A vegan medium for microorganisms suitable for use in the cosmetics industry

Abstract

The invention relates to a vegan culture medium and a production method thereof, which can meet the protein and carbohydrate nutritional requirements for the cultivation and growth of beneficial microorganisms suitable for use in particular in the cosmetic, food, pharmaceutical, agricultural and similar industries.

Description

[0001] A VEGAN MEDIUM FOR MICROORGANISMS SUITABLE FOR USE IN THE COSMETICS INDUSTRY TECHNICAL FIELD

[0002] The invention relates to a vegan culture medium capable of meeting the protein and carbohydrate nutritional requirements for the cultivation and growth of beneficial microorganisms suitable for use in particular the cosmetic, food, pharmaceutical, agricultural and similar industries, and a production method thereof.

[0003] State of the Art:

[0004] The term 'vegan' refers to products or formulations that contain no animal-derived components; they consist solely of plant-based, mineral, or microbial components. Within this scope, milk, eggs, honey, gelatin, meat, fish, seafood, animal fats, or additives derived from these are not used. Vegan ingredients are sourced from nonanimal sources such as grains, legumes, vegetables, fruits, oilseeds, plant extracts, vitamins, minerals, and microbial additives. Vegan cosmetics refer to cosmetic products that do not contain any animal-derived ingredients or ingredients indirectly obtained from animals (such as milk, eggs, honey) in their formula. There is also a difference between vegan and vegetarian products. Vegetarian cosmetic products contain ingredients derived indirectly from animals in theirformulas. However, products in both product groups are cruelty-free, meaning they are not tested on animals during the development process.

[0005] A culture medium is a nutrient-rich environment prepared to support the growth and metabolic activities of specific microorganisms or cell lines; it contains carbohydrates, proteins, vitamins, minerals, and other necessary growth factors. It can be prepared in liquid, semi-solid, or powder form depending on the application purpose. It can be used for a wide variety of purposes, such as the development, isolation, identification, counting, susceptibility testing, sterility testing, examination of clinical samples, food, water and environmental controls, obtaining biological products, antibiotic and vitamin analyses, industrial analyses, etc. Culture media can be prepared and used in many different ways. For example, milk is a "general culture medium" and contains all the nutrients necessary for the growth of microorganisms and sufficient water for their utilisation. Although grape juice also contains the nutrients and water necessary for growth, its high acidity makes it suitable only for the growth of microorganisms (certain bacteria and especially yeasts) that can thrive in such an acidic environment.

[0006] In the cosmetics industry, beneficial microorganisms such as probiotics, prebiotics, and postbiotics, along with their associated components, have emerged in recent years as bioactive agents that support skin health. These microorganisms or their metabolites are considered active ingredients in functional cosmetic product formulations, providing effects such as balancing the skin microbiome, improving skin barrier function, reducing inflammation, delaying signs of ageing (anti-ageing) and maintaining skin moisture levels. The increasing demand for microbiome-friendly products is highlighting the importance of solutions based on such biological agents in the cosmetics industry.

[0007] The skin microbiome plays a critical role in maintaining healthy skin. A healthy microbiome prevents the colonisation of pathogenic microorganisms and strengthens the skin barrier. Probiotics and the postbiotics they produce protect skin health and prevent various skin problems by increasing the diversity of the skin microbiome. Probiotics and postbiotics reduce oxidative stress by neutralising free radicals and delay signs of ageing. They also increase skin elasticity by promoting collagen production and reducing the appearance of fine lines. Furthermore, probiotics increase the skin's moisture retention capacity, reducing water loss and keeping the skin more hydrated and healthy. At the same time, they strengthen the skin barrier, providing protection against environmental factors. Probiotics and postbiotics reduce inflammation in the skin, alleviating redness and irritation. These microorganisms increase skin calmness by producing soothing components.

[0008] Some disadvantages are also associated with known culture media in the field. The quality of animal-derived components used in the field may vary depending on sources and production processes, thereby affecting the consistency of the culture medium. The use of animal components is not preferred by vegan and vegetarian consumers. Animal-derived components carry the risk of contamination with unwanted microorganisms. This complicates the sterilisation process of the culture medium and ultimately affects the purity of the resulting culture. Another disadvantage of animal derived components is their high cost. Animal-derived components are generally more expensive and increase costs, especially in large-scale production.

[0009] Another disadvantage of the media used in the previous technique is supply chain issues. Some animal components are difficult to source, or regional supply issues may arise. Providing a stable and consistent food source can be difficult, which affects the growth and production efficiency of cultures. Animal production processes generally require high water and energy consumption, which is detrimental to environmental sustainability. The management of waste generated during the processing of animal components creates additional challenges. The use of animal-derived components may be subject to strict legal regulations. This can complicate production processes and incur additional costs. The safety of animal products in technology requires additional testing and certification.

