Probiotic bifidobacterium adolescentis strain producing GABA and uses thereof

Abstract

The present invention relates to Bifidobacterium adolescentis strain NCC275 as deposited with the Collection Nationale de Cultures de Microorganismes (CNCM) under the Accession Number CNCM I-6090 on June 17, 2024. The invention further encompasses compositions comprising Bifidobacterium adolescentis strain NCC275 and, optionally, a source of glutamate. Further encompassed are therapeutic and non-therapeutic uses of such compositions.

Description

[0001] PROBIOTIC BACTERIA PRODUCING GABA AND USES THEREOF

[0002] BACKGROUND TO THE INVENTION

[0003] The present invention relates to the field of probiotics and, more specifically, to a novel probiotic strain that exhibits enhanced production of gamma-aminobutyric acid (GABA). GABA is a major inhibitory neurotransmitter of the vertebrate central nervous system. It is involved in the regulation of cardiovascular conditions such as blood pressure and heart rate, and plays a role in the sensation of pain and anxiety.

[0004] For example, Abdou et al. (Biofactors (2006) 26(3):201-8) studied the effect of orally administrated GABA on relaxation and immunity during stress in humans. GABA intake increased alpha waves and decreased beta waves, indicating relaxation and reduced anxiety. Additionally, GABA maintained higher salivary IgA levels in stressed subjects compared to a placebo group, suggesting enhanced immunity. Yoto et al. (Amino Acids (2012) 43(3) :1331-7) reported that oral GABA intake reduces stress in adults. In a randomized, placebo-controlled study with 63 participants, 100 mg of GABA mitigated the decrease in EEG alpha and beta brain waves caused by mental stress tasks. This stress reduction was also reflected in improved Profile of Mood States (POMS) scores. Yamatsu et al. (Japanese Pharmacology and Therapeutics (2013) 41(10):985-988.) reported that GABA reduces stress, improves sleep quality, and decreases night urination in elderly people. In a four-week study with 38 participants, those given 100 mg GABA chocolates per day had smaller increases in cortisol levels, better sleep, and reduced night urination compared to a placebo group.

[0005] The effects of GABA on sleep quality were further reported by Okada et al. (Nippon Shokuhin Kagaku Kogaku Kaishi (2000) 47(8):596-603). In a double-blind study with 20 post-menopausal female patients, defatted rice germ enriched with GABA (26.4 mg / day) was administered three times daily. Results showed improvements in sleeplessness, depression, and autonomic disorders in over 65% of patients, with overall improvement observed in 75% of participants. In particular, an improvement ofthe insomnia score ofthe Kupperman Menopause Index after 4 weeks of intervention was reported. Yamatsu et al. (Journal of Nutritional Science and Vitaminology (2015) 61(2):182-187) investigated the effects of GABA on sleep improvement in humans. GABA (100 mg) shortened sleep latency by 5.3 minutes. Questionnaires confirmed these benefits. Yamatsu et al. (Food Sci Biotechnol (2016) 25(2):547-551) also reported that GABA improved sleep by shortening sleep latency and increasing non-REM sleep time in a randomized, placebo-controlled study.

[0006] Probiotics are live microorganisms that, when administered in adequate amounts, confer health benefits to the host. They are widely used in nutritional supplements and functional foods due to their ability to positively influence gut health and overall well-being.

[0007] The ability of probiotic strains to produce GABA can be attributed to the enzyme glutamate decarboxylase (GAD), which catalyzes the conversion of glutamate into GABA. In bacteria, GAD is typically encoded by the GAD operon consisting of the genes gadR, gadCand gadB, wherein gadB encodes the GAD enzyme, gadC encodes the glutamate / GABA antiporter GadC and gadR encodes the transcriptional activator GadR.

[0008] The identification and development of probiotic strains capable of producing bioactive compounds like GABA offer promising therapeutic potential for managing stress, anxiety, and other neurological conditions.

[0009] GABA-producing probiotics have been described by Pokusaeva et al. (Neurogastroenterol Motil (2017) 29(l):el2904). Therein, it was reported that daily oral administration of a specific GABA-producing Bifidobacterium strain (B. dentium ATCC 27678) reduced visceral hypersensitivity and modulated sensory neuron activity in a rat model. This study suggests that microbial-derived GABA can desensitize colonic afferents and offers a new approach for treating abdominal pain by targeting the microbiome-gut-brain axis. WQ2020 / 157297 claims that a strain of L. lactis NCDO2118 is able to produce GABA. Chronic gavage of rats with this strain was shown to inhibit visceral hypersensitivity triggered by partial restraint stress. Nobile et al. (Neuropsychobiology (2022) 81(2):116-126) have conducted an uncontrolled open-label trial with a mix of two strains (L. reuteri PBS072 and B. breve BB077) showing a reduction of stress in a group of 30 stressed students (decrease in salivary cortisol and trait anxiety, improved sleep). From a mechanistic standpoint, the authors have shown that exposure of a neuronal cell line to these two strains triggers secretion of GABA. However, the authors have not assessed the potential production of GABA by the strains themselves, thus it remains unclear whether GABA originates from the bacteria, or neurons, or both. In two preclinical studies, Patterson et al. (Sci Rep (2019) 9(1):16323) have shown that two GABA-producing L. brevis DPC6108 and DSM32386 improve depressive-like behavior and metabolic abnormalities associated with metabolic syndrome in mice. Ko et al. (Process Biochemistry (2013) 48(4):559-568) showed the production of GABA upon fermentation of black soybean milk by L. brevis FPA3709, resulting in decreased depressive-like behavior triggered by forced swimming in rats. Laroute et al. (Elife (2022) 21:ll:e77100) reported that the microbial production of y-aminobutyric acid (GABA) by Lactococcus lactis NCDO2118, due to its glutamate decarboxylase (GAD) activity, significantly reduced stress-induced visceral hypersensitivity in rats, highlighting its potential therapeutic use for managing visceral pain and anxiety in irritable bowel syndrome (IBS) patients.

