Preparation of microbial functional proteins with reduced lipid and / or nucleic acid content

Abstract

The present invention relates to a method for preparing native proteins of microorganisms, comprising lysing the microorganisms, separating lipids from the supernatant, filtering the supernatant, and optionally isolating nucleic acids, disinfecting the supernatant, and / or drying the supernatant. The native proteins can be used to prepare protein preparations that are applied in the manufacture of foods or dietary supplements. The present invention also relates to food products or dietary supplements that contain the protein preparations.

Description

[Technical field]

[0001] FIELD OF THE PRESENT APPLICATION The present invention relates to a method for preparing native microbial proteins having reduced lipid and / or nucleic acid content. The obtained protein preparations can be used in the manufacture of food or nutritional supplements. [Background technology]

[0002] 2. Background of the Invention High-protein foods derived from microorganisms such as fungi, bacteria or algae are grouped under the term single cell protein (SCP). Compared to plant or animal protein sources, they have the advantage that they do not require large cultivation areas or high water consumption, and moreover, production is not dependent on seasonal or geographical factors. The fast growth rate of microorganisms also allows large amounts of microbial proteins to be produced in a short period of time. Although the use of microorganisms as food has existed for some time, currently commercially available single cell protein (SCP) products consist mostly of unprocessed microorganisms themselves.

[0003] Despite the high protein content of microorganisms, mainly non-functional protein concentrates of microbial origin are currently available. This is due to the fact that for the preparation of proteins procedures are used in which the proteins are irreversibly damaged and lose their physiological properties. As a result, the proteins lose functional properties such as solubility and textural properties, making their use in the production of alternative food systems unattractive. Protein functionality is crucial for the physicochemical properties of proteins in food systems and affects their behavior during preparation, processing, storage and consumption, and contributes to the sensory and textural properties of the food system. Protein denaturation leads to a decrease in functional properties and therefore the resulting proteins are no longer usable in many areas of the food industry.

[0004] Another aspect is that the presence of lipids shortens the shelf life of food products, as they become rancid and produce unpleasant taste.In addition, microorganisms usually have high concentrations of nucleic acids.Excessive consumption, as well as consumption of high purine foods such as meat, sausages and offal, can lead to high uric acid levels, which can cause pathological effects such as arthritis (gout), tophi or urinary stones.

[0005] According to known methods, the extraction of lipids from microbial cell lysates is usually carried out with organic solvents (e.g. hexane, methanol and tetrahydrofuran). The extraction is time-consuming, laborious and involves huge costs, since the solvents used are toxic, volatile and highly flammable, and therefore pose high health and fire risks. Furthermore, the removal of solvent residues is carried out by distillation at high temperatures, which leads to the denaturation of proteins and thus affects their functional properties. The efforts required for the analysis of residual solvents should not be underestimated, since solvents are only found in small residual amounts in food products. For example, US 4,206,243 describes the extraction of lipids from microbial cell mass with ammonia or ammonium hydroxide and isopropanol or organic solvents, such as alcohol. DE 2 328 628 describes a process for obtaining microbial proteins, in which lipid components are extracted with alcohol.

[0006] Supercritical fluid extraction method (SFE supercritical CO 2 The method offers an alternative to conventional extraction of fatty acids. However, it remains to be investigated its commercial feasibility on an industrial scale for the extraction of lipids from microbial cell lysates.

[0007] Conventional methods for reducing nucleic acid content can be divided into chemical, enzymatic, and ion exchange methods.

[0008] Chemical methods involve elevated temperature or high / low pH, which leads to denaturation of proteins, thereby impairing their functional properties. Furthermore, the nutritional safety of the isolated proteins is compromised due to the formation of potentially toxic compounds such as lysinoalanine. Another chemical method to deplete nucleic acids is precipitation with polymers such as polyethyleneimine, but this method leads to high protein losses of about 30%.

[0009] Enzymatic treatments due to activation of endogenous ribonucleases at high temperatures require microorganisms to remain active and also lead to protein denaturation. In addition, the protein loss by this method is about 33-35%.

[0010] Furthermore, GB2 101 606 describes column chromatography with anion exchange for the removal of nucleic acids from microbial homogenates. However, column chromatography of cell homogenates is limited by clogging of the packed columns by the unpurified viscous samples and the associated failures due to low flow rates during the process. Moreover, protein losses of 30-45% must be assumed.

[0011] The above methods for the reduction of nucleic acids generally have high production costs and also high protein losses, since the chemicals used cannot be regenerated or can only be regenerated uneconomically, or require large amounts of anion exchange material in the columns, which affects the economic viability of such processes.

[0012] Furthermore, WO2020 / 127951 describes a method for preparing a functional protein concentrate. However, the method does not mention lipid and / or nucleic acid separation. Similarly, US2022 / 071231A1 and WO2022 / 05287A1 describe methods for preparing protein preparations from S. cerevisiae and baker's yeast, without any reference to lipid reduction and / or nucleic acid reduction steps.

[0013] Therefore, there is a need for methods of preparing microbial proteins from microorganisms that have reduced lipid content and / or reduced nucleic acid content, while maintaining the functional properties of the proteins.

[0014] It is an object of the present invention to provide a method for preparing native microbial proteins having high yields and reduced lipid content, thereby improving the shelf life and flavor of the protein preparation.

[0015] A further object of the present invention is to provide a method for preparing natural microbial proteins with improved functional properties for the production of food or nutritional supplements, in particular in the field of vegan food production.

[0016] It is a further object of the present invention to provide a method for preparing natural microbial proteins having reduced nucleic acid content, thereby improving the quality of the human diet.

[0017] The object of the invention is achieved by the subject matter of the independent claims. Preferred embodiments are the subject matter of the dependent claims. Summary of the Invention

[0018] The present invention relates to a method for preparing a natural protein of a microorganism, comprising the steps of: a) providing a microorganism and, optionally, subjecting said microorganism to one or more pretreatment steps; b) comprising a step b1) of lysing the microorganisms, thereby obtaining a lysate comprising an aqueous liquid fraction containing lipids and dissolved native proteins of the microorganisms, and further clarifying the lysate, preferably by centrifugation or filtration, c) separating lipids from the aqueous liquid fraction using mechanical means, said lipid separation being carried out by a three-phase centrifuge, thereby obtaining an aqueous liquid fraction, said aqueous liquid fraction being a lipid-reduced aqueous liquid fraction comprising the lysed native proteins of the microorganism; d) filtering the aqueous liquid fraction, thereby obtaining a solution containing purified dissolved native proteins of the microorganism and an aqueous solvent, preferably water or a physiological saline solution; e) optionally removing at least a portion of the aqueous medium; f) optionally sterilizing the solution, thereby obtaining a sterile solution containing purified dissolved native proteins of the microorganism and an aqueous solvent; and g) optionally removing at least a portion of the aqueous medium of the sterilization solution; The present invention relates to the method comprising the steps of:

[0019] The present invention further relates to a protein preparation obtainable by the method according to the invention.

[0020] The present invention further relates to a gel-forming ability of the protein preparation at about 1% to 10% by weight of the total solution of the protein preparation and water after heat treatment, preferably without syneresis, and optionally the following: a) at least about 70% (w / w), preferably at least about 75% (w / w), more preferably at least about 85% (w / w), and most preferably at least about 95% (w / w) protein by dry weight of the protein preparation; b) less than about 110 mg / g, preferably less than about 50 mg / g, more preferably less than about 40 mg / g, and most preferably less than about 20 mg / g lipid, by dry weight of the protein preparation; c) a water binding capacity of about 4 g / g or more, preferably about 5 g / g or more, more preferably about 6.5 g / g or more of the dry weight of the protein preparation after heat treatment; d) the ability to form a gel at about 2%, about 3%, about 5%, or about 5.5% of the protein preparation by weight of the total solution of the protein preparation and water after heat treatment, preferably without syneresis, and / or e) less than about 10% (w / w), more preferably less than about 5.5% (w / w), and more preferably less than about 2.5% (w / w) nucleic acid by dry weight of the protein preparation; The present invention relates to a protein preparation derived from a microorganism, preferably a unicellular microorganism, comprising

[0021] The present invention further provides the following: (a) providing a protein preparation according to any one of claims 9 to 11, (b) mixing the protein preparation with an aqueous dispersion medium; and (c) heating the mixture to a temperature of at least about 55° C. to provide a protein gel; The present invention relates to a method for preparing a protein gel, comprising:

[0022] The present invention also relates to the use of a protein preparation according to the invention for preparing a food product or a dietary supplement, preferably for human or animal use.

[0023] Furthermore, the present invention relates to a dietary supplement or food product comprising the protein preparation of the present invention.

[0024] Furthermore, the present invention relates to a method for obtaining a natural protein of a microorganism, comprising the steps of: a) providing a microorganism and, optionally, subjecting said microorganism to one or more pretreatment steps; b) comprising a step b1) of lysing the microorganisms, thereby preparing a lysate comprising an aqueous liquid fraction containing the nucleic acids and lysed native proteins of the microorganism, and further clarifying the lysate, preferably by centrifugation or filtration, c) The following: i) adding to the aqueous liquid fraction a nucleic acid adsorbent immobilized on a solid support, preferably a free-floating solid support; ii) optionally stirring or shaking, and iii) separating the nucleic acids bound to the nucleic acid adsorbent immobilized on the solid support, preferably by precipitation and optionally filtration; separating the nucleic acids from the aqueous liquid fraction, comprising anion exchange chromatography and / or anion mixed mode chromatography comprising: d) filtering the aqueous liquid fraction, thereby obtaining a solution containing purified dissolved native proteins of the microorganism and an aqueous solvent, preferably water or a physiological saline solution; e) optionally removing at least a portion of the aqueous medium; f) optionally sterilizing the solution, thereby obtaining a sterile solution containing purified dissolved native proteins of the microorganism and an aqueous solvent; and g) optionally removing at least a portion of the aqueous medium of the sterilization solution of step f); Including, The present invention relates to a method, preferably further comprising a step of separating lipids from the aqueous liquid fraction. Detailed Description of the Invention

[0025] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs.

[0026] It should be noted that the use of the indefinite article "a" or "an" means one or more, unless otherwise stated. Also, the term "about" when used herein means + / - 10% unless otherwise stated.

[0027] The term "comprising" as used herein means "including, but not limited to." The term is intended to be open-ended and to specify the presence of any stated features, elements, integers, steps, or components, but not to preclude the additional presence of one or more other features, elements, integers, steps, components, or groups thereof. Thus, the term "comprising" encompasses the more restrictive terms "consisting of" and "consisting essentially of." In one embodiment, the term "comprising" may be replaced by the term "consisting of" as used throughout the application, and particularly the claims.

[0028] The present invention relates to a method for producing natural proteins from microorganisms.

[0029] The inventors have been able to show that a mild processing method produces microbial proteins that are best suited for use in food products. In particular, the method involves a safe and cost-effective step of isolating lipids and / or nucleic acids while at the same time maintaining functional properties and improving the tase, shelf life and health aspects of the microbial protein preparation, thereby allowing versatile use of the protein preparation in the manufacture of food and nutraceutical products. In particular, the proteins of the invention have high water binding properties, powder solubility, emulsification and foaming properties compared to conventional plant proteins, as well as gel forming ability comparable to egg white, making them particularly suitable as a replacement or equivalent in vegan, i.e. non-animal foods.

[0030] In one aspect, the present invention relates to a method for preparing native, i.e., non-denatured, proteins of a microorganism, comprising the steps of: a) providing a microorganism and, optionally, subjecting said microorganism to one or more pretreatment steps; b) comprising a step b1) of lysing the microorganisms, thereby obtaining a lysate comprising an aqueous liquid fraction containing lipids and dissolved native proteins of the microorganisms, and further clarifying the lysate, preferably by centrifugation or filtration, c) separating lipids from the aqueous liquid fraction using mechanical means, said lipid separation being carried out by a three-phase centrifuge, thereby obtaining an aqueous liquid fraction, said aqueous liquid fraction being a lipid-reduced aqueous liquid fraction comprising the lysed native proteins of the microorganism; d) filtering the aqueous liquid fraction, thereby obtaining a solution containing purified dissolved native proteins of the microorganism and an aqueous solvent, preferably water or a physiological saline solution; e) optionally removing at least a portion of the aqueous medium; f) optionally sterilizing the solution, thereby obtaining a sterile solution containing purified dissolved native proteins of the microorganism and an aqueous solvent; and g) optionally removing at least a portion of the aqueous medium of the sterilization solution; The method according to the present invention comprises the steps of: a) extracting a protein from a sample by subjecting the sample to a purification step; b) extracting a protein from a sample by subjecting the sample to a purification step; c) extracting a protein from a sample by subjecting the sample to a purification step; and e) extracting a protein from a sample by subjecting the sample to a purification step; f) extracting a protein from a sample by subjecting the sample to a purification step; and g) extracting a protein from a sample by subjecting the sample to a purification step. The method according to the present invention comprises the steps of:

[0031] The term "microbial protein" refers to a protein present in a microorganism. It includes, but is not limited to, a specific protein type, such as a metabolic, transport, storage, or structural protein. Furthermore, a microbial protein may refer to an endogenous protein of a microorganism. A microbial protein may also refer to a protein expressed by a genetic modification of a microorganism, such as a protein that enhances the value of a protein preparation in food production. In one embodiment, a microbial protein is an endogenous protein of a microorganism.

[0032] A native protein of the present invention is a protein that maintains its functional properties. In one embodiment, the protein maintains its native physical properties, such as solubility, water binding, oil binding, emulsifying or foaming properties. In one embodiment, the protein maintains its native structural properties.

[0033] The type of microorganism used in the present invention is not particularly limited. In one embodiment, the microorganism is a eukaryotic microorganism. In a further embodiment, the microorganism is a eukaryotic microorganism selected from the group consisting of fungi, yeasts, and algae.

[0034] In one embodiment, the microorganism is a fungus, preferably an Aspergillus spp., preferably a fungus selected from the group consisting of Aspergillus flavus, Aspergillus niger, Aspergillus ochraceus or Aspergillus oryzae, Rhizopus chinensis, Trichoderma harzianum, Cladosporium cladosporioides and Chrysonilia sitophilia. In a more preferred embodiment, the fungus is Aspergillus niger.

[0035] In another embodiment, the microorganism is an alga, preferably Aphanizomenon flos-aquae; Aphanothece microscopica; Arthrospira spp., preferably Arthrospira maxima (Spirulina maxima) or Arthrospira platensis (Spirulina platensis); Chlorella spp., preferably Chlorella vulgaris; Chlorella pyrenoidosa or Chlorella sorokiana; Euglena gracilis; and Scenesdesmus obliquus. In a more preferred embodiment, the algae is selected from the group consisting of Arthrospira maxima (Spirulina maxima), Arthrospira platensis (Spirulina platensis), Chlorella vulgaris, and Euglena gracilis.

[0036] In a preferred embodiment, the microorganism is a yeast. In one embodiment, the yeast is an alcohol producing yeast. In one embodiment, the yeast is selected from the group consisting of Saccharomyces spp., preferably Saccharomyces pastorianus, Saccharomyces cerevisiae, Saccharomyces carlsbergensis, Saccharomyces bayanus, Saccharomyces ellipsoides, Saccharomyces uvarum, and Saccharomycodes ludwigii; Pichia spp., preferably Pichia pastoris or Pichia anomala. anomala (Wickerhamomyces anomalus); Debaryomyces hansenii; Schizosaccharomyces spp., preferably Schizosaccharomyces pombe; Hansenula spp.; Schwanniomyces occidentalis; Zygosaccharomyces rouxii; Amoco Torula; Torulaspora delbruecki; Saccharomycopsis fibuligera; Debaryomyces hansenii hansenii; Brettanomyces bruxellensis; Candida spp.), preferably Candida intermedia, Candida arborea, Candida guilliermondii, Candida halophila, Candida krusei, Candida langeronii, Candida lipolytica, Candida parapsilosis, Candida pararugosa, Candida tropicalis, Candida novellas, or Candida utilis; Rhodotorula glutinis glutinis; Cyberlindnera jadinii; Hanseniaspora uvarum; Kluyveromyces fragilis; Kluyveromyces marxianus; Lipomyces spp.; Torulopsis spp.; and Yarrowia lipolytica.

[0037] In particularly preferred embodiments, the yeast is selected from the group consisting of Saccharomyces spp., preferably S. cerevisiae, S. carlsbergensis, S. bayanus, S. ellipsoides, S. uvarum, S. ludwigii, or S. pastorianus, Pichia spp., preferably P. pastoris; Hansenula spp.; Candida spp., preferably C. utilis; Torulopsis spp.; and Yarrowia lipolytica.

[0038] In a most preferred embodiment, the microorganism is a yeast of the genus Saccharomyces, more preferably S. cerevisiae, S. pastorianus or S. carlusbergensis.

[0039] In another embodiment, the microorganism is a prokaryotic microorganism.

