Composition containing probiotic supernate

Abstract

The present invention relates to beverage or food compositions or formulations comprising culture supernatant components from probiotic microorganisms, such as bacteria and fungi. The invention also relates to methods of making and using such compositions, for example as dietary supplements, medicaments or stimulants.

Description

Technical Field

[0001] The present invention relates to beverage or food compositions or formulations comprising ingredients from culture supernatants of probiotic microorganisms (such as bacteria and fungi), and also to methods of making and using such compositions, for example as dietary supplements, medicines or stimulants.

[0002] In particular, the present invention relates to beverage and food compositions or formulations comprising molecules secreted by probiotic microorganisms, molecules shed from their surfaces, and a water component from the culture supernatant, which corresponds to the cell-free portion of the probiotic culture. Such compositions, or simply the supernatant (including the secreted or shed molecules, which may or may not contain water), can be widely used in a variety of fields, such as improving consumer hydration, metabolism, and overall health, as well as recovering water-rich byproducts (i.e., supernatants) from the probiotic industry. However, it should be understood that the present invention is not limited to the aforementioned specific areas of use. Background Art

[0003] Background on water waste

[0004] The following discussion of the prior art is intended to place the present invention in an appropriate technical context and enable the advantages of the present invention to be more fully understood. However, it should be understood that any discussion of the prior art in this specification should not be taken as an admission, express or implied, that the prior art is widely known, or is common technical knowledge in the field in Australia or any other jurisdiction, or that the prior art can be reasonably combined and used by a person skilled in the art.

[0005] Approximately 70% of the Earth's surface is covered by water, but only about 3% is freshwater, and 65% of this remains frozen in high mountains and polar regions. Water resources available for drinking, agricultural irrigation, personal hygiene, and services are extremely scarce, making water shortages a major global concern. Consequently, international organizations have warned that by 2025, over 65% of the world's population could face water shortages. To make matters worse, the global population continues to grow at an alarming rate, and surrounding ecosystems are under pressure from rapidly changing climate patterns. Once-fertile regions are now experiencing chronic water shortages and droughts.

[0006] Globally, 1.1 billion people lack access to clean water, and another 2.7 billion face water shortages for at least one month each year. This situation contributes to a wide range of problems, including malnutrition, social injustice, conflict, and the spread of disease, which affects 2.4 billion people and kills at least one million people annually, primarily children.

[0007] Therefore, there is a pressing need for improved methods of recycling wastewater into consumable products.

[0008] Background related to probiotic cells and molecules

[0009] Microbial probiotics, which can be consumed through fermented foods or supplements, primarily function to restore and maintain the balance of the intestinal microbiome in humans and animals. This role of probiotics has been linked to a variety of health benefits, while an imbalance or absence of probiotics is increasingly being linked to the development of disease in both humans and animals.

[0010] Globally, probiotics are commonly consumed through food, beverages, and supplements. Currently, the industry is investigating the potential use of therapeutic probiotics as pharmaceuticals, with strong clinical efficacy claims supported by scientific evidence. Against this backdrop, the probiotic industry is working to accelerate and diversify the commercialization of probiotic mixtures, which have traditionally been available in the form of live probiotic cells or inactive, non-replicating probiotic extracts.

[0011] Probiotic cells are widely believed to have a variety of health benefits, including weight loss, digestive health, immune and endocrine function, etc. Literature research further shows that the main health benefits associated with probiotics include:

[0012] 1) Helps ensure and / or restore the balance of beneficial bacteria in the digestive systems of humans and animals. If this balance is imbalanced, the intestines or other organs and tissues tend to be colonized and / or occupied by pathogenic microorganisms and their molecules, which is usually caused by factors such as disease, antibiotic intake, and poor diet.

[0013] 2) Helps prevent and treat diarrhea and reduce its severity, especially as a common side effect of antibiotic use that causes an imbalance in intestinal flora. In addition, studies have found that probiotics can also help relieve other types of diarrhea, such as diarrhea caused by infection, traveler's diarrhea, or diarrhea caused by other factors in children and adults.

[0014] 3) Improve certain mental health conditions. This is due to the direct link between mental health and gut health, which has been validated in animal and human studies. In fact, supplementing with probiotic strains for just one month has been found to help alleviate anxiety, depression, autism, obsessive-compulsive disorder (OCD), and memory loss. Furthermore, another six-week study conducted among chemical workers showed that participants who consumed probiotic yogurt or took probiotic capsules daily showed significant improvements in overall well-being, reduced depression and anxiety, and stress relief.

[0015] 4) Certain probiotics can promote heart health, for example by lowering low-density lipoprotein (LDL) levels, cholesterol, and blood pressure. These benefits have been shown to be related to lactic acid bacteria breaking down bile in the intestines, thereby preventing bile from being reabsorbed as cholesterol in the intestines and blood.

[0016] 5) Reduce the severity of certain allergic reactions and eczema, especially in children and infants who consume formula supplemented with probiotics (compared to infants who do not consume probiotics). In addition, infants of women who took probiotics during pregnancy had a greater than 80% lower risk of developing eczema within two years of birth. Specific probiotic strains have also been shown to reduce inflammatory responses in people allergic to milk and / or dairy products.

[0017] 6) Probiotics can also boost the immune system in humans and animals, inhibiting the growth of harmful intestinal bacteria and modulating immune responses through stimulatory mechanisms, inducing the production of IgA-secreting cells, T lymphocytes, and natural killer cells. In fact, this effect has been shown to reduce the incidence and duration of respiratory infections, as well as the risk of urinary tract infections (UTIs) in women.

[0018] 7) Finally, probiotics can help promote the burning of body fat or prevent its accumulation. This effect is achieved by preventing the absorption of dietary fat in the intestines. Unabsorbed fat is then excreted in the feces instead of being stored in the body. Probiotics are also believed to help consumers extend the time between meals, thereby increasing calorie expenditure, reducing intake, and thus reducing fat storage. Notably, this is partly due to the upregulation of hormones (such as GLP-1 in the human body).

[0019] Despite extensive research evidence, there are currently no studies or commercial products specifically targeting the probiotic secretome (which we refer to here as the supernatant, postbiotic functional mixture, or water-rich fraction isolated from probiotic cells).

[0020] Background on dietary protein

[0021] Since the early 2000s, the muscle-building benefits of consuming a high-protein diet have become increasingly well-documented in the literature. Despite this, dietary protein intake patterns among adults in the United States, Europe, and Australia have remained largely unchanged. This may be due to several factors, such as:

[0022] Misconceptions associated with high-protein diets;

[0023] Incorrect or inefficient intake methods;

[0024] Lower protein quality; and

[0025] Misconceptions about the potential negative effects of high-protein diets.

[0026] The global market has clearly demonstrated a demand for increased protein supply, with traditional protein sources including animal and plant proteins. However, both protein sources are associated with greenhouse gas emissions, water consumption, land use, and environmental dependence, impacting their sustainability. Furthermore, a diet consisting solely of plant protein has been shown to reduce essential amino acid intake (compared to animal protein), which ultimately reduces protein synthesis in the body and increases the risk of hypoproteinemia due to insufficient methionine and lysine intake.

[0027] New technologies currently under consideration include edible insects and lab-grown meat, but these still require further economic optimization and scientific validation. Furthermore, mycoprotein has been commercially trialed for two decades (e.g., under the Korn brand), but the proportion of fungal protein used in pre-packaged meals is only 15% to 43%, meaning that the global protein supply remains largely dependent on traditional agricultural systems.

[0028] The object of the present invention is to overcome or improve one or more disadvantages of the prior art, or at least provide a beneficial alternative. Summary of the Invention

[0029] The present invention relates to beverage or food compositions or formulations comprising ingredients from culture supernatants of probiotic microorganisms (such as bacteria and fungi). Also disclosed are methods for making and using the compositions, for example as dietary supplements, medicines or stimulants.

[0030] Thus, in a broad sense, the present invention relates to a composition, such as a beverage or food composition, comprising water derived at least in part from at least one culture supernatant of at least one probiotic microorganism, and a base.

[0031] In a first aspect, the present invention relates to a composition comprising:

[0032] Water, wherein the water is at least partially derived from at least one culture supernatant of at least one probiotic microorganism; and

[0033] A substrate.

[0034] Suitably, the base comprises a protein source, a carbohydrate source, a lipid source and / or a sweetener.