[0010] Despite these developments, there is still a need in the art for novel probiotic strains that efficiently produce GABA when comprised in the human microbiome.

[0011] SUMMARY OF THE INVENTION

[0012] The present invention is characterized in the herein provided embodiments and claims. In particular, the present invention relates, inter alia, to the following embodiments:

[0013] 1. Bifidobacterium adolescentis strain NCC275 as deposited with the Collection Nationale de Cultures de Microorganismes (CNCM) at the Institut Pasteur under the Accession Number CNCM 1-6090 on June 17, 2024.

[0014] 2. A composition, preferably a nutritional composition comprising Bifidobacterium adolescentis strain NCC275 (CNCM 1-6090).

[0015] 3. A composition comprising (i) Bifidobacterium adolescentis strain NCC275 (CNCM 1-6090) and (ii) at least one excipient, and / or wherein the Bifidobacterium adolescentis strain NCC275 is spray-dried or freeze-dried.

[0016] 4. A nutritional composition or product comprising Bifidobacterium adolescentis strain NCC275 (CNCM 1-6090) and a source of glutamate.

[0017] 5. The nutritional composition or product according to embodiment 4, wherein the source of glutamate comprises or consists of purified glutamate, or wherein the source of glutamate is a natural product, in particular a fermentation product or a cell extract.

[0018] 6. The nutritional composition or product according to embodiment 4or embodiment 5, wherein the source of glutamate is obtainable through prokaryotic fermentation of a suitable substrate with one or more of the following bacteria: Corynebacterium glutamicum, Corynebacterium ammoniagenes, Corynebacterium casei, Corynebacterium efficiens, Brevibacterium lactofermentum and Bacillus subtilis.

[0019] 7. The nutritional composition or product according to embodiment 6, wherein the substrate is a carbon and / or nitrogen source.

[0020] 8. The nutritional composition or product according to embodiment 6 or embodiment 7, wherein the substrate is obtained by enzymatic hydrolysis of a plant material and / or by enzymatic hydrolysis of starch.

[0021] 9. The nutritional composition or product according to embodiment 8, wherein the plant is one or more of: wheat, corn, tapioca and rye; and the starch is the starch of one or more of the above mentioned plants.

[0022] 10. The nutritional composition or product according to any one of embodiments 6 to 9, wherein the substrate is corn starch or enzymatically hydrolyzed corn starch.

[0023] 11. The nutritional composition or product according to embodiment 4, wherein the source of glutamate is a fermented starch, in particular a fermented corn starch, in particular wherein the starch or the corn starch has been fermented with one or more of the following bacteria: Corynebacterium glutamicum, Corynebacterium ammoniagenes, Corynebacterium casei, Corynebacterium efficiens, Brevibacterium lactofermentum and Bacillus subtilis.

[0024] 12. The nutritional composition or product according to any one of embodiments 4to 11, wherein the source of glutamate is a non-purified fermentation product.

[0025] 13. The nutritional composition or product according to any one of embodiments 5 to 12, wherein the fermentation product is a dried fermentation product.

[0026] 14. The nutritional composition or product according to embodiment 5, wherein the cell extract is a yeast extract.

[0027] 15. The nutritional composition or product according to any one of embodiments 4 to 14, wherein the source of glutamate comprises between 0.01 to 70 % of glutamate, preferably between 5 and 50% of glutamate.

[0028] 16. The nutritional composition or product according to any one of embodiments 4 to 15, wherein the source of glutamate further comprises between 0.01 to 70 % of inosine monophosphate (IMP) and guanosine monophosphate (GMP), preferably wherein the content of IMP and / or GMP is between 0.01 and 10 %.

[0029] 17. The nutritional composition or product according to any one of embodiments 4 to 16, wherein the source of glutamate comprises further amino acids, in particular wherein the further amino acids are one or more of: alanine, aspartic acid, glutamine, glycine, leucine, lysine, methionine, tryptophan and valine.

[0030] 18. The nutritional composition or product according to any one of embodiments 4 to 17, wherein the source of glutamate further comprises polysaccharides, in particular wherein the polysaccharides are present between 0 and 20 %, in particular, wherein the polysaccharides are one or more of: cellulose, pectin, starch and maltodextrins. 19. The nutritional composition or product according to any one of embodiments 4to 18, wherein the source of glutamate further comprises organic acids, in particular wherein the organic acids are one or more of: lactic acid, citric acid, acetic acid and malic acid.

[0031] 20. The nutritional composition or product according to any one of embodiments 4 to 19, wherein the source of glutamate further comprises minerals, in particular wherein the minerals are one or more of: sodium, potassium, chlorine, calcium, phosphorus, magnesium, iron, copper, zinc, iodine, manganese, selenium, chromium, molybdenum and fluorine.

[0032] 21. The nutritional composition or product according to any one of embodiments 4 to 20, wherein the source of glutamate further comprises fatty acids and / or sugars.

[0033] 22. The nutritional composition or product according to any one of embodiments 4 to 21, wherein the Bifidobacterium adolescentis strain NCC275 is spray-dried or freeze-dried.

[0034] 23. The nutritional composition or product according to any one of embodiments 4 to 22, wherein the nutritional composition or product further comprises inulin.

