Granules comprising fungal biomass particles

Abstract

The present invention relates to a particulate protein composition comprising granules having a protein content of at least 30 wt.% and comprising at least 80 wt.% of fungal biomass particles wherein the particle size distribution of the granules satisfies the following conditions: D50-gml > = 50 [mu] m; dx-gml represents the weight percent of granules having a diameter of less than x [mu] m; wherein the particle size distribution of fungal biomass particles contained in the granules meets the following conditions: D10 <-> prtcl is less than or equal to 30 microns; 2 [mu] m < = D50-gprtcl < = 60 [mu] m; d90-prtcl < = 150 [mu] m; dx-prtcl represents the weight percentage of the fungal biomass particles contained in the granules having a diameter of less than x [mu] m; and wherein the ratio D50-grnl: D50-prtcl is greater than or equal to 2.0. The function of the proteinaceous fungal biomass is enhanced by I) greatly reducing the particle size of the fungal biomass and (ii) agglomerating the fine fungal biomass particles so obtained into granules.

Description

Technical Field

[0001] The present invention relates to a granulated protein composition comprising at least 80 wt.% of granules having a protein content of at least 30 wt.% and comprising at least 80 wt.% of fungal biomass particles.

[0002] The present invention also relates to a method for preparing a granulated protein composition, the method comprising:

[0003] Providing fungal biomass;

[0004] subjecting the fungal biomass to a size reduction process to produce a powder or suspension comprising particles of the fungal biomass; and

[0005] Agglomeration of fungal biomass particles. Background Art

[0006] The demand for edible products that can provide high protein content obtained from non-animal sources is increasing. Driven by the growing awareness of personal health, edible products that contain components (such as protein and fiber) of non-animal origin are considered to be healthier alternatives to products based on animal proteins. In particular, there is a growing demand for edible meat substitutes that mimic meat in their composition and texture but are composed of non-animal components, which can reduce dependence on animals (such as cows) and reduce the carbon footprint caused by such animals.

[0007] Single cell protein (SCP) is an interesting alternative to meat protein and as a protein source in many other food applications such as breakfast cereals, bread, pasta, dairy products, ice cream, chocolate and soups. When produced from fungi, SCP can be produced from sugar-rich crops in a more sustainable way than meat protein production, as SCP produces more tons of protein per hectare and has lower nitrogen emissions than meat production.

[0008] One method for producing a protein dietary source for human food or animal feed is to produce "single cell protein" (SCP) by fermentation (Suman et al., 2015, Int J. Curr. Microbiol. Appl. Sci [International Journal of Contemporary Microbiological Applications], Volume 4, Issue 9, Pages 251-262). In this regard, fermentation is understood to be the microbial conversion of a carbon-rich raw material into a protein-rich product consisting of microbial cells (such as bacteria, yeast or fungi). The use of SCP as an animal feed and food ingredient brings the additional advantage that the microbial cells have a high content of essential amino acids. In addition, fungal cells in particular can be very rich in trace elements and vitamins that make the fermented feed very nutritious.

[0009] SCP has been used in food products for human consumption. For example, Quorn TM Contains mycoprotein produced as SCP by fermentation of the fungus Fusarium venenatum. TM Available in various forms such as sausages, steaks, burgers, pies and meat sticks.

[0010] The fermentation production of fungal biomass typically produces a fermentation broth with a dry matter content of about 5wt.%. This fermentation broth needs to be concentrated and optionally dried to make it suitable for transportation and storage under ambient conditions and to make it suitable for a wide range of food products. Concentrated or dried fungal biomass can be suitably applied to food products to provide valuable protein and dietary fiber. However, unlike some other protein sources (such as soy protein isolate, pea protein isolate and gluten), it does not impart a noticeable texture.

[0011] US 2008 / 226811 describes a granular pea protein composition having a protein content of at least 70 wt.% and an average diameter of 150 to 300 μm.

[0012] US2011 / 311599 describes a granulated powder comprising at least one vegetable protein and at least one vegetable fiber, wherein the granulated powder has:

[0013] The average diameter D of the laser volume is between 10 μm and 500 μm. 4,3 ,as well as

[0014] A dry matter content of greater than 80% was determined after baking at 130°C for 2 hours.

[0015] WO 2018 / 029353 describes a method for producing a single cell protein (SCP), the method comprising the following steps:

[0016] a) growing a thermophilic fungus in a culture medium containing a fermentable carbon-rich feedstock; wherein the fungus is grown in a submerged culture under non-sterile conditions at a temperature above 45° C. and a pH of less than 3.8; and,

[0017] b) recovering the SCP from the culture medium in the form of biomass of the thermophilic fungus grown in step a).

