Method for producing food product using Crab-negative yeast

Abstract

The invention relates to a method for obtaining edible biomass from by-products of the food industry. To the edible biomass thus obtained; to the use of the edible biomass for the manufacture of food products; or to a food product comprising an edible biomass obtained by the above process.

Description

Technical Field

[0001] The present invention relates to the field of food science or food technology. Specifically, the present invention relates to a method for obtaining edible biomass from by-products of the food industry; to the edible biomass obtained thereby; to the use of the edible biomass for producing food; or to a food containing the edible biomass obtained by the above method. Background Art

[0002] Projections indicate that feeding the world's nearly 10 billion people by 2050 will require a 70% increase in total food production. Currently, this food gap is linked to unsustainable agricultural and food practices. At the same time, there is an urgent need to reduce greenhouse gas (GHG) emissions from agricultural production and halt the conversion of remaining forests to agricultural land. Consequently, there is strong interest in developing new food sources that are sustainable, nutritious, and healthy.

[0003] While other non-animal alternatives are available, they come with some drawbacks, such as organoleptic properties, deforestation (soy), and being low in certain amino acids we need in our diet (namely lysine).

[0004] Against this backdrop, plant-based meat, dairy, and egg alternatives currently face numerous formulation challenges, which involve mimicking the sensory and functional properties of animal-based proteins and thus replacing them. Market trends are primarily concentrated in three areas: plant-based proteins, fermentation, and animal cultures.

[0005] The use of microorganisms as an alternative source of SCP (single-cell protein) has great potential due to their numerous advantages, such as their unaffected seasonal or climatic factors and their limited use in very small areas, which reduces the deforestation caused by other alternatives (plant and / or animal). Furthermore, compared to animal protein, the amount of water consumed and the amount of CO2 emitted into the atmosphere are reduced (approximately 98% and 85%, respectively). Furthermore, they have a high nutritional profile, containing all the amino acids recommended by the FAO, as well as other compounds associated with immunomodulatory effects, such as vitamins, ω-fatty acids, and beta-glucans.

[0006] One of the limitations of SCP is its poor process performance and the need to remove nucleic acids. The nucleic acid (RNA) content in SCP can be problematic for human health because high purine intake can increase uric acid concentrations (hyperuricemia), which is a precursor to diseases such as gout or kidney stones. A preventive strategy regarding nucleic acids is to select appropriate microorganisms that are typically present in low concentrations, for example algae are preferred over fungi and bacteria. Other methods for reducing nucleic acids have also been evaluated, including activating enzymes (ribonucleases) through heat treatment or adding salts, acids or hydroxides. For ribonuclease treatment, temperatures between 68°C and 74°C are used for optimal degradation, but with biomass losses of 30 to 40%. Other treatments combine high temperatures with ethanol solutions or solutions at extreme pH (acidic or alkaline solutions), which can affect protein quality and require washing steps that result in significant losses of biomass and protein material.

[0007] By 2050, the world will need to produce 1.25 billion tons of meat and dairy products annually to meet global demand for animal-based protein at current consumption levels. However, due to inefficiencies in converting feed into meat and dairy products, increasing meat and dairy production alone cannot sustainably meet the growing protein demand. New solutions are needed. Single-cell proteins (SCPs), proteins produced in microbial and algal cells, offer a promising option. Recent interest in SCPs has focused on using microorganisms to increase their protein content, thereby enhancing the value of by-products, which can then be used in animal feed.

[0008] Protein can also be provided by culturing various microorganisms and algae, preferably those whose biomass contains more than 30% protein dry matter and can provide a healthy balance of essential amino acids. Currently commercially available SCPs are based on mycoprotein, fungal cells, which are limited to meat analogs and have concerns about mycotoxins.

[0009] Due to climate change and the diversity of by-products around the world, it is difficult to find similar ingredients with high nutritional profiles in different regions, and the scarcity of some of these ingredients is indeed worrying.