[0035] 24. The nutritional composition or product according to any one of embodiments 4 to 23, wherein the nutritional composition or product is or comprises a powder, a solid, a semisolid, a liquid or a semi-liquid.

[0036] 25. The nutritional composition or product according to any one of embodiments 4 to 24, wherein the nutritional composition or product is a powder, in particular a reconstitutable powder.

[0037] 26. The nutritional composition or product according to any one of embodiments 4 to 25, wherein the nutritional composition or product is a nutritional supplement, a dietary supplement, or a food for special medical purpose (FSMP). 27. The Bifidobacterium adolescentis strain according to embodiment 1, the composition according to embodiments 2 or 3, or the nutritional composition or product according to any one of embodiments 4to 26for use in therapy.

[0038] 28. The Bifidobacterium adolescentis strain according to embodiment 1, the composition according to embodiment 2 or 3, or the nutritional composition or product according to any one of embodiments 4 to 26 for use in reducing one or more symptoms of stress in a subject.

[0039] 29. TThe Bifidobacterium adolescentis strain according to embodiment 1, the composition according to embodiment 2 or 3, or the nutritional composition or product according to any one of embodiments 4 to 26 for use in the dietary management of stress in a subject.

[0040] 30. A method for reducing one or more symptoms of stress in a subject, comprising administering to said subject the Bifidobacterium adolescentis strain according to embodiment 1, the composition according to embodiments 2 or 3 , or the nutritional composition or product according to any one of embodiments 4 to 26.

[0041] 31. A method for the dietary management of stress in a subject, comprising administering to said subject the Bifidobacterium adolescentis strain according to embodiment 1, the composition according to embodiments 2 or 3 , or the nutritional composition or product according to any one of embodiments 4 to 26.

[0042] 32. The Bifidobacterium adolescentis strain, the composition or the nutritional composition or product for use according to embodiment 28 or 29, or the method according to embodiment 30 or 31, wherein reducing the symptoms of stress and / or the dietary management of stress involves one or more of: increasing relaxation, increasing calmness, reducing morning drowsiness, restoring mental calmness, and / or supporting mental relaxation in said subject.

[0043] 33. The Bifidobacterium adolescentis strain according to embodiment 1, the composition according to embodiment 2 or 3, or the nutritional composition or product according to any one of embodiments 4 to 26 for use in reducing one or more symptoms of premenstrual syndrome in a subject. The Bifidobacterium adolescentis strain according to embodiment 1, the composition according to embodiment 2 or 3, or the nutritional composition or product according to any one of embodiments 4 to 26 for use in the dietary management of premenstrual syndrome in a subject. A method for reducing one or more symptoms of premenstrual syndrome in a subject, comprising administering to the subject the Bifidobacterium adolescentis strain according to embodiment 1, the composition according to embodiments 2 or 3 , or the nutritional composition or product according to any one of embodiments 4 to 26. A method for the dietary management of premenstrual syndrome in a subject, comprising administering to the subject the Bifidobacterium adolescentis strain according to embodiment 1, the composition according to embodiments 2 or 3 , or the nutritional composition or product according to any one of embodiments 4 to 26. TThe Bifidobacterium adolescentis strain according to embodiment 1, the composition according to embodiment 2 or 3, or the nutritional composition or product according to any one of embodiments 4 to 26 for use in reducing pain in a subject. The Bifidobacterium adolescentis strain according to embodiment 1, the composition according to embodiment 2 or 3, or the nutritional composition or product according to any one of embodiments 4 to 26 for use in the dietary management of pain in a subject. A method for reducing pain in a subject, comprising administering to the subject the Bifidobacterium adolescentis strain according to embodiment 1, the composition according to embodiments 2 or 3 , or the nutritional composition or product according to any one of embodiments 4 to 26. A method for the dietary management of pain in a subject, comprising administering to the subject the Bifidobacterium adolescentis strain according to embodiment 1, the composition according to embodiments 2 or 3 , or the nutritional composition or product according to any one of embodiments 4 to 26. The Bifidobacterium adolescentis strain, the composition or the nutritional composition or product for use according to embodiment 37 or 38, or the method according to embodiment 39 or 40, wherein the pain is visceral pain, chronic pain, neuropathic pain or inflammatory pain. The Bifidobacterium adolescentis strain, the composition or the nutritional composition or product for use according to embodiment 37, 38 or 41, or the method according to embodiment 39, 40 or 41, wherein the pain is associated with menstruation and / or endometriosis; or wherein the pain associated with colic in infants or toddlers. The Bifidobacterium adolescentis strain according to embodiment 1, the composition according to embodiment 2 or 3, or the nutritional composition or product according to any one of embodiments 4 to 26 for use in improving sleep in a subject. The Bifidobacterium adolescentis strain according to embodiment 1, the composition according to embodiment 2 or 3, or the nutritional composition or product according to any one of embodiments 4 to 26 for use in the dietary management of sleep in a subject. A method for improving sleep in a subject, comprising administering to the subject the Bifidobacterium adolescentis strain a according to embodiment 1, the composition according to embodiments 2 or 3, or the nutritional composition or product according to any one of embodiments 4 to 26. A method for the dietary management of sleep in a subject, comprising administering to the subject the Bifidobacterium adolescentis strain according to embodiment 1, the composition according to embodiments 2 or 3, or the nutritional composition or product according to any one of embodiments 4 to 26. The Bifidobacterium adolescentis strain according to embodiment 1, the composition according to embodiment 2 or 3, or the nutritional composition or product according to any one of embodiments 4 to 26 for use in the prevention or delay of age-related decline in physiological and / or health functions, or conditions associated with aging in a subject. TThe Bifidobacterium adolescentis strain according to embodiment 1, the composition according to embodiment 2 or 3, or the nutritional composition or product according to any one of embodiments 4 to 26 for use in the dietary management of age-related decline in physiological and / or health functions, or conditions associated with aging in a subject. A method for preventing or delaying age-related decline in physiological and / or health functions, or conditions associated with aging in a subject, comprising administering to the subject the Bifidobacterium adolescentis strain according to embodiment 1, the composition according to embodiments 2 or 3 , or the nutritional composition or product according to any one of embodiments 4 to 26. A method for the dietary management of age-related decline in physiological and / or health functions, or conditions associated with aging in a subject, comprising administering to the subject the Bifidobacterium adolescentis strain according to embodiment 1, the composition according to embodiments 2 or 3 , or the nutritional composition or product according to any one of embodiments 4 to 26. 51. The Bifidobacterium adolescentis strain, the composition or the nutritional composition or product for use according to embodiment 47 or 48, or the method according to embodiment 49 or 50, wherein the age-related decline in physiological and / or health functions involves one or more of: decreased cognitive function, decreased neuronal activity, increased secretion of inflammatory cytokines and / or histamine, immune system dysregulation, decreased autophagy, increased blood pressure, decreased blood cell levels, decreased thyroid hormone levels, increased glucagon levels and / or increased insulin levels.