[0035] In certain embodiments, the protein source is or includes protein of non-animal origin. In specific embodiments, the carbohydrate source is or includes carbohydrates of non-animal origin. In various embodiments, the lipid source is or includes lipids of non-animal origin. To this end, the protein source, carbohydrate source and / or lipid source are suitably at least partially derived from plants. More specifically, the protein source, carbohydrate source and / or lipid source can be at least partially derived from: plant extracts; fruit extracts, such as fruit powder or juice; flower extracts, such as pollen; and / or seed extracts, such as seed powder, seed paste or seed oil (e.g., essential oil, flavor oil).

[0036] Suitably, the protein source, carbohydrate source and / or lipid source is selected from the group consisting of cocoa powder, cocoa bean powder, whey powder, milk powder, cocoa butter, vegetable oil, coconut milk, lemon powder, honey powder, orange powder, berry powder, roselle pollen, Australian native bush food (fruit and flower) powder, tropical fruit and flower powder, and any combination thereof, wherein the powdered ingredient may be replaced by fruit puree, mixed jam, extract, essence, tincture, liquid, juice and / or concentrate.

[0037] In certain embodiments, the composition is a substantially vegetarian and / or vegan composition. To this end, the composition is suitably substantially free of ingredients of animal origin.

[0038] Suitably, the composition comprises from about 0.1% to about 99% by weight of at least one culture supernatant from at least one probiotic microorganism.

[0039] Suitably, the composition comprises from about 0.1% to about 99% by weight of the substrate.

[0040] Suitably, the composition comprises from about 0.1% to about 99% by weight of a protein source, a carbohydrate source and / or a lipid source.

[0041] Suitably, the composition comprises from about 0.1% to about 30% by weight of the sweetener.

[0042] In certain embodiments, the composition further comprises one or more of the following ingredients: pH adjusters, preservatives, solvents, emulsion stabilizers, colorants, flavors, vitamins, and minerals.

[0043] In certain embodiments, the composition and / or supernatant and / or supernatant molecule can be in the form of a supplement or used as a supplement. More specifically, the composition can be a protein supplement, a vitamin supplement, a carboxylic acid supplement, an amide supplement, an amine supplement, or other products comprising the composition and / or supernatant and / or supernatant molecule.

[0044] In certain embodiments, the composition and / or supernatant and / or supernatant molecules can be in the form of a drug or stimulant, or used in a drug or stimulant. More specifically, the composition can stimulate gene expression, modulate hormone production, or alter a molecular profile in a consumer.

[0045] In certain embodiments, the composition and / or supernatant and / or supernatant molecules may be in the form of an ingredient, or used as an ingredient. More specifically, the composition may comprise soft or clinical efficacy claims validated by scientific evidence and literature mining.

[0046] In certain embodiments, the composition and / or supernatant and / or supernatant molecules can be in the form of, or used as, an additive or processing aid. More specifically, the composition can extend the shelf life of foods, beverages, pharmaceuticals, cosmetics, and other products containing the composition and / or supernatant and / or supernatant molecules.

[0047] In various embodiments, the composition is substantially free of gluten.

[0048] In certain embodiments, the composition is substantially free of lactose.

[0049] In a preferred embodiment, the composition of this aspect of the invention comprises: secreted molecules and shed molecules derived at least in part from at least one culture supernatant of at least one probiotic microorganism and / or probiotic cell structures that do not constitute living cells; and water, wherein the water is at least partially derived from at least one culture supernatant of at least one probiotic microorganism; and a substrate.

[0050] In a second aspect, the present invention relates to a method for preparing a composition comprising the steps of mixing water (wherein the water is at least partially derived from at least one culture supernatant of at least one probiotic microorganism) with a base, thereby preparing the composition.

[0051] In a preferred embodiment, the present invention provides a method for preparing a composition, comprising the steps of:

[0052] Water, wherein the water is at least partially derived from at least one culture supernatant of at least one probiotic microorganism; and

[0053] a supernatant, wherein the supernatant comprises molecules and water derived at least in part from at least one culture of at least one probiotic microorganism; and

[0054] a supernatant molecule, wherein the molecule is isolated and / or purified and / or extracted and is at least partially derived from at least one culture supernatant of at least one probiotic microorganism; and

[0055] a substrate;

[0056] Thus, a composition is prepared.

[0057] Suitably, the composition is the composition described in the first aspect.

[0058] In a third aspect, the present invention provides a composition prepared according to the method of the second aspect.

[0059] Suitably, the compositions of the first and third aspects are used for:

[0060] (i) Supplement;

[0061] (ii) improve health; and / or

[0062] (iii) preventing and / or treating a disease, disorder or condition in a subject.

[0063] In certain embodiments, the disease, disorder or condition is selected from the group consisting of hypertension, heart disease, anxiety, obesity, neurodegenerative diseases, Alzheimer's disease, cancer, immunodeficiency, premature aging, wrinkles, inflammation, gastrointestinal dysfunction, arthritis, osteoarthritis, allergies, molecular and gene regulatory abnormalities, gut microbiota imbalance, hormone imbalance, vitamin deficiency, and any combination thereof.

[0064] In a fourth aspect, the present invention relates to a method for supplementing an individual, comprising administering to the individual a therapeutically effective amount of the composition of the first aspect and the third aspect, thereby achieving supplementation of the individual.

[0065] In a fifth aspect, the present invention relates to a method for improving or maintaining the health of an individual, comprising administering to the individual a therapeutically effective amount of the composition of the first and third aspects, thereby improving or maintaining the health of the individual.

[0066] In certain embodiments, improving individual health comprises at least one of the following: reducing body weight, controlling blood sugar, increasing the number of intestinal villi per unit area, increasing the average length of intestinal villi, promoting a healthier intestinal mucosal barrier, enhancing resistance to mechanical stress, preventing loss of lean body mass and / or promoting weight gain during growth, improving efficiency in converting food into energy and muscle, preventing or reducing abdominal fat accumulation, normalizing blood biochemical parameters through the intake of probiotics, and / or improving the intestinal biological environment (optionally, even after the intake of foods rich in gluten and fat).

[0067] In certain specific embodiments, the present invention may demonstrate the potential of probiotics to contribute to global sustainable protein growth, which is particularly important since dietary protein has been shown to be essential for health, particularly during growth and aging.

[0068] In certain embodiments, probiotic cells may also be present to enhance the effects of supernatant and / or secreted / shed molecules contained in the supernatant, wherein the concentration of the cells in the composition, ingredient, supplement, medicament, food additive, or other application thereof is: 1×10 5 , 1×10 6 , 1×10 7 , 1×10 8 , 1×10 9 , 1×10 10 , 1×10 11 , 5×10 5 , 5×10 6 , 5×10 7 , 5×10 8 , 5×10 9 , 5×10 10 , 5×10 11 , or any value between 1×10 5 With 5×10 11 CFU / recommended dose.

[0069] Suitably, administration of the composition results in a significant change in the microbiome. Preferably, the significant change in the microbiome comprises an increase in at least one of the following microorganisms: Anaerococcus, Romboutsia, Lachnospiraceae, Lactobacillus or Bacillus, but is not limited to these microorganisms. Preferably, the significant change in the microbiome comprises a decrease in at least one of the following microorganisms: Escherichia coli, Shigella, Clostridium, Salmonella, and other pathogenic bacteria, including coliform and non-coliform microorganisms.

[0070] In a sixth aspect, the present invention provides a method for treating and / or preventing a disease, disorder or condition in an individual, the method comprising administering to the individual a therapeutically effective amount of the composition described in the first and third aspects, thereby treating and / or preventing the disease, disorder or condition in the individual.

[0071] Suitably, the disease, disorder or condition is selected from the group consisting of hypertension, heart disease, anxiety, obesity, neurodegenerative disease, Alzheimer's disease, cancer, immunodeficiency, premature aging, wrinkles, inflammation, gastrointestinal dysfunction, arthritis, osteoarthritis and any combination thereof.

[0072] In a seventh aspect, the present invention provides a method for extending the shelf life of a beverage, food, drink, cosmetic or other product containing the content of the present invention, the method comprising adding an effective amount of the composition, supernatant or supernatant molecule described in the first aspect, thereby preventing and / or reducing and / or slowing down and / or delaying and / or preventing deterioration.

[0073] Suitably, the spoilage is caused by a group of biological agents comprising bacteria, fungi, viruses, parasites, invertebrates, algae or protozoa and any combination thereof.

[0074] Suitably, deterioration is caused by a group of abiotic factors including light, gases, temperature, surfaces, molecules and any combination thereof.

[0075] In relation to each of the above aspects, the individual is suitably a human or other animal.

[0076] Unless otherwise indicated, the terms "comprises," "comprising," and their variations are used in this specification in an inclusive, rather than exclusive, manner, so that the element or group of elements described may include one or more other elements or groups of elements not stated, such as other additives, components, elements, or steps. In contrast, the terms "consisting of," "consisting only of," and their variations are used in an exclusive manner, meaning that the element or group of elements described is required or mandatory, and no other elements are allowed to be present.