[0010] In traditional media, bacteria metabolise glucose to produce lactic acid, causing the pH level of the environment to drop to low values such as 3.8-4.2. Such low pH levels can adversely affect the growth of bacteria, causing them to enter a stationary phase and then a death phase, leading to a decrease in colony counts. Furthermore, the final pH value resulting from this process is generally unsuitable for the stability of probiotic cultures.

[0011] Patent application number KR102470309 relates to a non-irritating vegan cosmetic composition and a method for producing it.

[0012] Patent application number JP2006238706 relates to a medium and culture method for bifidobacteria. More specifically, it uses only plant-based components, without any animal-derived components. The patent is for a method of culturing bifidobacteria using a culture medium and for providing a food product to which bifidobacteria cultured using this method are added.

[0013] Considering these challenges, it has become necessary to develop innovative approaches in vegan and plant-based culture medium formulations to eliminate the aforementioned disadvantages.

[0014] Brief Description of the Invention

[0015] The present invention relates to a culture medium, in other words a vegan culture medium, containing vegan and plant-based components that are suitable for the cultivation and development of beneficial microorganisms, as well as the production method of this culture medium and its applications, particularly in sectors such as cosmetics, food, pharmaceuticals, agriculture and livestock farming.

[0016] One of the objectives of the invention is to develop a culture medium formulation that allows for the selective growth of target microorganisms while also limiting the risk of microbial contamination.

[0017] Another objective of the invention is to offer a culture medium formulation based on plant-derived and sustainable ingredients that eliminates the disadvantages of animal-derived components, such as inconsistent quality, contamination risk, high cost, and unacceptability to vegan / vegetarian users.

[0018] Another objective of the invention is to eliminate the multifaceted disadvantages of animal-derived culture medium components, such as supply chain issues, negative impacts on environmental sustainability, subjection to legal regulations, waste management difficulties, and quality / consistency deficiencies, to provide an alternative, plant-based culture medium that is readily available, more environmentally sustainable, and more compliant with regulations.

[0019] Another objective of the present invention is to provide a culture medium formulation that provides a suitable pH range for the effective cultivation and growth of microorganisms. The initial pH range of some known media in the art is generally between 5.5 and 5.7, which does not provide sufficient growth conditions for all Lactobacillus species. Furthermore, these bacteria convert glucose into lactic acid during their metabolic activities, lowering the pH of the environment; this causes the pH value to drop to levels between 3.8 and 4.2. Such low pH levels can halt bacterial growth, causing them to enter a stationary phase and subsequently a death phase, leading to a decrease in colony count. In this context, the invention aims to develop a culture medium that both provides an appropriate initial pH range and maintains pH balance at the end of the fermentation process, thereby sustaining microorganism viability.

[0020] Detailed Description of the Invention

[0021] The present invention concerns a culture medium composition made entirely from vegan and plant-based components, used specifically for the cultivation and multiplication of beneficial microorganisms. The invention covers the structural properties of the vegan culture medium, its components, functional effects, production method, and its potential in a wide range of applications and different application areas. The invention described in this detailed description relates to a vegan culture medium that meets the protein and carbohydrate nutritional requirements for the cultivation and growth of microorganisms useful for use, particularly in the cosmetics industry.

[0022] The vegan culture medium of the invention is formulated to provide a suitable environment for the cultivation of various probiotic bacteria, primarily Lactobacillus species, as well as microorganisms such as Bifidobacterium, Streptococcus, Bacillus, Saccharomyces, and Kluyveromyces. The formulation presented in the invention not only supports the growth of these microorganisms but also reduces the risk of contamination, maintains pH balance, promotes metabolic activity, and possesses properties suitable for industrial production. Lactobacillus bacteria are preferably used in the present invention. However, for the purpose of providing similar technical solutions, the use of different bacterial species when using the vegan culture medium included in the invention is also covered by the scope of protection of the invention. The vegan medium discussed in the invention aims to provide the nutrients necessary for the cultivation and culture of all described bacterial species and to eliminate the risk of contamination.

[0023] The present invention, by examining the proteins, carbohydrates, and other chemicals found in MRS and LBS media, offers a vegan medium derived entirely from different and natural sources that can meet the nutritional needs of probiotics.

[0024] The medium composition includes the following basic components:

[0025] Cereal Derivatives

[0026] The vegan medium in question contains cereals. The medium in question contains at least one cereal derivative, which acts as a source of carbohydrates and protein for microorganisms. Therefore, the aforementioned cereals and cereal derivatives serve as a nutrient source for the bacterial species to be cultivated and cultured. At least one grain selected from the group consisting of wheat, barley, rye, maize, rice, oats, millet, quinoa, and buckwheat is used as the grain. Preferably, at least one grain from the group consisting of wheat, barley, rye, or maize is used as the grain within the scope of the present invention. The aforementioned cereals and cereal derivatives may be used in the form of flour, extract, or hydrolysate.