[0044] 52. The Bifidobacterium adolescentis strain, the composition or the nutritional composition or product for use according to embodiment 47or 48, or the method according to embodiment 49 or 50, wherein the conditions associated with aging include: Neurodegenerative diseases, in particular Alzheimer's disease; neuropathic pain; depression; inflammation, in particular low-grade chronic systemic inflammation and inflammatory pain; diabetes, in particular type 1 diabetes (T1D) and type 2 diabetes (T2D); hypertension; heart rhythm disorders; vision decline; bone and / or cartilage degeneration; respiratory dysfunction; renal dysfunction; anemia; thyroid dysfunction; muscle atrophy; hearing loss; cancer; metabolic syndrome; glucose intolerance; hyperglycemia; insulin resistance; obesity and obesity-related inflammation; pancreatic dysregulation; and liver dysfunction.

[0045] 53. A (non-therapeutic) use of the Bifidobacterium adolescentis strain according to embodiment 1, the composition according to embodiments 2 or 3 , or the nutritional composition or product according to any one of embodiments 4 to 26 for reducing one or more appearance of aging.

[0046] 54. The (non-therapeutic) use of embodiment 53, wherein reducing the one or more appearance of aging is characterized by: reducing skin wrinkles, decreasing skin thickness, increasing skin moisture, and / or increasing skin elasticity.

[0047] 55. A (non-therapeutic) use of the Bifidobacterium adolescentis strain according to embodiment 1, the composition according to embodiments 2 or 3 , or the nutritional composition or product according to any one of embodiments 4 to 26 for reducing one or more symptoms of UV exposure, sun exposure and / or sun damage on skin. The (non-therapeutic) use of embodiment 55, wherein reducing the one or more symptoms of UV exposure, sun exposure and / or sun damage on skin is characterized by: reducing skin wrinkles, decreasing skin thickness, increasing skin moisture, and increasing skin elasticity. The Bifidobacterium adolescentis strain according to embodiment 1, the composition according to embodiment 2 or 3, or the nutritional composition or product according to any one of embodiments 4 to 26 for use in reducing one or more symptoms of aging. A method for reducing one or more symptoms of aging in a subject, comprising administering to the subject the Bifidobacterium adolescentis strain according to embodiment 1, the composition according to embodiments 2 or 3, or the nutritional composition or product according to any one of embodiments 4 to 26. The Bifidobacterium adolescentis strain, the composition or the nutritional composition or product for use according to embodiment 57 or the method according to embodiment 58, wherein reducing the one or more symptoms of aging is characterized by: reducing skin wrinkles, decreasing skin thickness, increasing skin moisture, and increasing skin elasticity. The Bifidobacterium adolescentis strain according to embodiment 1, the composition according to embodiment 2 or 3, or the nutritional composition or product according to any one of embodiments 4 to 26 for use in reducing one or more symptoms of UV exposure, sun exposure and / or sun damage on skin. A method for reducing one or more symptoms of UV exposure, sun exposure and / or sun damage on skin in a subject, comprising administering to the subject the Bifidobacterium adolescentis strain according to embodiment 1, the composition according to embodiments 2 or 3, or the nutritional composition or product according to any one of embodiments 4 to 26.

[0048] 62. The Bifidobacterium adolescentis strain, the composition or the nutritional composition or product for use according to embodiment 60or the method of embodiment 61, wherein reducing the one or more symptoms of UV exposure, sun exposure and / or sun damage on skin is characterized by: reducing skin wrinkles, decreasing skin thickness, increasing skin moisture, and increasing skin elasticity.

[0049] 63. A (non-therapeutic) use of the Bifidobacterium adolescentis strain according to embodiment 1, the composition according to embodiments 2 or 3, or the nutritional composition or product according to any one of embodiments 4 to 26 for improving recovery after exercise and / or for improving exercise tolerance.

[0050] 64. The Bifidobacterium adolescentis strain according to embodiment 1, the composition according to embodiment 2 or 3, or the nutritional composition or product according to any one of embodiments 4 to 26 for use in improving recovery after exercise and / or for improving exercise tolerance.