[0010] In addition, there is a demand and consumer preference for clean label products and non-animal derived ingredients and additives. These ingredients exhibit certain processing properties, such as gelling, emulsification and foaming properties, and are added to dairy and meat analogs to replace animal proteins such as casein or albumin. Currently, modified plant proteins (although this approach does not address environmental impact) or synthetic polymers are used as methylcellulose (a non-natural ingredient).

[0011] The present invention is dedicated to solving this problem and describes an innovative method for obtaining edible biomass from by-products of the food industry. As described above, the proposed method utilizes by-products of the food industry, so it helps to solve an important problem by producing edible biomass without using the earth's limited biological and physical resources. In addition, according to the method proposed in this article, a single edible biomass final product with consistent quantitative and qualitative components can be obtained from various by-products of the food industry, thereby simplifying, facilitating and optimizing the industrial process of biomass production. Therefore, a variety of agricultural food by-products can be used, such as by-products from sugar, potatoes, grains, vegetables, etc. This fact results in a sustainable process with minimal impact on the environment. Obtaining similar ingredients from different by-products will allow local production, so that the food chain will not rely on imports from other countries, thereby reducing the risk of ingredient shortages due to climate change or economic, political and social problems. Summary of the Invention

[0012] Summary of the Invention

[0013] The present invention relates to a method for obtaining edible biomass from by-products of the food or feed industry (wherein the biomass product comprises, for example, one or more of single-cell protein, cell lysate, protein concentrate, protein isolate, biomass extract and biomass hydrolysate); to the edible biomass obtained thereby; to the use of the edible biomass for producing food; or to a food comprising the edible biomass obtained by the above method.

[0014] The method of the present invention is based on the fermentation of byproduct solutions or suspensions (e.g., dissolved carbon, nitrogen, or other micronutrients) in the presence of specific microorganisms, particularly Crabtree-negative yeasts. By utilizing byproducts of the food industry, the inventors of the present invention are addressing an important problem, as they are able to produce edible biomass from byproducts without using the Earth's limited biological and physical resources. Furthermore, according to the method proposed herein, starting from various byproducts of the food industry, a single edible biomass end product with a consistent quantitative and qualitative composition can be obtained, thereby simplifying, facilitating, and optimizing the industrial process of biomass production.

[0015] Therefore, a first embodiment of the present invention relates to a method for obtaining edible biomass (hereinafter referred to as the method of the present invention), which comprises: a) subjecting a by-product of the food industry to conditions suitable for its dissolution and / or resuspension (in other words, dissolving and / or resuspending the by-product of the food industry), b) fermenting the by-product solution or suspension in the presence of Crabtree-negative yeast, and c) inactivating the Crabtree-negative yeast to obtain edible biomass.

[0016] In a preferred embodiment, step c) comprises inactivating Crabtree negative yeast and removing nucleic acids in one step by setting the temperature to 75 to 130° C. for 5 to 60 minutes, preferably 70 to 90° C. for 5 to 15 minutes, to obtain edible biomass. In a preferred embodiment, the method of the present invention is characterized in that the method does not include a washing step after inactivation and removal of nucleic acids, thereby improving the yield. It is noted that the use of these temperatures and times is sufficient to degrade unwanted nucleic acids without the need for a washing step to remove them after inactivation and removal of nucleic acids, thereby significantly reducing biomass losses. In fact, as shown in Example 12, less than 10% of the biomass is lost in this method (ranging from 2% to 10%).

[0017] In the context of the present invention, "conditions suitable for the solubilization and / or resuspension of by-products" are considered to be known to those skilled in the art. Generally, according to the present invention, by-products are solubilized or resuspended in aqueous solution at varying pH values (pH = 0.5 to 14), depending on their source, in the presence or absence of enzymes and in combination with physical (high temperature or ultrasound) or mechanical (high shear homogenization) treatment. If necessary, the culture medium can be supplemented with other macronutrients and micronutrients to promote yeast growth.

[0018] The harvested biomass is inactivated by thermal or non-thermal treatment. Furthermore, the biomass can be lysed, for example, by autolysis, enzymatic hydrolysis, homogenization, high pressure, ultrasound, or other lysis procedures. Various methods have been described for extracting certain components of yeast cells in order to obtain functional ingredients with specific process properties, such as emulsifying and gelling properties.