[0077] The term "consisting essentially of" means that the stated element or group of elements is necessary or mandatory, but other components that do not interfere with or affect the activity or function of the element or group of elements are optional.

[0078] In this specification, the indefinite article "a" or "an" may refer to one entity or multiple entities and should not be construed as being limited to a single entity. For example, "an individual" may refer to one individual, one or more individuals, or multiple individuals.

[0079] In this specification, the term "about" indicates a variation or tolerance to the stated value, concentration, ratio or proportion, and is preferably defined as not more than 10%, 5%, 2% or 1% above or below the stated value, concentration, ratio or proportion. BRIEF DESCRIPTION OF THE DRAWINGS

[0080] Preferred embodiments of the present invention will now be described, by way of example only, with reference to the accompanying drawings, in which:

[0081] Figure 1-1The benefits of the secretome of a probiotic fungus belonging to the family Sclerodermataceae, achieved by a composition comprising a supernatant and an additional 5% whey protein, further enhanced by the presence of living microbial cells that continuously produce molecules of the supernatant and by the presence of spores that render eukaryotic cells more responsive to treatment:

[0082] a) Animal weight;

[0083] b) animal body fat and blood sugar;

[0084] Figure 2 - Independent histological analysis of small intestinal villi:

[0085] a) Untreated rats;

[0086] b) Rats treated with a dose of 5×10 7 Probiotic fungus (Truffleaceae strain TF18), magnification 40X.

[0087] Figure 3-1 The secretome of a probiotic lactic acid bacterium (Lactococcus lactis strain QF16) promotes weight gain and is further enhanced by the presence of living cells that continuously secrete molecules that constitute the supernatant:

[0088] a) and b) low-concentration treatment groups increased body weight by 6.5% with all animals consuming the same amount of feed;

[0089] c) The body weight gain rate of the low concentration treatment group was also faster. DETAILED DESCRIPTION

[0090] The present inventors have created a composition comprising molecules, molecular structures, and water obtained from the culture supernatant used to cultivate probiotic microorganisms. Thus, the composition can effectively reduce water waste in the probiotic industry while utilizing the beneficial molecules secreted by the probiotic microorganisms and contained in the culture supernatant. The compositions described herein can be designed to supplement nutrition, improve health, and / or treat and / or prevent a range of diseases, disorders, or conditions in individuals. The compositions are suitably in the form of dietary supplements, fortified foods, pharmaceuticals, and / or stimulants.

[0091] In a first aspect, the present invention relates to a composition comprising, consisting of, or consisting essentially of:

[0092] Water, wherein the water is at least partially derived from a culture supernatant of at least one probiotic microorganism; and

[0093] A substrate.

[0094] The term "culture supernatant" refers to the cell-free liquid portion remaining after cells are grown in culture and separated by, for example, centrifugation, filtration, sedimentation, or other methods known in the art.

[0095] Typically, when a probiotic microorganism (such as the microorganism described herein) is cultured in a culture medium, specific substances or molecules (e.g., proteins, polysaccharides, lipids) produced by the probiotic microorganism and present in the cell are released or secreted. Therefore, the culture supernatant may contain a mixture of soluble components or metabolites present in the culture medium. It is foreseeable that the culture supernatant may be taken from one or more growth stages (e.g., "lag phase," "logarithmic phase" (i.e., "exponential phase"), "stationary phase," or "decline phase" (also known as "logarithmic decline phase")) during the probiotic batch culture process. In certain embodiments, the composition comprises a culture supernatant from the exponential growth phase of the probiotic batch culture process.

[0096] When the supernatant of a probiotic culture is used, the supernatant can be used directly or can be subjected to one or more purification steps before being added to the composition, for example to concentrate or isolate active additives or metabolites therein.

[0097] As generally used in this specification, the term "probiotic" or "probiotic microorganism" refers to one or more live microorganisms that can confer a health benefit on an animal when administered at an appropriate dose. Such health benefit is generally due to the probiotic beneficially regulating the gastrointestinal microbial balance or flora of the animal. Suitably, the probiotic microorganism is a bacterium or a fungus. In a broad sense, the probiotic microorganism can be selected from the following genera: Lactococcus, Sclerodermataceae, Lactobacillus, Bifidobacterium, Enterococcus, Streptococcus, Bacillus, Propionibacterium, Pediococcus, Clostridium, Aspergillus, Candida, Saccharomyces and Megasphaera, but is not limited thereto. In a preferred embodiment, the probiotic microorganism is Lactococcus lactis strain QF16 or Sclerodermataceae strain TF18.

[0098] Lactococcus lactis strain QF16 is a probiotic bacterium that is a safe lactic acid producer and traditionally used by the dairy industry. The strain described in the present invention is innovative because it contains natural antioxidant and metabolic genes.

[0099] Truffle Fungus strain TF18 is a probiotic fungus. This strain is a natural filamentous fungus with anti-cancer properties.

[0100] As used herein, the term "base" refers to a basic composition that can be used to prepare a beverage or food. For example, the base can be a fruit, vegetable or dairy base, to which ingredients such as culture supernatant and other additives can be added to form the composition.

[0101] Suitably, the substrate comprises a protein source, a carbohydrate source and / or a lipid source. In certain embodiments, the substrate is or comprises: a protein source; a carbohydrate source; a lipid source; a protein source and a carbohydrate source; a protein source and a lipid source; a carbohydrate source and a lipid source; or a protein source, a carbohydrate source and a lipid source. It should be understood that two or three of the protein source, carbohydrate source and lipid source can come from a single source, such as those described herein. For example, fruit juice or extract can be used as the source of protein, carbohydrate and lipid in the substrate. Alternatively, the protein source, carbohydrate source and lipid source can also come from different sources, respectively. In addition, the protein source, carbohydrate source and / or lipid source can also come from multiple sources (e.g., two or more fruit juices, powders or extracts).

[0102] As used herein, the term "protein" includes any amino acid polymer composed of D- or L-amino acids, including natural or unnatural amino acids (as known in the art). Proteins, peptides and / or amino acids can be present in extracts, hydrolysates, purified products, chemically synthesized products, enriched products or partially purified forms.

[0103] In certain embodiments, the protein source is obtained, extracted, obtainable, or extractable from a natural source (eg, a plant and / or animal).

[0104] Animal sources include, but are not limited to, vertebrates such as mammals, reptiles, fish, and / or birds, and invertebrates such as shellfish, crab, lobster, and / or insects. Non-limiting examples include, but are not limited to, whey protein concentrate, whey protein isolate, hydrolyzed whey, casein, milk protein isolate, egg white protein, beef, lamb, pork, veal, venison, bison, goat, kangaroo, chicken, turkey, duck, emu, rabbit, fish, shellfish, crab, lobster, and / or crocodile protein.

[0105] Plant sources include, but are not limited to, trees, vines, plant seeds, bark, roots, leaves, bulbs, tubers, nuts and / or fruits. Plants may include, but are not limited to, grasses, grains, beans, seaweed, leafy vegetables, cruciferous vegetables and fungi. Non-limiting examples include soy protein, tofu, tempeh, pea protein, rice protein, bean protein, corn protein, chickpeas, broad beans, mung beans, chia seed protein, quinoa protein, spirulina protein, lentils, nut protein, wheat germ protein, sorghum protein, and proteins obtained, extracted, obtainable or extractable from vegetables and / or fruits, such as spinach, watercress, kale, broccoli, Brussels sprouts, cauliflower, mushrooms, parsley, lettuce, zucchini, cucumber, bell pepper, artichoke, cabbage, celery, eggplant, tomato, onion, beet, pumpkin, squash, potato, yam and / or sweet potato.

[0106] The term "carbohydrate" (or sugar) includes monosaccharides, disaccharides, oligosaccharides and polysaccharides. Generally speaking, smaller (low molecular weight) carbohydrates, such as monosaccharides and disaccharides, are often referred to as sugars.

[0107] As used herein, the term "lipid" refers to a class of organic compounds with lipophilicity or amphiphilicity, including but not limited to fats, fatty oils, essential oils, essential oils, waxes, steroids, sterols, phospholipids, glycolipids and fatty acids. The term "lipid" includes naturally occurring lipids and synthetically produced lipids.