[0040] In one embodiment the microorganism is a bacterium, preferably a Bacillus spp., preferably Bacillus cereus, Bacillus licheniformis, Bacillus pumilis, Bacillus subtilis or Bacillus megaterium; a Lactobacillus spp., preferably Lactobacillus casei, Lactobacillus salivarius, Lactobacillus bulgaricus, Lactobacillus delbrueckii, Lactobacillus helveticus, helveticus, Lactobacillus pentisus, Lactobacillus plantarum, Lactobacillus curvatus or Lactobacillus sake; Pediococcus spp., preferably Pediococcus acidilactici or Pediococcus pentosaceus; Lactococcus spp., preferably Lactococcus lactis; Leuconostoc mesenteroides; Oenococcus oeni oeni; Pseudomonas fluorescens; Corynebacterium spp.), preferably Corynebacterium ammoniagenes or Corynebacterium glutamicum; Cupriavidus necator; Methylomonas spp.; Rhizospheric diazotrophs; Rhodopseudomonas palustris; Aeromonas hydrophila; Methylococcus capsulatus; Ralstonia spp.; Brevibacillus agri; Aneurunibacillus spp. spp.); Achromobacter Calcoacenticus; Spirulina spp., preferably Spirulina maxima or Spirulina platensis; Xanthomonas spp., preferably Xanthomonas campestris; Acetobacter spp.; Gluconobacter spp.; Staphylococcus spp., preferably Staphylococcus carnosus or Staphylococcus xylosus; and Ideonella sakaiensis. In a preferred embodiment, the bacterium is selected from the group consisting of Bacillus subtilis, Lactobacillus spp., Corynebacterium glutamicum, Methylomonas spp., and Xanthomonas spp.

[0041] In one embodiment, the microorganism is a unicellular organism.

[0042] In the first step of the method, the microorganisms are provided in any form, including but not limited to, microorganisms in suspension. In a simple embodiment, the suspension of the microorganisms is the cell-containing medium used to cultivate the microorganisms, for example waste from the beer brewing process, preferably yeast cake. The cell-containing medium can be used directly at the discretion of step b). Alternatively, the cell-containing medium can be subjected to one or more pre-treatment steps, including filtration, sieving, washing, and / or centrifugation. For example, the microorganisms can be harvested from the culture medium by centrifugation. The centrifugation step may be preceded by a filtration step to remove the cells from the medium components. Subsequently, the harvested microorganisms may be subjected to one or more washing steps to remove residual cell medium components, optionally followed by resuspension in a suitable buffer or water. Preferably, the one or more pre-treatment steps, in particular the washing steps, are carried out at a temperature not exceeding 45°C, preferably at a temperature of about 30°C to 40°C, preferably at about 37°C.

[0043] In one embodiment, the suspension is preferably an aqueous suspension. In principle, the method of the present invention can be carried out in any volume from laboratory scale, e.g., 1-10 liters to industrial scale. In one embodiment, the suspension has a volume of about 1 liter or more, about 4 liters or more, about 5 liters or more, about 10 liters or more, about 20 liters or more, about 50 liters or more, about 100 liters or more, about 200 liters or more, about 300 liters or more, about 400 liters or more, about 500 liters or more, about 600 liters or more, about 700 liters or more, about 800 liters or more, about 900 liters or more, about 1000 liters or more, about 5000 liters or more, about 10000 liters or more. In a further embodiment, the suspension is adjusted to a dry matter content of about 5% to 20%, preferably about 10% to 15%, more preferably about 12% to 14%, per weight percent of the total mass of the suspension. Dry matter content can be measured by any method known in the art, including commercially available halogen moisture analyzers, for example, MB 35 Halogen OHAUS Europe GmbH (105° C.±2° C.). After measuring the dry matter content, the suspension can be diluted or concentrated to achieve the ranges described above.

[0044] In a particular preferred embodiment, the suspension of microorganisms is a waste product of the beer brewing process, preferably yeast cake. In one embodiment, the pretreatment step comprises filtering or sieving the suspension. Filtering or sieving can be useful to remove residual hops. Filtering or sieving can be carried out using a sieve, for example a nylon sieve or a vibrating sieve, preferably a stainless steel vibrating sieve. Filtering can also be carried out using a filter bag. The mesh size for filtering or sieving is about 110 μm to about 140 μm, preferably about 120 μm to about 130 μm, more preferably about 125 μm. The pretreatment step may further comprise a step of centrifuging the suspension to remove the yeast cake. Centrifugation is advantageously carried out at 2000 g to 4000 g, preferably at 3000 g. The pretreatment step further comprises contacting and preferably incubating the microorganism with a polysorbate solution, such as Tween® 80, more preferably an alkaline polysorbate solution. This step may be useful to remove yeast cake, thereby improving the taste of the protein. Preferably, the contacting is carried out at a temperature of about 35° C. to about 40° C., preferably about 37° C. Subsequently, the microorganism is washed, preferably the washing is repeated until the pH of the suspension reaches a pH of 5.5 to 7.0, preferably a pH of 6.4.

[0045] Step b) of the method of the invention involves lysing the microorganism, thereby obtaining a lysate comprising an aqueous liquid fraction comprising lipids and dissolved native proteins of the microorganism. More precisely, the aqueous liquid fraction comprises a lipid fraction and an aqueous fraction comprising dissolved native proteins of the microorganism. The skilled person will understand that the aqueous liquid fraction further comprises nucleic acids. The aqueous liquid fraction may further comprise a solid fraction comprising, for example, residual cell debris not removed by clarification of the lysate. In one embodiment, the pH is set to about 6.3-8.5, preferably about 6.4, before lysis. It will be understood by the skilled person that the specific method for lysis will generally depend on the particular microorganism. Microorganisms useful in the context of the invention, such as, for example, yeast, fungi, algae or bacteria, have cell walls and plasma membranes, both of which need to be disrupted to release the proteins. Another factor to be considered is that the lysis method must be chosen to maintain the native structure of the microbial proteins. In this regard, in one embodiment of the method of the invention, steps b) to g), preferably steps b) to d), in particular steps b) and d) are carried out at a temperature of about 40° C. or less (up to a temperature of about 2° C. to 8° C.), preferably about 30° C. or less, such as about 30° C. to 2° C., preferably about 30° C. to 8° C., more preferably about 30° C. to 20° C. Carrying out lysis within this temperature range avoids undesired protein degradation, which would lead to a loss of functional properties of proteins and / or a reduced activity of proteases that are also released from the microorganisms upon lysis. Thus, in a preferred embodiment of the method of the invention, lysis comprises mechanical lysis, e.g. high pressure homogenization or bead milling, or physical lysis, e.g. sonoporation and / or electroporation.

[0046] The beads may be made of steel, ceramic, rubber, or glass. For the purposes of the present invention, the use of ceramic beads, such as zirconia / silicon carbide beads or glass beads, has been found to be particularly useful. Furthermore, the beads, such as ceramic or glass beads, may have a size of about 0.05 mm to 0.7 mm, preferably about 0.5 mm to 0.6 mm, more preferably about 0.5 mm. Furthermore, the bead filling volume may range from about 40% to about 90%, preferably about 50% to 80%, more preferably about 60% to about 70%. To increase the effectiveness of the grinding, it is preferable to use a crushing medium, i.e., an acceleration device that accelerates the beads. Furthermore, an energy input of 0.01 to 0.2 kWh / kg of a slurry, such as a microbial suspension, has been found to be useful.

[0047] The effect of cell disruption can be observed by microscopic contrast, e.g., the phase contrast method; or by the protein content in the supernatant after centrifugation, e.g., (Pierce™ BCA Protein Assay Kit, Thermo Scientific). These methods are known to those skilled in the art. After 95% cell disruption, the protein content in the supernatant is about 40 mg / ml to 80 mg / ml, preferably about 50 mg / ml to about 70 mg / ml, more preferably about 55 mg / ml to 60 mg / ml.

[0048] Prior to separating the lipids in step c), the suspension of disrupted cells, i.e. the lysate, is clarified. This step separates insoluble cell debris, e.g. chromosomal DNA or cell walls, to obtain an aqueous liquid fraction containing lipids and dissolved native proteins of the microorganism. Thus, the method of the invention comprises a further step b1) of clarifying the lysate. Clarification can be carried out by various methods, including but not limited to centrifugation or filtration. In one embodiment, clarification is carried out by centrifugation at about 2000 g to 25000 g, about 2000 g to 20000 g, preferably about 5000 g to 19000 g, more preferably about 6000 g to 17000 g, about 17000 g. Clarification may also be carried out stepwise by centrifugation at about 2000 g to 7000 g, followed by a further centrifugation of the supernatant at about 5000 g to 19000 g, preferably about 6000 g to 17000 g, more preferably about 17000 g. Such clarification (centrifugation or filtration) steps do not reduce the lipid content in the aqueous liquid fraction.

[0049] Step c) of the method of the present invention comprises separating lipids from the aqueous liquid fraction, thereby obtaining an aqueous liquid fraction that is a lipid-reduced aqueous liquid fraction that contains dissolved native proteins of the microorganism. It is advantageous to separate lipids that include lipophilic substances, since fat-soluble (lipophilic) substances have a strong influence on flavor, especially by causing rancid taste. Microorganisms, especially yeasts (Saccharomyces spp.), have a high content of unsaturated fatty acids, such as oleic acid, palmitoleic acid and linoleic acid. In addition, yeasts contain so-called lipid particles, mainly non-polar lipids and sterols, which serve as building blocks for membrane lipid synthesis. Fatty acid residues, especially of unsaturated fatty acids, are particularly susceptible to oxidation processes and therefore tend to become rancid very quickly, which has a negative impact on the shelf life and taste of food products or dietary supplements produced with the protein. The term "lipid" as used herein is a collective term referring to biomolecules that are soluble in non-polar solvents such as hydrocarbons (e.g., hexane). It may also refer to lipids, lipid fractions, or lipid-containing fractions. In living organisms, lipids are primarily used as structural components in cell membranes, as energy stores, or as signaling molecules. Most biological lipids are amphiphilic, i.e., they have lipophilic hydrocarbon residues and a polar hydrophilic head group, which is why they form micelles or membranes in polar solvents such as water. The term fat is often used as a synonym for lipid, but fats only represent one subgroup of lipids, namely the triglyceride group. Lipids also encompass molecules such as fatty acids and their derivatives (including tri-, di-, monoglycerides, and phospholipids), as well as other sterol-containing metabolites, such as cholesterol. Lipids can be classified into seven groups: fatty acids, triacylglycerides (oils and fats), waxes, phospholipids, sphingolipids, lipopolysaccharides, and isoprenoids (steroids, carotenoids, etc.). Non-natural or synthetic molecules are not typically referred to as lipids.

[0050] For the purposes of the present invention, lipids are separated using mechanical means, because conventional lipid separation methods using organic solvents involve toxic solvents that are not acceptable for providing protein preparations for food production. Furthermore, regulations for food production limit the amount of organic solvents that require removal of residual solvents by distillation. Distillation is carried out at high temperatures, which leads to denaturation of proteins, thereby affecting their functional properties. Moreover, mechanical methods are an effective approach, since they are less dependent on the type of microorganisms processed and less likely to cause contamination. More specifically, the mechanical means is a centrifuge that separates lipids from the aqueous liquid fraction. This separation is based on the different densities of lipids and the lipid-reduced aqueous liquid fraction, which contains the dissolved native proteins of the microorganisms. Thus, the centrifuge is a three-phase centrifuge. According to the method of the present invention, the step of separating lipids is therefore carried out by a centrifugal (three-phase) separator, such as a skimming separator or a three-phase decanter. The term "mechanical means" as used herein does not refer to and excludes extraction with organic solvents. The inventors have surprisingly found a safe and cost-effective method for reducing lipids from a microbial cell lysate or an aqueous liquid fraction thereof while at the same time maintaining the functional properties of the proteins, which is based on the differential density between the lipids and the lipid-reduced aqueous liquid fraction containing the lysed native proteins of the microorganisms.

[0051] Those skilled in the art will understand that a three-phase centrifuge (e.g., skimming separation) is required to reduce the lipid content, and that it is different from a separator using a two-phase separator (clarifier, sedimentation centrifuge). A separator using two-phase separation, such as a centrifugal clarifier, can be used to separate a suspension consisting of a solid fraction (sludge) and a liquid fraction (two-phase separation). The suspension is placed in a centrifuge, and centrifugal force separates the solid particles from the liquid. The solid particles settle to the bottom of the centrifuge by gravity, and the clear liquid is poured out from the top. In a separator using three-phase separation, particularly using a skimming separator, a fat / lipid-containing solution consisting of a light liquid fraction, a heavy liquid fraction, and a solid fraction (sludge) (three-phase separation) can be separated. The fat / lipid-containing solution is placed in a bowl, and centrifugal force separates the two liquids and the solids by gravity. The two liquids are then discharged from the drum through a specific channel. The solids are either retained in the drum or, alternatively, are discharged discontinuously.

[0052] The term "skimming separator" as used herein refers to a three-phase disc stack separator. It contains a disc stack with a number of disc plates arranged parallel to each other. The disc plates are specially equipped with riser holes arranged in the center of the disc plates. The liquid is introduced into the main separation zone through the riser channel. From there, the light liquid phase (fat / lipid solution) flows towards the axis of rotation, whereas the heavy liquid phase (defatted protein solution) moves towards the bowl wall. An additional impeller disc on the disc stack prevents the liquid phases from mixing after separation. The disc stack not only separates the light and heavy liquid phases, but also the solid particles. The centrifugal force causes the solid particles to be separated in the disc stack and slide down the underside of the discs into the solids compartment of the bowl. The disc stack introduces more settling area. This increased the surface area which exponentially accelerates the separation process. In contrast, three-phase decanters use a screw conveyor for separation. Both decanters and disk stack separators (skimming separators) are centrifuges. In the context of lipid separation from aqueous liquid fractions in the method of the present invention, the centrifuge is a three-phase centrifuge (e.g., a three-phase decanter or a skimming separator). Centrifugation is one of the separation techniques that allows high-speed separation of immiscible (non-mixable) liquids by gravity, and it is also applied by a three-phase centrifuge. The term "centrifuge" as used herein refers to a three-phase centrifuge and does not include and is different from classical centrifuges, which typically include a rotating bowl (preferably including a means such as a disk stack or a screw conveyor) and use a container (e.g., a tube or a bucket) containing the sample or fluid to be separated, i.e., placed in the rotor.

[0053] According to the present invention, the lipid separation from the aqueous liquid fraction by mechanical means in step c) is carried out by a centrifuge, in particular a three-phase centrifuge. A preferred centrifuge is a centrifuge comprising a disk stack designed with vertically arranged riser holes (i.e. skimming separator). The aqueous liquid fraction containing lipids and dissolved native proteins of the lysate or microorganisms is made to flow through the vertical riser holes. Thereby, under the influence of different densities and centrifugal forces, the lipids of the lysate or aqueous liquid fraction can be separated from the aqueous liquid fraction containing the lysate or dissolved native proteins of the microorganisms. Since the lipids have a lower density, they flow inward in the axial direction of rotation. Thus, the lipids can be separated via an axially arranged outlet, so that the total lipid content can be reduced. The centrifugal (three-phase) separator can be oriented vertically or horizontally. In one embodiment, the feed rate is about 0.5 L / min to 50 L / min, about 1 L / min to 20 L / min, about 5 L / min to 10 L / min. In another embodiment, the feed rate is from about 100 l / h to about 20000 l / h, preferably from about 500 l / h to about 15000 l / h, more preferably from about 1000 l / h to about 10000 l / h. Furthermore, for the purposes of the present invention, it has been found that for the separation of lipids, temperatures below 40° C. are preferred (up to a temperature of about 2° C. to 8° C.), in particular below 30° C., for example from 30° C. to 2° C., preferably from 30° C. to 8° C., preferably from 30° C. to 20° C. In a particularly preferred embodiment, the separation of lipids is carried out using a skimming separator or a three-phase decanter, for example a Tricanter® (Flottweg). A three-phase decanter separates lipids and, advantageously, if present, also a solid fraction from the aqueous liquid fraction comprising the dissolved native proteins of the microorganism, thereby improving the purity of the aqueous liquid fraction comprising the dissolved native proteins of the microorganism.