[0027] The cereal contains more carbohydrates than MRS and LBS media; as for protein, it contains plant proteins at levels similar to other media. The cereals in the vegan medium contain at least 75 g / l carbohydrates and at least 10 g / l protein. These contents are the main sources for bacterial growth. Proteins are broken down by enzymes and contribute to protein synthesis within the bacteria. Carbohydrates generally enter the glycolysis pathway to produce energy. The cereal component also contains natural prebiotic fibres (e.g. beta-glucan, inulin, arabinoxylan), supporting the growth of microorganisms for a longer period of time.

[0028] Additionally, minerals such as manganese, magnesium, zinc, and calcium from grains, along with vitamins E and B, help provide the nutrients required for the ideal growth of Lactobacillus bacteria in the vegan culture medium used in the present invention. In some applications, cereals are pre-treated with enzymes such as amylase and / or protease to make them more digestible. This enzymatic pre-treatment allows microorganisms to adapt and grow more rapidly by breaking down the complex polysaccharides and proteins in cereals into simpler forms.

[0029] Surfactants

[0030] Plant-derived surfactants are used in the culture medium to support the growth of microorganisms and increase the homogeneity of the environment.

[0031] Traditionally, culture media contain only Tween 80 as a surfactant. The culture medium subject to the present invention contains Coco Glucoside, obtained from coconut, as a surfactant. The basic nature of the Coco Glucoside component also creates a buffering effect in the environment, eliminating the need to add extra buffer solution as in other culture media. This buffering effect prevents sudden pH changes in the medium. Consequently, non-ionic, coconut-based surfactants such as Coco Glucoside are preferably used in the medium subject to the invention. This component provides stability without harming microorganisms due to its low toxicity profile and also prevents sudden pH fluctuations by exhibiting a basic buffering effect in the medium. The vegan culture medium composition of the invention contains 0.8 to 1.4% by weight of the Coco Glucoside component. Organic Acids and pH Regulators

[0032] The natural components of the flour mixtures (wheat, barley, rye, maize) used in the current invention's vegan medium provide a more suitable initial pH for Lactobacillus species. This allows the bacteria to adapt and grow rapidly. Although the pH in MRS medium is usually adjusted for bacterial growth, the appropriate pH in a natural vegan medium helps Lactobacillus species pass the lag phase more quickly.

[0033] The medium defined within the scope of the invention provides pH control by containing organic acids such as citric acid, lactic acid, and malic acid, and helps prevent unwanted microorganism contamination. These organic acids can be used alone or in combination of two or more. The citric acid content, for example, can be adjusted between 0.1% and 0.5%.

[0034] The initial pH value of the culture medium can be designed to be in the range of 2.5-9.0, and the final pH value in the range of 2.5-7.0. Preferably, the initial pH value is set to approximately 6.0, thereby enabling many probiotic microorganisms, primarily Lactobacillus species, to adapt quickly and pass the lag phase in a shorter time. Since starch-based complex carbohydrates are preferred as the carbohydrate source used in the invention, the medium pH does not fall below 4.5 during fermentation. This contributes to the preservation of cell viability and production efficiency.

[0035] According to a preferred embodiment of the present invention, citric acid is preferably included in the vegan culture medium composition to achieve target pH values and prevent other bacterial contamination. According to this embodiment, the initial pH of the vegan culture medium is 6.0. This pH is a more suitable starting pH for all Lactobacillus species than other media, as it is closer to neutral pH. This pH helps bacteria adapt more quickly and reduces the stress they experience in the early stages of growth, enabling them to pass through the lag phase more rapidly. Lactobacillus species produce lactic acid as a by-product during fermentation, which causes the pH to drop over time. Starting at pH 6.0 provides a longer buffering period before the environment becomes too acidic, preventing early cessation of growth. Furthermore, starting fermentation applications at a higher pH reduces early acid stress on the bacteria. Although Lactobacillus species are acid-resistant, minimising exposure to lower pH environments in the early stages increases the overall viability and functionality of the cells. Bacteria that enter the stationary phase transition to the death phase, and colony counts also decrease. In the present invention, lactic acid production is limited due to the vegan medium containing more starch than buffer content and ready glucose. For this reason, the final pH does not fall below 4.5. Furthermore, the pH buffering capacity can be increased by supplementing with natural mineral salts, preferably calcium carbonate and magnesium phosphate. This provides resistance to the acidic stress of the environment. This situation increases the logarithmic growth phase (exponential phase) more than competitors in the same period of time, ensuring that the commercially targeted CFU (colony-forming unit) count is reached more quickly. In cosmetic products using postbiotics or fermented components, rather than a specific CFU count, the beneficial compounds obtained during the fermentation process, rather than live bacteria, come to the fore. However, for the postbiotic content to be high, the bacterial count must also be high. For this purpose, the target amount in fermented products can be expressed as 10A9 CFU / mL (1 billion colony-forming units per millilitre).