[0019] In a preferred embodiment, the method of the invention comprises the use of a Crabtree-negative yeast selected from the group consisting of Candida utilis, Kluyveromyces marxianus, Debaryomyces hansenii or Yarrowia lipolytica. Other Crabtree-negative yeasts that can be used are selected from the group consisting of: Candida cylindrace, Saccharomyces cabtree negative species, Hansenula spp., Pichia spp., Kluyveromyces spp., Brettanomyces spp., Kamogataella spp., Ogatea angusta, Schizosaccharomyces pombe, Lindnera jardinii or Wickerhamomyces anomalus, among others.

[0020] Therefore, this first embodiment also relates to the use of Crabtree-negative yeast for obtaining edible biomass from by-products of the food industry, preferably the use of Crabtree-negative yeast selected from the group consisting of Candida utilis, Kluyveromyces marxianus, Debaryomyces hansenii, or Yarrowia lipolytica. Other Crabtree-negative yeasts that can be used are selected from the group consisting of Candida columnaris, cabtree-negative yeast species, Hansenula, Pichia, Kluyveromyces, Brettella, Kamogataella, Ogatea angusta, Schizosaccharomyces pombe, Lindnera jardinii, or Wickham's yeast.

[0021] In a preferred embodiment of the present invention, the method of the present invention is characterized in that, starting from various by-products of the food industry, a single edible biomass end product with a consistent quantitative and qualitative composition is obtained.

[0022] In a preferred embodiment of the present invention, the edible biomass comprises a protein dry matter content of at least 30%, a fiber dry matter content of at least 10% and / or a fat dry matter content of at least 1% (w / w).

[0023] In a preferred embodiment of the present invention, the edible biomass final product comprises: a protein dry matter content of 30% to 80%, a fiber dry matter content of 10% to 70% and / or a fat dry matter content of 1% to 60% (w / w).

[0024] In a preferred embodiment of the present invention, the by-product is a by-product of non-animal origin.

[0025] In a preferred embodiment of the present invention, the by-product is a plant-derived by-product.

[0026] A second embodiment of the present invention relates to a method for obtaining a food product, comprising: a) obtaining edible biomass according to the method defined above, and b) fractionation to obtain the food product.

[0027] In a preferred embodiment, classification is carried out by inactivation and / or lysis. After lysis, different cell fractions are treated to produce final components with similar nutritional structures and different process properties. This procedure allows a variety of components to be obtained to be included in a wide range of applications. Specifically, the first step includes inactivation and / or lysis of the yeast. The process includes autolysis, chemical lysis (solvents, detergents, osmotic shock), physical destruction (heat treatment - including autoclaving, ultrasonic treatment, freeze-thaw, hot water or steam, ultrasound), mechanical lysis (high pressure, homogenization) or enzyme treatment. Afterwards, the sample can be classified directly or after a centrifugation step (cell wall). The classification step includes alkaline, acidic or neutral treatment, buffer citrate treatment or enzyme treatment with different incubation times, combined or not combined with other physical or mechanical treatments. The optional third step includes separation, which can be completed by performing at least one of the following procedures: physical separation (filtration or centrifugation), acid precipitation and organic solvent precipitation. Finally, the final component can be dried. These ingredients are characterized by their nutritional profile and process properties related to gelling, foaming, and emulsifying activity, as well as solubility, water absorption, and oil absorption. In these processes, no specific purification steps are used; instead, different fractions are enriched or enhanced to produce several functional ingredients. Minimal modifications are made to the downstream process to obtain ingredients with a similar nutritional profile and a wide range of functional properties to the starting biomass. Depending on the combination of these three steps (lysis, fractionation, and separation), the final ingredients exhibit different process properties.