[0054] The success of lipid separation can be determined, for example, by recording the UV spectrum (200-350 nm) after solvent extraction of the aqueous liquid fraction (lipid-reduced aqueous liquid fraction and / or aqueous liquid fraction before lipid separation) or lysate. This method is rapid and provides fast results regarding the reduction of unsaturated fatty acids and other lipophilic substances. The spectral (UV absorption spectrum) difference between the untreated and lipid-reduced aqueous fraction or lysate is used. The success of lipid reduction can then be compared and determined based on the appearance of the characteristic diene and triene fatty acid bands (200-350 nm) in the UV spectrum. Thus, in one embodiment, the method of the present invention further comprises the following: i) contacting the aqueous liquid fraction with a lipophilic solvent, preferably hexane, thereby obtaining a lipophilic phase; ii) measuring the absorbance of the lipophilic phase, preferably in the UV wavelength range; iii) comparing the absorbance measured in the lipophilic phase with a reference absorbance measured in lipids; and iv) quantifying lipids in the lipophilic phase; quantification of lipids in the aqueous liquid fraction (e.g., the lipid-reduced aqueous liquid fraction and / or the aqueous liquid fraction prior to separation), including

[0055] Clarification can also be carried out after separation of lipids.Thus, in one embodiment the method of the invention comprises a further step c1) of clarifying the aqueous liquid fraction, which is a lipid-reduced aqueous liquid fraction containing the lysed native proteins of the microorganisms.

[0056] In step d) of the method of the present invention, the aqueous liquid fraction is filtered to remove particles having a size smaller than 1 kDa to about 100 kDa, preferably about 3 kDa to about 50 kDa, more preferably about 5 kDa to about 30 kDa, most preferably smaller than about 10 kDa. The particles removed by filtration are preferably smaller than 10 kDa, more preferably smaller than 5 kDa. Step d) comprises filtering the aqueous liquid fraction, preferably the aqueous liquid fraction of step c), more preferably the lipid-reduced aqueous liquid fraction comprising the lysed native proteins of the microorganism, thereby obtaining a solution comprising purified lysed native proteins of the microorganism and an aqueous solvent, preferably wherein the aqueous solvent is water or a physiological saline solution. The steps of the method are performed in the order listed in the method of the present invention.

[0057] In one embodiment, the filtration is step d) and is carried out by ultrafiltration. The separation principle of ultrafiltration is based on a membrane that allows the passage of particles and solvents whose size is smaller than the membrane pore size, while particles or molecules whose size is larger than the membrane pore size are retained. Thus, the filtration step results in two fractions, the permeate (solvent containing particles or molecules smaller than the membrane pore size) and the retentate (solvent containing particles or molecules larger than the membrane pore size). The membranes used for ultrafiltration, preferably hydrophilic membranes, have a molecular weight cut-off in the range of about 1 kDa to about 100 kDa, preferably about 3 kDa to about 50 kDa, more preferably about 5 kDa to about 15 kDa, most preferably about 10 kDa. For the purposes of the present invention, ultrafiltration is applied as tangential flow filtration. In one embodiment, the membrane is a hollow fiber membrane with a molecular weight cut-off of about 10 to 20 kDa.

[0058] In a further embodiment of step d), ultrafiltration is combined with diafiltration to change the liquid. During diafiltration, a solvent is continuously applied to the retentate in the ultrafiltration process until the desired degree of exchange with the solvent is achieved. Thus, in the present invention, diafiltration is applied to replace at least a part of the liquid of the aqueous liquid fraction by a solvent, preferably an aqueous solvent, more preferably water or a buffer, such as a physiological saline solution. The physiological saline solution is a mixture of salt and water. In one embodiment, the salt is sodium chloride, ammonium sulfate, potassium phosphate, or ammonium chloride. In a preferred embodiment, the physiological saline solution is preferably a sodium chloride solution of about 0.01% to about 5%, preferably about 0.5% to about 2%, more preferably about 0.9% to about 1.5% (w / v). In one embodiment, the liquid of the aqueous liquid fraction is substantially, preferably completely, changed by the solvent. After diafiltration, the solution is preferably concentrated to a factor within the range of about 1.5 to about 4.5, preferably about 2 to about 3.5, more preferably about 3. The magnification is defined as the total volume at the start of filtration / the volume of the retentate. In one embodiment, the diavolume is in the range of about 0.0 to 5.0, preferably 0.3 to 5.0, more preferably 0.7. The diavolume is a relative volume and is therefore defined as the total volume of product introduced into the run during diafiltration / the volume of the retentate.

[0059] Optionally, the aqueous solvent of the solution obtained after filtration in step d) is at least partially removed in step e) to further concentrate the solution, preferably at least about 50% of the solvent, preferably at least about 75%, more preferably at least about 90%, most preferably at least about 94% or even up to 98% of the solvent. In one embodiment, the solution is dried to obtain a powder of the native protein of the microorganism. Essentially, the removal of at least a portion of the solvent in step e) and / or step g) may involve any method, including but not limited to spray drying, vacuum drying, drum drying, fluidized bed drying or freeze drying, preferably spray drying. These methods are conventional and known to those skilled in the art. In one embodiment, the product of step e) has a solvent content of about 4% to 40%, preferably about 4% to 30%, more preferably about 4% to 20%, based on the dry weight of the total product.

[0060] In an even further embodiment of the method of the invention, the solution of step d) or step e), if present, may optionally be sterilized in step f).

[0061] For the purposes of the present invention, it is important that the sterilization does not result in degradation or denaturation of the protein, which may result in the loss of its functional properties. Suitable sterilization methods are known in the art and the skilled person is aware of them, for example, sterile filtration, preferably close to 1 second, ultra-high temperature treatment (UHT), ultraviolet (UV) treatment, and pulsed electric field (PEF) treatment. In a preferred embodiment, the sterilization is carried out by sterile filtration. The principle is based on filtration using a membrane with a pore size suitable for the exclusion of bacteria and fungi. In general, any membrane filter system known in the art of sterile filtration can be used. These systems are known to the skilled person. Particularly useful in this respect are filtration membranes with a pore size of about 0.1 μm to 1 μm, preferably about 0.2 μm. For the purposes of the present invention, it has been found that sterile filtration using a heterogeneous membrane system, for example a polyethersulfone (PES) bilayer (0.8 μm) and a glass fiber membrane (0.2 μm), is particularly useful.

[0062] Optionally, the aqueous solvent of the solution obtained after sterilization in step f) is at least partially removed, preferably at least about 50% of the solvent, preferably at least about 75%, more preferably at least about 90%, most preferably at least about 94% or even up to 98% of the solvent, in order to further concentrate the solution. In one embodiment, the solution is dried to obtain a powder of the native protein of the microorganism. Essentially, the removal of at least a portion of the solvent in step e) and / or step g) may involve any method, including but not limited to spray drying, vacuum drying, drum drying, fluidized bed drying or freeze drying, preferably spray drying. These methods are conventional and known to those skilled in the art. In one embodiment, the product of step g) has a solvent content of about 4% to 40%, preferably about 4% to 30%, more preferably about 4% to 20%, based on the dry weight of the total product.

[0063] In a specific embodiment, the method of the invention further comprises a step of isolating nucleic acids, such as ribonucleic acid (RNA) or deoxyribonucleic acid (DNA). In one embodiment, the separation of nucleic acids is performed after step b) or b1), i.e. the nucleic acids are separated from the lysate (or clarified lysate). In another embodiment, the separation of nucleic acids is performed after step d), i.e. the nucleic acids are separated from the solution of step d). In a preferred embodiment, the separation of nucleic acids is performed after step c), preferably from the lipid-reduced aqueous liquid fraction containing the lysed native proteins of the microorganism. Thus, the aqueous liquid fraction of step d) may be a lipid-reduced aqueous liquid fraction or a lipid-reduced and nucleic acid-reduced aqueous liquid fraction.

[0064] For the purposes of the present invention, the separation of nucleic acids is carried out at temperatures of about 40° C. or less, 30° C. or less, 20° C. or less, 10° C. or less, preferably about 30° C. or less, and / or without the use of low or high pH conditions to avoid denaturation of proteins and loss of functional properties of proteins. According to a preferred embodiment, the separation of nucleic acids comprises chromatography. Chromatography can be carried out in any volume, from laboratory scale to industrial scale. For example, chromatography may be carried out in a volume of about 0.5 liter to 2 liter. Chromatography may also be carried out in a volume of about 1 liter or more, 10 liter or more, 20 liter or more, 50 liter or more, 100 liter or more, 200 liter or more, 500 liter or more, 1000 liter or more, 2000 liter or more, 5000 liter or more, 7000 liter or more, or 10000 liter or more.

[0065] In principle, any type of chromatography that is suitable for the separation of nucleic acids is possible. Those chromatographic methods are known in the art and include anion exchange chromatography and / or anion exchange mixed mode chromatography. According to the present invention, the separation of nucleic acids from the aqueous liquid fraction comprises anion exchange chromatography and / or mixed mode chromatography. In anion exchange chromatography, the nucleic acid adsorbent is immobilized on a solid support, for example a ceramic or a resin, for example styrene-DVB. The nucleic acid adsorbent comprises a positively charged functional group, for example a quaternary ammonium compound. Nucleic acids with a negatively charged backbone can bind to the nucleic acid adsorbent, thereby separating the nucleic acid from the aqueous liquid fraction or solution. In anion exchange mixed mode chromatography, the solid support comprises a nucleic acid adsorbent with a positively charged functional group and a nucleic acid adsorbent with a further functional group having another type of interaction with the nucleic acid. The functional group and the further functional group may be present in the same nucleic acid adsorbent. For example, the positively charged functional group can be combined with a functional group for hydrophobic interaction. An example for a mixed mode nucleic acid adsorbent is hydroxylapatite (Ca 5 (PO 4 ) 3 OH) 2The solid support may be packed in a column. Alternatively, the solid support may be a floating support (wherein the particles of the solid support move freely within a container), e.g., batch binding chromatography. Thus, in one embodiment of the method of the invention, the chromatography comprises column chromatography. In another embodiment, the chromatography comprises expanded bed adsorption chromatography. In a preferred embodiment, the chromatography comprises batch binding chromatography.

[0066] In one embodiment, the separation of nucleic acids in the method of the invention comprises batch binding chromatography, which comprises: a) adding a nucleic acid sorbent immobilized on a solid support, preferably a floating solid support; b) preferably stirring or shaking, more preferably shaking with an overhead shaker or agitator; and c) separating the nucleic acids bound to the nucleic acid sorbent immobilized on the solid support. In one embodiment, the nucleic acid sorbent is added to the lysate or clarified lysate of the method of the invention. In one embodiment, the nucleic acid sorbent is added to the aqueous liquid fraction, preferably the lipid-reduced aqueous liquid fraction, of the method of the invention. In one embodiment, the nucleic acid sorbent is added to a solution obtained by filtering the (lipid-reduced) aqueous liquid fraction of the method of the invention. In one embodiment, the separation of nucleic acids bound to the nucleic acid sorbent immobilized on the solid support comprises filtering the nucleic acid bound to the nucleic acid sorbent immobilized on the solid support. In another embodiment, the separation of nucleic acids bound to the nucleic acid sorbent immobilized on the solid support comprises centrifugation of the nucleic acid bound to the nucleic acid sorbent immobilized on the solid support, preferably at about 1000 g to 4000 g. In a preferred embodiment, the separation of the nucleic acids bound to the nucleic acid adsorbent immobilized on the solid support comprises precipitation of the nucleic acids bound to the nucleic acid adsorbent immobilized on the solid support, and d) optionally filtering the fraction, preferably by dead-end filtration. The method according to the invention does not comprise a step of combining previously separated fractions.

[0067] In batch binding mode chromatography, the solid support particles (stationary phase) are added directly to the sample and not in the separation column in the case of column chromatography or expanded bed adsorption chromatography. The binding of the nucleic acids to the stationary phase due to their affinity for the stationary phase is carried out by transport, dispersion and adsorption of the active mass in a well-mixed container. The solid support particles are floating, e.g., they are evenly spread in the sample (mobile phase) by mixing, stirring or shaking, e.g., by an overhead shaker, so that they can be separated from the sample by sedimentation and / or filtration after the completion of the mixing step. In contrast to column chromatography, in batch binding mode chromatography, particles with larger diameters are advantageous due to their faster sedimentation properties. Subsequently, the nucleic acids bound to the solid support can be eluted from the solid support. The elution can be carried out in stages so that the various bound nucleic acids are recovered separately. After elution of the bound nucleic acids and subsequent equilibration, the chromatographic material, e.g., the solid support, can be regenerated for further use. Batch binding chromatography allows for the selective binding of nucleic acids from unpurified, viscous biological samples, thereby avoiding the costly and labor-intensive preparation procedures of lysates. Thus, batch binding chromatography can be applied directly to lysates and overcome limitations of column chromatography such as blockage of packed columns by viscous biological samples and associated bottlenecks due to low flow rates during the process.

[0068] Typically, the nucleic acid concentration in the lysate in step b) of the method of the present invention is about 10% to 15% based on the dry matter of the lysate. The separation of nucleic acids described herein results in a reduction of the nucleic acid concentration in the lysate by at least about 40%, preferably at least about 50%, more preferably at least about 65%, more preferably at least about 75%, more preferably at least about 80%, while maintaining the functional properties of the protein.

[0069] In a further aspect, the present invention provides a protein preparation obtainable by a method according to the method of the present invention. In a particular embodiment, the protein preparation is characterized in that it comprises, after heat treatment, preferably without syneresis, a gel-forming capacity of about 1% to 10% of the protein preparation per total weight of a solution consisting of the protein preparation and water.

[0070] In a preferred embodiment, the protein preparation of the present invention is in a dry form, preferably in the form of a powder.

[0071] In yet another aspect, the invention provides a protein preparation from a microorganism, preferably a unicellular microorganism, comprising at least about 70% (w / w), or at least about 72%, preferably at least about 75% (w / w), more preferably at least about 85% (w / w), and most preferably at least about 95% (w / w) protein by dry weight of the protein preparation. In one embodiment, the protein preparation comprises about 70%-80% (w / w) protein by dry weight of the protein preparation. In a preferred embodiment, the protein preparation comprises native protein.

[0072] In a preferred embodiment, the protein preparations of the invention contain lipids (fat) of about 110 mg / g or less, preferably about 50 mg / g or less, more preferably about 40 mg / g or less, even more preferably about 20 mg / g or less, and even more preferably about 15 mg / g or less by dry weight of the protein preparation (assessed using the Determination of Total Fat Content of Cereal Products After Acid Hydrolysis by Extraction and Gravimetric Measurement Method according to § 64 LFGB L16.00-5: 2017-10), and optionally contain about 10% (w / w) or less, preferably about 5.5% (w / w) or less, more preferably about 2.5% nucleic acid by dry weight of the protein preparation. In certain embodiments, the lipid (fat) content by dry weight (assessed using a method according to §64 LFGB L16.00-5: 2017-10) is reduced by about 20% or more (e.g., 25% or more, 30% or more, 35% or more, 40% or more, or 50% or more) after lipid reduction compared to the lysate or aqueous liquid fraction before lipid reduction. Moreover, the fat-soluble components, as measured by the UV method described herein, are reduced by about 50-60% or more after lipid reduction compared to the lysate or aqueous liquid fraction before lipid reduction. The total lipid (fat) content can be measured, for example, using the method for determining the total fat content of a cereal product after acid hydrolysis by extraction and gravimetric measurement (according to §64 LFGB L16.00-5: 2017-10). Instruction § 64 LFGB L16.00-5: 2017-10 describes a method to be carried out according to DIN standards by the DAkks accredited laboratory (holding a certificate of accreditation from the Deutsche Akkreditierungsstelle), which can be found in the BVL method collection for food, e.g. at Beuth Verlag GmbH.