[0036] The vegan culture medium composition subject to the invention contains citric acid at a rate of 0.1 to 0.5 per cent by weight.

[0037] Vitamins and Minerals

[0038] The culture medium formulation is designed to contain trace minerals such as manganese, magnesium, zinc, and calcium, which are essential for the metabolism of microorganisms, as well as vitamin E and B vitamins. These components are naturally derived from cereal derivatives, increasing the nutritional value of the growth medium without the need for additional chemical supplements.

[0039] Additional Functional Components

[0040] The medium may also contain cereal / plant extracts rich in phenolic compounds and flavonoids. These components impart antioxidant capacity to the formulation.

[0041] Preferably, maltodextrin or starch-based carriers can be added to the formulation to ensure the viability of probiotics is maintained after fermentation and drying processes. Plant extracts with antimicrobial properties, such as rosemary, thyme, garlic, tea tree, mint, eucalyptus, ginger, clove, lavender, cinnamon, sage, sage, fennel, and turmeric, can be added to the culture medium, either alone or in mixtures containing two or more, to reduce the risk of pathogenic microbial contamination. The present invention also encompasses a production method for the production of the vegan feed medium described above. This method includes technical steps for processing the cereal components, the fermentation process, and preparing the resulting product in a suitable form.

[0042] The cereal components used within the scope of the invention (e.g. wheat, barley, rye, maize, rice, oats, etc.) are first subjected to enzymatic hydrolysis with amylase and / or protease enzymes. This process breaks down the complex polysaccharide and protein structures found in cereals into smaller molecules. Thus:

[0043] - Carbohydrates are converted into simpler sugars, making them more readily and easily available for microorganisms.

[0044] - Proteins are broken down into peptides and amino acids, contributing to the protein metabolism of probiotics.

[0045] This enzymatic pre-treatment increases the growth rate and biomass production of probiotics, particularly Lactobacillus and Bifidobacterium species.

[0046] The enzymatically hydrolysed mixture is then subjected to a controlled fermentation process. During this stage:

[0047] - Fermentation conditions such as pH, temperature, and duration are optimised to enhance the viability and metabolic activity of probiotic microorganisms.

[0048] - Beneficial metabolites produced by microorganisms during the fermentation process (e.g., organic acids, short-chain fatty acids, antimicrobial peptides, postbiotic compounds) accumulate in the environment.

[0049] - Controlled fermentation minimises the risk of contamination and allows for the selective growth of target microorganisms.

[0050] The product obtained as a result of fermentation is subjected to a drying process for industrial-scale use. Drying converts the culture medium from liquid to powder form, and one of the following methods can be used for this purpose:

[0051] Spray drying: Provides economical and rapid drying for high-volume production.

[0052] Freeze-drying (lyophilisation): Preserves the viability of probiotics and the integrity of active components to a high degree. Vacuum drying: Allows water to be removed at low temperatures and is suitable for heat-sensitive components.

[0053] Spray drying is based on converting the liquid culture medium solution into very small droplets using an atomiser or nozzle and spraying it into a hot air stream. When the droplets come into contact with the hot air, the water evaporates rapidly, leaving behind a powdered product. This method is highly suitable for industrial scale as it allows large volumes to be processed in a very short time. The resulting powder particles are free-flowing, homogeneous and easy to store. Spray drying converts vegan nutrients into a form with a long shelf life that is portable and suitable for redissolution.

[0054] In the freeze-drying (lyophilisation) method, the product is first frozen, then the water in its frozen state is removed from the environment through sublimation, directly transitioning to the vapour phase under low pressure. During this process, most of the water in the product is removed while its structural integrity is preserved. This process, which takes place at low temperatures, preserves the viability of probiotic microorganisms and the stability of biologically active components to a high degree. The product obtained by this method has a porous structure and is easily redissolvable in water. Freeze-drying preserves the biological value of the vegan culture medium, allowing it to be stored for long periods and used in various applications.

[0055] Vacuum drying is a method that allows the product to be dried at lower temperatures under low pressure. As the boiling point of water decreases under vacuum conditions, the water in the product is removed at lower temperatures. This ensures product stability by preserving the vitamins, phenolic compounds, and other functional components in the feed. The powdered feed obtained through vacuum drying is advantageous in terms of both transport and storage and can be easily dissolved during application. This method contributes to making vegan feed suitable for industrial use, particularly by preserving functional contents.

[0056] As a result, the powdered vegan feed obtained from the drying process has a long shelf life and facilitates storage and transportation. Furthermore, it can be easily redissolved during application. Therefore, the vegan nutrient solution in question can be prepared in liquid, semi-solid or powder form. As mentioned, techniques such as spray drying, freeze drying or vacuum drying can be applied for the powder form. The dried product offers advantages in terms of transport and storage, while also providing ease of use through redissolution.

[0057] According to the present invention, the advantages provided by the production method of the vegan culture medium are as follows, without being limited to these:

[0058] - Enzymatic hydrolysis increases the bioavailability of nutrients and shortens the growth time of microorganisms.