[0108] It is foreseeable that the lipid source can include any edible oil, butter or the like, including natural or synthetic oils or butters, as long as it is suitable for human consumption. Exemplary lipid sources can include vegetable oils and vegetable butters, such as soybean, safflower, sunflower, sesame, peanut, corn, olive, rice bran, rapeseed oil, coconut oil, pomegranate seed oil, palm oil, cottonseed oil, rapeseed, carrot, evening primrose, borage, mustard, citrus seeds, flaxseed, chia seeds, avocado, sugarcane, macadamia, Brazil nuts, almonds, beech nuts, hazelnuts, cashews, pecans, pine nuts, Mongolian nuts, pistachios, walnuts, pumpkin seeds, grapefruit seeds, melon seeds, gourd seeds, blackcurrant, evening primrose, cumin seeds, amaranth seed oil, quinoa oil, almond seed oil, apple seeds, argan, babassu, borage, guava ... The present invention can be carried out using oils such as thuja oil, chestnut oil, chinut oil, carob oil, cocoa oil, tribulus terrestris oil, coriander seed oil, date seed oil, dicca oil, grape seed oil, kapok oil, kenaf oil, larramanthi oil, mefula oil, marula oil, black sesame oil, nutmeg oil, papaya oil, perilla seed oil, persimmon seed oil, pequeño oil, pili nut oil, poppy seed oil, prakasi oil, plum kernel oil, ramtir oil, royal oil, Inca peanut oil, zapote oil, saer oil, shea butter oil, talamira oil, teser oil, thistle oil, Jerusalem artichoke oil, tobacco seed oil, tomato seed oil, wheat germ oil, brassica oil, panax sylvestris oil, kuju oil, neem oil, ohun fruit oil, passion fruit oil, rose hip oil, sea buckthorn oil, viburnum oil, tonga bean oil, ukukhuba oil and meadow foam oil. In addition, the present invention can be carried out using these oils, butters and mixtures thereof or mixtures of some of their components.

[0109] Suitably, one or more of the protein source, carbohydrate source and lipid source may be derived from a non-animal source. In a broad sense, carbohydrates, proteins and lipids of non-animal origin may be obtained or extracted in whole or in part from any non-animal source, such as a microbial source (e.g., bacteria, fungi or algae) and / or a plant source, such as a plant extract.

[0110] The terms "extract", "ingredient" and "derivative" refer to any product that can be obtained from a source such as a plant by downstream processing techniques known in the art, such as a series of extraction and purification methods.

[0111] In certain embodiments, one or more of the various components of the composition, such as the carbohydrate source, protein source, and / or lipid source, may be isolated or substantially purified. For purposes of the present invention, "isolated" or "purified" refers to a substance that has been removed from its natural state or has been artificially manipulated. An isolated or purified substance may be substantially or essentially free of components that normally accompany it in its natural state, or may have been manipulated to be in an artificial state with components that normally accompany it in its natural state. An isolated substance may be in a natural form, a chemically synthesized form, or a recombinant form.

[0112] For example, plant-derived or plant-based carbohydrate, protein and lipid sources can be obtained or extracted from any plant or plant part. Plant-based sources include trees, vines, plant seeds, flowers, bark, roots, leaves, bulbs, tubers, nuts and / or fruits, but are not limited thereto. Plants may include fruits, grasses, grains, legumes, seaweed, leafy vegetables, cruciferous vegetables, mosses and fungi, etc., but are not limited thereto. More specifically, the protein source, carbohydrate source and / or lipid source may be derived at least in part from one or more of the following: plant extracts, vegetable extracts, root extracts, fruit extracts (e.g., fruit powder or juice); flower extracts (e.g., pollen); and / or seed extracts (e.g., seed powder, seed oil or seed paste).

[0113] Thus, in various embodiments, the substrate is or includes a fruit extract and / or a vegetable extract. The fruit and / or vegetable extract can be added to the composition as a liquid form (e.g., juice, liquid concentrate, puree, powder, or other modified form). The liquid in the fruit and / or vegetable can be filtered before use. The fruit extract can include juices and powders from tomatoes, berries, citrus fruits, melons, and / or tropical fruits. A single fruit juice or extract can be used, or a juice / extract mixture can be used. Vegetable extracts can include a variety of vegetable juices, powders, etc. Exemplary fruit and vegetable extracts (e.g., fruit or vegetable juices, fruit or vegetable powders) are available or extracted from various berries, currants, apricots, peaches, nectarines, plums, cherries, apples, pears, oranges, grapefruits, coconuts, lemons, limes, tangerines, mandarins, bananas, pineapples, grapes, tomatoes, rhubarb, prunes, figs, pomegranates, passion fruit, guava, kiwi fruit, kumquats, mangoes, avocados, various melons, papayas, turnips, rutabagas, carrots, cabbage, cucumbers, pumpkins, celery, radishes, native Australian fruits / legumes / vegetables, bean sprouts, alfalfa sprouts, bamboo shoots, beans, and / or seaweed. It will be appreciated that one or more fruits, one or more vegetables, and / or a combination of fruits and vegetables may be added to the composition, depending on the desired flavor.

[0114] In a specific embodiment, the protein source, carbohydrate source and / or lipid source is selected from the group consisting of cocoa powder, cocoa bean powder, whey powder, milk powder, cocoa butter, vegetable oil, coconut milk, lemon powder, honey powder, orange powder, berry powder, hibiscus powder, Australian native wild plant (fruit and flower) powder, tropical fruit and flower powder, and any combination thereof, wherein the powder component can be replaced by fruit puree, paste, extract, essence, wine, liquid, juice and / or concentrate.

[0115] In certain embodiments, the composition is a substantially vegetarian and / or vegan composition. To this end, the composition may be substantially free of animal-derived ingredients.

[0116] As used herein, the term "vegetarian" refers to a product, composition, or formulation, including but not limited to foods, beverages, and supplements, that does not contain, or is not used in the production of, products derived from animals that died, were slaughtered, or were consumed. This can include farmed, wild, or domesticated animals, including but not limited to livestock, poultry, game, fish, shellfish, crustaceans, amphibians, tunicates, echinoderms, molluscs, and insects. For example, if a product contains dairy and eggs, it can still be considered vegetarian.

[0117] As used herein, the term "vegan" refers to a product, including but not limited to foods and supplements, that is produced without the use of animals or animal products (including products derived from live animals) as an auxiliary ingredient.

[0118] In certain embodiments, the compositions of aspects of the present invention are in the form of a supplement, such as a protein supplement. As used herein, the term "supplement" refers to a product intended to supplement a diet of food and water, where the diet is sufficient to sustain life. Supplements can include vitamins, minerals, herbs or other plant-based ingredients, amino acids, enzymes, organ tissues, glandular metabolites, or combinations thereof (e.g., fortified food products).

[0119] In certain embodiments, the composition is in a single-dose form, a single-unit form, or a multi-serving product form. In other embodiments, the composition can be in a concentrated form, such as a syrup, a juice drink, or a powder, which requires the addition of additional water for dilution before ingestion by the subject.

[0120] The terms "concentration", "ratio" or "mass" generally used in this specification, when referring to any specific component or ingredient in a composition, may refer to the concentration, ratio or mass of the specific component or ingredient when used to prepare the composition, or may refer to the concentration, ratio or mass of the component or ingredient in the final product.

[0121] It should also be noted that those skilled in the art can easily adjust the concentration, ratio or mass of each component or ingredient. In addition, it should be understood that the concentration, ratio or mass range expressed in the form of "xy" includes all values and sub-ranges within the range.

[0122] The composition suitably comprises a culture supernatant of at least one probiotic microorganism at a weight percent (wt %) concentration of about 0.1% to about 99%, or any subrange within this range, such as, but not limited to, about 1% to about 75%, about 2% to about 30%, or about 5% to about 20%. In a specific embodiment, the weight percent concentration of the culture supernatant of at least one probiotic microorganism is about 1, 1.25, 1.5, 1.75, 2, 2.25, 2.5, 2.75, 3, 3.25, 3.5, 3.75, 4, 4.25, 4.5, 4.75, 5, 5.25, 5.5, 5.75, 6, 6.25, 6.5, 6.75, 7, 7.25, 7.5, 7.75, 8, 8.25, 8.5 , 8.75, 9, 9.25, 9.5, 9.75, 10, 10.25, 10.5, 10.75, 11, 11.25, 11.5, 11.75, 12, 12.25, 12.5, 12.75, 13, 13.25, 13.5, 13.75, 14, 14.25, 14.5, 14.75, 15, 15.25, 15.5, 15.75, 16, 16.25, 16.5, 1 6.75, 17, 17.25, 17.5, 17.75, 18, 18.25, 18.5, 18.75, 19, 19.25, 19.5, 19.75, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50 98, 99, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100

[0123] The composition suitably comprises a matrix in a weight percent (wt %) concentration of about 1% to about 99%, or any subrange therein, such as, but not limited to, about 1% to about 75%, about 2% to about 50%, or about 5% to about 40%. In certain embodiments, the matrix is present in a weight percent concentration of about 1, 1.25, 1.5, 1.75, 2, 2.25, 2.5, 2.75, 3, 3.25, 3.5, 3.75, 4, 4.25, 4.5, 4.75, 5, 5.25, 5.5, 5.75, 6, 6.25, 6.5, 6.75, 7, 7.25, 7.5, 7.75, 8, 8.25, 8.5, 8.75, 9, 9. 25, 9.5, 9.75, 10, 10.25, 10.5, 10.75, 11, 11.25, 11.5, 11.75, 12, 12.25, 12.5, 12.75, 13, 13.25, 13.5, 13.75, 14, 14.25, 14.5, 14.75, 15, 15.25, 15.5, 15.75, 16, 16.25, 16.5, 16.75, 17, 17.25, 17.5, 17.75, 18, 18.25, 18.5, 18.75, 19, 19.25, 19.5, 19.75, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52 , 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99%, and any interval therein. In a particularly preferred embodiment, the composition comprises from about 10 wt% to about 90 wt% of a culture supernatant of at least one probiotic microorganism.