[0028] In a preferred embodiment, the fractionation step b) is performed to obtain a single food product comprising a consistent percentage content of protein, fiber and / or fat selected from the following ranges: at least 30% protein dry matter content (w / w), at least 10% fiber dry matter content (w / w) and / or at least 1% fat dry matter content (w / w); preferably 30% to 80% protein dry matter content (w / w), 10% to 70% fiber dry matter content (w / w) and / or 3% to 60% fat dry matter content (w / w). This means that the components of the final product (protein, fiber and / or fat) are broken down but not separated (they form part of the same product), resulting in a single product biomass with a consistent quantitative and qualitative composition (i.e., a consistent percentage range of protein, fiber and / or fat). Thus, the same single product has different technical and / or nutritional properties due to the presence of a consistent percentage range of protein, fiber and / or fat. Please note that obtaining a single product with a "consistent percentage content of protein, fiber and / or fat" means that, according to the method of the present invention, a specific percentage of each component (protein, fiber and / or fat) can be selected from the above-mentioned percentage ranges to be present in the same final product and this percentage can be maintained in a reproducible manner. Thus, the final product is versatile (the percentage content of each component can be selected and adjusted according to the final intended use) and reproducible (the selected percentage can be maintained in a reproducible manner over time).

[0029] Furthermore, it is worth noting that the edible biomass obtained according to this method is characterized by improved processing properties compared to other yeast-based SCPs (Razzaq 2020), such as high oil absorption capacity and foaming stability (Table 10). In addition, the treatment of the biomass (KZ12F) resulted in a significant improvement in the foaming and emulsification-related abilities of the ingredient, maintaining its nutritional structure (Table 9).

[0030] A third embodiment of the present invention relates to edible biomass obtained by the method of the present invention, characterized in that the edible biomass is a single product comprising a consistent percentage content of protein, fiber and / or fat selected from the following ranges: at least 30% protein dry matter content (w / w), at least 10% fiber dry matter content (w / w) and / or at least 1% fat dry matter content (w / w); preferably 30 to 80% protein dry matter content (w / w), 10 to 70% fiber dry matter content (w / w) and / or 3 to 60% fat dry matter content (w / w).

[0031] A fourth embodiment of the present invention relates to the use of the edible biomass defined above for producing food products.

[0032] A fifth embodiment of the present invention relates to a food product obtainable by the method described in the second embodiment of the present invention.

[0033] A sixth embodiment of the present invention relates to a food product comprising the edible biomass defined above.

[0034] In a preferred embodiment, the food product of the present invention is characterized in that the food product is a single food product comprising a consistent percentage content of protein, fiber and / or fat selected from the following ranges: at least 30% protein dry matter content (w / w), at least 10% fiber dry matter content (w / w) and / or at least 1% fat dry matter content (w / w); preferably 30 to 80% protein dry matter content (w / w), 10 to 70% fiber dry matter content (w / w) and / or 3 to 60% fat dry matter content (w / w).

[0035] For the purposes of this invention, the following terms are defined:

[0036] The term "including" means including, but not limited to, whatever follows the word "including". Thus, the use of the term "including" means that the listed elements are required or mandatory, but other elements are optional and may or may not be present.

[0037] • The term “consisting of means including, but not limited to, whatever follows the phrase “consisting of.” Thus, the phrase “consisting of” means that the listed elements are required or mandatory, and no other elements may be present. BRIEF DESCRIPTION OF THE DRAWINGS

[0038] Figure 1 Kluyveromyces marxianus was grown in different byproduct media (Medium-1 and Medium-2) and rich medium (YEPD). DETAILED DESCRIPTION

[0039] The present invention is illustrated by the following examples, which are not intended to limit the scope of protection thereof.

[0040] Example 1. Byproduct treatment

[0041] Culture medium preparation

[0042] The by-products are weighed, resuspended in different aqueous solutions (neutral, alkaline or acidic), and treated with temperature to dissolve most of the components (room temperature or 80°C). Afterwards, the soluble components are neutralized and used directly for culture medium preparation. Avoiding the removal of acid and alkali and the drying steps, the method is more scalable and more suitable for industrial applications. Table 1 shows some by-products processed by the present invention. Representatives of each group of by-products have been selected: sugar-rich by-products (from the sugar industry, containing monosaccharides and disaccharides), carbohydrate-rich by-products, fiber-rich by-products, and lipid- or protein-rich by-products. If necessary, other macronutrients and micronutrients can be added to the culture medium. The culture medium is sterilized by autoclaving or heating the culture medium to 100°C. When necessary, some nutrients are sterilized by filtration.