[0059] - Controlled fermentation enables high cell density (e.g. >109CFU / mL) and rich postbiotic content to be obtained.

[0060] - The drying process increases the stability of the product, enabling its safe use in various sectors (cosmetics, food, pharmaceuticals, agriculture, livestock).

[0061] EXAMPLE

[0062] Raw Material Preparation and Target Values

[0063] Target initial pH: 6.0 (±0.2)

[0064] Final fermentation pH: > 4.5

[0065] Carbohydrate > 75 g / L, protein > 10 g / L

[0066] Surfactant: Coco Glucoside 0.8-1.4% (by weight)

[0067] Organic acid (pH regulator): Citric acid 0.1 -0.5% (by weight)

[0068] Buffer enhancer: Calcium carbonate or magnesium phosphate (0.1 -0.3% w / w) Prebiotic fibre: Inulin / beta-glucan / arabinooligosaccharide (5-10 g / L) Antimicrobial plant extracts: rosemary / thyme / garlic; 0.05-0.20% w / w, total Antioxidant extract: rich in phenolics / flavonoids; 0.05-0.2% w / w

[0069] Carrier: Maltodextrin or starch 20-40 g / L

[0070] Grain mixture: Wheat + maize + oat flour (e.g. 50:30:20, w / w)

[0071] Natural mineral / vitamin source: Naturally derived from grains.

[0072] Note: If a total grain solids content of ~120-140 g / L is targeted for a 10 L batch, typical nutrient values of > 75 g / L carbohydrates and > 10 g / L protein are provided. Preparation and Solution Formation

[0073] Water preparation: 10 L of deionised water is heated to 25-30 °C.

[0074] Grain dispersion: Slowly add a total of 1.3 kg of grain flour mixture and stir for 30-45 minutes (300-400 rpm).

[0075] Prebiotic fibre (optional): 50-100 g of inulin / beta-glucan is added and stirred until dissolved.

[0076] Citric acid pre-adjustment: Add at a level of 0.1 -0.2% (w / w) and adjust the pH to 6.0 (±0.2).

[0077] Buffer enhancer (optional): 10-30 g CaC03or Mg-phosphate suspension is added. Coco Glucoside and plant extracts are not added at this stage; they are reserved for aseptic addition in subsequent steps.

[0078] Enzymatic Pre-Treatment

[0079] Heating: The mixture is heated to 55 °C.

[0080] Amylase addition: 10-30 U / g starch (or 0.3-0.8% w / w) based on total starch.

[0081] Protease addition: 5-15 U / g protein (or 0.2-0.5% w / w) based on total protein.

[0082] Duration: 60-120 min, stirring at 55 °C (300 rpm).

[0083] Enzyme inactivation: Heat at 85-90 °C for 10 min, then rapidly cool to 40 °C.

[0084] Enzymatic hydrolysis makes complex carbohydrates / proteins more accessible, accelerating the growth of Lactobacillus, etc.

[0085] Aseptic Component Additions

[0086] Coco Glucoside: Target 1.0% (w / w); 0.8-1.4% range acceptable. Pass through a sterile 0.22 pm filter and add while <40 °C.

[0087] Antimicrobial plant extracts (optional): Total 0.05-0.2%. Added after sterile filtration. Antioxidant extract (optional): 0.05-0.2%, added after sterile filtration.

[0088] pH control: After additions, the pH is readjusted to 6.0 ± 0.2 (if necessary, a very small amount of citric acid / sodium citrate; if sodium salt is not desired, the balance can be maintained with citric acid + buffer minerals). Inoculation and Controlled Fermentation

[0089] Sterilisation: The medium is sterilised at 121 °C for 15 minutes (or HTST equivalent). Coco Glucoside and its extracts are added later via sterile filtration and are therefore not exposed to heat.

[0090] Inoculation: Lactobacillus spp. (e.g. L. plantarum I L. rhamnosus, etc.) are inoculated with an initial inoculum of 1-3% v / v.

[0091] Conditions:

[0092] Temperature: 34-37 °C

[0093] Duration: 12-24 hours (to achieve a target pH > 4.5)

[0094] Oxygen: Microaerophilic / anaerobic; closed fermenter + inert gas purging (optional). Agitation: 100-200 rpm

[0095] Monitoring: pH (continuous), optical density, sample CFU (every 4-6 hours). CFU target: > 109CFU / mL.

[0096] Carrier (optional for drying): Add maltodextrin 20-40 g / L and homogenise for 15 minutes.

[0097] The complex KH / starch and buffer capacity in the formulation help maintain the final pH at > 4.5 by limiting the pH drop rate.

[0098] Separation and Conditioning

[0099] If desired, the cell / supernatant ratio can be adjusted: the complete suspension can be dried directly, or the solid content can be increased by light separation (e.g. 3-5,000 g, 10 min).