[0073] In a preferred embodiment, the protein preparation of the present invention comprises a water binding capacity of about 4 g / g or more, about 4.5 g / g or more, about 5 g / g or more, about 6.5 or more, for example, about 4.5-20 g / g, about 4.5 g / g-10 g / g, about 6-10 g / g, about 6-7.5 g / g, about 6.0-7.0 g / g, etc., by dry weight of the protein preparation, after heat treatment, preferably at about 80°C. Water-binding capacity can be measured by any method known in the art, including, but not limited to, the method used herein in the Examples, or as described in Kneifel, W. and Seiler, A. (1993) "Water-holding Properties of Milk Protein Products - A Review", Food Structure: Vol. 12: No. 3, Article 3; or as described in Wang, JS, Wang, AB, Zang, XP, Tan, L., Xu, BY, Chen, HH, et al. (2019). "Physicochemical, functional and emulsion properties of edible protein from avocado (Persea americana Mill.) oil processing by-products.", Food Chemistry, 288 (February), 146-153; or S. Thammakiti, M. Suphantharika, T. Phaesuwan, C. Verduyn: "Preparation of spent brewer's yeast β-glucans for potential applications in the food industry, An example of such a method is the method described in Int. J. Food Sci.Technol. 39 (2004) 21-29.This method has been adapted from Vlatka Petravic-Tominac, Vesna Zechner-Krpan, Katarina Berkovic, Petra Galovic, Zoran Herceg, Sinisa Srecec, Igor Spoljaric: “Rheological Properties, Water-Holding and Oil-Binding Capacities of Particulate β-Glucans Isolated from Spent Brewer's Yeast by Three Different Procedures”, January 2011, Food Technology and Biotechnology 49(1):56-64) (incorporated herein by reference in its entirety). In a particularly preferred embodiment, the water-binding capacity is as follows: i) preparing a solution (w / v) containing 0.5 g of the protein preparation to be tested in 4 mL or 5 mL of demineralized water; ii) Mix the solution for 20 seconds; iii) Mixing was repeated seven times with an interval of 10 minutes; iv) The solution is allowed to stand at 80°C for 10 minutes; v) Allow the solution to cool to room temperature; vi) Centrifuge in a container at 2000 g for 25 minutes at 20° C. vii) discard the supernatant and obtain a wet sample; viii) Remove any remaining water by placing the container at a 20° angle for 10 minutes; and ix) Calculating the water binding capacity (WBC) by (weight of water wet sample-weight of container-mass of protein) / mass of protein; The method includes:

[0074] In a preferred embodiment, the protein preparation of the present invention comprises a gel forming ability, preferably without syneresis, of about 0.5%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5% or 10% or more, preferably about 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5% or 5% or more of the protein preparation based on the total weight of the solution consisting of the protein preparation and water, after heat treatment, preferably at about 80° C. Preferably, gel formation is observed in about 1%-20%, 5%-20%, preferably about 1-10%, about 2%-10%, about 5-10%, about 1-7% or about 7%-10% of the protein preparation based on the total weight of the solution consisting of the protein preparation and water, after heat treatment. In certain embodiments, the protein preparations of the invention already comprise a gel forming ability, preferably without syneresis, at about 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, and / or about 10% (w / w) of the protein preparation, preferably at (only) about 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, or 5.5% (w / w) of the protein preparation, based on the total weight of the solution consisting of the protein preparation and water, after heat treatment. Gel forming ability can be measured by any method known in the art, including but not limited to the method used herein in the Examples or the method described in Langton et al., “Gelation of faba bean proteins - Effect of extraction method, pH and NaCl”, Food Hydrocolloids (2020) 103, 105622:1-8, all of which are incorporated herein by reference. In a particularly preferred embodiment, the gel forming ability is one of the following: i) preparing a solution (w / w) containing 5% of the protein preparation to be tested in demineralized water and stirring the solution for 20 minutes; ii) exposing the solution to 80°C for 20 minutes; iii) determining the condition of the test preparation (wherein there is gel formation if the condition of the test preparation is equivalent to that of a reference preparation containing 5% egg white protein); The method includes:

[0075] In one embodiment, the protein preparation of the invention, after heat treatment, preferably at about 80° C., comprises an oil binding capacity of about 0.3 g / g or more, preferably about 0.5 g / g or more, about 2 g / g or more, more preferably 3 g / g or more, or from about 0.3 g / g to 4 g / g, from about 0.5 g / g to 4 g / g, from about 0.5 g / g to 3 g / g, from about 0.5 g / g to 2 g / g, or from about 0.5 to 0.7 g / g of dry weight of the protein preparation.The oil-binding capacity can be measured by any method known in the art, including, but not limited to, the method used herein in the Examples or the method described in Wang, JS, Wang, AB, Zang, XP, Tan, L., Xu, BY, Chen, HH, et al. (2019). “Physicochemical, functional and emulsion properties of edible protein from avocado (Persea americana Mill.) oil processing by-products.”, Food Chemistry, 288 (February), 146-153; or the method described in Vlatka Petravic et al. “Rheological Properties, Water-Holding and Oil-Binding Capacities of Particulate β-Glucans Isolated from Spent Brewer's Yeast by Three Different Procedures”, January 2011, Food Technology and Biotechnology 49(1):56-64; or the method described in V. PETRAVI-TOMINAC et al.: “Properties of b-Glucans from Brewer's Yeast”, Food Technol. Biotechnol. 49 (1) 56-64 (2011), or Zayas, JF: Oil and Fat Binding Properties of Proteins. In: Functionality of Proteins in Food, Springer Verlag Berlin Heidelberg, 1997 (all of which are incorporated herein by reference). In a particularly preferred embodiment, the oil binding capacity is determined by the following: i) preparing a dispersion (w / v) to be tested containing 0.5 g of protein preparation in 4 mL or 5 mL of sunflower oil; ii) Mix the dispersion for 20 seconds; iii) Mixing was repeated seven times with an interval of 10 minutes; iv) The dispersion is allowed to stand at 80° C. for 10 minutes, v) Cool the dispersion to room temperature; vi) centrifuging the dispersion in a container at 2000 g for 25 minutes at 20° C.; vii) discard the supernatant and obtain the oil-wet sample; viii) Remove any remaining oil by placing the container at a 20° angle for 10 minutes; and ix) Calculating the Oil Binding Capacity (OBC) by (Weight of oil wet sample-Weight of container-Mass of protein) / Mass of protein; The method includes:

[0076] In one embodiment, the protein preparation of the present invention comprises a powder solubility of about 74% or more, about 75% or more, about 78% or more, about 80% or more, about 82% or more, about 85% or more of the initial concentration of the protein preparation suspended in water by total weight of the suspension. Powder solubility can be measured by any method known in the art, including but not limited to the method used herein in the Examples or the method described in US 4,465,702 or the method described in Elif Ezgi Ozdemir, Ahmet Gorguc, Esra Gencdag, Fatih Mehmet Yilmaz., “Physicochemical, functional and emulsifying properties of plant protein powder from industrial sesame processing waste as affected by spray and freeze drying.”, LWT Food Science and Technology, 154 (2022) 112646 (all of which are incorporated herein by reference). In a preferred embodiment, powder solubility is measured as disclosed in US 4,465,702, which is incorporated herein by reference. In a particularly preferred embodiment, powder solubility is measured according to the following: i) preparing a test solution containing 2% of the protein preparation (w / v) in 50 ml of demineralized water; ii) Mix the solution using a magnetic stirrer, preferably at 800 rpm, for 30 minutes; iii) centrifugation at 2000 g and 20° C. for 25 minutes; iv) 25 ml of the supernatant was transferred into an aluminum shell; v) The supernatant was dried at 160° C. for 1.5 hours to obtain a sample; vi) Cool the aluminum shell containing the sample in a desiccator. vii) Weighing the aluminum shell containing the sample; viii) Calculate the powder solubility (%) by (weight of sample with aluminum shell - empty weight of aluminum shell) / (weight of protein in solution x concentration of protein preparation) x 2 x 100%; The method includes:

[0077] In one embodiment, the protein preparation of the present invention comprises, after centrifugation, about 54% or more, about 55% or more, about 56% or more, about 57% or more, about 58% or more, about 59% or more, about 60% or more emulsifying activity per total volume of emulsion consisting of 25 ml of sunflower oil and 25 ml of solution consisting of 5% protein preparation in water, where the emulsifying activity is defined as emulsion layer (ml) / total volume (ml) x 100%.Emulsifying activity can be measured by any method known in the art, including, but not limited to, the method used herein in the Examples, or the turbidimetric measurement described in Pearce et al., “Emulsifying properties of proteins: evaluation of a turbidimetric technique”, J. Agric. Food, Chem., 198, 26:716-723, or the method described in Elif Ezgi Ozdemir, Ahmet Gorguc, Esra Gencdag, Fatih Mehmet Yilmaz, “Physicochemical, functional and emulsifying properties of plant protein powder from industrial sesame processing waste as affected by spray and freeze drying”, LWT Food Science and Technology, 154 (2022) 112646, or the method described in Lam and Nickerson, Food proteins: A review on their emulsifying properties using a structure-function approach. Food Chemistry 141 2013: 975-984, or the method described in Hasenhuettl and Hartel, Food emulsifiers and their applications: Second edition, 2008, Springer Science + Business Media, LLC (all of which are incorporated herein by reference). In a preferred embodiment, the emulsifying activity is measured by the method disclosed in Ozdemir et al. (all of which are incorporated herein by reference). In a particularly preferred embodiment, the emulsifying activity is measured by the following method: i) preparing an emulsion to be tested consisting of a 5% solution (w / v) of the protein preparation powder in 25 ml of water and 25 ml of sunflower oil; ii) homogenizing the emulsion; iii) Immediately centrifuge at 1200 g for 5 minutes; iv) Measure the emulsion layer and total volume; v) Calculating the emulsifying activity by emulsion layer (ml) / total volume (ml) x 100%; The method includes:

[0078] In one embodiment, the protein preparation of the present invention comprises an emulsion stability of about 97% or more, preferably about 98% or more, more preferably about 100% per total volume of emulsion consisting of 25 ml of sunflower oil and 25 ml of a solution consisting of 5% protein preparation in water after exposing the emulsion to 80° C. for 30 minutes followed by centrifugation, wherein the emulsion stability is defined as emulsion layer (ml) / total volume (ml)×100%. Emulsion stability can be measured by any method known in the art, including, but not limited to, the method used herein in the Examples, or the turbidimetric measurement described in Pearce et al., "Emulsifying properties of proteins: evaluation of a turbidimetric technique", J. Agric. Food, Chem., 198, 26:716-723, or the method described in Elif Ezgi Ozdemir, Ahmet Gorguc, Esra Gencdag, Fatih Mehmet Yilmaz, "Physicochemical, functional and emulsifying properties of plant protein powder from industrial sesame processing waste as affected by spray and freeze drying", LWT Food Science and Technology, 154 (2022) 112646, all of which are incorporated herein by reference. In a preferred embodiment, emulsion stability is measured as disclosed in Ozdemir et al., which are incorporated herein by reference. In a particularly preferred embodiment, emulsifying activity is measured as follows: i) preparing an emulsion to be tested consisting of a 5% solution (w / v) of the protein preparation powder in 25 ml of water and 25 ml of sunflower oil; ii) homogenizing the emulsion; iii) Immediately centrifuge at 1200 g for 5 minutes; iv) Exposing the emulsion to 80°C for 30 minutes; v) Allow the emulsion to cool; vi) Centrifuge at 1200 g for 5 minutes; vii) Measure the emulsion layer and total volume; viii) Calculating emulsion stability by emulsion layer (ml) / total volume (ml) x 100%; The method includes:

[0079] In one embodiment, a protein preparation of the invention comprises a foaming capacity of about 40% or more, about 41% or more, about 42% or more, about 43% or more, about 44% or more, about 45% or more, about 46% or more, about 47% or more, about 48% or more, about 49% or more, about 50% or more, about 51% or more, about 52% or more per total volume of a solution consisting of 100 mg of protein preparation in 10 ml of water, where the foaming capacity is defined as foam volume (ml) / total volume (ml) x 100%. Foaming ability can be measured by any method known in the art, including but not limited to the method used herein in the Examples, or by turbidimetric measurements as described in Pearce et al., “Emulsifying properties of proteins: evaluation of a turbidimetric technique”, J. Agric. Food, Chem., 198, 26:716-723, or by the method described in Elif Ezgi Ozdemir, Ahmet Gorguc, Esra Gencdag, Fatih Mehmet Yilmaz, “Physicochemical, functional and emulsifying properties of plant protein powder from industrial sesame processing waste as affected by spray and freeze drying”, LWT Food Science and Technology, 154 (2022) 112646, or by the method described in Richard K Owusu-Apenten, “Testing protein functionality”, April 2004, in “Proteins in food processing”: 217-244, 1.Edition, Publisher: Woodhead Publishing Ltd. 17124 N), editor: Forschungskreis der Ernahrungsindustrie eV, Bonner Universitats-Buchdruckerei, Bonn or MEBAK online. Methode B-420.09.100. Schaumhaltbarkeit nach ROSS und CLARK. Rev. 2020-10. Analysenkommission (MEBAK(R)) eV, Freising. https: / / www.mebak.org / methode / b-420-09-100 / schaumhaltbarkeit-nach-ross-und-clark / 711 (all of which are incorporated herein by reference). In a preferred embodiment, the foaming capacity is as follows: i) preparing a dispersion consisting of 100 mg of the protein preparation to be tested in 10 ml of distilled water; ii) homogenizing the dispersion, preferably at 11000 rpm for 30 seconds; iii) Transfer the dispersion into a graduated cylinder; iv) Measure the foam volume after 30 seconds; iv) Calculate the foaming capacity by foam volume (ml) / total volume (ml) x 100%; The method includes:

[0080] In one embodiment, the protein preparations of the invention comprise a foam stability of about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 75% or more, about 76% or more foam volume per total volume of a solution consisting of 100 mg of protein preparation in 10 ml of water after 60 minutes, where the foam stability is defined as foam volume after 60 minutes (ml) / initial volume (ml) x 100%. Foam stability can be measured by any method known in the art, including, but not limited to, the method used herein in the Examples, or the turbidimetric measurements described in Pearce et al., “Emulsifying properties of proteins: evaluation of a turbidimetric technique”, J. Agric. Food, Chem., 198, 26:716-723, or the methods described in Elif Ezgi Ozdemir, Ahmet Gorguc, Esra Gencdag, Fatih Mehmet Yilmaz, “Physicochemical, functional and emulsifying properties of plant protein powder from industrial sesame processing waste as affected by spray and freeze drying”, LWT Food Science and Technology, 154 (2022) 112646, or Richard K Owusu-Apenten, “Testing protein functionality”, April 2004, in “Proteins in food processing”: 217-244, 1.Edition, Publisher: Woodhead Publishing Ltd. 17124 N), editor: Forschungskreis der Ernahrungsindustrie eV, Bonner Universitats-Buchdruckerei, Bonn or MEBAK online. Methode B-420.09.100. Schaumhaltbarkeit nach ROSS und CLARK. Rev. 2020-10. Analysenkommission (MEBAK(R)) eV, Freising. https: / / www.mebak.org / methode / b-420-09-100 / schaumhaltbarkeit-nach-ross-und-clark / 711 (all of which are incorporated herein by reference). In a preferred embodiment, foam stability is achieved by the following: i) preparing a dispersion consisting of 100 mg of the protein preparation to be tested in 10 ml of distilled water; ii) homogenizing the dispersion, preferably at 11000 rpm for 30 seconds; iii) Transfer the dispersion into a graduated cylinder; iv) Measure the foam volume after 60 minutes; iv) Calculate the foam stability by: foam volume measured after 60 minutes (ml) / initial volume (initial volume is the volume measured immediately after transferring the dispersion into a graduated cylinder) (ml) x 100%; The method includes:

[0081] In a preferred embodiment of the inventive protein preparation, the inventive protein preparation is present in dry form, preferably in the form of a powder.

[0082] In an even further preferred embodiment of the protein preparation of the present invention, the microorganism is a fungus, preferably a fungus as described herein, more preferably Aspergillus niger.

[0083] In an even further preferred embodiment of the protein preparation of the present invention, the microorganism is an algae, preferably an algae as described herein, more preferably an algae selected from the group consisting of Arthrospira maxima (Spirulina maxima), Arthrospira platensis (Spirulina platensis), Chlorella vulgaris, and Euglena gracilis.

[0084] In certain preferred embodiments of the protein preparation of the invention, the microorganism is a yeast, preferably a yeast as described herein, more preferably a yeast selected from the group consisting of Saccharomyces spp., preferably S. cerevisiae, S. carlsbergensis, S. bayanus, S. ellipsoides, S. uvarum, S. ludwigii or S. pastorianus, Pichia spp., preferably P. pastoris; Hansenula spp.; Candida spp., preferably C. utilis; Torulopsis spp.; and Yarrowia lipolytica.

[0085] In another aspect, the present invention provides a method for producing a composition comprising the steps of: (a) providing a protein preparation according to the present invention; (b) mixing the protein preparation with an aqueous dispersion medium; and (c) heating the mixture to a temperature of at least about 55° C. to provide a protein gel; The present invention provides a method for preparing a protein gel comprising:

[0086] In one embodiment, the protein preparation is provided in a solution. In a preferred embodiment, the protein preparation is provided in a dry form, e.g., as a powder. In one embodiment, the aqueous dispersion medium includes, but is not limited to, water or an aqueous dispersion medium, e.g., a buffer. In one embodiment, the protein preparation is present at about 5% (w / w) or more, about 6% or more, about 7% or more, about 8% or more, about 9% or more, about 10% or more, about 20% or more, about 30% or more, about 40% or more, about 50% or more, preferably about 5% to about 20%, more preferably about 5% to 10% by weight of the total volume of the aqueous dispersion medium. Mixing of the aqueous dispersion medium and the protein preparation can be performed by stirring or shaking. In one embodiment, the mixture is heated to a temperature of at least about 55°C, at least about 60°C, at least about 65°C, at least about 70°C, at least about 75°C, or at least about 80°C. At this temperature, the protein denatures and forms a gel. Heating is performed in a water bath.