[0043] Table 1. By-products used and their nutritional profile

[0044]

[0045] In the future, byproducts from the sugar industry will be diluted directly in water. Other byproducts will undergo other treatments. The most promising treatments involve a combination of high-temperature treatment and alkali or acid treatment (preferably acid), as shown in Table 2. These treatments can dissolve more than 80% of the byproducts.

[0046] 2. Processing by-products and components

[0047]

[0048] Treatment: 1: HCl at room temperature, 2: HCl + 80 ° C, 30 minutes; 3: HCl at room temperature and high pressure sterilization; 4:

[0049] HCl+80℃, 30 minutes + autoclave.

[0050] By these treatments, more than 80% of the by-products and carbon sources (starch, monosaccharides and disaccharides) were solubilized. By some treatments, partial starch saccharification was obtained.

[0051] After processing, other macronutrients and micronutrients are added to the by-products during the conditioning process. These compounds include other by-products, inorganic nitrogen sources, organic nitrogen sources, MgSO4, phosphates, trace salts or other desired growth factors (such as vitamins).

[0052] The culture medium was sterilized by autoclaving or heating the medium to 100° C. When necessary, some nutrients were sterilized by filtration.

[0053] Example 2. Growth of Crabtree-negative yeast in by-products

[0054] Several yeast strains, such as Kluyveromyces marxianus, Candida utilis, Yarrowia lipolytica, and Debaryomyces hansenii, were used to evaluate the feasibility of byproduct-based growth media. Several strains were used, and one was selected for further optimization. Growth rate was monitored by OD of the culture, as Figure 1 As shown, similar or even higher growth rates were obtained using the by-product based medium (Medium-2) compared to the commercial rich medium (YEPD).

[0055] The increase of carbon source resulted in an increase in biomass concentration as described for Crabtree negative yeasts (Table 3).The efficiency of the method resulted in conversion rates greater than 70% (Table 4).

[0056] Table 3. Biomass of Yarrowia lipolytica grown in different media

[0057] Sugar byproduct 2 Biomass (g / L) 40g / L 12.0 50g / L 15.3 60g / L 18.0 70g / L 19.7

[0058] Table 4. Conversion efficiency of biomass produced by Debaryomyces hansenii from dry bread crusts

[0059] culture medium By-product C Biomass (g / L) Conversion rate Medium-1 5g / L 4.6g / L 99.2 Medium-2 8g / L 7.30g / L 91.28 Medium-3 15g / L 13.5 73.1 Medium-5 20g / L 15.7 81.25 Medium-5 30g / L 23.3 78.7 <![CDATA[YEPD 1 ]]> <![CDATA[n / a 2 ]]> 17.96 n / a

[0060] 1 Rich medium (yeast extract peptone glucose); 2 not applicable

[0061] Example 3. Same by-product, different microorganisms and similar ingredients

[0062] Different microorganisms (Kluyveromyces marxianus and Candida utilis) are grown on the same byproduct (sugar-derived byproduct). The fermentation process consists of several steps: pre-inoculation in rich medium or byproduct medium for 8-24 hours. The culture temperature ranges from 20 to 45°C (depending on the strain).

[0063] The fermentation process involves either batch or fed-batch fermentation (depending on the microorganism) with a duration of 24 to 72 hours. The batch phase lasts until the carbon source is exhausted and fed-batch fermentation is initiated. Variations in specific fermentation conditions can result in different compositions.

[0064] The byproducts were diluted and a mixture of organic and inorganic nitrogen sources (yeast extract and (NH4)2SO4), phosphate, and Mg2SO4 were added. The culture media were autoclaved in bioreactors. An overnight culture of crabtree-negative yeast was added to each culture medium and incubated at 35°C with stirring and aeration for 24 to 48 hours.