[0124] Suitably, the composition comprises from about 0.1 wt % to about 99 wt % (e.g., about 0.1, 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 99, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100

[0125] Suitably, the composition comprises from about 0.1 wt % to about 99 wt % (e.g., about 0.1, 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51 , 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99%, and any intervals therein) of a carbohydrate source, such as those described above. In particularly preferred embodiments, the composition comprises from about 1 wt% to about 20 wt% of a carbohydrate source.

[0126] Suitably, the composition comprises from about 0.1 wt % to about 99 wt % (e.g., about 0.1, 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50 , 51 , 52, 53, 54, 55, 56, 57, 58, 59, 60, 61 , 62, 63, 64, 65, 66, 67, 68, 69, 70, 71 , 72, 73, 74, 75, 76, 77, 78, 79, 80, 81 , 82, 83, 84, 85, 86, 87, 88, 89, 90, 91 , 92, 93, 94, 95, 96, 97, 98, 99%, and any intervals therein) of a lipid source, such as those described above. In particularly preferred embodiments, the composition comprises from about 1 wt % to about 10 wt % of a lipid source.

[0127] In certain embodiments, the base further comprises a sweetener.

[0128] As generally used in this specification, the term "sweetener" refers to a natural or artificial compound used to enhance the sweetness of the compositions of the present invention. It should be understood that sweeteners include carbohydrate sweeteners (i.e., sugars and other carbohydrate sweeteners) as well as non-carbohydrate sweeteners. Therefore, a sweetener can also be considered a carbohydrate source, and vice versa. In this specification, the term "sugar" refers to sucrose and its components (e.g., glucose and / or fructose), syrups, malt syrup, maple syrup, starch syrup, glucose syrup, high fructose syrup (e.g., high fructose corn syrup), honey, molasses, and other carbohydrates that can serve as sweeteners or their sources. The term "other carbohydrate sweeteners" refers to, for example, sugar alcohols such as xylitol, maltitol, lactitol, glycerol, and sorbitol. Examples of non-carbohydrate sweeteners suitable for use in the present invention include, but are not limited to, stevia, aspartame (phenylalanine), acesulfame potassium (Ace-K), saccharin, cyclohexane salts, and sucralose. Thus, the sweetener may include low-calorie sweeteners, natural sweeteners, non-nutritive sweeteners and / or artificial sweeteners. In addition, the sweetener may be of natural or synthetic origin.

[0129] Suitably, the sweetener or combination of sweeteners is present in the composition at a concentration of about 0.01 to about 30 wt % (e.g., about 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.25, 1.5, 1.75, 2, 2.25, 2.5, 2.75, 3, 3.25, 3.5, 3.75, 4, 4.25, 4.5, 4.75, 5, 5.25, 5.5, 5.75, 6, 6.25, 6.5, 6.75, 7. %. .5, 6.75, 7, 7.25, 7.5, 7.75, 8, 8.25, 8.5, 8.75, 9, 9.25, 9.5, 9.75, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30 wt %, and any range therein), more preferably about 0.05 to about 20 wt %, or more preferably about 0.1 to about 15 wt %.

[0130] In certain embodiments, the composition further comprises one or more pH adjusting agents, preservatives, solvents, emulsion stabilizers, coloring agents, flavoring agents, amino acids, vitamins, prebiotic fibers, and minerals. In certain embodiments, the composition is substantially free of artificial sweeteners, artificial flavoring agents, and / or artificial coloring agents.

[0131] Suitably, the amino acid can be any amino acid known in the art, including essential and non-essential amino acids, and derivatives or variants thereof, such as amino acid salts. Non-limiting examples include leucine, isoleucine, valine, lysine, glutamine, threonine, phenylalanine, methionine, tryptophan, histidine, taurine, arginine, serine, proline, glycine, alanine, ornithine, citrulline, uroquinone, asparagine, and tyrosine.

[0132] In this embodiment, the composition can include one or more amino acids at a concentration of about 0.1% to about 20% by weight (e.g., about 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.25, 1.5, 1.75, 2, 2.25, 2.5, 2.75, 3, 3.25, 3.5, 3.75, 4, 4.25, 4.5, 4.75, 5, 5.25, 5.5, 5.75, 6, 6.25, 6.5, 6.75, 7, 7.25, 7.5, 7.75, 8, 8.25, 8.5, 8.75, 9, 9.25, 9.5, 9.75, 10, 19.25, 19.5, 19.75, 20), or any range therein, such as, but not limited to, about 1% to about 15%, or about 2% to about 5%.

[0133] It will be appreciated that the coloring agent can be any coloring agent known in the art. For this reason, the food industry uses multiple food-grade coloring agents to enhance the visual appearance of food, dietary supplements and similar products. In known food-grade coloring agents, only a few are of natural origin, such as betalain or chlorophyll. Most food-grade coloring agents are synthetic analogs (i.e., so-called "natural equivalents") or fully synthetic compounds of naturally occurring substances.

[0134] In accordance with the foregoing, the compositions of the present invention may include one or more colorants (e.g., those described above) in a weight percent (wt %) concentration of about 0.1% to about 20% (e.g., about 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.25, 1.5, 1.75, 2, 2.25, 2.5, 2.75, 3, 3.25, 3.5, 3.75, 4, 4.25, 4.5, 4.75, 5, 5.25, 5.5, 5.75, 6, 6.25, 6.5, 6.75, 7, 7.25, 7.5, 7.75, 8, 8.25, 8.5, 8.75, 9, 9.25, 9.5 , 17.25, 17.5, 17.75, 18, 18.25, 18.5, 18.75, 19, 19.25, 19.5, 19.75, 20), or any range therein, for example, from about 1% to about 15%, or from about 2% to about 12%.

[0135] The flavoring agent of the present invention can be any natural or non-natural substance that can impart or enhance flavor. Flavoring agents are generally composed of one or more flavoring substances and their complexes, which can be extracted from nature or artificially produced in liquid or powder form and are used to impart a specific flavor to a product.

[0136] In this embodiment, the composition can include one or more flavoring agents, such as those described above, at a concentration of about 0.1% to about 20% by weight (e.g., about 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.25, 1.5, 1.75, 2, 2.25, 2.5, 2.75, 3, 3.25, 3.5, 3.75, 4, 4.25, 4.5, 4.75, 5, 5.25, 5.5, 5.75, 6, 6.25, 6.5, 6.75, 7, 7.25, 7.5, 7.75, 8, 8.25, 8.5, 8.75, 9, 9.25, 9.5, 9.75, , 18.25, 18.5, 18.75, 19, 19.25, 19.5, 19.75, 20), or any subrange therein, for example, about 1% to about 15%, or about 2% to about 10%.

[0137] Suitably, the composition of the present embodiment may further include one or more vitamins and / or minerals, such as electrolytes. Exemplary vitamins include vitamin A (e.g., vitamin A1 retinol, anti-dry eye hormone, α-carotene, β-carotene, γ-carotene), B vitamins (e.g., B1 vitamins, including thiamine, anti-neuritis hormone, thiamine pyrophosphate, carboxylase; B2 vitamins, including riboflavin, vitamin G, lactoflavin, heparin, lutein, chlorophyll, riboflavin monophosphate, FMN, riboflavin diphosphate, FAD; vitamin B3 (niacin), vitamin B6 (pyridoxine, pyridoxal-5-phosphate), vitamin B7 (biotin), vitamin B12 (cobalamin, methylcobalamin)), vitamin C (ascorbic acid, scorbutic vitamin, dehydroascorbic acid), vitamin D (anti-rickets vitamin, vitamin D2, D3, cholecalciferol, etc.), vitamin E, vitamin K, etc. Non-limiting examples of minerals include sodium, selenium, zinc, magnesium, calcium, iron, manganese, copper, chromium, phosphorus, iodine, potassium, and molybdenum.