[0087] In yet another aspect, the present invention relates to the use of the protein preparation of the present invention for preparing a food product or a dietary supplement, preferably for human or animal use. In one embodiment, the protein preparation serves as an equivalent or substitute for methylcellulose, especially in meat substitutes, ice cream, bakery products, cake creams, mayonnaise, ready-to-eat meals or frozen products. In another embodiment, the protein preparation serves as an equivalent or substitute for vegetable proteins, egg proteins, preferably egg yolk proteins and / or egg white proteins, meat proteins, gluten proteins, and / or milk proteins. In a preferred embodiment, the protein preparation serves as an equivalent or substitute for egg proteins, preferably egg yolk proteins and / or egg white proteins, for example in bakery products, pasta, savory systems, such as scrambled eggs, omelettes, mayonnaise, or dressings. In a preferred embodiment, the protein preparation serves as an equivalent or substitute for meat proteins, for example in nuggets, steaks, minced meat, burger patties, kebabs, or gyros. In a preferred embodiment, the protein preparation serves as an equivalent or substitute for milk protein, for example in milk, fermented beverages, dairy products, spoonable products, such as yoghurt, mousse, cream, cake cream, quark or ice cream. In a preferred embodiment, the protein preparation serves as an equivalent or substitute for gluten protein, for example in bakery or dough products, such as pasta. The protein preparation of the present invention may also be used in high protein systems, such as muesli, protein bars, bread, bakery or dough products. The protein preparation of the present invention may also be used in combination with other proteins, such as non-animal proteins, for example plant proteins, such as rice proteins, pea proteins, sunflower proteins, soybean proteins, hemp proteins, broad bean proteins, egg proteins or potato proteins, or animal proteins, such as meat proteins, fish proteins, insect proteins, egg proteins or milk proteins.In further embodiments, the protein preparation acts as a gelling agent, foaming agent, texturizing agent, binder, thickener, stabilizer, and / or emulsifier. Functional properties such as water binding capacity, gel forming capacity, powder solubility, oil binding capacity, emulsifying activity or stability, foaming activity or stability are described elsewhere herein and in the Examples. In a preferred embodiment, the protein preparation acts as a gelling agent, preferably having a water binding capacity of about 4 g / g or more, preferably about 5 g / g or more, more preferably about 6-7 g / g of dry weight of the protein preparation after heat treatment. In a further preferred embodiment, the protein preparation of the invention is used to prepare a food product or dietary supplement, which is a vegan or non-animal derived food product or dietary supplement. In a further preferred embodiment, the protein preparation of the invention is used to prepare a food product or dietary supplement, which is a food product and / or dietary supplement that does not have a rancid tase.

[0088] In a further aspect, the present invention provides a dietary supplement comprising the protein preparation of the present invention, hi one embodiment, the dietary supplement is in the form of a tablet, pill, powder, granules, or flakes.

[0089] In a further aspect, the present invention provides a food product comprising the protein preparation of the present invention or the dietary supplement of the present invention.In one embodiment, the food product is a meat substitute, an egg substitute, a fish substitute, an insect substitute or a dairy substitute, preferably a non-animal-based substitute.In another embodiment, the food product is a nugget, a burger patty, a kebab, a steak, a minced meat, a gyros, milk, a fermented drink, a dairy product, a spoonable product such as yogurt, a mousse, a cream, a cake cream, a quark or an ice cream; a bakery product, a dough product, a pasta, a savory product such as scrambled eggs, an omelette, a mayonnaise or a dressing; a muesli or a protein bar.

[0090] In a further aspect, the present invention provides a method for the preparation of a method for treating a pulmonary circulation comprising the steps of: a) providing a protein preparation according to the present invention; b) optionally mixing the protein preparation with one or more further ingredients or supplements of the product; and c) preparing a food product of the invention or a dietary supplement of the invention; The present invention provides a method for preparing a food product or dietary supplement product that does not taste rancid, comprising:

[0091] In this embodiment, the rancid taste of the food product or dietary supplement is reduced or absent because the protein preparation of the invention is devoid of lipids, preferably unsaturated lipids, such as oleic acid, palmitoleic acid, and linoleic acid. These unsaturated lipids are particularly susceptible to oxidation processes and therefore become rancid more quickly than saturated fatty acids.

[0092] In one embodiment, the further ingredients may include bakery, dairy, dough, egg or meat ingredients.

[0093] In a further aspect, the present invention provides a method for preparing a food product comprising the use of a protein preparation of the present invention, wherein the protein preparation is a) a gelling agent, preferably having gel-forming ability as described herein, b) a substitute for methylcellulose, c) a substitute for a vegetable protein, d) a substitute for a meat protein, e) a substitute for a gluten protein, f) a substitute for a milk protein, g) a substitute for a fish protein or f) a substitute for an egg protein, preferably a substitute for an egg yolk protein and / or a substitute for an egg white protein.

[0094] In a further aspect, the present invention provides a method for preparing a dietary supplement comprising the use of a protein preparation of the present invention, wherein the protein preparation is a) a gelling agent, preferably having gel-forming ability as described herein, or b) a substitute for methylcellulose.

[0095] In a further aspect, the present invention provides a method for the preparation of a method for treating a pulmonary circulation comprising the steps of: Furthermore, the present invention relates to a method for obtaining a natural protein of a microorganism, comprising the steps of: a) providing a microorganism and, optionally, subjecting said microorganism to one or more pretreatment steps; b) lysing the microorganism, thereby preparing a lysate comprising an aqueous liquid fraction containing nucleic acids and lysed native proteins of the microorganism; c) The following: i) adding to the aqueous liquid fraction a nucleic acid adsorbent immobilized on a solid support, preferably a free-floating solid support; ii) optionally stirring or shaking, preferably with an overhead shaker or agitator; and iii) separating the nucleic acids bound to the nucleic acid adsorbent immobilized on the solid support, preferably by precipitation and optionally by filtration, preferably dead-end filtration; separating the nucleic acids from the aqueous liquid fraction, thereby obtaining an aqueous liquid fraction, said aqueous liquid fraction being a nucleic acid-reduced aqueous liquid fraction; d) filtering the (nucleic acid-reduced) aqueous liquid fraction, thereby obtaining a solution containing purified dissolved native proteins of the microorganism and an aqueous solvent, preferably water or a physiological saline solution; and e) optionally removing at least a portion of the aqueous medium; f) optionally sterilizing the solution, thereby obtaining a sterile solution containing purified dissolved native proteins of the microorganism and an aqueous solvent; and g) optionally removing at least a portion of the aqueous medium of the sterilization solution of step f); The present invention provides a method for obtaining a natural protein of a microorganism comprising the steps of:

[0096] As far as the application of the particular embodiments disclosed herein in relation to the method of the present invention is concerned, in particular the particular embodiments of steps a), b), c), d), e), f) and g) relating to the separation of lipids disclosed herein are also applicable to this aspect of the present invention.

[0097] The type of microorganism used in the present invention is not particularly limited. In one embodiment, the microorganism is a eukaryotic microorganism. In a further embodiment, the microorganism is a eukaryotic microorganism selected from the group consisting of fungi, yeast, and algae. In a preferred embodiment, the microorganism is a fungus, preferably Aspergillus niger. In a further preferred embodiment, the microorganism is an alga, preferably an alga selected from the group consisting of Arthrospira maxima (Spirulina maxima), Arthrospira platensis (Spirulina platensis), Chlorella vulgaris, and Euglena gracilis. In a particularly preferred embodiment, the yeast is selected from the group consisting of Saccharomyces spp., preferably S. cerevisiae, S. carlsbergensis, S. bayanus, S. ellipsoides, S. uvarum, S. ludwigii, or S. pastorianus, Pichia spp., preferably P. pastoris; Hansenula spp.; Candida spp., preferably C. utilis; Torulopsis spp.; and Yarrowia lipolytica. In a most preferred embodiment, the microorganism is a yeast of the genus Saccharomyces, more preferably S. cerevisiae, S. pastorianus, or S. carlsbergensis. In another embodiment, the microorganism is a prokaryotic microorganism. In a preferred embodiment, the microorganism is a bacterium, preferably selected from the group consisting of Bacillus subtilis, Lactobacillus spp., Corynebacterium glutamicum, Methylomonas spp., Spirulina spp., and Xanthomonas spp. In one embodiment, the microorganism is a unicellular organism.

[0098] In a further embodiment, the method of the invention comprises a step b1) of clarifying the lysate, preferably by centrifugation. Alternatively, the method of the invention comprises a step c1) of clarifying the aqueous liquid fraction, preferably the nucleic acid-reduced aqueous liquid fraction, preferably by centrifugation. In a preferred embodiment, the separation of nucleic acids in step c) is performed after step b), i.e. from the lysate, or after b1), i.e. clarifying the lysate. In a further embodiment, the step c) of separating nucleic acids is performed after step d), i.e. after filtering the aqueous liquid fraction from the solution. In one embodiment, a nucleic acid sorbent is added to the lysate. In one embodiment, the stirring or shaking is performed by an overhead shaker. In one embodiment, a nucleic acid sorbent is added to the solution of the method of the invention. In one embodiment, the separation of nucleic acids bound to the nucleic acid sorbent immobilized on the solid support comprises filtering the nucleic acids bound to the nucleic acid sorbent immobilized on the solid support. In another embodiment, separation of nucleic acids bound to the nucleic acid adsorbent immobilized on the solid support comprises centrifugation of the nucleic acids bound to the nucleic acid adsorbent immobilized on the solid support, preferably at about 1000 g to 4000 g. In a preferred embodiment, separation of nucleic acids bound to the nucleic acid adsorbent immobilized on the solid support comprises precipitation of the nucleic acids bound to the nucleic acid adsorbent immobilized on the solid support, and d) optionally filtration, preferably by dead-end filtration.

[0099] For the purposes of the present invention, the separation of nucleic acids is carried out at a temperature of about 40° C. or less, about 30° C. or less, about 20° C. or less, or about 10° C. or less (to a temperature of about 2° C. to 8° C.), preferably about 30° C. or less, and / or does not involve the use of low or high pH conditions to avoid denaturation of proteins and loss of functional properties of proteins. In a preferred embodiment, steps b) to g), preferably steps b) to d), of the method are carried out at a temperature of about 40° C. or less, preferably about 30° C. or less, such as, for example, about 30° C. to 2° C., preferably about 30° C. to 8° C., more preferably about 30° C. to 20° C.

[0100] Nucleic acid separation can be performed in any volume, from laboratory scale to industrial scale. For example, chromatography may be performed in a volume of about 0.5 liter to 2 liter. Chromatography may also be performed in a volume of about 1 liter or more, 10 liter or more, 20 liter or more, 50 liter or more, 100 liter or more, 200 liter or more, 500 liter or more, 1000 liter or more, 2000 liter or more, 5000 liter or more, 7000 liter or more, or 10000 liter or more.

[0101] In a preferred embodiment, the separation of nucleic acids from the aqueous liquid fraction is carried out by chromatography, preferably anion exchange chromatography or anion exchange mixed mode chromatography. In anion exchange chromatography, a nucleic acid adsorbent is immobilized on a solid support, such as a ceramic or a resin, such as styrene-DVB. The nucleic acid adsorbent comprises a positively charged functional group, such as a quaternary ammonium compound. Nucleic acids with a negatively charged backbone can bind to the nucleic acid adsorbent, thereby separating the nucleic acid from the aqueous liquid fraction or solution. In anion exchange mixed mode chromatography, the solid support comprises a nucleic acid adsorbent with a positively charged functional group and a nucleic acid adsorbent with a further functional group having another type of interaction with nucleic acids. The functional group and the further functional group may be present in the same nucleic acid adsorbent. For example, a positively charged functional group can be combined with a functional group for hydrophobic interaction. An example for a mixed mode nucleic acid adsorbent is hydroxylapatite (Ca 5 (PO 4 ) 3 OH) 2 In a preferred embodiment, the solid support is a floating support (wherein the particles of the solid support move freely within a container), e.g., batch binding chromatography.

[0102] In a further preferred embodiment, the method of the invention further comprises a step of separating lipids from the aqueous liquid fraction as described herein in the context of the method of the invention. The specific embodiments of lipid separation disclosed herein are fully applicable to the method of separating nucleic acids. In this embodiment, the further step of separating lipids is carried out before the step c) of separating nucleic acids, i.e. from the lysate or the clarified lysate of step b1); after step c), i.e. from the nucleic acid-reduced aqueous liquid fraction containing the lysed native proteins of the microorganism or after the step c1) of clarifying the aqueous liquid fraction.