[0065] Afterwards, the biomass was harvested, inactivated by heat and dried using a spray dryer. As shown in Table 5, the biomass derived from different microorganisms was characterized according to its nutritional structure.

[0066] By adjusting the fermentation strategy, we can target the composition of certain compounds in order to obtain similar compositions from different by-products (Table 5).

[0067] Table 5. Nutritional structure of two different biomasses derived from two crabtree-negative yeasts

[0068] Parameters (g / 100g) <![CDATA[Kluyveromyces marxianus 1 > Candida utilis Moisture 3.8 3.7 Fat 4.1 9.9 protein 38.2 40.8 Ash 8.6 7.6 Total carbohydrates 12.6 7.0 sugar 0.9 2.0 Dietary fiber 31.9 31.0

[0069] 1 8 batches of culture medium

[0070] Example 4. Different by-products, same microorganisms and similar ingredients

[0071] Different by-products were used for the growth of a microorganism, Debaryomyces hansenii. Both by-products contained different macronutrients (Table 6), with the main differences being in carbohydrate and fiber content.

[0072] Table 6. Nutritional structure of the tested by-products

[0073] Parameters (g / 100g) dry bread crust wheat bran Moisture 10 10 carbohydrate 75 24 protein 10.9 17 Total fat 6.7 5 Total dietary fiber 5.8 37 Salt / Ash 1.8 6

[0074] Both byproducts were treated with an alkaline solution at 80°C, neutralized, and autoclaved to achieve a solubility of >80%. Salt and other micronutrients were added. An overnight culture of crabtree-negative yeast was added to each medium and incubated in baffled flasks at 25°C with stirring for 24 to 48 hours. Afterwards, the biomass was recovered, heat-inactivated, and air-dried. Table 7 summarizes the composition of the biomass obtained from the two bioprocesses.

[0075] Table 7. Debaryomyces hansenii biomass obtained from two different by-products

[0076] Parameters (g / 100g) Debaryomyces hansenii 1 Debaryomyces hansenii 2 Moisture 8.02 7.25 Fat 2.37 3.12 protein 43.93 45.01 Ash 9.98 8.95 Total carbohydrates 5.30 5.6 sugar nd nd Dietary fiber 31.4 29.6

[0077] Nd: Not conducted

[0078] Example 5. Different by-products, different microorganisms and similar ingredients

[0079] Another microorganism, Yarrowia lipolytica, was used to increase the value of another sugar by-product. The by-product-based culture medium was prepared as follows. First, the by-product was dissolved, autoclaved, and other pre-sterilized compounds (MgSO4·7H2O, (NH4)2SO4 and / or yeast extract and phosphate) were added. The incubation temperature was 25+ / -1°C and the stirring speed was 500-800rpm. 2.5M H3PO4 was used as acid and 25% KOH as base to control the pH. The aeration rate was stabilized at 1vvm. After 48 hours, the biomass was collected by centrifugation and freeze-dried. As shown in Table 8, the nutritional composition of the biomass was similar.

[0080] Table 8. Nutritional structure of biomass derived from different microorganisms and their by-products

[0081] Parameters (g / 100g) Yersinia lipolytica 1 Yersinia lipolytica 2 Debaryomyces hansenii 1 Moisture 13.38 1.89 8.02 Fat 4.68 5.54 2.37 protein 47.41 49.99 43.93 Ash 4.05 8.27 9.98 Total carbohydrates 8.58 3.71 5.30 sugar 3.4 3.6 nd Dietary fiber 21.9 30.6 31.4

[0082] Example 6. Food Application: Sauce

[0083] Edible biomass derived from Kluyveromyces yeast has good solubility, emulsification, and partial water and oil retention capabilities. A variety of food applications have utilized this ingredient to replace animal-derived ingredients, bringing added value to the final product. One example is pesto sauce, in which Parmigiano-Reggiano cheese is replaced with this edible biomass and offers additional beneficial claims: as a source of protein, vegetarian, a source of vitamins B1 and B2, and rich in vitamins B8 and B9. The process of making the sauce involves grinding nuts, spices, vinegar, and seasonings in a cutter until thoroughly combined. The final step is to add oil to the cutter to create and emulsify the sauce.