[0051] 65. A method for improving recovery after exercise and / or for improving exercise tolerance in a subject, comprising administering to the subject the Bifidobacterium adolescentis strain according to embodiment 1, the composition according to embodiments 2 or 3 , or the nutritional composition or product according to any one of embodiments 4 to 26.

[0052] 66. The use according to embodiment 63, the Bifidobacterium adolescentis strain, the composition or the nutritional composition or product for use according to embodiment 64 or the method according to embodiment 64, wherein improving recovery after exercise and / or improving exercise tolerance is characterized by improved muscle recovery and / or improved muscle growth, in particular wherein muscle recovery and / or muscle growth is improved through increased production of immunoreactive and / or immunofunctional growth hormones in response to GABA production by the probiotic bacteria according to the invention. 66. The Bifidobacterium adolescentis strain according to embodiment 1, the composition according to embodiment 2 or 3, or the nutritional composition or product according to any one of embodiments 4 to 26 for use as a medicament.

[0053] The present invention meets the need in the art for novel probiotic strains that efficiently produce GABA by providing Bifidobacterium adolescentis strain NCC275. It is shown in appended Example 1 that B. adolescentis NCC275 efficiently produces GABA in an ex vivo Systemic Intestinal Fermentation Research (SIFR®) study that simulates human gut fermentation.

[0054] SIFR® is a high-throughput, ex vivo technology that is used to predict clinical outcomes of prebiotics and / or probiotics within 1-2 days (see Van den Abbeele et al., Frontiers in Microbiology (2023) 14:1131662). It evaluates the impact of these agents on specific gut microbes and their metabolic activities, maintaining high reproducibility and similarity to in vivo conditions. SIFR® bridges the gap between preclinical and clinical research, improving the development and success of microbiome-modulating therapies.

[0055] It was surprisingly found that B. adolescentis NCC275 produced high amounts of GABA in a simulated human gut fermentation. Of note, the amount of GABA that was produced by B. adolescentis NCC275 in this study was significantly higher than that of the Lactococcus lactis strain NCDO2118, a strain that was previously reported to produce GABA in the gastrointestinal tract of rats (Laroute et al., Elife (2022) ll:e77100).

[0056] Accordingly, in one aspect the invention relates to the Bifidobacterium adolescentis strain NCC275 as deposited with the Collection Nationale de Cultures de Microorganismes (CNCM) at the Institut Pasteur under the Accession Number CNCM 1-6090 on June 17, 2024.

[0057] Compositions comprising Bifidobacterium adolescentis strain NCC275

[0058] Another aspect of the present invention relates to compositions comprising Bifidobacterium adolescentis strain NCC275. The composition may be a nutritional composition. The composition according to the present invention may be any type of composition comprising Bifidobacterium adolescentis strain NCC275, provided that the Bifidobacterium adolescentis strain NCC275 remains stable in said composition for an adequate amount of time.

[0059] Bifidobacterium adolescentis strain NCC275 may be present in the compositions of any aspect or embodiment of the present invention in varying amounts, such as greater than 5 million, 10 million, 15 million, 20 million, 25 million, 30 million, 35 million, 45 million, 50 million, 75 million, 100 million, 250 million, 500 million, 750 million, 1 billion, or 2 billion bacteria per dosage form. For example, the probiotic may be present in amounts ranging from 5 million to 2.5 billion bacteria per dosage form, including specific ranges such as 5 million to 2.5 billion, 10 million to 2.5 billion, 30 million to 2.5 billion, 50 million to 2.5 billion, 50 million to 1 billion, 75 million to 2.5 billion, 75 million to 1 billion, 100 million to 2.5 billion, 100 million to 1 billion, 250 million to 2.5 billion, 250 million to 1 billion, 500 million to 2.5 billion, 500 million to 1 billion, 750 million to 2.5 billion, 750 million to 1 billion, 1 billion to 2.5 billion, and 1.5 billion to 2.5 billion bacteria per dosage form.

[0060] In the context of this invention, the term "dosage form" refers to the specific physical form in which a composition is prepared and administered to deliver active ingredients to a subject. This includes, but is not limited to, tablets, capsules, powders, liquids, gels, and chewables. The dosage form ensures the precise and consistent delivery of the active ingredients, tailored to the intended route of administration.

[0061] The compositions of the present invention may be formulated to provide a daily dose of probiotic bacteria in a range from, for example, 10e3 to 10el4, 10e4 to 10el2, 10e5 to 10el2, 10e6 to 10el2, 10e7 to lOell, and particularly 10e7 to lOelO colony forming units (cfu).

[0062] In the context of this patent application, the term "cfu" stands for "colony forming units," which is a measure of viable bacterial cells. It quantifies the number of individual cells that are capable of forming colonies when cultured under suitable conditions. This unit is commonly used to express the concentration of probiotics in nutritional supplements and other compositions, indicating the number of live microorganisms that can confer health benefits to the host.

[0063] The term "composition" as used herein refers to a combination or a mixture of two or more different ingredients, components, or substances. That is, the present invention relates to a composition comprising Bifidobacterium adolescentis strain NCC275 and at least one additional ingredient, component or substance. The composition of the invention can be any type of composition in which probiotic bacteria can be incorporated, such as a food product, a beverage, an animal feed product, a dietary or nutritional supplement for human or animal, a pharmaceutical composition or a cosmetic composition.