[0103] The present invention is further characterized by the following: 1. below: a) providing a microorganism and, optionally, subjecting said microorganism to one or more pretreatment steps; b) comprising a step b1) of lysing the microorganisms, thereby obtaining a lysate comprising an aqueous liquid fraction containing lipids and dissolved native proteins of the microorganisms, and further clarifying the lysate, preferably by centrifugation or filtration, c) separating lipids from the aqueous liquid fraction using mechanical means, thereby obtaining an aqueous liquid fraction that is a lipid-reduced aqueous liquid fraction that contains lysed native proteins of the microorganism; d) filtering the aqueous liquid fraction, thereby obtaining a solution containing purified dissolved native proteins of the microorganism and an aqueous solvent, preferably water or a physiological saline solution; e) optionally removing at least a portion of the aqueous medium; f) optionally sterilizing the solution, thereby obtaining a sterile solution containing purified dissolved native proteins of the microorganism and an aqueous solvent; and g) optionally removing at least a portion of the aqueous medium of the sterilization solution; 2. A method for preparing a natural microbial protein, comprising: 2. The method according to claim 1, wherein the separation of lipids from the aqueous liquid fraction using mechanical means in step c) is based on the different densities of the lipids and the lipid-reduced aqueous liquid fraction containing the lysed native proteins of the microorganism (i.e., wherein the separation of lipids is carried out using a three-phase centrifuge). 3. 3. The method according to claim 1 or 2, wherein the separation of lipids from the aqueous liquid fraction by mechanical means in step c) is carried out by a centrifuge (i.e. a three-phase centrifuge), preferably a skimming separator and / or a three-phase decanter. 4. The method according to any one of items 1 to 3, wherein the filtration in step d) is ultrafiltration, preferably diafiltration / ultrafiltration, having a molecular weight cut-off preferably in the range of about 1 kDa to about 100 kDa, preferably about 3 kDa to about 50 kDa, more preferably about 5 kDa to about 15 kDa, and most preferably about 10 kDa. 5. 5. The method according to any one of items 1 to 4, wherein the microorganism is a eukaryotic microorganism, preferably the eukaryotic microorganism is selected from the group consisting of a fungus, preferably Aspergillus niger, a yeast, and an alga, preferably Arthrospira maxima (Spirulina maxima), Arthrospira platensis (Spirulina platensis), Chlorella vulgaris or Euglena gracilis. 6. 6. The method according to any one of items 1 to 5, wherein the microorganism is selected from the group consisting of Saccharomyces spp., preferably S. cerevisiae, S. carlsbergensis, S. bayanus, S. ellipsoides, S. uvarum, S. ludwigii, or S. pastorianus, Pichia spp., preferably P. pastoris; Hansenula spp.; Candida spp., preferably C. utilis; Torulopsis spp.; and Yarrowia lipolytica. 7. 5. The method according to any one of items 1 to 4, wherein the microorganism is a prokaryotic microorganism, in particular a bacterium selected from the group consisting of Bacillus subtilis, Lactobacillus spp., Corynebacterium glutamicum, Methylomonas spp., Spirulina spp., and Xanthomonas spp. 8. 8. The method according to any one of items 1 to 7, wherein the microorganism is a unicellular organism. 9. 9. The method according to any one of items 1 to 8, wherein said lysis in step b) comprises mechanical lysis, preferably mechanical lysis including high pressure homogenization and / or bead milling. 10. 9. The method according to any one of items 1 to 8, wherein the lysis in step b) comprises physical lysis, preferably physical lysis including sonoporation and / or electroporation. 11. 11. The method according to any one of items 1 to 10, wherein the method comprises a step b1) of clarifying the lysate by centrifugation. 12. 11. The method according to any one of items 1 to 10, wherein the method further comprises a step c1) of clarifying the aqueous liquid fraction, preferably by centrifugation. 13. 13. The method according to any one of items 1 to 12, wherein steps b) to g), preferably steps b) to d), are carried out at a temperature of about 40° C. or less, preferably at a temperature within the range of about 30° C. to about 20° C. 14. 14. The method according to any one of items 1 to 13, wherein the one or more pretreatment steps in step a) are selected from the group consisting of filtering, sieving, washing, and centrifuging the microorganisms. 15. 15. The method according to any one of items 1 to 14, wherein the sterilization in step f) is carried out by sterile filtration. 16. 16. The method according to any one of items 1 to 15, wherein the removal of at least a portion of the solvent in step e) and / or step g) comprises removal of at least about 50%, preferably at least about 75%, more preferably at least about 90%, most preferably at least about 94% of the solvent. 17. 17. The method according to any one of items 1 to 16, wherein removing at least a portion of the solvent in step e) and / or step g) comprises spray drying, vacuum drying, drum drying, fluidized bed drying or freeze drying. 18. 18. The method according to any one of items 1 to 17, wherein the method comprises the further step of separating nucleic acids from the aqueous liquid fraction, preferably the lipid-reduced aqueous liquid fraction, comprising the lysed native proteins of the microorganisms of step c) or from the solution. 19. 19. The method of claim 18, wherein the separation of nucleic acids comprises chromatography, wherein the chromatography is anion exchange chromatography and / or anion exchange mixed mode chromatography. 20. 20. The method according to claim 19, wherein the chromatography comprises using a nucleic acid adsorbent immobilized on a solid support, preferably wherein the nucleic acid adsorbent comprises a quaternary ammonium compound as a functional group. twenty one. 21. The method according to item 19 or 20, wherein the chromatography comprises a) column chromatography, preferably in expanded bed adsorption mode, or b) batch binding chromatography. twenty two. 22. The method according to claim 21, wherein said chromatography performed in batch binding mode comprises: a) adding a nucleic acid adsorbent immobilized on a solid support, preferably a floating solid support; b) preferably stirring or shaking; and c) separating the nucleic acid bound to the nucleic acid adsorbent immobilized on the solid support, preferably by sedimentation and optionally by filtration. twenty three. 23. A protein preparation obtainable by the method according to any one of items 1 to 22, wherein preferably the protein preparation comprises, after heat treatment, preferably without syneresis, a gel-forming ability of about 1% to 10% of the protein preparation per total weight of a solution consisting of the protein preparation and water, and optionally the following: a) at least about 70% (w / w), preferably at least about 75% (w / w), more preferably at least about 85% (w / w), and most preferably at least about 95% (w / w) protein by dry weight of the protein preparation; b) less than about 110 mg / g, preferably less than about 50 mg / g, more preferably less than about 40 mg / g, even more preferably less than about 20 mg / g, or even more preferably less than 15 mg / g lipid, by dry weight of the protein preparation; c) a water binding capacity of about 4 g / g or more, preferably about 5 g / g or more, more preferably about 6.5 g / g of dry weight of the protein preparation after heat treatment; d) an oil-binding capacity of about 0.3 g / g or more, preferably about 0.5 g / g or more, more preferably 0.5 to 0.7 g / g of dry weight of the protein preparation after heat treatment; e) a powder solubility of about 74% or more, preferably about 80% or more, of the initial concentration of the protein preparation suspended in water, by total weight of the suspension; f) the ability to form a gel after heat treatment at about 2%, about 3%, about 4%, about 5%, or about 5.5% of the protein preparation based on the total weight of the solution consisting of the protein preparation and water, preferably without syneresis; g) an emulsifying activity of about 55% or more, preferably about 56% or more, more preferably about 57% or more, per total volume of emulsion consisting of 25 ml of sunflower oil and 25 ml of a solution of 5% protein preparation powder solution in water (w / v) after centrifugation, wherein said emulsifying activity is defined as emulsion layer (ml) / total volume (ml) x 100%; h) an emulsion stability of about 97% or more, preferably about 98% or more, more preferably about 100% or more per total volume of emulsion consisting of 25 ml of sunflower oil and 25 ml of a solution of 5% protein preparation powder solution in water (w / v) after exposing the emulsion to 80° C. for 30 minutes and subsequent centrifugation, wherein the emulsion stability is defined as emulsion layer (ml) / total volume (ml)×100%; i) a foaming capacity of about 40% or more, preferably about 48% or more, more preferably about 50% or more of foam volume per total volume of a dispersion of 100 mg of protein preparation in 10 ml of water, where the foaming capacity is defined as foam volume (ml) / total volume (ml) x 100%; j) a foam stability of about 20% or more, preferably about 40% or more, and more preferably about 75% or more foam volume per total volume of a dispersion of 100 mg of protein preparation in 10 ml of water after 60 minutes, where the foam stability is defined as foam volume (ml) measured after 60 minutes / initial volume (ml) x 100%; and / or k) less than about 10% (w / w), more preferably less than about 5.5% (w / w), and even more preferably less than 2.5% (w / w) nucleic acid by dry weight of the protein preparation; The protein preparation comprising: twenty four. 24. The protein preparation according to item 23, wherein the protein preparation is in dry form, preferably in powder form. twenty five. A protein preparation derived from a microorganism, preferably a unicellular microorganism, preferably comprising native proteins and, after heat treatment, preferably without syneresis, comprising a gel-forming ability at about 1% to 10% of the protein preparation per total weight of a solution consisting of the protein preparation and water, and optionally comprising: a) at least about 70% (w / w), preferably at least about 75% (w / w), more preferably at least about 85% (w / w), and most preferably at least about 95% (w / w) protein by dry weight of the protein preparation; b) less than about 110 mg / g, preferably less than about 50 mg / g, more preferably less than about 40 mg / g, even more preferably less than about 20 mg / g, or even more preferably less than 15 mg / g lipid, based on the dry weight of the protein preparation; c) a water binding capacity of about 4 g / g or more, preferably about 5 g / g or more, more preferably about 6.5 g / g or more of dry weight of the protein preparation after heat treatment; d) after heat treatment, an oil-binding capacity of about 0.3 g / g or more, preferably about 0.5 g / g or more, about 2 g / g or more, more preferably about 3 g / g or more, or about 0.3-4 g / g, about 0.5 g / g-4 g / g, about 0.5 g / g-3 g / g, about 0.5 g / g-2 g / g, or 0.5-0.7 g / g, based on the dry weight of the protein preparation; e) a powder solubility of about 74% or more, preferably about 80%, of the initial concentration of the protein preparation suspended in water, by total weight of the suspension; f) gel-forming ability, preferably without syneresis, after heat treatment at about 2%, about 3%, about 4%, about 5%, or about 5.5% of the protein preparation based on the total weight of the suspension consisting of the protein preparation and water; g) an emulsifying activity of about 54% or more, preferably about 55% or more, more preferably about 56% or more, per total volume of emulsion consisting of 25 ml of sunflower oil and 25 ml of a solution of 5% protein preparation powder solution in water (w / v) after centrifugation, wherein said emulsifying activity is defined as emulsion layer (ml) / total volume (ml) x 100%; h) an emulsion stability of about 97% or more, preferably about 98% or more, more preferably about 100% or more per total volume of emulsion consisting of 25 ml of sunflower oil and 25 ml of a solution of 5% protein preparation powder solution in water (w / v) after exposing the emulsion to 80° C. for 30 minutes and subsequent centrifugation, wherein the emulsion stability is defined as emulsion layer (ml) / total volume (ml)×100%; i) a foaming capacity of about 40% or more, preferably about 48% or more, more preferably about 50% or more of foam volume per total volume of a dispersion of 100 mg of protein preparation in 10 ml of water, where the foaming capacity is defined as foam volume (ml) / total volume (ml) x 100%; j) a foam stability of about 20% or more, preferably about 40% or more, and more preferably about 75% or more foam volume per total volume of a dispersion of 100 mg of protein preparation in 10 ml of water after 60 minutes, where the foam stability is defined as foam volume (ml) measured after 60 minutes / initial volume (ml) x 100%; and / or k) less than about 10% (w / w), more preferably less than about 5.5% (w / w), and even more preferably less than about 2.5% (w / w) nucleic acid by dry weight of the protein preparation; The protein preparation comprising: 26. 26. The protein preparation according to item 25, wherein the protein preparation is in dry form, preferably in powder form. 27. below: (a) providing a protein preparation according to any one of items 23 to 26; (b) mixing the protein preparation with an aqueous dispersion medium; and (c) heating the mixture to a temperature of at least about 55° C. to provide a protein gel; 1. A method for preparing a protein gel, comprising: 28. 27. Use of a protein preparation according to any of items 23 to 26 for preparing a food product, or a dietary supplement, preferably for human or animal use. 29. 29. Use of a protein preparation according to item 28, which serves as an equivalent or substitute for methylcellulose. 30. 30. Use of a protein preparation according to items 28 or 29 to serve as an equivalent or substitute for vegetable proteins, egg proteins, preferably egg yolk proteins and / or egg white proteins, meat proteins, gluten proteins, fish proteins, and / or milk proteins. 31. 31. Use of a protein preparation according to any one of items 28 to 30, which acts as a gelling agent, foaming agent, texturizing agent, binder, thickener, stabilizer and / or emulsifier. 32. 32. The use according to any one of items 28 to 31, wherein the food product or dietary supplement is a vegan or non-animal-derived alternative. 33. 27. Use of a protein preparation according to any one of items 23 to 26 for providing a food product and / or a dietary supplement that does not have a rancid tase. 34. 27. A dietary supplement comprising the protein preparation according to any one of items 23 to 26. 35. A food product comprising the protein preparation according to any one of items 23 to 26, or the dietary supplement according to item 34. 36. 36. The food product according to item 35, wherein the food product is a meat substitute, an egg substitute, a fish substitute, or a dairy substitute, preferably a non-animal based substitute. 37. 37. The food product according to item 35 or 36, wherein the food product is a product such as a pate, a nugget, a steak, a bakery product, a dough product, a spoonable product, a cereal product, a dairy product, a sausage product, a fish product or a minced meat product. 38. 35. The dietary supplement product according to item 34, wherein the dietary supplement is in the form of a tablet, pill, powder, granule, or flake. 39. below: a) providing a protein preparation according to any one of items 23 to 26, b) optionally mixing the protein preparation with one or more further ingredients or supplements of the product; and c) preparing food products or dietary supplements; 1. A method for preparing a food product or dietary supplement product that does not taste rancid, comprising: 40. 27. A method for preparing a food product comprising use of a protein preparation according to any one of items 23 to 26, wherein the protein preparation is a) a gelling agent, b) a substitute for methylcellulose, c) a substitute for vegetable protein, d) a substitute for meat protein, e) a substitute for gluten protein, f) a substitute for milk protein, g) a substitute for fish protein or f) a substitute for egg protein, preferably a substitute for egg yolk protein and / or a substitute for egg white protein. 41. 27. A method for preparing a dietary supplement comprising the use of a protein preparation according to any one of items 23 to 26, wherein the protein preparation is a) a gelling agent or b) a substitute for methylcellulose. 42. below: a) providing a microorganism and, optionally, subjecting said microorganism to one or more pretreatment steps; b) comprising a step b1) of lysing the microorganisms, thereby preparing a lysate comprising an aqueous liquid fraction containing the nucleic acids and lysed native proteins of the microorganism, and further clarifying the lysate, preferably by centrifugation or filtration, c) separating the nucleic acids from the aqueous liquid fraction, comprising chromatography, wherein the chromatography is anion exchange chromatography and / or anion mixed mode chromatography: i) adding to the aqueous liquid fraction a nucleic acid adsorbent comprising a positively charged functional group immobilized on a solid support, preferably a free-floating solid support; ii) optionally stirring or shaking, and iii) separating the nucleic acids bound to the nucleic acid adsorbent immobilized on the solid support, preferably by precipitation and optionally filtration; thereby obtaining an aqueous liquid fraction, said aqueous liquid fraction being a nucleic acid-reduced aqueous liquid fraction; d) filtering the aqueous liquid fraction, thereby obtaining a solution containing purified dissolved native proteins of the microorganism and an aqueous solvent, preferably water or a physiological saline solution; and e) optionally removing at least a portion of the aqueous medium; f) optionally sterilizing the solution, thereby obtaining a sterile solution containing purified dissolved native proteins of the microorganism and an aqueous solvent; and g) optionally removing at least a portion of the aqueous medium of the sterilization solution of step f); A method for obtaining a natural protein of a microorganism, comprising: 43. Item 43. The method according to item 42, wherein the nucleic acid adsorbent comprises a quaternary ammonium compound or a hydroxylapatite compound as a functional group. 44. 44. The method according to claim 42 or 43, further comprising separating lipids from the aqueous liquid fraction. 45. 45. The method according to any one of items 42 to 44, wherein the method further comprises a step b1) of clarifying the lysate by centrifugation. 46. The step c) further comprises the steps of: i) contacting the aqueous liquid fraction with a lipophilic solvent, preferably hexane, thereby obtaining a lipophilic phase; ii) measuring the absorbance of the lipophilic phase, preferably in the UV wavelength range; iii) comparing the absorbance measured in the lipophilic phase with a reference absorbance measured in lipids; and iv) quantifying lipids in the lipophilic phase; 23. The method according to any one of items 1 to 22, comprising quantifying lipids in the aqueous liquid fraction. List of Figures: [Brief description of the drawings]

[0104] [Figure 1] Decrease in absorbance maximum (A, C) and peak area (B, D) of hexane-extracted brewer's yeast cell lysate before and after use of a skimming separator, for the indicated peak and peak area (A and B) using the original peak setting, and for the more stable later peak setting (C and D). [Diagram 2](A) Comparison of water binding capacity [g / g] of proteins from pea, broad bean and sunflower with (right bar) or without (left bar) heat treatment with PD yeast protein of the invention, and (B) comparison of proteins from pea, broad bean, sunflower, rice and egg white with heat treatment with two different batches of baker's yeast protein and brewer's yeast protein of the invention. [Diagram 3] Maximum positive force (top) and positive area (bottom) for measurements using various protein concentrations on the Texture Analyser system. [Figure 4] Comparison of percent powder solubility of various proteins with the PD yeast protein of the present invention. [Diagram 5] Comparison of emulsifying activity of various proteins with the PD yeast protein of the invention. [Figure 6] Comparison of foaming capacity [%] (left bar graph) and foam stability [%] after 60 min (right bar graph) of various proteins with the PD yeast protein of the invention. [Figure 7] Comparison of storage stability of protein powders with and without skimming separator treatment, assessed by taste rating scale 1-6. Samples were stored as powders (A) at 23°C (<75% humidity) protected from light, or (B) under accelerated storage conditions at 60°C, for up to 182 days. [Figure 8]Comparison of lipid content in samples tested with and without skimming and / or diafiltration treatment. (A) Lipid soluble components detected in the hexane phase are shown as absorbance at the indicated wavelengths, (B) Lipid soluble components detected in the hexane phase are shown as peak areas, and (C) Total fat [g / 100g]. Untreated CF / DF (first bar): Clarifying centrifugation followed by CF / DF and spray drying (no skimming); Skimming treated CF / DF (second bar): Clarifying centrifugation followed by skimming, CF / DF, and spray drying: Untreated (third bar): Clarifying centrifugation and spray drying (no skimming and CF / DF); Skimming treated (fourth bar): Clarifying centrifugation followed by skimming and spray drying. [Figure 9] Comparison of lipid content in samples with and without skimming separator treatment, including fatty acids, showing (A) total fat [g / 100g] separator, (B) fatty acid subgroups indicated, and (C and D) fat-soluble components detected in the hexane phase by (C) absorbance and peak area (D). [Figure 10] Comparison of lipid content in tested samples with and without skimming separator treatment. Reduction [%] of total fat and fatty acid subgroups measured using gas chromatography. [Figure 11] On the left is a muffin made using the PD protein of the present invention (recipe, vegan muffin), and on the right is a conventional muffin.

[0105] The present invention is further illustrated by the following examples. EXAMPLES

[0106] Example 1 - Production of a functional brewer's yeast protein preparation (Saccharomyces spp.) Raw materials: Yeast biomass (Saccharomyces spp.) was obtained from Kaiser Brauerei GmbH, Geislingen an der Steige, DE.

[0107] Hop filtration / sieving: 4 liters of Brewer's yeast (Saccharomyces carrusbergensis) TS 15% (w / w), pH 5.3, stored as yeast cake as delivered by the brewery, was sieved using a vibrating sieve or a filter bag with a mesh size of 125 μm (120 US Mesh) to remove residual hops.

[0108] Debittering process: The yeast cake was separated from the sieved cell suspension (9-15% w / w) by a centrifuge / separator (3000g, 5 min, 4°C). The resulting beer-free cell mass was then transferred to warm debittering solution (0.5% polysorbate 80, 0.2% NaOH, pH 9.1) 1:2 (w / w) at 37°C and incubated for 10 min (possible range approx. 10-120 min). The spent debittering solution was then removed by centrifugation and the debittered cell mass was washed with water. The washing process was repeated until the pH of the cell suspension reached a pH of 5.7-6.5 (pH 6.4).

[0109] Cell disruption: The cell suspension was adjusted to 12-14% (w / w) dry mass. The cells were then lysed using a Dyno®-Mill Research Lab of Willy A. Bachofer AG (Muttenz, CH) ball mill (glass beads 0.5 mm, 70% loading, circulation mode: 2.5 L, circulation 45 min; 3.500 rpm) at 4°C-8°C. The effect of cell disruption was measured by microscopic contrast (phase contrast) and protein content (Pierce™ BCA Protein Assay Kit, Thermo Scientific), measured in the supernatant after centrifugation. The protein content at 95% cell disruption was approximately 55 mg ml -1 It was.