[0084] Example 7. Food Application: Pasta

[0085] The pasta contains the same biomass (9.73%), replacing eggs and offering additional product claims: as a source of protein, a source of fiber, a vegetarian diet, a source of vitamin B6, and a rich source of vitamins B1, B2, B8, and B9. The process of making fresh pasta involves mixing water, flour or semolina, salt, and MOA protein to form a dough. After the dough rests for one hour, it is formed into the desired shape.

[0086] Example 8. Food Application: Cheese Analogs

[0087] The same biomass (16%) is used to make cheese analogs, which carry relevant claims: as a source of protein, a source of fiber, vegetarian, a source of vitamin B6, a rich source of vitamins B1, B2, B8, and B8, as well as a rich source of phosphorus and iron. The process of making cheese analogs involves grinding nuts and mixing them with starter cultures, salt, and moa protein. Once the cheese is set, it is left at room temperature for 4 to 7 days. After the fermentation period, the cheese is covered with spices and salt.

[0088] Example 9. Fractionation to obtain gelling, emulsifying and foaming properties (prototype KS22AcEt)

[0089] Yeast cells are lysed by physicochemical treatment using a weakly alkaline (pH 8 to 11) sodium hexametaphosphate buffer, and the suspension is heated to 65°C for 2 hours. The cell walls are then removed and the yeast is resuspended in citrate buffer and treated at 121°C for 20 minutes. These treatments are followed by two consecutive precipitation steps, neutralization of the solution to a pH of 4.5 to 5.5, and addition of anhydrous ethanol. The samples are then dried, and the obtained process properties are gelling activity under thermal conditions, as well as emulsifying and foaming activity, while the nutritional properties remain unchanged.

[0090] Example 10. Fractionation to obtain gelling and emulsifying properties (prototype KD12F)

[0091] Yeast cells were lysed by mechanical disruption using pressure. The cell walls were then removed and resuspended in citrate buffer and treated at temperatures between 80°C and 121°C for 20 minutes. The samples were then directly dried. The resulting nutritional profile was similar, and the process properties achieved were gelling and emulsifying activity (Table 9). Cold gelling properties were also achieved if a centrifugation step and citrate buffer treatment were added after lysis.

[0092] Example 11. Fractionation to obtain gelling, emulsifying and foaming properties (KZ12F)

[0093] The yeast cells were lysed by enzymatic treatment for 4 hours. Afterwards, the cell walls were removed and resuspended in citrate buffer and treated at 121°C for 20'. The samples were then directly dried. The technological properties achieved were gelling activity under thermal conditions as well as emulsification and foaming activity, while the nutritional structure was improved due to the higher protein content (Table 9). Compared to other yeast-based SCPs (Razzaq 2020), the edible biomass obtained according to this method is characterized by improved technological properties, such as high oil absorption capacity and foaming stability (Table 10). In addition, the treatment of the biomass (KZ12F) resulted in a significant improvement in the foaming and emulsification-related abilities of the ingredient, maintaining its nutritional structure (Table 9).

[0094] Table 9. Nutritional and process properties of different fractions from the same biomass

[0095] parameter <![CDATA[Prototype 0 1 > KD12F KZ12F Moisture 3.8 2.4 2.5 Fat 4.1 4.3 3.9 protein 38.2 36.9 36.9 Ash 8.6 12.6 3.9 Total carbohydrates 12.6 16.5 13.1 sugar 0.9 nd nd Dietary fiber 31.6 27.5 27.6

[0096] 1 Heat-killed biomass (no extraction process)

[0097] Table 10. Comparison of properties of the edible biomass of Example 11 with previously described edible biomass representatives.

[0098]

[0099]

[0100] Example 12. RNA Removal

[0101] After the fermentation step, the biomass was harvested and processed at various temperatures. RNA was then extracted, quantified using ultraviolet light (260 nm), and expressed as the percentage of RNA per unit dry cell weight (Table 11). It is recommended that humans consume less than 2% of the RNA. Depending on the processing, biomass losses ranged from 2% to 10%.