[0138] In certain embodiments, the composition further includes a pH adjusting agent. The pH adjusting agent can be any reagent, substance or molecule that helps to obtain and / or maintain the optimal pH value of the composition. The pH adjusting agent can be a buffer, an acid or a base, for achieving acidic, alkaline or neutral pH (i.e., about pH 7). Typically, the pH adjusting agent is an acid, such as an organic acid. The organic acid can be any food acid suitable for human consumption, such as a carboxylic acid, and includes salts of such organic acids. Non-limiting examples include tartaric acid, citric acid, lactic acid, benzoic acid, acetic acid, malic acid, ascorbic acid, succinic acid, fumaric acid, oxalic acid and salts thereof (e.g., tartrate, citrate, lactate, etc.). Preferred organic acids include citric acid, malic acid and ascorbic acid.

[0139] Suitably, the total weight concentration of the pH adjuster is about 1-10% (e.g., 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or any value within the foregoing range), more preferably about 2-7% (by total weight).

[0140] The composition may further comprise one or more emulsion stabilizers (co-emulsifiers) for preventing phase separation and stabilizing the composition. Examples of emulsion stabilizers include gums such as xanthan gum, guar gum, sodium alginate, gum arabic, ester gum, and beet pectin; phospholipids such as soybean lecithin; and modified food starch.

[0141] The preservative can be any substance known in the art. Exemplary preservatives include benzoates, such as sodium benzoate, potassium benzoate, and calcium benzoate; sorbates, such as sodium sorbate, potassium sorbate, and calcium sorbate; citrates, such as sodium citrate and potassium citrate; polyphosphates, such as sodium hexametaphosphate; antioxidants, such as ascorbic acid (vitamin C), ethylenediaminetetraacetic acid (EDTA), β-hydroxy acid, butylated hydroxytoluene (BHT), tert-butylhydroquinone (TBHQ), dehydroacetic acid, dimethyl carbonate, ethoxyquinoline, and heptylparaben.

[0142] In various embodiments, the composition is substantially free of gluten. In this regard, the composition can be substantially free of gluten-containing products, such as gluten-containing flours and starches (e.g., having a gluten content of no more than about 0.5%, 0.2%, 0.1%, or 0.05% by weight).

[0143] In certain embodiments, the composition is substantially free of lactose. To this end, the composition can be substantially free of milk or dairy-derived or dairy-derived ingredients (e.g., having a milk or dairy-derived or derived lactose content of no more than about 0.5%, 0.2%, 0.1%, or 0.05% by weight).

[0144] In a specific embodiment of the present invention, the composition is a protein supplement comprising:

[0145] (i) at least one culture supernatant from at least one probiotic bacterium;

[0146] (ii) a protein source, a carbohydrate source, and a lipid source, wherein the lipid source comprises a seed extract and / or a fruit extract;

[0147] (iii) a sweetener; and

[0148] (iv) Optionally include pH adjusters, preservatives, solvents, emulsion stabilizers, colorants, flavorings, amino acids, prebiotic fibers, vitamins, and minerals.

[0149] In another embodiment of another aspect of the present invention, the composition comprises:

[0150] (i) at least one culture supernatant from at least one probiotic microorganism;

[0151] (ii) a protein source, a carbohydrate source, and a lipid source, wherein the lipid source comprises a fruit extract and / or a flower extract;

[0152] (iii) a sweetener; and

[0153] (iv) may optionally include pH adjusters, preservatives, solvents, emulsion stabilizers, colorants, flavorings, amino acids, vitamins, and minerals.

[0154] In view of the ingredients included, one skilled in the art will appreciate that the composition can be used to:

[0155] (i) Nutritional supplementation, such as protein supplementation;

[0156] (ii) improving health conditions, such as gastrointestinal health; and / or

[0157] (iii) preventing and / or treating diseases, disorders or conditions;

[0158] For main body use.

[0159] In certain embodiments, the disease, disorder or condition is selected from the group consisting of hypertension, heart disease, anxiety, obesity, neurodegenerative diseases, Alzheimer's disease, cancer, immunodeficiency, premature aging, wrinkles, inflammation, gastrointestinal dysfunction, arthritis, osteoarthritis, allergies, molecular and gene regulatory abnormalities, gut microbiota imbalance, hormone imbalance, vitamin deficiency, and any combination thereof.

[0160] With respect to improving gastrointestinal health, administration of a composition as described herein can modulate one or more microbial species or genera in at least a portion of a subject's gastrointestinal tract. Thus, the composition can be considered a modbiotic.

[0161] In a further aspect, the present invention relates to a method for preparing a composition comprising the steps of mixing:

[0162] water, wherein the water is derived at least in part from at least one culture supernatant from at least one probiotic microorganism; and

[0163] a substrate;

[0164] Thus, the composition was prepared.

[0165] Suitably, the composition is the composition described in the first aspect.

[0166] In some embodiments, the method further comprises mixing one or more pH adjusters, preservatives, solvents, emulsion stabilizers, colorants, flavorings, amino acids, prebiotic fibers, vitamins, and minerals.

[0167] Suitably, the composition will be as described herein.

[0168] In another aspect, the present invention provides a composition prepared according to the above method.

[0169] It is expected that the compositions described above may be useful for providing nutritional supplementation, such as protein supplementation, to a subject in need thereof.

[0170] Therefore, in another aspect, the present invention relates to a method of supplementing a subject, comprising administering to the subject a therapeutically effective amount of the composition described herein, thereby supplementing the subject.

[0171] "Administering" or "administering" refers to introducing a composition (e.g., a composition comprising, consisting of, or consisting essentially of, comprising water derived at least in part from a probiotic microorganism culture supernatant and a substrate) into a subject by a selected route, particularly oral administration.

[0172] The term "therapeutically effective amount" refers to a specific dosage sufficient to achieve the desired effect in the subject being treated. For example, the dosage can be an amount of the above-mentioned composition used to: (a) provide sufficient supplementation; (b) promote health; and / or (c) reduce, alleviate and / or prevent the disease, disorder or condition in the subject. In certain embodiments, a "therapeutically effective amount" is sufficient to reduce or eliminate the symptoms of the disease, disorder or condition, or at least partially meet the recommended daily intake of, for example, proteins, amino acids, vitamins or minerals. In other embodiments, a "therapeutically effective amount" is a dosage sufficient to achieve the desired biological effect.

[0173] Ideally, a therapeutically effective amount is a dose sufficient to induce the desired effect without causing significant cytotoxic effects in the subject. For example, the effective dose for reducing, alleviating and / or preventing a disease, disorder or condition will depend on the subject being treated, the type and severity of the relevant disease, disorder and / or condition, and the mode of administration of the therapeutic composition.

[0174] In one embodiment, the dosage of the composition (e.g., oral) can be administered as a single dose or multiple doses over a specific time period (e.g., for the entire period that the subject requires treatment). The dosing regimen can be carried out at predetermined time intervals, examples of which include hourly, daily, weekly, biweekly, or monthly dosing schedules.

[0175] The above-mentioned composition can be administered in a manner compatible with the dosage form, and the dosage should be a pharmaceutically or therapeutically effective amount. In the context of the present invention, the dosage administered to the subject should be sufficient to produce a beneficial response in the subject (e.g., to alleviate or relieve the symptoms of the disease, disorder or condition) within an appropriate time period. The amount of the administered dosage may depend on the condition of the subject to be treated, including factors such as the subject's age, sex, weight, and overall health, all of which should be determined by the judgment of a practitioner with ordinary skill.

[0176] In certain embodiments, the present methods are intended to supplement the nutritional or protein intake of a subject engaged in bodybuilding and protein supplementation, meal replacement, athletic recovery, and / or weight gain.

[0177] In other embodiments, the present methods are intended to replenish nutrient or protein intake in subjects affected by age-related and / or disease-related muscle loss, such as sarcopenia and muscle loss associated with rheumatoid arthritis, diabetes, hormonal imbalances, peripheral arterial disease, chronic obstructive pulmonary disease (COPD), congestive heart failure, end-stage renal disease, cirrhosis, cancer, and HIV.

[0178] In a related aspect, the present invention relates to a method of improving the health of a subject, comprising administering to the subject a therapeutically effective amount of a composition described herein, thereby improving the health of the subject.