[0064] In certain embodiments, the composition according to the invention comprises Bifidobacterium adolescentis strain NCC275 and at least one excipient. In the context of the present invention, an "excipient" refers to an inactive substance that serves as a vehicle or carrier for the active probiotic ingredients, i.e., the B. adolescentis strain NCC275. Excipients may be used to facilitate the manufacturing process, enhance the stability, bioavailability, and delivery of the probiotics, and / or improve the overall formulation's taste, texture, and shelflife. Common excipients in probiotic compositions may include, inter alia, fillers, binders, disintegrants, preservatives, and flavoring agents. While excipients do not have physiological or therapeutic effects themselves, they may play a crucial role in ensuring the efficacy and usability of the probiotic product.

[0065] Accordingly, in a particular embodiment, the invention relates to a composition comprising Bifidobacterium adolescentis strain NCC275 (CNCM 1-6090) and at least one excipient.

[0066] In certain embodiments, the composition according to the invention comprises spray-dried and / or freeze-dried Bifidobacterium adolescentis strain NCC275. That is, Bifidobacterium adolescentis strain NCC275 may be added to the composition in spray-dried and / or freeze- dried form.

[0067] Bacteria, specifically probiotics, may be spray dried to convert them from a liquid suspension into a stable, dry powder form. This process typically involves the following steps: (i) preparation of a liquid bacterial suspension, optionally containing protective agents and / or stabilizers; (ii) atomizing the liquid suspension into fine droplets using a nozzle or rotary atomizer to create a mist of small droplets containing the probiotic bacteria; (iii) drying the atomized droplets in a chamber with a stream of hot air to rapidly evaporate the moisture from the droplets, resulting in the formation of dry particles; and (iv) collecting the dried probiotic powder from the drying chamber. The resulting powder contains viable probiotic bacteria in a stable, dry form.

[0068] Methods for spray-drying bacteria (probiotics) are known to the person skilled in the art and have been previously described in the art, for example, without limitation, in WO 2017 / 001590, which is fully incorporated herein by reference.

[0069] In certain embodiments, the spray-dried powder comprising Bifidobacterium adolescentis strain NCC275 comprises at least one polysaccharide, at least one antioxidant and / or at least one amino acid. In certain embodiments, the spray-dried powder comprising Bifidobacterium adolescentis strain NCC275 comprises a combination of two or more amino acids. In certain embodiments, the spray-dried powder comprises two or more amino acids selected from cysteine, lysine, alanine and arginine. In certain embodiments, the combination of the two or more amino acids may comprise: cysteine and alanine; cysteine, lysine and alanine; lysine and arginine; cysteine and arginine; cysteine, lysine and arginine; lysine, alanine and arginine; and cysteine, arginine and alanine, preferably wherein the amino acids lysine, alanine and arginine are each present in an amount of 8 to 20 wt.% based on total dry weight of the spray-dried powder and wherein cysteine is present in an amount of 2 to 10 wt.% based on the total dry weight of the spray-dried powder. The total amino acid concentration may be in the range from 3.5 to 36.5 wt%, based on the total dry weight of the spray-dried powder. The polysaccharide may be selected from maltodextrin, dextrin, cyclodextrin, starch, an oligosaccharide or cellulose. Preferably, the polysaccharide is maltodextrin. The concentration of the polysaccharide may be in the range of 20-60%, preferably 30-56% by weight, based on the total dry weight of the spray-dried powder. The antioxidant may be vitamin C, vitamin E, glutathione, coenzyme Q10, p-carotene, lycopene or vitamin A or a derivative thereof. Preferably, the antioxidant is sodium ascorbate (vitamin C). The antioxidant concentration may be in the range of 13-50%, preferably 20-40% by weight, based on the total dry weight of the spray-dried powder. Freeze-dried bacteria, specifically probiotics, refer to beneficial microorganisms that have undergone a lyophilization process to convert them from a liquid suspension into a stable, dry powder form. This process typically involves the following steps: (i) preparation of a liquid bacterial suspension, optionally containing cryoprotectants and / or other stabilizers to protect the bacteria during the freezing and drying processes; (ii) rapidly freezing the bacterial suspension at very low temperatures, typically below -40°C to solidify the suspension, (iii) primary drying (sublimation) of the frozen bacterial suspension in a vacuum chamber, where the pressure is reduced, and the temperature is slightly increased to remove the majority of the water content from the frozen suspension; (iv) secondary drying (desorption) at increasing temperatures to remove any residual moisture that remains bound to the bacterial cells to ensure that the final product has a very low moisture content; and (v) collecting the resulting freeze-dried probiotic powder.

[0070] Methods for obtaining freeze-dried bacteria are known to the person skilled in the art and have been previously described in the art.

[0071] Accordingly, in a particular embodiment, the invention relates to a composition comprising (i) Bifidobacterium adolescentis strain NCC275 and (ii) at least one excipient, and / or wherein the Bifidobacterium adolescentis strain NCC275 is spray-dried or freeze-dried.

[0072] It is preferred that the compositions disclosed herein comprise spray-dried or lyophilized (freeze-dried) Bifidobacterium adolescentis strain NCC275.

[0073] In certain embodiments, the composition according to the invention may consist of a spray- dried or freeze-dried powder of Bifidobacterium adolescentis strain NCC275. In certain embodiments, the composition may further comprise one or more excipients. In certain embodiments, the composition may comprise additional ingredients, such as, without limitation, a source of glutamate as described herein below.

[0074] The probiotic bacteria present in the composition of the present invention must be live probiotic bacteria, as their beneficial effects are associated with production of GABA by the bacteria in the gastrointestinal tract of a subject consuming the composition. Bacteria are considered as "live" when they are able to multiply under controlled culture conditions and form colonies or suspensions or when the microorganism's metabolic activity and / or membrane integrity can be established using methods known to the person skilled in the art, such as for example flow cytometry.