[0018] US2020 / 0093155 describes a method for producing a food ingredient, the method comprising the following steps:

[0019] a. cultivating filamentous fungi in a growth medium;

[0020] b. harvesting filamentous fungal biomass;

[0021] c. optionally, processing the harvested filamentous fungal biomass;

[0022] d. adjusting the size of the biomass to form particles; and

[0023] e. Drying the particles to form a food ingredient. Summary of the invention

[0024] The inventors have surprisingly found that the functionality of protein fungal biomass can be enhanced by substantially reducing the particle size of the fungal biomass and by agglomerating the fine fungal biomass particles thus obtained into granules. These granules typically contain at least 80 wt.% of fungal biomass particles. The protein content of the granules is typically at least 30 wt.%.

[0025] Thus, the present invention relates to a granulated protein composition comprising at least 80 wt.% of granules having a protein content of at least 30 wt.% and comprising at least 80 wt.% of fungal biomass particles, wherein the particle size distribution of the granules satisfies the following conditions:

[0026] ·D 50-grnl ≥50μm;

[0027] D x-grnl represents the weight percentage of granules having a diameter less than x μm;

[0028] The particle size distribution of the fungal biomass particles contained in the granules satisfies the following conditions:

[0029] ·D 10-prtcl ≤30μm;

[0030] ·2μm≤D 50-gprtcl ≤60μm;

[0031] ·D 90-prtcl ≤150μm;

[0032] D x-prtcl represents the weight percentage of fungal biomass particles contained in the granules having a diameter less than x μm; and

[0033] The ratio D 50-grnl :D 50-prtcl ≥2.0.

[0034] The granules according to the invention offer the advantage that they provide better gelling and foaming functions, as well as a firmer texture when applied in food products such as meat analogs, compared to non-granular particles of fungal biomass of equal size.

[0035] The present invention also provides a method for preparing a granulated protein composition, the method comprising:

[0036] Providing fungal biomass;

[0037] subjecting the fungal biomass to a size reduction process to produce a powder or suspension comprising fungal biomass particles; wherein the particle size distribution of the fungal biomass particles satisfies the following conditions:

[0038] ○D 10-prtcl ≤30μm;

[0039] ○2μm≤D 50-gprtcl ≤60μm;

[0040] ○D 90-prtcl ≤150μm;

[0041] Agglomeration of fungal biomass particles. DETAILED DESCRIPTION

[0042] A first aspect of the present invention relates to a granulated protein composition comprising at least 80 wt.% granules having a protein content of at least 30 wt.% and comprising at least 80 wt.% fungal biomass particles, wherein the particle size distribution of the granules satisfies the following conditions:

[0043] ·D 50-grnl ≥50μm;

[0044] D x-grnl represents the weight percentage of granules having a diameter less than x μm;

[0045] The particle size distribution of the fungal biomass particles contained in the granules satisfies the following conditions:

[0046] ·D 10-prtcl ≤30μm;

[0047] ·2μm≤D 50-gprtcl ≤60μm;

[0048] ·D 90-prtcl ≤150μm;

[0049] D x-prtcl represents the weight percentage of fungal biomass particles contained in the granules having a diameter less than x μm; and

[0050] The ratio D 50-grnl :D 50-prtcl ≥2.0.

[0051] As used herein, the term "fungal biomass" refers to organic matter derived from fungi.

[0052] The term "protein" as used herein refers to a molecule comprising a chain of at least 20 amino acids.

[0053] The term "granule" as used herein refers to particles that are agglomerates of smaller particles, notably smaller particles of fungal biomass. Granules (granules / granulates) can be produced by agglomeration techniques such as, for example, fluidized bed agglomeration, compaction, extrusion and spray drying agglomeration.

[0054] The term "fat" as used herein refers to lipids containing one or more fatty acid residues. Unless otherwise indicated, the terms "fat" and "oil" as used herein should be considered synonyms.

[0055] As used herein, the term "dietary fiber" refers to polysaccharides that are not digested by human digestive enzymes. Chitin, chitosan, and β-glucan are examples of dietary fibers found in fungi.

[0056] As used herein, the term "polysaccharide" refers to a polymer comprising long, optionally branched chains of monosaccharides, the total number of monosaccharides in the polysaccharide being at least ten.