[0179] In a specific embodiment, the method is directed to improving gastrointestinal health in the subject.

[0180] In another aspect, the present invention provides a method for treating and / or preventing a disease, disorder or condition in a subject, the method comprising administering to the subject a therapeutically effective amount of a composition as described herein, thereby treating and / or preventing the disease, disorder or condition in the subject.

[0181] In another aspect, the present invention provides a method for extending the shelf life of a beverage, food or cosmetic, comprising adding an effective amount of the composition or ingredient described in this specification to the product to prevent spoilage.

[0182] In a specific embodiment, the method is intended to ensure that the product has a refrigerated shelf life of at least 2.5 months and a room temperature shelf life of at least 1 month.

[0183] As used herein, "treatment," "therapeutic method," or "treatment" refers to a therapeutic intervention, course of action, or regimen that at least alleviates the symptoms of a disease, disorder, or condition that has at least begun to develop. As used herein, "prevention," "preventative method," or "prevention" refers to a therapeutic intervention, course of action, or regimen that is initiated before a disease, disorder, or condition and / or its symptoms develop, to prevent, inhibit, or slow the development or progression of the disease, disorder, or condition or its symptoms. In this regard, "prophylactic" treatment refers to treatment given to a subject who has not yet developed symptoms of a disease, disorder, or condition, or who is displaying only early symptoms, to reduce the subject's risk of developing the disease, disorder, or condition.

[0184] Suitably, the disease, disorder or condition is selected from the group consisting of hypertension, heart disease, anxiety, obesity, neurodegenerative disease, Alzheimer's disease, cancer, immunodeficiency, premature aging, wrinkles, inflammation, gastrointestinal dysfunction, arthritis, osteoarthritis and any combination thereof.

[0185] As used herein, the term "subject" includes human subjects and veterinary subjects. For example, administering to a subject may include administering to a human subject or a veterinary subject. Preferably, the subject is a human. However, the therapeutic uses of the present invention may also be applicable to mammals, such as domestic and companion animals, performance animals (e.g., horses), livestock, and laboratory animals.

[0186] All patents and scientific literature cited herein are incorporated by reference in their entirety into this specification.

[0187] In order to make the present invention easier to understand and put into practice, the following non-limiting examples are provided for reference by those skilled in the art.

[0188] Example

[0189] The present invention will now be described with reference to the following examples, which are to be considered in all respects as illustrative and non-restrictive.

[0190] Example 1

[0191] Example 1 illustrates a hydrating beverage composition supplemented with dietary protein, dietary fiber, and vitamins, which has a high antioxidant content due to the presence of carboxylic acids and low concentrations of biogenic amines and amides.

[0192] Example 1 (Drink Version: Cocoa Super Protein)

[0193] Composition and weight percentage

[0194] Probiotic bacterial culture supernatant - 10%

[0195] Cocoa powder - 1.5%

[0196] Cocoa bean powder - 1.5%

[0197] Cocoa butter - 0.5%

[0198] Coconut milk - 2%

[0199] Lemon powder - 0.2%

[0200] Manuka Honey Powder - 20%

[0201] Glycerin - 1%

[0202] Stevia powder - 0.015%

[0203] Soy lecithin - 0.3%

[0204] Water - 62.985%

[0205] Preparation and production methods

[0206] The following methods utilize aseptic techniques to help prevent and control contamination.

[0207] 1. Weigh powder ingredients and measure liquid ingredients;

[0208] 2. Add powder and liquid ingredients to the container;

[0209] 3. Add filtered water until all powder and liquid ingredients are covered;

[0210] 4. Stir at 30°C to ensure uniform mixing and full dissolution of all ingredients;

[0211] 5. Filter the probiotic bacterial culture supernatant to 0.22 microns, sonicate, or pasteurize (as needed) before adding to the base formula;

[0212] 6. Continue adding filtered water until the final required volume is reached;

[0213] 7. Dispense the final mixture into bottles;

[0214] 8. After bottling, package, palletize, and store in a refrigerator at 4°C;

[0215] 9. Randomly sample bottled products for quality control testing;

[0216] 10. The colony forming units (CFU / g) of coliform bacteria and non-coliform bacteria are less than 10 2 Only qualified batches can be shipped to commercial sales locations.

[0217] Example 2

[0218] Example 2 provides another example of a hydrating beverage composition that acts as a dietary fiber and vitamin supplement while having a high antioxidant content due to the presence of carboxylic acids and low concentrations of biogenic amines and amides.

[0219] Example 2 (Beverage Version: Australian Berries)

[0220] Composition and weight percentage

[0221] Probiotic bacterial culture supernatant—7%

[0222] Orange powder—0.5%

[0223] Mixed berry powder – 3%

[0224] Glycerin—2%

[0225] Hibiscus pollen – 0.5%

[0226] Lemon powder – 0.3%

[0227] Manuka honey – 10%

[0228] Xanthan gum—0.01%

[0229] Water – 76.69%

[0230] Preparation and production methods

[0231] Same as described in Example 1.

[0232] Although the present invention has been described with reference to specific embodiments, it will be understood by those skilled in the art that the present invention may be embodied in a variety of other forms, and the specific features of any one of the described embodiments may be used in any combination in other embodiments. Various modifications and variations of the present invention will be apparent to those skilled in the art and do not depart from the scope and spirit of the invention. It will be understood that the present invention is not intended to be improperly limited by the exemplary embodiments and examples described herein, which are provided as examples only, and the scope of the present invention shall be limited only by the following claims.

[0233] Example 3

[0234] Example 3 presents preliminary experimental data on the beneficial effects of the probiotics of the present invention.

[0235] Probiotics

[0236] Lactococcus lactis isolate QF16 and Sclerodermaceae isolate TF18 have been approved by the Queensland Department of Agriculture and Fisheries (DAF-QLD) (approval number SA 2020 / 08 / 750) for use in animal science research.

[0237] Phase 1

[0238] Ten-year-old dogs were fed a diet consisting of dog food pellets (containing fat and carbohydrates), processed meat, food scraps, tap water, a 5% protein supplement, and a probiotic mixture containing strains 1a and 1b (Lactococcus lactis isolate QF16 and Sclerodermaceae isolate TF18, respectively) for 30 days.

[0239] Reports indicate that dogs have improved mobility and activity, with no signs of pain and / or toxicity.

[0240] Phase 2

[0241] The composition was administered orally to 4-week-old experimental mice for 30 days, once every other day for the first two weeks and once a week for the next two weeks.

[0242] The composition includes dog food pellets (containing fat and carbohydrates), organic fruit, tap water, 5% protein supplement and probiotics. The probiotics in the first group are commercial probiotics [Blackmores PAW DigestiCare]. The probiotics in the second group are fungal cells of the Sclerodermaceae isolate TF18, at a concentration of 5×10 7 The probiotics in the third group were fungal cells of the Scleroderma isolate TF18, with a concentration of 5×10 8 The control group did not contain probiotics.

[0243] The results showed that accepting 5×10 7 The body weights of animals treated with cell doses were similar to those of untreated controls (see Figure 1 A), but these animals were leaner and had better blood sugar control (see Figure 1 B) It was particularly surprising and unexpected that the Sclerodermaceae isolate TF18 was significantly more effective than the commercial probiotic.

[0244] Phase 3

[0245] After the 30-day dosing period, intestinal samples were collected from the experimental mice. These samples were then analyzed in the pathology laboratory. According to the established intestinal histology test protocol, the results showed that the intestinal tract was significantly different from that of the untreated control group ( Figure 2 ), the Sclerodermaceae isolate TF18 showed:

[0246] Intestinal villus density increased by 83%;

[0247] The average length of intestinal villi increased by 62%;

[0248] A healthier intestinal mucosal barrier, which is crucial for the precise transport of water, nutrients, and molecules between cells (e.g., gut-brain-endocrine responses);

[0249] Increased resistance to mechanical stress.

[0250] Phase 4

[0251] Chickens in industrial chicken cages were orally dosed once a week for 43 days.

[0252] The compositions used included a whey protein supplement and probiotics. The probiotic in the first group was Lactococcus lactis QF16 at a concentration of 1 × 10^8 or 8 × 10^7 CFU / ml. The probiotic in the second group was the same strain at a concentration of 1 × 10^9 CFU / ml. Control groups contained no probiotics. Some control groups contained only the culture medium mixture without probiotic cells or their supernatant; the results were identical to those of the untreated control group (data not shown).