[0075] It is preferred herein that the composition of the present invention is formulated for administration to the gastrointestinal tract of a subject such that GABA can be produced by B. adolescentis NCC275 within the gastrointestinal tract, in particular in the small and large intestine, of said subject.

[0076] Nutritional compositions comprising a source of glutamate

[0077] Within the present invention, it is particularly preferred that the composition comprising Bifidobacterium adolescentis strain NCC275 is a nutritional composition or a nutritional product. A "nutritional composition" broadly refers to any composition of matter suitable for consumption by a mammalian subject. The term "nutritional product" refers to an end product that comprises the nutritional composition according to the invention and is ready for consumer use. The nutritional compositions and products according to the invention are generally suitable for oral consumption by a human.

[0078] The nutritional composition or product according to the invention comprises further ingredients in addition to the Bifidobacterium adolescentis strain NCC275. In preferred embodiments, the nutritional composition or product according to the invention comprises a source of glutamate.

[0079] Glutamate is the anionic form of the amino acid glutamic acid. The glutamate comprised in the compositions or products of the invention may be present in any salt form, including, without limitation, monosodium glutamate (MSG), monopotassium glutamate, calcium glutamate, and magnesium glutamate. Accordingly, the term "glutamate" as used herein encompasses any salt of glutamic acid. The term "source of glutamate" refers to any ingredient that comprises or consists of glutamate or glutamic acid (the protonated form). In particular, when the composition according to the invention is a liquid composition, the source of glutamate may comprise glutamic acid (the protonated form).

[0080] Accordingly, in a particular embodiment, the invention relates to a nutritional composition or product comprising Bifidobacterium adolescentis strain NCC275 (CNCM 1-6090) and a source of glutamate.

[0081] In certain embodiments, the source of glutamate may be pure or essentially pure glutamate. In certain embodiments, the source of glutamate may be any composition comprising at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 98%, or at least 99% glutamate.

[0082] In certain embodiments, the source of glutamate may be a product that is rich in glutamate, i.e., a product that comprises at least 0.01%, at least 0.1%, at least 1% at least 5%, at least 20%, at least 25%, at least 30%, at least 35%, or at least 40% in glutamate. Such products may be of natural or synthetic origin. Natural products that may be used as source of glutamate in the context of the present invention include, without limitation, fermentation products and cell extracts. Accordingly, in a particular embodiment, the invention relates to the nutritional composition or product according to the invention, wherein the source of glutamate is a natural product, in particular a fermentation product or a cell extract.

[0083] The source of glutamate may be present in the nutritional composition or product according to the invention in an amount ranging from about 0.01 % to about 90%, from about 0.04% to about 90%, from about 0.1% to 90%, from about 0.2% to about 90%, from about 0.5% to about 90%, or from about 1% to about 90%. More specifically, the source of glutamate may be present in the nutritional composition or product according to the invention in amounts ranging from about 0.01% to about 10%, about 0.1% to 10%, about 1% to about 10%, about 10% to about 20%, about 20% to about 30%, about 30% to about 40%, about 40% to about 50%, about 50% to about 60%, about 60% to about 70%, about 70% to about 80%, or about 80% to about 90%. Fermentation as the source

[0084] In certain embodiments, the source of glutamate is a fermentation product. A "fermentation product" refers to any substance or composition that is produced through the process of fermentation, which is a metabolic process where microorganisms such as bacteria, yeast, or fungi convert organic substrates (typically carbohydrates) into simpler compounds, often in the absence of oxygen. Fermentation products may be rich in glutamate due to the metabolic activities of the microorganisms involved in the fermentation process.

[0085] In certain embodiments, the source of glutamate is a prokaryotic (bacterial) fermentation product. Thus, in certain embodiments, the invention relates to the nutritional composition or product according to the invention, wherein the source of glutamate is obtainable through prokaryotic fermentation of a suitable substrate. In other embodiments, the source of glutamate is a yeast or fungal fermentation product.

[0086] In particular embodiments, the source of glutamate is a fermentation product, preferably a prokaryotic fermentation product, comprising between 0.01 to 70 % of glutamate, preferably 0.04 to 70% of glutamate, preferably 0.2 to 70% of glutamate , preferably between 5 and 50% of glutamate. That is, the fermentation product comprised in the nutritional composition or product according to the invention may comprise between 0.01 to 70 % of glutamate, between 0.04 to 70 % of glutamate, between 0.2 to 70 % of glutamate, between 1% and 70% of glutamate, between 5% and 70% of glutamate, between 10% and 70% of glutamate, between 20% and 70% of glutamate, between 20% and 60% of glutamate, or between 30% and 50% of glutamate.

[0087] In certain embodiments, the fermentation product comprised in the nutritional composition or product according to the invention comprises about 0.01% glutamate, about 0.04% glutamate, about 0.05% glutamate, about 0.1% glutamate, about 0.2% glutamate, about 0.5% glutamate, about 1% glutamate, about 2% glutamate, about 3% glutamate, about 4 % of glutamate about 5 % of glutamate, about 10 % of glutamate, about 15 % of glutamate, about 20 % of glutamate, about 25 % of glutamate, about 30 % of glutamate, about 35 % of glutamate, about 40 % of glutamate, about 45 % of glutamate, about 50 % of glutamate, about 55 % of glutamate, about 60 % of glutamate, about 65 % of glutamate, or about 70 % of glutamate.