[0057] The terms "a" or "an", as used herein, are defined as "at least one", unless otherwise stated.

[0058] The term "or", as used herein, is defined as "and / or" unless stated otherwise.

[0059] Unless otherwise indicated, all percentages mentioned herein are to be interpreted as percentages by weight.

[0060] Unless otherwise indicated, the particle size distribution referred to herein is determined by laser diffraction analysis using a Mastersizer 3000 (from Malvern Panalytical). The technique is based on passing a laser beam through dispersed particles. The angular variation of the scattered light intensity provides information about the particle size. Larger particles scatter light at smaller angles, and smaller particles scatter light at larger angles. The scattered intensity data can be used to calculate the particle size distribution.

[0061] The particle size distribution of the granules in the granulated protein composition of the present invention can be determined using a Mastersizer 3000 equipped with an Aero S dry powder dispenser. In addition, the particle size distribution of the fungal biomass particles contained in the granules can be appropriately determined using this setup. The Aero S disperses the dry sample by accelerating the particles through a venturi using compressed air. The particles are then sucked into the measuring cell of the Mastersizer 3000 using a vacuum source. The air pressure drop through the venturi is controlled to achieve complete sample dispersion, and the air pressure drop can be controlled within + / - 0.1 bar. The particle size distribution of the granules can be appropriately determined by the Mastersizer 3000 by using a dispersion pressure of 1 bar in the Aero S dispenser. The particle size distribution of the fungal biomass particles contained in the granules can be appropriately determined by the Mastersizer 3000 by using a dispersion pressure of 4 bar in the Aero S dispenser. Using a maximum dispersion pressure of 4 bar ensures that the granules are deagglomerated before measurement.

[0062] The granulated protein composition of the present invention preferably does not contain any meat or meat-derived components.Preferably, the composition is a vegetarian composition, more preferably a vegan composition.

[0063] Granules having a protein content of at least 30 wt.% and comprising at least 80 wt.% of fungal biomass particles preferably constitute at least 90 wt.%, more preferably at least 95 wt.% of the granulated protein composition of the present invention. In addition to these granules, the granulated protein composition may contain other components such as desiccants, glidants, micronutrients, flavoring agents and coloring agents.

[0064] According to a particularly preferred embodiment, the granulated protein composition consists of granules comprising particles of fungal biomass.

[0065] The protein content of the granules is preferably at least 35 wt.%, more preferably 40 to 70 wt.% and most preferably 42 to 60 wt.%.

[0066] The biomass particles in the granules preferably have a protein content of at least 35 wt.%, more preferably 40 to 70 wt.% and most preferably 42 to 60 wt.%.

[0067] Dietary fibres are preferably contained in the fungal biomass particles in a concentration of 35-42 wt.%, more preferably 30-40 wt.%, calculated on the weight of dry matter.

[0068] Preferably, at least 35 wt.%, more preferably 40-75 wt.% of the dietary fiber in the fungal biomass particles is selected from chitin, chitosan and combined amino polysaccharides.

[0069] Chitin preferably constitutes 15-50 wt.%, more preferably 20-40 wt.%, most preferably 25-35 wt.% of the dietary fiber in the fungal biomass particles.

[0070] Chitosan preferably constitutes 15-50 wt.%, more preferably 20-40 wt.%, most preferably 24-34 wt.% of the dietary fiber in the fungal biomass particles.

[0071] The dietary fiber in the fungal biomass particles preferably contains 10-70 wt.%, more preferably 20-50 wt.% polyglucuronic acid.

[0072] The combination of aminopolysaccharides and polyglucuronic acid preferably constitutes at least 50 wt%, more preferably at least 70 wt% of the dietary fiber in the fungal biomass particles.

[0073] The combination of fungal protein and aminopolysaccharide preferably constitutes at least 40 wt.%, more preferably 50-70 wt.% of the dry matter contained in the fungal biomass particles.

[0074] The fungal biomass particles preferably contain 0-15 wt.%, more preferably 0-10 wt.% and most preferably 0.5-5 wt.% digestible carbohydrates calculated on the weight of dry matter.

[0075] The fat content of the fungal biomass particles is preferably in the range of 6-20 wt.%, more preferably 6.5-12 wt.%, calculated on the weight of dry matter.

[0076] The combination of protein, fat and dietary fiber preferably constitutes at least 80 wt.%, more preferably 88-99.8 wt.% of the dry matter contained in the fungal biomass particles.