[0110] Isolation of the soluble protein fraction: The lysed cell suspension was centrifuged at 17,000 g and 4° C. for 20 min using an Avanti J20 XP Beckman Coulter (Brea, US) to separate the supernatant from the yeast cell walls. The resulting supernatant has the following characteristics:

[0111] Reduction of lipid fraction: The soluble protein fraction was centrifuged using a skimming separator (10500 rpm, bowl diameter 365 mm, 12 disks) and a feed rate of about 1 L / min. The effectiveness of lipid separation was measured by UV spectroscopy (200-350 nm). The liposoluble components detected in the hexane phase were reduced by 50-60% (see Figures 1A and B). In the later examples, the measurement of the absorption maximum at 222 nm and the corresponding peak areas are replaced by the absorption maximum at 260 nm (measured peaks: 260 nm, 271 nm, 282 nm and 294 nm; integrated peak areas: 244-265 nm, 265-276 nm, 276-289 nm and 289-310 nm), because this peak was found to give more stable results. The results for the same samples measured using the new settings are shown in Figures 1C and D. In separate experiments, a reduction in the total lipid (fat) content of about 20% was observed using the method for determining the total fat content of cereal products after acid hydrolysis by extraction and gravimetric determination (according to §64 LFGB L16.00-5: 2017-10). The instruction §64 LFGB L16.00-5: 2017-10 describes a method that is carried out according to DIN standards by the DAkks accredited laboratory (holding a certificate of approval from the Deutsche Akkreditierungsstelle). The method can be found in the BVL method collection for food products, e.g. at Beuth Verlag GmbH.

[0112] Reduction of nucleic acid content: Chromatographic material was diluted to 80 mg ml -1 A chromatographic material concentration of 100 ml was added to 150 ml of lipid-reduced supernatant after appropriate pretreatment (equilibration). The supernatant was then incubated with shaking (level 6, 40 rpm for 30 min) in an overhead shaker (STR4 Haberle LABORTECHNIK GmbH+Co.KG). After binding, the chromatographic material was separated from the sample by centrifugation (3345 g, 18° C., 1 min).

[0113] Taste Optimization with Ultrafiltration / Diafiltration (UF / DF): The lipid-reduced and nucleic acid-reduced supernatant was diafiltered (DF=0.667-2 filtration unit Akta Flux S Cytiva) and filtered using a hydrophilic membrane (MWCO 10 kDa, filter area 0.02 m 2 ) at 12°C. 2 O was added as the diafiltration buffer. After diafiltration (DF=0.667-2) was completed, the supernatant was concentrated three-fold (CF=3). Table 1 - Characteristics of Brewer's Yeast Protein Concentrate [Table 1]

[0114] Drying: The brewer's yeast protein concentrate was dried by means of a spray dryer (Spray Dryer B-290 from Buchi Labortechnik GmbH) using an input temperature of 180-155°C and a resulting output temperature of 75°C-80°C.

[0115] Storage / sterile filtration: Sterile filtration of brewer's yeast protein concentrate was carried out in a separate experiment by separating unwanted particles using a heterogeneous PES bilayer + glass fiber membrane (0.8 μm + 0.2 μm). Example 2 - Water binding capacity of protein preparations method: 0.5 g of sample (powder from example 1) was mixed with 4 mL of demineralized water for 30 seconds. For samples with higher water absorption, the amount of water was increased to 5 mL. The mixture was shaken with a test tube vibrator for 20 seconds. The shaking was repeated 7 times at 10 minute intervals. Measurement after heat treatment: The mixture was heated to 80°C, kept at 80°C for 10 minutes, and then cooled to room temperature. The mixture was centrifuged at 2000g for 25 minutes at 20°C. The supernatant was decanted. To drip off any remaining water: The test tube was placed at a 20° angle for 10 minutes. ·Calculation

number

[0116] The results are further illustrated in Figure 2A. Based on the high solubility of the PD yeast protein of the present invention, it is clear that the water binding capacity cannot be measured without heat treatment. After heat treatment, a very high water binding capacity of 6-6.9 g / g was found. The method also provided information about the native state of the protein. Table 3 - Comparison of water binding ability of various proteins of the invention (baker's yeast and brewer's yeast) with various proteins [Table 3]

[0117] The results are further illustrated in FIG. 2B. The baker's yeast proteins of the invention were present in a protein concentration range of 50-65% and the brewer's yeast proteins of the invention were in a protein concentration range of 70-80% protein. The vegetable and egg white proteins were at a protein content of 80% concentrate, except for the broad bean and sunflower proteins (60%). In all cases it could be demonstrated that the water binding capacity of the PD proteins of the invention was greater than 4.5 g / g. The highest value was detected for the brewer's yeast proteins, about 7 g / g, compared to 2.2 g / g-3.8 g / g for the conventional vegetable proteins. The results confirm that the proteins of the invention have an improved water binding capacity compared to the conventional vegetable proteins. In particular, the value of the proteins of the invention was closer to that of the egg white proteins of 9.6 g / g, confirming that the proteins of the invention are particularly useful as egg substitutes or equivalents. Example 3 - Gel-forming ability of protein preparations Method 1: Yeast protein (powder of brewer's yeast protein 1 from Example 1) was dispersed in water. The dispersion was stirred for 20 minutes. The dispersion was spread in a 15 ml Sarstedt tube. The dispersion was treated in a water bath at 80°C for 20 minutes. · Rating: Coagulated: No water is lost when the tube is turned upside down; Noncoagulated: Water is lost. Table 4 - Results for PD Brewer's Yeast Protein 1 [Table 4] Method 2: A 5% dispersion of the protein preparation in water was prepared. The dispersion was stirred for 20 minutes. The dispersion was spread in a 15 ml Sarstedt tube. The dispersion was treated in a water bath at 80°C for 20 minutes. Rating: visual rating using scores 0-5; 0=poor to 5=excellent. Table 5 - Results [Table 5]

[0118] It was demonstrated that the protein preparation of the present invention, containing native, functional proteins, formed excellent gels at a protein concentration of 5% after heat treatment, comparable to gels obtained with conventional egg white protein at the same concentration. In contrast, a yeast protein preparation containing non-functional proteins at a protein concentration of 80% (yeast protein 1 in Table 5, Proteissimo, Lesaffre) did not show gel-forming ability. Method 3: Measurement with a texture analyzer

[0119] Various protein solutions (100 mL) were prepared by dissolving the powder in water and the solution was stirred on a stir plate at room temperature. Foam formation must be avoided. Six solutions with concentrations of 2.5%, 5%, 7.5%, 10%, 12.5% ​​and 15% (w / w%) were prepared. After complete dissolution of the powder, the pH was adjusted to pH 7, stirred for 10 min and measured / adjusted again. Approximately 30 mL of protein solution was filled into each tube (height: 3.6 cm, diameter 3.3 cm, volume: 30 mL) such that the tube was filled to the top and foam formation was avoided. Four tubes were filled for each concentration for triplicate measurements and additional temperature standard samples. The tubes were placed in a water bath at 90° C. for 15 min and completely submerged, cooled to room temperature and stored in the refrigerator (4° C.) overnight.

[0120] The gels were measured by compression test on the texture analyzer in tubes at a core temperature of 20° C. The core temperature was measured using a thermometer in an additional temperature standard sample.

[0121] For the compression test, 5 kg of cells were loaded into a texture analyzer (Stable Micro Systems; Texture Analyser Model XT2i HR) and a probe with a diameter of 1.1 cm (series number: SMS P / 1KS; area 1 cm^2) was used. Measurements were performed with the following test settings: test speed: 1.00 mm / sec; post test speed: 10.00 mm / sec; target mode: distance; distance: 14.000 mm; trigger type: button; stop plot at start position; no temperature detection. Before starting the measurement, the probe must be carefully placed on the gel surface.

[0122] Results: In one batch, gel strength was measured as a function of protein concentration. As shown in Figure 3, gel strength (maximum positive force) is dependent on protein concentration. Measurable gels were produced at protein concentrations >2%.

[0123] Measurement of another batch using the above method at 20° C. gave a gel strength of 0.959 N (standard deviation 0.017 N). Example 4 - Oil binding capacity of protein preparations

[0124] The same method as in Example 2 was used, except that the demineralized water for preparing the 5% protein preparation (powder of PD Brewer's Yeast Protein 2 in Example 1) was replaced by sunflower oil (other vegetable oils such as rapeseed can also be used) and the oil binding capacity was calculated by (weight of oil-wet sample-weight of test tube-mass of protein) / mass of protein. The oil binding capacity of the protein of the present invention was found to be 0.5-0.7 g / g. Example 5 - Powder solubility of protein preparations: method: Source: U.S. Patent, Patent Number: 4,465,702 (Eastman et al.): · In a 100 mL beaker, a solution was prepared by adding protein (2%) to 50 mL of demineralized water. The solution was stirred with a magnetic stirrer (800 rpm) for 60 min. · Samples were transferred to 50mL centrifuge tubes. · Samples were centrifuged for 25 minutes at 2000g and 20°C. 25 mL of the supernatant was transferred into an aluminum shell. The samples were placed in a dry oven at 160°C for 1 hour and 40 minutes. The aluminum shell was allowed to cool in a desiccator. The aluminum shell containing the dried contents was weighed. ·Calculation ·Solubility

number

[0125] Figure 4 shows a comparison of the percent powder solubility of various proteins in comparison with the PD yeast proteins of the invention. The baker's yeast proteins of the invention were in the protein concentration range of 50-65% and the brewer's yeast proteins of the invention were in the protein concentration range of 70-80%. The vegetable proteins were 80% protein concentrates, except for broad bean and sunflower proteins (60%) and potato protein (20%). In all cases it could be demonstrated that the solubility of the PD proteins of the invention was more than 75% with respect to the initial amount of protein used in the samples. The highest value of the proteins of the invention was detected for the baker's yeast proteins with about 82%. The egg white protein had a powder solubility of 87.1%. Example 6 - Emulsifying properties of protein preparations

[0126] Emulsifying properties were measured using turbidity measurements by Elif Ezgi Ozdemir, Ahmet Gorguc, Esra Gencdag, Fatih Mehmet Yilmaz, “Physicochemical, functional and emulsifying properties of plant protein powder from industrial sesame processing waste as affected by spray and freeze drying”, LWT Food Science and Technology, 154 (2022) 112646. Method 1: Emulsifying activity · 25ml of sunflower oil was added to a 5% solution of protein powder in 25ml of water. Homogenization with IKA T18 at 11000 rpm for 30 seconds. The emulsion was then immediately centrifuged at 1200 g for 5 minutes. The amount of emulsion layer and the total volume were recorded. Emulsifying activity (EA) [%] = emulsion layer (mL) / total volume (mL) x 100 Method 2: Emulsion stability · 25ml of sunflower oil was added to a 5% solution of protein powder in 25ml of water. Homogenization with IKA T18 at 11000 rpm for 30 seconds. The emulsion was then immediately centrifuged at 1200 g for 5 minutes. The emulsion was kept in a water bath at 80°C for 30 minutes and then cooled rapidly. The samples were centrifuged at 1200g for 5 minutes. Emulsion stability (ES) [%] = remaining emulsion layer (ml) / total volume (ml) x 100 Table 7 - Results [Table 7]

[0127] The results are illustrated in Figure 5 and show a comparison of the emulsifying activity of various proteins in comparison with the PD yeast protein of the invention. The baker's yeast proteins of the invention were in the concentration range of 50-65% and the brewer's yeast proteins of the invention were in the range of 70-80% protein. The vegetable proteins had a concentrated protein content of 80%, except for the broad bean and sunflower proteins (60%) and the potato protein (20%). In all cases it could be demonstrated that the emulsifying activity of the PD proteins of the invention was greater than 56%. The emulsifying stability in this test for the PD proteins of the invention was 100%. Example 7 - Foaming properties of protein preparations

[0128] Foaming properties were measured using turbidity measurements by Elif Ezgi Ozdemir, Ahmet Gorguc, Esra Gencdag, Fatih Mehmet Yilmaz, “Physicochemical, functional and emulsifying properties of plant protein powder from industrial sesame processing waste as affected by spray and freeze drying”, LWT Food Science and Technology, 154 (2022) 112646. method: · 20ml of distilled water was added to 200mg of protein powder. Homogenization for 30 seconds at 11,000 rpm, e.g. with an IKA T18. The foam was transferred to a measuring cylinder. After 30 seconds the foam volume was recorded. Foaming capacity [%] was calculated as foam volume (ml) / total volume of mixture × 100. The foam was allowed to stand for an additional 60 minutes to determine foam stability. The foam volume was measured after 60 minutes. The foam stability [%] was calculated as the foam volume after 60 minutes / initial foam volume. Table 8 - Results [Table 8]

[0129] The results are also shown in Figure 6. It could be shown that the PD proteins of the invention from different origins can be widely tuned with regard to their foaming properties. In particular, the foam stability can be tuned. Example 8 - Effect of lipid reduction on storage stability of protein preparations

[0130] The storage stability of protein powders can be compromised by factors such as lipid oxidation, which leads to changes in taste and reduced functionality. One possible way to improve the long-term stability of protein powders is to reduce the lipid content in them.

[0131] Two different powders were made essentially as described in Example 1 with minor modifications, where a portion of the protein solution was treated with a skimming separator (10500 rpm, bowl diameter 365 mm, 12 disks) to reduce the lipid content and the other portion was left untreated.

[0132] Briefly, 20 liters of brewery-derived TS 15% (w / w), pH 5.3, stored as yeast cake and delivered by the brewery, was sieved using a vibrating sieve or filter bags with mesh sizes between 125 μm and 50 μm (270 US Mesh to 120 US Mesh) to remove residual hops.

[0133] The yeast meal was adjusted to 7-15% w / w dry mass and subjected to a debittering process essentially as described in Example 1, with minor modifications.

[0134] The cell suspension was adjusted to 12-14% (w / w) dry mass. The cells were then lysed using a LabStar Discus Mill NETZSCH-Feinmahltechnik GmbH (70% loading, pass-through mode). The effectiveness of cell disruption was measured by microscopic contrast (phase contrast) and protein content (Pierce™ BCA Protein Assay Kit, Thermo Scientific), measured in the supernatant after centrifugation. The protein content at 95% cell disruption is approximately 55 mg ml -1 It was.

[0135] The lysed cell suspension was centrifuged at 17,000 g and 4° C. for 20 min using an Avanti J20 XP Beckman Coulter (Brea, US) to separate the supernatant from the yeast cell walls.

[0136] The resulting supernatant was divided into two parts: one part of the supernatant was treated with a skimming separator after centrifugation (Figure 7 = skimming separator treatment (10500 rpm, bowl diameter 365 mm, 12 disks, feed rate of about 1 L / min) and was thereby subjected to lipid reduction (separator skimming treatment), whereas the other part was processed as it was after centrifugation without lipid reduction (Figure 7 = untreated).

[0137] Taste optimization by diafiltration (UF / DF): Both unprocessed and skimming separator processed samples were filtered at 12°C with a hydrophilic membrane (MWCO 10 kDa) and H2O as diafiltration buffer. 2 The supernatant was then subjected to diafiltration (DF=0.667-2 SW18 HFC filtration unit-MMS AG) using O. After diafiltration (DF=0.667-2) was completed, the supernatant was concentrated three-fold (CF=3).

[0138] Drying: Both the unprocessed and skimming separator processed samples, as well as the concentrated samples, were dried by a spray dryer (Spray Dryer B-290 from Buchi Labortechnik GmbH) using an input temperature of 180-155°C and a resulting output temperature of 75°C-80°C.

[0139] Two different powders (skimmed and untreated) were stored under defined storage conditions for a duration of one month, including a temperature of 23°C, a relative humidity of less than 75%, and protection from light (see Figure 7A). Additionally, accelerated storage studies were conducted to evaluate the long-term stability of two different powder samples (skimmed and untreated). Two target shelf-life periods of 91 and 182 days were selected, and a temperature of 60°C (TAA = accelerated aging temperature) was applied with a Q10 of 2, whereas real-time ambient conditions (TRT) were maintained at 23°C (Figure 7B). Accelerated aging was calculated by determining the accelerated aging time (AAT) using the formula AAT = desired real-time (RT) Q10 [(TAA-TRT) / 10].

[0140] Finally, the sensory properties of the different powder samples during storage were evaluated by a group of five independent individuals in a blinded manner who rated their assessment of the taste changes in the powder samples on a scale of 1 (very good, no rancidity) to 6 (very bad, very strong rancidity).

[0141] The data show that lipid reduction using skimming separation significantly improves the long term stability of the protein powder and results in a large reduction in taste change during storage. Example 9 - Importance of skimming separation in protein solution processing

[0142] As part of Example 8, the following further experiments were carried out:

[0143] After lysing the yeast cell suspension, the suspension was centrifuged at 17,000 g and 4° C. for 20 min using an Avanti J20 XP Beckman Coulter (Brea, US) to separate the supernatant from the yeast cell walls. The resulting supernatant was divided into four parts. Two parts of the supernatant were processed in a skimming separator (10500 rpm, drum diameter 365 mm, 12 pieces of disks, flow rate of about 1 L / min) after centrifugation of the cell walls, thereby subjecting them to lipid reduction (skimming separator processing), whereas the other two parts were not subjected to lipid reduction after centrifugation of the cell walls.