[0100] Solid content adjustment: For spray drying, a total solid content of 12-20% is targeted; for vacuum / lyophilisation, 8-15% is targeted.

[0101] Viscosity: A range of 100-800 mPa s is ideal for nozzle spraying (dilute if necessary).

[0102] Drying

[0103] A) Spray Drying

[0104] Inlet air: 160-180 °C Exit temperature: 80-90 °C

[0105] Feed flow rate: 0.5-1.0 L / h (example for a 10 L batch)

[0106] Atomisation: 0.7-1.5 mm nozzle or disc atomiser; air / liquid ratio adjusted to ensure stable flow.

[0107] Target final moisture content: < 4%; water activity aw < 0.30.

[0108] B) Freeze-Drying (Lyophilisation)

[0109] Pre-freezing: -40 °C, 2-4 hours

[0110] Main drying: -20 °C, 100-200 mTorr, 18-24 s

[0111] Secondary drying: +20 °C, 100-200 mTorr, 6-8 s

[0112] Target moisture: < 3%; porous structure, rapid rehydration.

[0113] C) Vacuum Drying

[0114] Temperature: 45-55 °C

[0115] Pressure: 100-200 mbar

[0116] Duration: 8-12 s (depending on batch thickness)

[0117] Target moisture content: < 4%; high colour / active content retention.

[0118] Final Product Conditioning and Packaging

[0119] Sieve analysis (optional): Adjusted to an 80-200 mesh range (depending on application).

[0120] Packaging: Multi-layered, moisture-barrier bags purged with N2; 10-25 kg.

[0121] Storage: 15-25 °C; RH < 50%; protected from direct sunlight.

[0122] Quality Control and Performance Criteria

[0123] Microbiology: Total viable count (CFU / g or mL); target > 109CFU / mL (liquid) or equivalent CFU / g (powder) depending on formulation.

[0124] pH profile: Initial 6.0 ± 0.2; end of fermentation > 4.5.

[0125] Moisture / aw: Moisture < 3-4%; aw < 0.30. Chemical composition: KH and protein; phenolic / flavonoid (optional), minerals (Mn, Mg, Zn, Ca).

[0126] Solubility / Rehydration: Complete dissolution in 1:10 (w / v) at 20-25 °C within < 2 min. Stability (accelerated): 40 °C / 75% RH, 1-3 months; CFU and moisture monitoring. Variations

[0127] Grain alternatives: Rice, millet, quinoa, buckwheat.

[0128] Organic acid combination: Citric + lactic + malic

[0129] Surfactant equivalent: Coco Glucoside equivalent non-ionic plant-based surfactants. Target microorganisms: Bifidobacterium / Streptococcus / Bacillus / Saccharomyces / Kluyveromyces

[0130] Buffer salt level: Can be optimised within the range of 0.05-0.5% w / w.

[0131] This formulation can be used not only in the cosmetics industry, but also in various sectors such as food supplements, functional foods, pharmaceuticals, agriculture and animal husbandry for the production of fermentation substrates, biostimulants, fertilisers that support soil microbiomes, or postbiotics.

[0132] Consequently, the invention offers a functionally rich vegan nutrient solution that eliminates the problems caused by animal-derived ingredients, is more environmentally and economically sustainable, and has broad microbial compatibility. The grains present in the vegan medium of the current invention provide bacteria with energy for a longer period due to their rich polysaccharide and fibre content. This allows bacteria to grow and develop more steadily. While simple sugars such as glucose in MRS media are a quick source of energy, the complex carbohydrates in your vegan medium are a more sustained source of nutrition. This results in a higher number of bacterial colonies per unit of time.

[0133] The vegan feed medium in question contains grains and easily digestible and absorbable plant proteins. It has been observed that the protease and peptidase enzymes produced by Lactobacillus bacteria effectively hydrolyse plant proteins in both ruminants and humans, thereby enabling their metabolism by bacteria. While animal protein sources are present in MRS feed, the different structure and digestion of various proteins in the vegan feed subject to the invention help Lactobacillus adapt more quickly.

[0134] The cereals contained in the vegan medium of the invention have natural prebiotic properties. This supports the growth of Lactobacillus species and helps them produce beneficial metabolites.

[0135] The components of the vegan culture medium of the invention contain fewer stress factors than MRS. While culture media such as MRS contain chemical salts of minerals and vitamins, the minerals and vitamins in the vegan culture medium of the invention are naturally present. This causes Lactobacillus bacteria to experience less stress during their growth and adaptation processes and, consequently, to develop more rapidly.

[0136] The different applications of the invention are detailed below and are defined in the claims; they should not be understood as limited to the examples given in the detailed description section of the invention.

[0137] A vegan culture medium for the cultivation and growth of beneficial microorganisms, characterised by comprising,

[0138] - at least one grain or grain derivative to provide the necessary nutrients for microorganisms,

[0139] - at least one surfactant, and

[0140] - at least one organic acid as a pH regulator and contaminant inhibitor.