[0077] The particulate protein composition according to the present invention preferably has a water content of 1-15 wt%, more preferably 2-10 wt% and most preferably 3-9 wt%.

[0078] Granules of the invention may suitably be prepared by fluid bed agglomeration, compaction or extrusion. Fluid bed agglomeration typically produces relatively fine granules, whereas compaction and extrusion are mainly used to produce rather coarse granules.

[0079] Preferably, the particle size distribution of the granules is such that D 50-grnl In the range of 60 to 1,500 μm, more preferably in the range of 70 to 1,200 μm and most preferably in the range of 80 to 1,000 μm.

[0080] In a preferred embodiment, the particle size distribution of the granules is such that D 10-grnlAt least 10 μm, more preferably in the range of 15 to 1,000 μm and most preferably in the range of 20 to 800 μm.

[0081] Advantageously, the particle size distribution of the granules is such that D 90-grnl At least 100 μm, more preferably in the range of 150 to 3,000 μm and most preferably in the range of 200 to 2,000 μm.

[0082] In a preferred embodiment, the granules of the present invention are fine granules having a particle size distribution such that D 50-grnl In the range of 60 to 500 μm, more preferably in the range of 70 to 300 μm and most preferably in the range of 80 to 250 μm.

[0083] In a preferred embodiment, the particle size distribution of the fine granules is such that D 10-grnl At least 10 μm, more preferably in the range of 15 to 100 μm and most preferably in the range of 20 to 80 μm.

[0084] Advantageously, the particle size distribution of the fine granules is such that D 90-grnl It is at least 100 μm, more preferably in the range of 150 to 1,000 μm and most preferably in the range of 200 to 800 μm.

[0085] According to another preferred embodiment, the granules in the granular composition have a span of less than 8, wherein the span is equal to (D 90 -D 10 ) / D 50 More preferably, the granules have a span of less than 5, more preferably the span is in the range of 0.8 to 4 and most preferably the span is in the range of 1.5 to 3.

[0086] The fungal biomass particles in the granules of the present invention preferably have a particle size distribution such that D 10-prtcl Not more than 20 μm, more preferably in the range of 0.5 to 15 μm and most preferably in the range of 1 to 12 μm.

[0087] Preferably, the particle size distribution of the fungal biomass particles in the granules is such that D 50-grnl In the range of 2 to 40 μm, more preferably in the range of 3 to 30 μm and most preferably in the range of 4 to 25 μm.

[0088] In a preferred embodiment, the particle size distribution of the fungal biomass particles is such that D 90-prtcl In the range of 10 to 130 μm, more preferably in the range of 20 to 110 μm and most preferably in the range of 25 to 100 μm.

[0089] According to a particularly preferred embodiment, the granules of the present invention are relatively large agglomerates of fungal biomass particles, as indicated by the ratio D 50-grnl :D 50-prtcl More preferably, the ratio is at least 3.0. Most preferably, the ratio is in the range of 4.0 to 15.0.

[0090] The fungal biomass contained in the fungal biomass particles is preferably the biomass of one or more fungi belonging to the class Zoopagomyceta, the class Mucoromyceta or the class Symbiomyceta. Even more preferably, the fungal biomass is the biomass of one or more fungi belonging to the class Zoopagomyceta or the class Mucoromyceta.

[0091] In a preferred embodiment, the fungal biomass in the fungal biomass particles is biomass of a fungal strain selected from the group consisting of: Rasamsonia, Talaromyces, Penicillium, Acremonium, Humicola, Paecilomyces, Chaetomium, Rhizomucor, Rhizopus, Thermomyces, Myceliophthora, Thermoascus, Thielavia, Mucor, Stibella, Melanocarpus, Malbranchea, More preferably, the fungal biomass is the biomass of one or more fungi belonging to the species selected from the group consisting of: Lasamsonia compostii, Rosacea emersonii, Basketella emersonii The genus Rhizomucor, Rhizomucor miehei, Rhizomucor microsporus, Thermomucor indica-sudati, Thielava terrestris, Thermoascus and Rhizopus, among which the following strains are more preferred: Lasamsonella composting strain CBS141695, Rossella emersonii CBS143030, Thermomucor indica-sudati CBS143027 and CBS104.75, Rhizomucor miehei CBS143029, Rhizomucor microsporus CBS 143028, Thermoascus aurantiacus CBS 528.71, Thielavia terrestris CBS 546.86, Talaromyces emersonii CBS 393.64, Myceliophthora thermophila CBS117.65 and Rhizopus sp. CBS143160, among which strains CBS141695, CBS143030, CBS143027, CBS143029, CBS143160 and CBS 143028 are most preferred.

