass fraction was evaluated for its organoleptic properties, such as its taste and smell. The results of this organoleptic evaluation are shown in Table 4,

[0307] Table 4: Organoleptic characteristics of several microbial biomass fractions (MBF1 and MBF2) before (control) and after thermal treatment References

[0308] Konstantina Kyriakopoulou, Julia K. Keppler, and Atze Jan van der Goot. Functionality of Ingredients and Additives in Plant-Based Meat Analogues. Foods. 2021 Mar; 10(3): 600.

[0309] US3887431 US2908661

[0310] GB 2617170

[0311] EP3622828A1

[0312] US3887431A

[0313] US9249235B2

[0314] EP3670646A1

[0315] US4810509A NL2026504

Claims

53CLAIMS:

1. A composition comprising: a. at least one disintegrated microbial biomass, and / or at least one microbial biomass fraction (MBF) wherein the protein content of the disintegrated microbial biomass or microbial biomass fraction is at least 50%, and b. at least one plant based protein, and / or c. at least one plant based fibre.

2. A composition according to claim 1 wherein the disintegrated microbial biomass or microbial biomass fraction has a protein content of 65-75%.

3. A composition according to claim 1 or 2 wherein the plant based protein is selected from one more of potato protein, soya protein, lentil protein, chickpea protein, peanut protein, almond protein, spirulina, quinoa protein, chia seed protein, tofu protein, pea protein, pistachio protein, oat protein or wheat protein4. A composition according to any of claims 1 to 3 wherein the plant based fibre is selected from one or more of psyllium fibre, konjac fibre, oat fibre, lentil fibre, resistant starch, acacia gum, methylcellulose, cellulose, hydroxypropyl methylcellulose, citrus fibre, p-glucan or pectin.

5. A composition according to claim 4 wherein the plant based fibre is oat p-glucan6. A composition according to any preceding claim wherein the at least one disintegrated microbial biomass or microbial biomass fraction has a moisture content of less than 10%.

7. A composition according to any preceding claim further comprising one or more of a polysaccharide, fat, oil, enzyme or salt.

8. A composition according to any preceding claim further comprising flour, preferably wheat flour.

549. A composition according to any preceding claim wherein the ratio of microbial biomass fraction to plant based protein is approximately 25:75 (dry matter).

10. A composition according to any of claims 1 to 8 wherein the ratio of microbial biomass fraction to plant based protein is 1.7-2.3:1.11 . A composition according to any preceding claim wherein the ratio of microbial biomass fraction to plant based fibre is 1-2:1 (dry matter).

12. A composition according to any of claims 1 to 10 wherein the ratio of microbial biomass fraction to plant based fibre is 5-6:1 (dry matter).

13. A composition according to claim 8 wherein the composition comprises a blend of 0.2-0.5:1 wheat flour to microbial biomass fraction.

14. A composition comprising: a. at least one disintegrated microbial biomass, and / or at least one microbial biomass fraction (MBF) wherein the fibre content of the disintegrated microbial biomass or microbial biomass fraction is at least 20% and b. at least one plant based fibre, and / or c. at least one animal based protein.

15. A composition according to claim 14 wherein the animal based protein is selected from one or more of milk protein, egg protein, beef protein, pork protein, chicken protein, turkey protein, duck protein, rabbit protein, goat protein, venison protein, pheasant protein, casein, whey protein, lactoglobulin, lactoferrin, ovalbumin, ovotransferrin or ovomucoid.

16. A composition according to claim 15 wherein the animal based protein is milk protein.

17. A composition according to any of claims 14 to 16 wherein the plant based fibre is selected from one or more of psyllium fibre, konjac fibre, oat fibre, lentil fibre, resistant starch, acacia gum, methylcellulose, cellulose, hydroxypropyl methylcellulose, citrus fibre, p-glucan or pectin.5518. A composition according to claim 17 wherein the plant based fibre is oat p-glucan.

19. A composition according to any of claims 14 to 18 wherein the at least one disintegrated microbial biomass or microbial biomass fraction has a moisture content of less than 10%.

20. A composition according to any of claims 14 to 19 further comprising one or more of a polysaccharide, fat, oil, enzyme or salt.

21. A composition according to any of claims 14 to 20 further comprising flour, preferably wheat flour.

22. A composition according any of claims 14 to 21 wherein the ratio of microbial biomass fraction to animal based protein is approximately 25:75 (dry matter).

23. A composition according to any of claims 14 to 21 wherein the ratio of microbial biomass fraction to animal based protein is 1.7-2.3:1.

24. A composition according to any of claims 14 to 23 wherein the ratio of microbial biomass fraction to plant based fibre is 1-2:1 (dry matter).

25. A composition according to any of claims 14 to 23 wherein the ratio of microbial biomass fraction to plant based fibre is 5-6:1 (dry matter).

26. A composition according to claim 21 wherein the composition comprises a blend of 0.2-0.5:1 wheat flour to microbial biomass fraction.