[0141] Being a vegan medium, its characteristic is that the cereal or cereal derivatives are selected from a list containing at least one of wheat, barley, rye, maize, rice, oats, rice, oats, millet, quinoa or buckwheat in the form of flour, extract or hydrolysate.

[0142] It is a vegan-friendly product, characterised by the presence of Coco Glucoside as the plant-based surfactant in its composition at a concentration of 0.8% to 1.4% by weight, or the selection of other non-ionic surfactants or mixtures thereof that are functionally equivalent.

[0143] A vegan-friendly product whose defining characteristic is that it contains organic acids selected from a list comprising citric acid, lactic acid, and malic acid at a concentration of 0.1% to 0.5% by weight, or a combination of two or more acids selected from this list.

[0144] It is a vegan nutrient medium, characterised by an initial pH value in the range of 2.5-9.0 or an initial pH value of 6.

[0145] A vegan medium characterised by a final pH value remaining within the range of 2.5-7.0 or being 4.5 or higher.

[0146] A vegan nutrient solution characterised by the ability to be prepared in liquid, semisolid, or powder form.

[0147] A vegan feed medium characterised by containing one or more selected from a list of manganese, magnesium, zinc, calcium minerals and / or at least one vitamin from the E and B groups.

[0148] A vegan feed, characterised by containing a formulation of cereal-derived prebiotic components, plant-based surfactants, organic acids, and naturally occurring minerals and vitamins that are conducive to balancing the skin microbiome, strengthening the barrier, and exhibiting anti-ageing effects.

[0149] It is a vegan product, characterised by containing beta-glucan, inulin or arabinoxylan as a prebiotic fibre source.

[0150] It is a vegan product whose feature is that it contains one or more antimicrobial plant extracts selected from a list containing rosemary, thyme, garlic, tea tree, mint, eucalyptus, ginger, clove, lavender, cinnamon, sage, sage, fennel and turmeric to reduce the risk of pathogenic microbial contamination.

[0151] A vegan medium characterised by containing grain / plant extracts rich in phenolic compounds or flavonoids to increase antioxidant capacity.

[0152] It is a vegan culture medium, characterised by containing grains that have undergone enzyme pre-treatment, such as amylase and protease, to accelerate the growth of Lactobacillus.

[0153] A vegan medium whose characteristic is that it contains components that support the growth of probiotic microorganisms such as Bifidobacterium, Streptococcus or Bacillus species. A vegan culture medium whose characteristic is that it contains components that support the growth of yeast species such as Saccharomyces or Kluyveromyces.

[0154] A vegan medium characterised by containing maltodextrin or starch-based carriers to increase the viability of probiotics.

[0155] A vegan medium whose characteristic is that its pH buffering capacity is enhanced with natural mineral salts such as calcium carbonate or magnesium phosphate.

[0156] The production method of the vegan medium, characterised by:

[0157] - subjecting the cereal components to enzymatic hydrolysis using enzymes such as amylase and / or protease,

[0158] - subjecting the enzymatically hydrolysed mixture to controlled fermentation,

[0159] - drying the product obtained from fermentation using one of the spray drying, freeze drying or vacuum drying methods,

[0160] characterised by these process steps.

[0161] It is a production method characterised by the sterilisation of the vegan medium using at least one of the following methods: pasteurisation, autoclaving, UV light or membrane filtration.

[0162] The use of vegan culture medium as a fermentation substrate in biotechnological production processes.

[0163] The use of postbiotic content obtained as a result of fermentation of vegan culture medium as an active ingredient in cosmetics, food, food supplements, pharmaceuticals, agriculture or livestock products.

[0164] The use of vegan culture media for the production of probiotic products in the food, cosmetics, food supplement, agriculture and livestock sectors.

[0165] The use of vegan feedstock for the production of biofertilisers, plant probiotics and biostimulants in the agriculture sector.

[0166] The use of vegan feed as a probiotic or postbiotic feed additive in livestock farming. The use of vegan culture medium in cosmetic products to balance the skin microbiome, strengthen the barrier and / or provide anti-ageing effects. The use of vegan feed in the food sector in yoghurt, kefir, beverages or powdered supplement products.

[0167] The use of vegan feed in the agricultural sector as a foliar-applied bio stimulant or soil microbiome-supporting fertiliser.

[0168] The scope of protection of the invention is specified in the claims provided in the annex and cannot be limited to the examples described in this detailed description. It is clear that a person skilled in the art could devise similar structures in light of the above description without departing from the main theme of the invention.

Claims

CLAIMS1. A vegan culture medium for the cultivation and growth of beneficial microorganisms, comprising;- at least one grain or grain derivative to provide the necessary nutrients for microorganisms,- at least one surfactant, and- at least one organic acid as a pH regulator and contaminant inhibitor.