[0092] Another aspect of the present invention relates to a method for preparing a granulated protein composition, preferably a granulated protein composition according to the present invention, the method comprising:

[0093] Providing fungal biomass;

[0094] · Subject the fungal biomass to a size reduction treatment to produce a powder or suspension comprising fungal biomass particles; wherein the particle size distribution of the fungal biomass particles satisfies the following conditions:

[0095] ○ D 10-prtcl ≤ 30 μm;

[0096] ○ 2 μm ≤ D 50-gprtcl ≤ 60 μm;

[0097] ○ D 90-prtcl ≤ 150 μm;

[0098] · Agglomerate the fungal biomass particles.

[0099] The fungal biomass subjected to the size reduction treatment in the method of the present invention can be provided in the form of, for example, flocs or coarse powder. Preferably, the fungal biomass is provided in the form of a particulate composition, wherein at least 80 wt.% of the particles have a sieve pore size of at least 100 μm, more preferably at least 160 μm, and most preferably at least 200 μm. The weight percentage of particles having a sieve pore size of at least x μm is determined by using a sieve with an opening of x μm.

[0100] The fungal biomass subjected to the size reduction treatment preferably contains at least 10 wt.%, more preferably at least 30 wt.%, and most preferably at least 50 wt.% of intact fungal cells, and the weight percentage is calculated based on the weight of the total fungal biomass.

[0101] The fungal biomass preferably has a protein content of at least 30 wt.%, more preferably at least 35 wt.%, even more preferably 40 to 70 wt.%, and most preferably 42 to 60 wt.% calculated based on the weight of dry matter.

[0102] The fungal biomass provided in the method of the present invention is preferably the biomass of one or more fungi belonging to the Entomophthoromycetes, Mucoromycetes, or Mycorrhizal symbiota superphylum. Even more preferably, the fungal biomass is the biomass of one or more fungi belonging to the Entomophthorales or Mucorales.

[0103] In a preferred embodiment, the fungal biomass is biomass of a fungal strain selected from the group consisting of: Rosalia, Talaromyces, Penicillium, Acremonium, Humicola, Paecilomyces, Chaetomium, Rhizomucor, Rhizopus, Thermomyces, Myceliophthora, Thermoascus, Thielavia, Mucor, Sphaerotheca, Melanocarpa, Malbranchia, Aglaonema, Canary Sandia, Columnar Acremonium, Apostrophe, More preferably, the fungal biomass is a biomass of one or more fungi belonging to the species selected from the group consisting of: Lasamsonia composting, Rosacea emersonii, Talaromyces emersonii, Rhizomucor miehei, Rhizomucor microsporus, Thermomucor indica-sudati, Thielavia terrestris, Thielavia terrestris, Thermoascus and Rhizopus, wherein the following strains are more preferred: Lasamsonia composting strain CBS 141695, Rosella emersonii CBS143030, Thermomucor indica-sudati CBS143027 and CBS 104.75, Rhizomucor miehei CBS143029, Rhizomucor microsporus CBS143028, Thermoascus ascomycetes CBS528.71, Thielavia terrestris CBS 546.86, Talaromyces emersonii CBS 393.64 and Thermomycetes thermophila CBS117.65 and Rhizopus CBS143160, among which strains CBS141695, CBS143030, CBS143027, CBS143029, CBS143160 and CBS143028 are most preferred.

[0104] The size reduction process used to produce a powder or suspension comprising particles of fungal biomass preferably comprises milling or grinding.

[0105] In an advantageous embodiment of the method of the invention, the size reduction process produces a powder comprising particles of fungal biomass. Such a powder may suitably be agglomerated by fluidized bed agglomeration, compaction or extrusion.

[0106] In another embodiment, the fungal biomass is provided in the form of wet biomass having a water content of 75 to 99 wt.%, and the size reduction treatment produces a suspension of fungal biomass particles. The fungal biomass particles in the suspension may be suitably agglomerated by, for example, spray drying agglomeration.

[0107] According to a particularly preferred embodiment, the process of the invention produces a granulated protein composition as defined hereinbefore.

[0108] A further aspect of the invention relates to a food product comprising at least 1 wt.%, preferably 10-95 wt.%, calculated on weight of dry matter, of the particulate protein composition of the invention.