[0253] The results showed that chickens receiving 1×10^8 or 1×10^7 CFU of probiotics gained significant weight, even though their feed intake was the same as that of the untreated control group ( Figure 3 A and 3B). Surprisingly, the lower concentration of probiotics combined with protein supplementation resulted in faster weight gain than the higher concentration probiotic control group ( Figure 3 C).

[0254] Phase 5

[0255] After the 43-day dosing period, intestinal samples were collected from the chickens. These samples were then sent to a pathology laboratory for analysis. Intestinal histology, performed according to established standard procedures, showed no signs of toxicity, tissue damage, or cytopathological changes in the groups treated with Probiotic 1a compared to the untreated control group (see Table 1).

[0256] Phase 6

[0257] Feces collected from the above-mentioned chicken flock model were sent to an external microbiome sequencing site after 43 days, and the sequencing results were analyzed according to established bioinformatics protocols.

[0258] The results showed that the intestinal flora of chickens treated with low concentrations of Lactococcus lactis QF16 (1×10^8 or 8×10^7 CFU / ml) changed significantly compared with the untreated control group. The main microorganisms found in the intestines of the treated groups included:

[0259] Anaerococcus (increased 2.24-fold): This genus of bacteria breaks down sugars and produces natural butyrate. The increase in this genus was associated with better nutrient utilization and improved metabolism in poultry.

[0260] Romboutsia (4.39-fold increase): This genus of bacteria is associated with a healthy gut and has been clearly documented in healthy humans. Maintaining the Romboutsia community may help reduce or prevent intestinal abnormalities. This genus is commonly found in the gut and eggshell microbiota of poultry and is involved in energy metabolism and hormone regulation. In this study, this genus significantly increased with treatment and was associated with accelerated growth and weight gain.

[0261] Lachnospiraceae (population increase of approximately 10%): This genus of bacteria improves feed conversion efficiency, promoting greater weight gain with less food. This genus has been enriched in multiple animal studies and demonstrated positive feed conversion efficiency (FCR). Lachnospiraceae bacteria have been well-documented to capture energy from food, producing acetate for muscle growth and fat burning. They also release naturally occurring propionate and butyrate, and regulate FFAR3 gene expression, influencing fat deposition and gluconeogenesis. Lachnospiraceae also protects individual health by producing short-chain fatty acids, and their increase may be associated with improved lean meat growth and quality.

[0262] Lactobacillus (2.21-fold increase): This genus of bacteria can promote cell proliferation and increase serum testosterone levels in male mice. In this study, the L. reuteri population increased by 88%. Other studies have shown that mice treated with this genus strain appeared younger in physical assessments than their untreated counterparts, suggesting that microbial therapy may play a role in public health by improving hormone health and reproductive capacity. In this poultry trial, this genus was significantly increased by treatment with 1×10^8 or 8×10^7, which was associated with lean growth and active metabolism.

[0263] Bacillus (increased 2.30-fold): This genus of bacteria forms spores and is known for promoting beneficial gut microbial stimulation and immune responses. In this study, an increase in this genus was associated with improved gut structure and metabolic activity.

[0264] Escherichia and Shigella (3.84-fold reduction): These two species are known zoonotic pathogens, and their significant reduction was negatively correlated with the increase of beneficial gut flora.

[0265] in conclusion

[0266] This study recommends the use of probiotics as a method to promote protein accumulation in the body. Studies have shown that when the human body is exposed to the right combination of probiotics, it can more efficiently utilize nutrients, promote greater protein synthesis in muscle, and potentially contribute to global sustainable development by improving food metabolism.

[0267] The technology developed by this invention is rapidly scalable and can reduce pressure on agricultural systems, ultimately having a positive impact on the environment by reducing greenhouse gas emissions and water consumption.

Claims

1. A composition comprising: secreted and shed molecules and / or probiotic cell structures not constituting living cells, said molecules and structures being derived at least in part from the culture supernatant of at least one cellular organism; and water, the water being at least partially derived from a culture supernatant of at least one cellular organism; and a base component.

2. The composition according to claim 1, wherein the cell organism from which the culture supernatant is derived is a probiotic microorganism.

3. The composition according to claim 1 or 2, wherein the base component comprises a protein source, a carbohydrate source, a lipid source and / or a sweetener.

4. The composition of claim 3, wherein the protein source, carbohydrate source and / or lipid source is derived from one or more non-animal sources.

5. The composition according to claim 4, wherein the protein source, carbohydrate source and / or lipid source are at least partially derived from plants.

6. A composition according to any one of claims 3 to 5, wherein the protein source, carbohydrate source and / or lipid source is at least partly derived from plant extracts, fruit extracts (such as fruit powder and fruit juice), floral extracts (such as flower powder) and / or seed extracts (such as seed powder, seed oil and seed fat).

7. A composition according to any preceding claim, wherein the composition is a substantially vegetarian and / or vegan composition.

8. The composition of any preceding claim, wherein the composition comprises from about 0.1% to about 99% by weight of a culture supernatant of at least one probiotic microorganism.

9. The composition of any preceding claim, wherein the composition comprises from about 0.1% to about 99% by weight of the base component.

10. The composition of any one of claims 2 to 8, wherein the composition comprises from about 0.1% to about 30% by weight of a sweetener.

11. A composition according to any preceding claim further comprising one or more pH adjusters, preservatives, solvents, emulsion stabilizers, colorants, flavors, prebiotic fibers, vitamins and minerals.

12. A composition according to any preceding claim in the form of a supplement.

13. The composition according to claim 12, which is a protein supplement.

14. A method for preparing a composition comprising the steps of: Water, which is at least partially derived from a culture supernatant of at least one cellular organism, is mixed with a base component to prepare the composition. The method according to claim 14 , wherein the cell organism from which the culture supernatant is derived is a probiotic microorganism.

16. The method according to claim 14 or 15, wherein the prepared composition is the composition according to any one of claims 1 to 13.

17. A composition prepared according to the method of claims 14 to 16.

18. The composition according to any one of claims 1 to 13 and 17, for use in: (i) Supplement; (ii) improve and / or maintain health; and / or (iii) preventing and / or treating a disease, disorder or condition in a subject.

19. A method of supplementation comprising administering to a subject a therapeutically effective amount of the composition of any one of claims 1 to 13 and 17 to supplement the subject.

20. A method of improving the health of a subject, comprising administering to the subject a therapeutically effective amount of the composition of any one of claims 1 to 13 and 17 to improve the health of the subject.

21. The method of claim 20, wherein improving health comprises at least one of the following: reducing weight, controlling blood sugar, increasing intestinal villus density, increasing average intestinal villus length, promoting intestinal mucosal barrier health, and / or improving resistance to mechanical stress.

22. The method according to claim 20 or 21, wherein the concentration of the composition is 1×10 5 , 1×10 6 , 1×10 7 , 1×10 8 , 1×10 9 , 1×10 10 , 1×10 11 , 5×10 5 , 5×10 6 , 5×10 7 , 5×10 8 , 5×10 9 , 5×10 10 , 5×10 11 or between 1×10 5 to 5×10 11 Any CFU / recommended dose between.

23. The method of claim 20 or 21, wherein administration of the composition results in a significant change in microbial flora.

24. The method of claim 23, wherein the significant microbial community changes include an increase in at least one of the following microorganisms: Anaerococcus, Romboutsia, Lachnospiraceae, Lactobacillus, or Bacillus.

25. The method of claim 23, wherein the significant microbial community change comprises a decrease in at least one of the following microorganisms: Escherichia, Shigella, or other Gram-positive or -negative bacteria, fungi, parasites, or protozoa.

26. A method of treating and / or preventing a disease, disorder or condition in a subject, comprising administering to the subject a therapeutically effective amount of the composition of any one of claims 1 to 13 and 17 to treat and / or prevent the disease, disorder or condition.

27. The use of claim 16 or the method of claim 24, wherein the disease, disorder or condition is selected from the group consisting of hypertension, heart disease, anxiety, obesity, neurodegenerative diseases, Alzheimer's disease, cancer, immunodeficiency, premature aging, wrinkles, inflammation, gastrointestinal dysfunction, arthritis, osteoarthritis, allergies, molecular and gene regulatory disorders, gut microbiota dysfunction, hormone imbalance, vitamin deficiency, and any combination thereof.

28. The method or use according to any one of claims 16 to 25, wherein the subject is a human or an animal.

29. The composition of claim 1, wherein the result of use of the composition or culture supernatant or culture supernatant molecules or shed molecules or shed cell structures is a cosmetic.

30. The composition of claim 1, wherein the use of the composition or culture supernatant or culture supernatant molecules or shed molecules or shed cell structures results in extending the shelf life of a product stored at refrigeration and / or room temperature.

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