[0102] Table 11. RNA content in yeast biomass after different treatments (expressed as percentage)

[0103]

Claims

1. A method for obtaining edible biomass, comprising: a. Dissolving and / or resuspending by-products of the food industry, b. Fermentation of a by-product solution or suspension in the presence of Crabtree negative yeast, c. Inactivate the Crabtree-negative yeast and nucleic acid removal to obtain the edible biomass.

2. The method according to claim 1, characterized in that The method does not include a washing step after inactivation of the Crabtree-negative yeast and removal of nucleic acids.

3. The method according to any one of the preceding claims, characterized in that Less than 10% of the biomass is lost during the process.

4. The method of claim 1, wherein the Crabtree-negative yeast is selected from the group consisting of Candida utilis, Kluyveromyces marxianus, Debaryomyces hansenii, and Yarrowia lipolytica.

5. The method according to any one of the preceding claims, characterized in that Starting from various by-products of the food industry, a single edible biomass end product with a consistent quantitative and qualitative composition is obtained.

6. The method according to any one of the preceding claims, characterized in that The edible biomass is a single product comprising consistent percentage contents of protein, fiber and / or fat selected from the following ranges: at least 30% protein dry matter content (w / w), at least 10% fiber dry matter content (w / w) and / or at least 1% fat dry matter content (w / w); preferably 30% to 80% protein dry matter content (w / w), 10% to 70% fiber dry matter content (w / w) and / or 3% to 60% fat dry matter content (w / w).

7. The method according to any one of the preceding claims, characterized in that The by-products are non-animal-derived by-products.

8. The method according to any one of the preceding claims, characterized in that The by-products are plant-derived by-products.

9. Methods of obtaining food, including: a. obtaining edible biomass by the method according to claims 1 to 8, b. fractionating the edible biomass to obtain a single food product, the single food product comprising a product selected from the group consisting of Consistent percentages of protein, fiber, and / or fat within the following ranges: At least 30% protein dry matter content (w / w), at least 10% fiber dry matter content (w / w) and / or at least 1% fat dry matter content (w / w); preferably 30 to 80% protein dry matter content (w / w), 10 to 70% fiber dry matter content (w / w) and / or 3 to 60% fat dry matter content (w / w).

10. The method for obtaining food according to claim 9, characterized in that Fractionation is performed using crabtree-negative yeast or crabtree-positive yeast to obtain food products with improved gelling, emulsifying, water-absorbing and oil-absorbing abilities.

11. Use of Crabtree-negative yeast for obtaining edible biomass from by-products of the food industry.

12. Use of Crabtree-negative yeast according to claim 11 for obtaining edible biomass from by-products of the food industry, characterized in that The Crabtree-negative yeast is selected from the group consisting of Candida utilis, Kluyveromyces marxianus, Debaryomyces hansenii, and Yarrowia lipolytica.

13. Edible biomass obtained by the method of any one of claims 1 to 8.

14. Edible biomass obtained by the method according to any one of claims 1 to 8 according to claim 13, characterized in that The edible biomass is a single product comprising consistent percentage contents of protein, fiber and / or fat selected from the following ranges: at least 30% protein dry matter content (w / w), at least 10% fiber dry matter content (w / w) and / or at least 1% fat dry matter content (w / w); preferably 30% to 80% protein dry matter content (w / w), 10% to 70% fiber dry matter content (w / w) and / or 3% to 60% fat dry matter content (w / w).

15. Use of edible biomass according to claim 13 or 14 for producing food.

16. Food obtained by the method according to claim 9 or 10.

17. The food according to claim 16, characterized in that The food product is a single food product comprising consistent percentage amounts of protein, fiber and / or fat selected from the following ranges: at least 30% protein dry matter content (w / w), at least 10% fiber dry matter content (w / w) and / or at least 1% fat dry matter content (w / w); preferably 30 to 80% protein dry matter content (w / w), 10 to 70% fiber dry matter content (w / w) and / or 3 to 60% fat dry matter content (w / w).

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