[0109] Examples of food products in which the particulate protein compositions of the present invention may be suitably administered include texturized proteins (eg, TVP), meat analogs, meat products, soups, sauces, grain-based food products, and pet foods.

[0110] Yet another aspect of the present invention relates to a method of preparing the above-mentioned food product, said method comprising combining the granulated protein composition of the present invention with one or more other edible ingredients.

[0111] The present invention is further illustrated by the following non-limiting examples.

[0112] Examples

[0113] Example 1: Production of Rhizomucor illus biomass in cake or powder form

[0114] For pre-culture, Rhizomucor microplus strain CBS143028 was inoculated into 200 ml of a defined mineral medium containing 0.17 g / L KCl, 0.17 g / L KH 2 PO 4 1.3g / L, Na 2 HPO 4 0.4g / L, citric acid 0.5g / L, MgSO 4 .7aq 0.7g / L, FeSO 4 .7aq 0.03g / L, CaCl 2 .2aq 0.035g / L, ZnSO 4 .7aq0.04g / L, MnCl 2 .4aq 0.004, CuSO 4 .5aq 0.0005g / L, CoCl 2 .6aq 0.0005g / L, Na2B 4 O 7 .10aq0.003g / L, KI0.0003g / L, Na 2 MoO 4 .2aq 0.0005g / L, 11g dextrose / L, 4g(NH 4 ) 2 SO 4 / L; and 7.5 g tartaric acid / L. The preculture was cultured at 46°C in a 1 L Erlenmeyer flask with a baffled gas-permeable plug in an orbital shaker at 200 rpm for 24 hours. The preculture was then used to inoculate a fermentor containing a defined mineral medium as described above at a pH of 3.5 and containing 77 g dextrose / L as a C source; 1.4 g (NH 4 ) 2 SO 4 / L as a N source and supplemented with NH 3 As titrant. The fungus was grown in a fermenter in fed-batch mode with a doubling time of 12 h. Olive oil was fed continuously to maintain a concentration of 50 ppm.

[0115] The fermentation broth, which had reached a dry matter content ranging from 2 to 5 weight percent, was concentrated using a shaker to reach a minimum of 10% (w / w) dry matter. The biomass was then mixed with an antioxidant and pasteurized.

[0116] Next, the sieved biomass was compressed with a hydraulic press to obtain a microscopic Rhizomucor biomass in the form of a cake having a dry matter content of about 29% (w / w). A portion of the biomass cake was further freeze-dried and ground (6,000 rpm, 0.5 mm mesh size) to obtain a microscopic Rhizomucor biomass in the form of a powder having a dry matter content of about 96% (w / w).

[0117] The composition of Rhizomucor microsporus biomass in cake form and powder form was analyzed. The results are shown in Tables 1-3.

[0118] Table 1

[0119] Composition Biomass in cake form Biomass in powder form Moisture 71.1g / 100g 4g / 100g Ash 1g / 100g 3.2g / 100g

[0120] Table 2

[0121]

[0122] *Based on Kjeldahl method (N*6.25).

[0123] **Method for determination of total dietary fiber content; enzyme pretreatment, gravimetric method

[0124] Table 3

[0125]

[0126]

[0127] *Essential amino acids for humans.

[0128] It must be considered that the protein content of the Rhizomucor microcarpa biomass as shown in Table 2 is based on the Kjeldahl method, which is a standard method for analyzing the protein content of different food products. The Kjeldahl method is based on total nitrogen content and uses a standard conversion factor of 6.25 to estimate the protein content in the analyzed food products. However, a conversion factor of 6.25 may not be suitable for some food products.

[0129] In Rhizomucor microscopy biomass, conversion factors may be overestimated due to the presence of other nitrogen sources (e.g., RNA, chitin, and chitosan). For this reason, the true protein content of Rhizomucor microscopy biomass can be better estimated by amino acid analysis.

[0130] Example 2: Dietary fiber composition of Rhizomucor microcarpa biomass

[0131] The dietary fiber content and composition of Rhizomucor pusillus biomass were determined. The results are shown in Tables 4 and 5.

[0132] Table 4

[0133]

[0134] Table 5

[0135] Batch 1 Batch 2 Batch 3 Batch 4 average value unit Chitin 8.6 10.8 9.8 9.9 9.8 g / 100g Chitosan 9.2 9.5 8.9 9 9.2 g / 100g

[0136] The dietary fiber content of Rhizomucor microsporus biomass on a dry matter basis is extremely high. Interestingly, Rhizomucor microsporus biomass contains not only chitin as a fiber, but also an almost equal amount of the fiber chitosan. Chitosan is a deacetylated form of chitin, and the health benefits of chitosan in animals have been described.