2. The vegan culture medium according to claim 1 , characterised in that the cereal or cereal derivative is selected from the list comprising wheat, barley, rye, maize, rice, oats, millet, quinoa or buckwheat, in the form of flour, extract or hydrolysate.

3. The vegan culture medium according to anyone of the preceding claims, characterised by the presence of Coco Glucoside as the plant-based surfactant in the composition at a weight percentage of 0.8% to 1.4%, or the selection of other non-ionic surfactants or mixtures thereof that are functionally equivalent.

4. The vegan culture medium according to any one of the preceding claims, characterized in that it comprises an organic acid selected from the group comprising citric acid, lactic acid, and malic acid, or a combination of two or more thereof, in an amount ranging from 0.1 % to 0.5% by weight.

5. The vegan culture medium according to anyone of the preceding claims, characterised by an initial pH value in the range of 2.5 to 9.0 or an initial pH value of 6.

6. The vegan culture medium according to anyone of the preceding claims, characterised by a final pH value remaining within the range of 2.5 - 7.0 or being within a range of 4.5 or higher.

7. The vegan culture medium according to anyone of the preceding claims, characterized in that it is preparable in liquid, semi-solid, or powder form.

8. The vegan culture medium according to anyone of the preceding claims, characterised by containing one or more selected from the list of minerals includingmanganese, magnesium, zinc, calcium, and / or at least one vitamin from the E and B groups.

9. The vegan culture medium according to anyone of the preceding claims, characterised by containing a formulation of cereal-derived prebiotic components, plant-based surfactants, organic acids, and naturally occurring minerals and vitamins that are suitable for balancing the skin microbiome, strengthening the barrier, and exhibiting anti-ageing effects.

10. The vegan culture medium according to anyone of the preceding claims, characterised by containing beta-glucan, inulin or arabinoxylan as a prebiotic fibre source.

11. The vegan culture medium according to anyone of the preceding claims, characterized in that it comprises one or more antimicrobial agents selected from rosemary, thyme, garlic, tea tree, mint, eucalyptus, ginger, clove, lavender, cinnamon, sage, fennel, and turmeric, thereby reducing the risk of pathogenic microbial contamination.

12. The vegan culture medium according to anyone of the preceding claims, characterised by containing grain / plant extracts rich in phenolic compounds or flavonoids to increase antioxidant capacity.

13. The vegan culture medium according to anyone of the preceding claims, characterised by containing grains pre-treated with enzymes such as amylase and protease to accelerate Lactobacillus growth.

14. The vegan culture medium according to anyone of the preceding claims, characterized in that it comprises components that promote the growth of probiotic microorganisms, including species of Bifidobacterium, Streptococcus, or Bacillus.

15. The vegan culture medium according to anyone of the preceding claims, characterised by containing components that support the growth of yeast species such as Saccharomyces or Kluyveromyces.16 The vegan culture medium according to anyone of the preceding claims, characterised by containing maltodextrin or starch-based carriers to enhance the viability of probiotics.

17. The vegan culture medium according to any one of the preceding claims, characterized in that its pH buffering capacity is enhanced by the inclusion of natural mineral salts, such as calcium carbonate or magnesium phosphate.

18. A method for producing a vegan culture medium according to anyone of claims 1 to 17, comprising the steps:- subjecting the cereal components to enzymatic hydrolysis using enzymes such as amylase and / or protease,- subjecting the enzymatically hydrolysed mixture to controlled fermentation,- drying the product obtained from fermentation using one of the spray drying, freeze-drying or vacuum drying methods.

19. A method according to Claim 18, characterised in that the vegan culture medium is sterilised by at least one of pasteurisation, autoclaving, UV light or membrane filtration.

20. The use of vegan culture medium according to anyone of Claims 1 to 17 as a fermentation substrate in biotechnological production processes.

21. The use of a postbiotic composition obtained by fermentation of the vegan culture medium according to any one of claims 1 to 17 as an active ingredient in cosmetic, food, food supplement, pharmaceutical, agricultural, or livestock products.

22. The use of vegan culture medium according to anyone of claims 1 to 17 for the production of probiotic products in the food, cosmetic, food supplement, agriculture and livestock sectors.

23. The use of vegan culture medium according to anyone of claims 1 to 17 for the production of biofertilisers, plant probiotics and biostimulants in the agriculture sector.

24. The use of vegan culture medium according to anyone of claims 1 to 17 as a probiotic or postbiotic feed additive in livestock farming.

25. The use of vegan culture medium according to anyone of claims 1 to 17 in cosmetic products to balance the skin microbiome, strengthen the barrier and / or provide antiageing effects.

26. The use of vegan culture medium according to anyone of claims 1 to 17 in the food industry in yoghurt, kefir, beverages or powdered supplement products.

27. The use of vegan culture medium according to anyone of Claims 1 to 17 as a biostimulant applied to leaves in the agricultural sector or as a fertiliser supporting the soil microbiome.