[0144] One of the untreated and skimming separator-treated samples was dried to a powder without diafiltration (UF / DF), and the other sample was filtered through a hydrophilic membrane (MWCO 10 kDa, filtration area 0.16 m) at 12 °C. 2) and H as the diafiltration buffer. 2 The filtrate was filtered and concentrated using a SW18 HFC UF / DF system type MMS AG using O (DF=0.667-2, concentrated three times (CF=3)). These two samples were then also dried.

[0145] All four powders were analyzed by UV spectroscopy (200-350 nm) and the total lipid (fat) content was measured gravimetrically according to Weibull-Stoldt ASU L06.00-6 (2014-08). The analysis of the total lipid (fat) content was carried out by an external DAkks accredited (holding a certificate of accreditation from the Deutsche Akkreditierungsstelle) analytical laboratory. The analyzed samples were simplified as follows: Unprocessed: without skimming separator treatment, centrifuged at 17000 g and 4° C. for 20 min and dried using spray drying. Untreated CF / DF: Centrifuged at 17000g and 4°C for 20 min without skimming separator treatment, and filtered through a hydrophilic membrane (MWCO 10 kDa, filtration area 0.16 m 2The extract was filtered and concentrated using a SW18 HFC UF / DF system type MMS AG using 100% ethanol, and then dried using a spray dryer. Skimming separator treatment: followed by skimming separator treatment, centrifuging at 17000 g and 4° C. for 20 minutes, and then drying using a spray dryer. Skimming separator-treated CF / DF: Skimming separator-treated CF / DF was centrifuged at 17,000 g and 4° C. for 20 min, and then filtered through a hydrophilic membrane (MWCO 10 kDa, filtration area 0.16 m 2 The extract was filtered and concentrated using a SW18 HFC UF / DF system type MMS AG using 100% ethanol, and then dried using a spray dryer.

[0146] In this example, the fat-soluble components detected in the hexane phase were reduced by 25-30% (Figures 8A and B), which corresponds to a total fat reduction of 10-11% (Figure 8C). During UF-DF / CF, both the fat-soluble components detected in the hexane phase and the total fat reduced by the skimming separator were concentrated by filtration. Since the reduction profile of the fat-soluble components detected in the hexane phase and the total fat reduction of the non-concentrated sample were similar to the non-concentrated sample (the fat-soluble components detected in the hexane phase were reduced by 25-30%, and the total fat is 10-11%), it indicates that the fat reduction is caused by the skimming separation and that the filtration does not affect the fat reduction (Figures 8A and C). We note that the detected lipid reduction had a strong impact on the taste perception, especially after storage of the powder, as shown in Figure 7. Moreover, in other experiments, the lipid reduction was higher, and even higher lipid reduction is expected since industrial skimming separators were used and these can be performed more precisely. Example 10 - Lipid Reduction of Protein Solutions Using a Skimming Separator (UV Spectometry, Gravimetry, and Gas Chromatography)

[0147] Example 9 was repeated in an independent experiment, and samples were taken from the solution treated with the skimming separator to reduce the lipid content and from the portion left untreated. The lipid reduction achieved by the skimming separator compared to the untreated solution was measured by UV spectroscopy (200-350 nm), and the total lipid (fat) content was measured by gravimetric method according to Weibull-Stoldt ASU L06.00-6 (2014-08), and the fatty acid content was measured by gas chromatography (ISO 12966-2:2011 mod., GC / FID.).

[0148] In this example, the fat-soluble components detected in the hexane phase were reduced by 50-55% (Figures 9C and D), which corresponds to a total fat reduction of 29% (Figures 9A and B) and a specific fatty acid reduction of 22%-36% (Figure 9B). Another independent experiment demonstrated a 32% removal of the total fat content and a specific fatty acid reduction of 22%-39% (Figure 10). In particular, polyunsaturated fatty acids, which tend to oxidize more rapidly and thereby become rancid, were significantly reduced by up to 36%-39% by using the skimming separator (see Figure 10), which improves the shelf life and prevents the sensory off-flavor (rancidity) of the yeast preparation, as already demonstrated in experiment 7. In this example, the fat content (lipid content) of the light liquid phase removed by the skimming separator was also analyzed. The total lipid (fat) content in the removed light liquid phase was found to be approximately 260% higher than the heavy liquid phase (fat-reduced phase). Example 11 - Preparation of muffins using protein preparations

[0149] Muffins were prepared according to the following recipe: Table 9 [Table 9] Table 10 - Results [Table 10]

[0150] Figure 11 shows a comparison of a vegan muffin of the invention (left) and a conventional muffin (right), showing that most can be produced on the basis of the PD protein of the invention leading to sponge cakes (e.g. muffins) that are comparable or better than their egg-based alternatives in terms of taste, juiciness, texture (including shelf stability), pore structure and volume. Example 12 - Preparation of angelica cake with protein preparations

[0151] Angel cake is one of the best-known food models for simultaneously testing foaming and gelling of food proteins. The height, texture, and compressibility of the cake appear to be related to four basic characteristics: viscosity, foaming capacity (FC), foam stability (FS), and gelling (Kneifel, W. and Seiler, A. (1993) "Water-holding Properties of Milk Protein Products - A Review," Food Structure: Vol. 12: No. 3, Article 3).

[0152] Angelica cake was prepared according to the following recipe: Table 11 [Table 11] Recipe source: Kneifel et al.

[0153] Preparation of angelica cake: Protein dispersion in water or egg whites are whipped to form a thick foam. Sucrose is added. Then flour is added to make a cake batter, which is baked at 88°C for 30 minutes. It could be shown that the protein of the present invention (PD protein) can replace eggs. Example 13 - Preparation of scrambled eggs with protein preparations

[0154] Scrambled eggs were prepared according to the following recipe: Table 12 [Table 12] · Mixed the dry ingredients and added mustard and tomato paste to the water. The mass was mixed in a blender and the oil was gradually added. The chunks were deep fried in a pan with oil, allowed to set, and then the scrambled eggs were broken up using a silicone scraper. Result: the scrambled eggs of the present invention have a similar consistency to scrambled eggs made with eggs. Therefore, the protein preparation of the present invention is suitable as an egg substitute. Example 14 - Preparation of burger patties with protein preparations

[0155] Method 1: I used the following recipe: Table 13 [Table 13] Preparation: Starch was dispersed in 25% water and heated. · The fabric was soaked in 50% water for at least an hour. The protein was dispersed and, if necessary, hydrocolloidized using an additional 25% water. For methylcellulose (EMC): Methylcellulose, oil and a small amount of water were dispersed, followed by addition of the protein dispersion and cooling the mass (4°C overnight). The chunks were formed into patties and fried. Method 2: Our recipe (Table 14) [Table 14] The preparation was carried out according to the following protocol: · Mix all the "seasoning ingredients" with half a cup of water and soak the dough in it until it becomes soft (at least 45 minutes). · Dissolve or disperse the PD protein in the remaining half of the water. Heat the other half of the remaining water and dissolve the starch in it (viscous starch slurry). Mix all ingredients from steps 1-3 together with the oil and flour to form a thick paste. Shape and deep fry in oil over medium heat. Method 3: Based on the recipe of Method 2, different concentrations of yeast protein of the present invention, pea protein, and methylcellulose were used. Table 15 [Table 15] result: Method 1: The patties containing methylcellulose (EMC) held together when fried. The patties using the pea protein based recipe (E1) easily fell apart when fried. The chunks were easily formed. The patties based on PD2 and 3 were easily formed and stable when fried (comparable to EMC). The results confirm that the proteins of the invention can be used as substitutes for methylcellulose. Method 2 is the adapted recipe with spices (the final adapted recipe). Method 3: Testing has shown that a concentration of 2-3% provided comparable or even better results than 2% methylcellulose in the basic recipes evaluated at that time. The yeast proteins of the present invention can be used as partial substitutes for other proteins such as pea protein at lower concentrations. Example 15 - Protein preparations as dairy substitutes (examples of dairy substitutes) Table 16: Materials [Table 16]

[0156] The preparation was carried out according to the following protocol: 1. Mix the dry ingredients together and add the mustard and tomato paste to the water. 2. Mix ingredients from step 1, melt fat and gradually add. 3. Heat gently in pot at lowest level. 4. Whisk and cool. The protein content can also be increased.

Claims

1. below: a) Prepare a microorganism, and optionally subject the microorganism to one or more pretreatment steps, b) The process includes step b1), which involves lysing the microorganisms to obtain a lysate containing an aqueous liquid fraction with respect to the lipids and dissolved natural proteins of the microorganisms, and further purifying the lysate, preferably by centrifugation or filtration. c) Separating lipids from the aqueous liquid fraction using mechanical means, thereby obtaining an aqueous liquid fraction (which is a lipid-reduced aqueous liquid fraction containing dissolved natural proteins of microorganisms), where the separation of lipids is carried out by a three-phase centrifuge. d) Filter the aqueous liquid fraction to obtain a solution containing purified and dissolved natural proteins from microorganisms and an aqueous solvent, preferably water or physiological saline solution. e) Optionally, remove at least a portion of the aqueous solvent, f) Optionally, sterilize the solution to obtain a sterile solution containing purified microbial lysates, natural proteins, and an aqueous solvent, and then, g) Optionally, remove at least a portion of the aqueous solvent from the sterile solution. A method for preparing natural proteins from microorganisms, including [specific components].

2. The method according to claim 1, wherein the separation of lipids from the aqueous liquid fraction using mechanical means in step c) above is carried out by a skimming separator and / or a three-phase decanter.

3. The method according to claim 1, wherein the filtration in step d) is more preferably ultrafiltration, preferably dialysis filtration / ultrafiltration, having a fractional molecular weight in the range of about 1 kDa to about 100 kDa, preferably about 3 kDa to about 50 kDa, more preferably about 5 kDa to about 15 kDa, and most preferably about 10 kDa.

4. The method according to claim 1, wherein the microorganism is a eukaryotic microorganism, preferably a eukaryotic microorganism selected from the group consisting of fungi, preferably Aspergillus niger; preferably a species of the genus Saccharomyces, preferably S. cerevisiae, S. carlusbergensis, S. bayanus, S. ellipsoides, S. ubarum, S. ludwigii, or S. pastorianus; a species of the genus Pichia, preferably P. pastris; a species of Hansenula; a species of Candida, preferably C. uchilis; a species of Torlopsis; and a yeast selected from the group consisting of Yarowia liporitica; and an algae, preferably selected from the group consisting of Arthrospira maxima (Spirulina maxima), Arthrospira pratensis (Spirulina pratensis), Chlorella vulgaris, or Euglena gracilis.

5. The method according to claim 1, wherein the microorganism is a prokaryotic microorganism, and in particular a bacterium selected from the group consisting of Bacillus subtilis, Lactobacillus species, Corynebacterium glutamicum, Methylomonas species, Spirulina subspecies, and Xanthomonas species.

6. The method according to claim 1, wherein the method comprises the step of purifying the dissolved substance in step b1) by centrifugal separation.

7. The method according to claim 1, wherein steps b) to g), preferably steps b) to d), are carried out at a temperature of about 40°C or lower, preferably in the range of about 30°C to 20°C.

8. The method according to claim 1, wherein the method comprises a further step of separating nucleic acids from an aqueous liquid fraction, preferably a lipid-reduced aqueous liquid fraction, containing the lysed intrinsic proteins of the microorganisms of step c), or from the solution, wherein the step of separating the nucleic acids comprises anion exchange chromatography and / or anion mixed-mode chromatography.

9. A protein preparation that can be obtained by the method of any one of claims 1 to 8, wherein, preferably, the protein preparation contains gel-forming ability of about 1% to 10% of the protein preparation by total weight of the solution consisting of the protein preparation and water after heat treatment, preferably without syneresis, and optionally, the following: a) The protein preparation contains at least about 70% (w / w), preferably at least about 75% (w / w), more preferably at least about 85% (w / w), and most preferably at least about 95% (w / w) of protein by dry weight. b) Lipids in a dry weight of approximately 110 mg / g or less, preferably approximately 50 mg / g or less, more preferably approximately 40 mg / g or less, even more preferably approximately 20 mg / g or less, or even more preferably approximately 15 mg / g or less of protein preparations. c) After heat treatment, the water-binding capacity of the protein preparation is approximately 4 g / g or more, preferably approximately 5 g / g or more, and more preferably 6.5 g / g or more, by dry weight. d) Preferably without syneresis, after heat treatment, gel-forming ability with about 2%, about 3%, about 5%, or about 5.5% of the protein preparation by total weight of the solution consisting of the protein preparation and water, and / or e) nucleic acids, by dry weight of the protein preparation, comprising about 10% (w / w) or less, more preferably about 5.5% (w / w) or less, and more preferably about 2.5% (w / w) or less. The protein preparation comprising the above.

10. A protein preparation derived from a microorganism, preferably a single-celled microorganism, which preferably, without syneresis, after heat treatment, contains gel-forming ability with approximately 1% to 10% of the protein preparation per total weight of the solution consisting of the protein preparation and water, and optionally includes the following: a) The protein preparation contains at least about 70% (w / w), preferably at least about 75% (w / w), more preferably at least about 85% (w / w), and most preferably at least about 95% (w / w) of protein by dry weight. b) Lipids in a dry weight of approximately 110 mg / g or less, preferably approximately 50 mg / g or less, more preferably approximately 40 mg / g or less, even more preferably approximately 20 mg / g or less, or even more preferably approximately 15 mg / g or less of protein preparations. c) After heat treatment, the water-binding capacity of the protein preparation is approximately 4 g / g or more, preferably approximately 5 g / g or more, and more preferably 6.5 g / g or more, by dry weight. d) Preferably without syneresis, after heat treatment, gel-forming ability with about 2%, about 3%, or about 5%, or about 5.5% of the protein preparation by total weight of the solution consisting of the protein preparation and water, and / or e) nucleic acids in the dry weight of the protein preparation, which amount to about 10% (w / w) or less, more preferably about 5.5% (w / w) or less, and even more preferably about 2.5% (w / w) or less. The protein preparation comprising the above.

11. The protein preparation according to claim 9, wherein the protein preparation exists in a dry form, preferably in a powder form.

12. below: (a) Prepare the protein preparation according to claim 9, (b) Mix the protein preparation with an aqueous dispersion medium, and then, (c) Heat the mixture to a temperature of at least about 55°C to provide a protein gel. A method for preparing a protein gel, including [a specific component].

13. Preferably, the use of the protein preparation according to claim 9 for preparing a food product or nutritional supplement for human or animal use.

14. A nutritional supplement or food product comprising the protein preparation described in claim 9.

15. below: a) Prepare a microorganism, and optionally subject the microorganism to one or more pretreatment steps, b) comprising step b1) lysing the microorganisms, thereby preparing a lysate containing an aqueous liquid fraction with nucleic acids and lysed natural proteins of the microorganisms, and further purifying the lysate, preferably by centrifugation or filtration. c) The following: i) Add a nucleic acid adsorbent immobilized on a solid support, preferably a floating solid support, to the aqueous liquid fraction. ii) Optionally, stir or shake, and iii) Preferably by sedimentation and optionally by filtration, the nucleic acids bound to the nucleic acid adsorbent immobilized on the solid support are separated. This is used to obtain an aqueous liquid fraction (which is a nucleic acid-reduced aqueous liquid fraction). Including, Including anion exchange chromatography and / or anion mixed-mode chromatography, nucleic acids are separated from the aqueous liquid fraction. d) Filter the aqueous liquid fraction to obtain a solution containing purified and dissolved natural proteins from microorganisms and an aqueous solvent, preferably water or physiological saline solution. e) Optionally, remove at least a portion of the aqueous solvent, f) Optionally, sterilize the solution to obtain a sterile solution containing purified microbially dissolved natural proteins and an aqueous solvent, and g) Optionally, remove at least a portion of the aqueous solvent from the sterile solution in step f). A method for preparing a microbial natural protein, including Preferably, the process further includes the step of separating lipids from the aqueous liquid fraction. The aforementioned method.

16. The protein preparation according to claim 10, wherein the protein preparation exists in a dry form, preferably in a powder form.

17. The following: (a) Prepare the protein preparation according to claim 10, (b) Mix the protein preparation with an aqueous dispersion medium, and then, (c) Heat the mixture to a temperature of at least about 55°C to provide a protein gel. A method for preparing a protein gel, including [a specific component].

18. Use of the protein preparation according to claim 10 for preparing a food product or nutritional supplement, preferably for human or animal use.

19. A nutritional supplement or food product comprising the protein preparation described in Claim 10.

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