[0137] Example 3: Preparation of fine powder of Rhizomucor microplus biomass

[0138] The milled biomass powder was produced in the same manner as described in Example 1, except that in addition to the normal milled biomass powder (powder 1), a very finely milled biomass powder (powder 2) was produced (18,000 rpm, 0.12 mm mesh size). The particle size distribution of the milled powder was determined by laser diffraction using a Mastersizer 3000 equipped with an Aero S dry powder dispenser. The results are summarized in Table 6.

[0139] Table 6

[0140] Powder 1 Powder 2 <![CDATA[D 10 (in μm)]]> 6.6 2.1 <![CDATA[D 50 (in μm)]]> 70 11 <![CDATA[D 90 (in μm)]]> 240 80

[0141] Example 4: Rheological testing

[0142] The milled powder of Example 3 was combined with 25 mM Tris-HCl buffer at pH 8 at a ratio of 1:5 (w / w) and stirred for 2.5 hours. Next, the suspension was centrifuged at 5000 g and separated into a precipitate and a supernatant fraction. The supernatants thus obtained were each concentrated five times. The concentrated supernatants obtained had a protein content of about 2.5 mg / mL.

[0143] The concentrated supernatant was heated to 80° C. for 20 minutes and then cooled to 25° C. Next, the fracture stress and strain of the heated samples were measured using an MCR301 rheometer from Anton Paar using amplitude sweep measurements.

[0144] No fracture stress was detected in the heated sample produced from the supernatant of the normally milled biomass (powder 1). However, the heated sample of the supernatant of the finely milled biomass powder (powder 2) had a fracture stress of 55 Pa, a fracture strain of 1.18%, and a storage modulus of 8000 Pa.

[0145] Example 5: Application test

[0146] The milled biomass powder of Example 3 was applied to vegetarian burgers. The composition of the burgers is shown in Table 7.

[0147] Table 7

[0148]

[0149] 1 Tex, purchased from Loryma GmbH, Germany

[0150] Prepare the burgers as follows:

[0151] · The milled biomass powder and TVP were each hydrated with tap water at a volume ratio of 1:3 · The hydrated TVP and hydrated biomass powder were mixed together and egg white was added

[0152] • Mix the mixture with a hand mixer at the lowest setting (1) for 1 minute.

[0153] · Form into 100g burger

[0154] Leave the burgers to rest for 10 minutes

[0155] Steam the burgers at 100°C for 20 minutes

[0156] Freeze the burgers

[0157] Thaw the burgers on the day of tasting

[0158] Bake the burgers at medium / high heat for 5 minutes

[0159] The burgers were ready to eat and evaluated by an expert panel. The evaluation results are summarized in Table 8.

[0160] Table 8

[0161] Evaluation results Hamburger 1 Acceptable texture Hamburger 2 Less chewy, less firm, and more crispy than Burger 1 Hamburger 3 Good texture. Firmer and denser than Burger 1 Hamburger 4 Firmer than Burger 2 More hydrated than Burger 3

[0162] Example 6: Preparation of Rhizomucor microcarpon biomass granules

[0163] Powder 2 of Example 3 was granulated by fluidized bed agglomeration.

[0164] The granules thus obtained had a particle size distribution conforming to the specifications shown in Table 9.

[0165] Table 9

[0166] <![CDATA[D 10 (in μm)]]> 10-50 <![CDATA[D 50 (in μm)]]> 70-120 <![CDATA[D 90 (in μm)]]> 80-500

[0167] Example 7: Application Test

[0168] Powder 2 of Example 3 and biomass granules of Example 6 were applied to vegetarian burgers having the composition shown in Table 10.

[0169] Table 10

[0170]

[0171] 1 Tex, purchased from Loryma GmbH, Germany

[0172] The burgers were ready to eat and evaluated by a panel of experts. The burgers were found to be very similar and to have a very good firm texture.

[0173] Example 8: Preparation of Rhizomucor microcarpon biomass granules

[0174] Powder 2 of Example 3 was granulated in a Process 11 twin-screw extruder (Thermo Fischer Scientific, Karlsruhe, Germany). The extrusion conditions used are summarized in Table 11.

[0175] Table 11

[0176]

[0177] The particle size distribution of the granules was determined by laser diffraction using a Mastersizer 3000 equipped with an Aero S dry powder dispenser and the results so obtained are shown in Table 12.