27. A composition comprising a. at least one disintegrated microbial biomass, and / or at least one microbial biomass fraction (MBF) wherein the protein content of the disintegrated microbial biomass or microbial biomass fraction is at least 50% and b. at least one oil or fat.5628. A composition according to claim 27 wherein the at least one oil or fat is selected from one or more of coconut oil, sunflower oil, olive oil, canola oil, rice oil, vegetable oil, avocado oil, sesame seed oil, flaxseed oil, walnut oil, peanut oil, rice fat, bran fat or coconut fat.

29. A composition according to claim 27 or 28 further comprising one or more of a polysaccharide, enzyme or salt.

30. A composition according to any of claims 27 to 29 wherein the composition comprises a second disintegrated microbial biomass or microbial biomass fraction.

31. A composition according to claim 30 wherein the second disintegrated microbial biomass, or microbial biomass fraction has a fibre content of at least 20%.

32. A composition comprising a. at least one disintegrated microbial biomass, and / or at least one microbial biomass fraction (MBF) wherein the protein content of the disintegrated microbial biomass or microbial biomass fraction is at least 50%, or at least one disintegrated microbial biomass, and / or at least one microbial biomass fraction (MBF) wherein the fibre content of the disintegrated microbial biomass or microbial biomass fraction is at least 20%, and b. at least one salt.

33. A composition according to claim 32 wherein said salt is CaCh or sodium bicarbonate.

34. A composition comprising a. at least one disintegrated microbial biomass, and / or at least one microbial biomass fraction (MBF) wherein the protein content of the disintegrated microbial biomass or microbial biomass fraction is at least 50% and b. at least one enzyme.

35. A composition according to any preceding claim further comprising a thickening agent.

36. A composition according to any preceding claim wherein said composition is in an emulsion system.

37. A composition according to claim 36 wherein the emulsion system comprises at least 40% oil.

38. A composition according to any preceding claim wherein the at least one disintegrated microbial biomass, or microbial biomass fraction has been heat treated at a temperature in the range of approximately 50-200°C.

39. A composition according to claim 38 wherein the relative humidity during said heat treatment is at least 60%.

40. A composition according to any preceding claim for use in the preparation of a food product or a cosmetic and / or their precursors.41 . A food product comprising the composition of any of claims 1-39.

42. A cosmetic product comprising the composition of any of claims 1-39.

43. A method of preparing the composition of any of claims 1 to 13 wherein said method comprises combining: a. at least one disintegrated microbial biomass, and / or microbial biomass fraction (MBF), wherein the protein content of the disintegrated microbial biomass or microbial biomass fraction is at least 50%, and b. at least one plant based protein, and / or c. at least one plant based fibre in an aqueous system or an emulsion system to form a homogenous suspension.

44. A method of preparing the composition of any of claims 14 to 26 wherein said method comprises combining a. at least one disintegrated microbial biomass, and / or at least one microbial biomass fraction (MBF) wherein the fibre content of the disintegrated microbial biomass or microbial biomass fraction is at least 20% andb. at least one plant based fibre, and / or c. at least one animal based protein. in an aqueous system or an emulsion system to form a homogenous suspension.

45. A method of preparing the composition of any of claims 27 to 31 wherein said method comprises combining: a. at least one disintegrated microbial biomass, and / or microbial biomass fraction (MBF), wherein the protein content of the disintegrated microbial biomass or microbial biomass fraction is at least 50%, and b. at least one oil or fat, in an aqueous system or an emulsion system to form a homogenous suspension.

46. The method of any of claims 43 to 45 further comprising adding one or more of a polysaccharide or salt.

47. The method of any of claims 43 to 46 further comprising adding flour.

48. A method of preparing the composition of claim 34 wherein said method comprises combining: a. at least one disintegrated microbial biomass, and / or microbial biomass fraction, wherein the protein content of the disintegrated microbial biomass, or microbial biomass fraction is at least 50%, and b. at least one enzyme in an aqueous system or an emulsion system to form a homogenous suspension.

49. A method of preparing the composition of claim 32 or 33 wherein said method comprises combining: a. at least one and / or disintegrated microbial biomass, and / or microbial biomass fraction (MBF), wherein the protein content of the disintegrated microbial biomass or microbial biomass fraction is at least 50%, or at least one disintegrated microbial biomass, and / or at least one microbial biomass fraction (MBF) wherein the fibre content of the disintegrated microbial biomass or microbial biomass fraction is at least 20%, and b. at least one salt59 in an aqueous system or an emulsion system to form a homogenous suspension.

50. The method of any of claims 43 to 49 further comprising adding a thickening agent.51 . The method of any of claims 43 to 50 wherein the emulsion system comprises at least 40% oil.

52. The method of any of claims 43 to 51 further comprising mechanically homogenizing the homogenous suspension53. The method of any of claims 43 to 52 further comprising heat treating the at least one disintegrated microbial biomass, or microbial biomass fraction at a temperature in the range of approximately 50-200°C.

54. The method according to claim 53 wherein the relative humidity during said heat treatment is at least 60%.

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