[0178] Table 12

[0179] <![CDATA[D 10 (in μm)]]> 513 <![CDATA[D 50 (in μm)]]> 899 <![CDATA[D 90 (in μm)]]> 1460 <![CDATA[Span (D 90 -D 10 ) / D 50 > 1.06

Claims

1. A granulated protein composition comprising at least 80 wt.% granules having a protein content of at least 30 wt.% and comprising at least 80 wt.% fungal biomass particles, wherein the particle size distribution of the granules satisfies the following conditions: ·D 50-grnl ≥50μm; D x-grnl represents the weight percentage of granules having a diameter less than x μm; The particle size distribution of the fungal biomass particles contained in the granules satisfies the following conditions: ·D 10-prtcl ≤30μm; ·2μm≤D 50-gprtcl ≤60μm; ·D 90-prtcl ≤150μm; D x-prtcl represents the weight percentage of the fungal biomass particles contained in the granules having a diameter less than x μm; and The ratio D 50-grnl :D 50-prtcl ≥2.0; wherein the particle size distribution is determined using a Mastersizer 3000 equipped with an Aero S dry powder dispenser; and wherein the particle size distribution of the granules is determined by employing a dispersion pressure of 1 bar, and wherein the particle size distribution of the fungal biomass particles contained in the granules is determined by employing a dispersion pressure of 4 bar.

2. The granulated protein composition according to claim 1, in, These granules meet the following requirements: ·D 10-grnl ≥50μm。 3. The granulated protein composition according to claim 1 or 2, in, These granules meet the following requirements: ·D 90-grnl ≥100μm。 4. A granulated protein composition according to any one of the preceding claims, in, The composition has a water content of 1-15 wt.%.

5. A granulated protein composition according to any one of the preceding claims, in, The fungal biomass is biomass of one or more fungi belonging to the class Entomophyceae, the class Mucormycetes or the superphylum Symbiont.

6. The granulated protein composition according to claim 5, in, The fungal biomass is biomass of one or more fungi belonging to the phylum Entomophyceae or Mucormycetes.

7. A granulated protein composition according to any one of the preceding claims, in, The fungal biomass is biomass of a fungal strain selected from the group consisting of: Rosalia, Talaromyces, Penicillium, Acremonium, Humicola, Paecilomyces, Chaetomium, Rhizomucor, Rhizopus, Thermomyces, Myceliophthora, Thermoascus, Thielavia, Mucor, Fasciola, Melanocarp, Malbranch, Aglaonema, Canary Sandia, Columnar Acremonium, Asporus, Corymbia and Firmicutes.

8. The granulated protein composition according to claim 7, in, The fungal biomass is biomass of a strain of the genus Rhizomucor.

9. A method for preparing a particulate protein composition according to any one of the preceding claims, said method include: Providing fungal biomass; subjecting the fungal biomass to a size reduction process to produce a powder or suspension comprising particles of fungal biomass; The particle size distribution of these fungal biomass particles meets the following conditions: ○D 10-prtcl ≤30μm; ○2μm≤D 50-gprtcl ≤60μm; ○D 90-prtcl ≤150μm; agglomerating the fungal biomass particles; wherein the particle size distribution is determined using a Mastersizer 3000 equipped with an Aero S dry powder dispenser; and wherein the particle size distribution of the granules is determined by employing a dispersion pressure of 1 bar, and wherein the particle size distribution of the fungal biomass particles contained in the granules is determined by employing a dispersion pressure of 4 bar.

10. The method according to any one of claims 9, in, The fungal biomass has a D of at least 80 μm 50 or in the form of a granular composition comprising particles having a D of at least 80 μm. 50 The suspended material is provided in the form of a suspension.

11. The method according to any one of claims 9 or 10, in, The size reduction process includes milling or grinding.

12. The method according to any one of claims 9 to 11, in, The size reduction process produces a powder comprising the fungal biomass particles.

13. The method according to any one of claims 9 to 12, in, These fungal biomass particles are agglomerated by fluidized bed granulation, compaction, extrusion or spray drying agglomeration.

14. A food product comprising at least 1 wt.%, preferably 10-95 wt.%, calculated on the weight of dry matter, of the particulate protein composition according to any one of claims 1-8.

15. A method of preparing a food product according to claim 14, the method comprising combining the particulate protein composition according to any one of claims 1 to 8 with one or more other edible ingredients.

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