[0088] The skilled person is aware of fermentation processes that result in fermentation products comprising glutamate. An exemplary process and products thereof are disclosed in WO 2010 / 108542, which is fully incorporated herein by reference. Accordingly, in certain embodiments, the source of glutamate may be any of the fermentation products disclosed in WO 2010 / 108542.

[0089] The source of glutamate may be obtained by fermenting a suitable substrate with a prokaryotic organism. Particularly preferred prokaryotic organism for the production of glutamic acid from a suitable substrate include, without limitation, Corynebacterium glutamicum, Corynebacterium ammoniagenes, Corynebacterium casei, Corynebacterium efficiens, Brevibacterium lactofermentum and Bacillus subtilis.

[0090] Thus, in a particular embodiment, the invention relates to the nutritional composition or product according to the invention, wherein the source of glutamate is obtainable through prokaryotic fermentation of a suitable substrate with one or more of the following bacteria: Corynebacterium glutamicum, Corynebacterium ammoniagenes, Corynebacterium casei, Corynebacterium efficiens, Brevibacterium lactofermentum and Bacillus subtilis.

[0091] For the fermentation process, any substrate may be used that results in a fermentation product comprising glutamate. The substrate is preferably a natural substrate. The natural substrate used may be a carbon and / or nitrogen source of any kind, provided that it is utilizable for the prokaryotic organism employed. As for the carbon source, monosaccharides or oligosaccharides, like glucose, fructose, mannose, trehalose, sucrose, or maltose; or enzymatic hydrolysates of starch or molasses may be used separately or in combination of two or more. As for the nitrogen source, ammonia, urea, ammonium salts, like ammonium sulphate, amino acids, peptides, proteins, yeast extract, corn steep liquor, enzymatic hydrolysate of plant material or starch, meat or fish products may be used separately or in combination of two or more. In certain embodiments, the substrate used in the fermentation process is hydrolyzed plant material or hydrolyzed starch. That is, in a particular embodiment, the invention relates to the nutritional composition or product according to the invention, wherein the substrate is obtained by enzymatic hydrolysis of a plant material and / or by enzymatic hydrolysis of starch.

[0092] Enzymatic hydrolysis of plant material is a biochemical process where specific enzymes break down complex carbohydrates, proteins, and other macromolecules in plant biomass into simpler, more accessible forms. This process typically involves a pretreatment step to disrupt plant cell walls, followed by the addition of enzymes such as cellulases, hemicellulases, amylases, proteases, and pectinases under controlled conditions of temperature and pH.

[0093] The plant material may be derived from any suitable plant. Preferably, the plant material is derived from plants that are rich in starch. In certain embodiments, the plant material is derived from one or more of wheat, corn, tapioca and rye.

[0094] Enzymatic hydrolysis of starch is a biochemical process where specific enzymes, primarily alpha-amylase and glucoamylase, break down starch, a complex carbohydrate, into simpler sugars. The starch can be derived from plants. Preferably, the starch is derived from wheat, corn, tapioca and rye.

[0095] Accordingly, in a particular embodiment, the invention relates to the nutritional composition or product according to the invention, wherein the plant is one or more of wheat, corn, tapioca and rye; and the starch is the starch of one or more of the above-mentioned plants.

[0096] In a preferred embodiment, the source of glutamate is obtained through prokaryotic fermentation of corn starch or, more preferably, enzymatically hydrolyzed corn starch.

[0097] Alternatively, the source of glutamate may be obtained through prokaryotic fermentation of glucose and / or polymers thereof, such as amylose and amylopectin. Accordingly, in certain embodiments, the suitable substrate may be glucose and / or a polymer thereof, preferable wherein the polymer is amylose and / or amylopectin. In a particular embodiment, the invention relates to the nutritional composition or product according to the invention, wherein the source of glutamate is a fermented starch, in particular a fermented corn starch, in particular wherein the starch, has been fermented with one or more of the following bacteria: Corynebacterium glutamicum, Corynebacterium ammoniagenes, Corynebacterium casei, Corynebacterium efficiens, Brevibacterium lactofermentum and Bacillus subtilis.

[0098] It is to be understood that the starch, and in particular the corn starch, may be enzymatically hydrolyzed prior to fermentation to be more accessible for the (prokaryotic) microorganisms.

[0099] A fermentation process for obtaining the source of glutamate may comprise the processing steps described below:

[0100] (i) Fermentation on a suitable substrate using a microorganism, preferably one or more of the following bacteria: Corynebacterium glutamicum, Corynebacterium ammoniagenes, Brevibacterium lactofermentum, Corynebacterium casei, Corynebacterium efficiens and Bacillus subtilis, and

[0101] (ii) cell disruption to yield a crude extract including cell debris.

[0102] After fermentation, the fermentation broth may undergo a cooking step to kill the fermenting microorganisms and to induce a controlled Mai Ila rd reaction.

[0103] This process may further comprise a step of removing cells or cell debris by filtration and / or centrifugation as known in the art.

[0104] However, in certain embodiments, the fermentation product is a non-purified fermentation product. That is, the crude extract may not undergo any purification steps to remove the cell debris. Accordingly, in a particular embodiment, the invention relates to the nutritional composition or product according to the invention, wherein the source of glutamate is a nonpurified fermentation product.

[0105] Fermentation may be carried out for 20 to 72 hours, at a pH between 5 and 9 and at a temperature between 25 and 40°C. Alternatively, fermentation may be carried out for 3 to 6 days, at a pH between 5 and 9 and at a temperature comprised 25 and 40°C. The skilled person is aware that the exact fermentation conditions depend on the type of microorganism and / or the type of substrate.

[0106] After the fermentation process, sodium chloride (NaCI) may be added to the fermentation broth. In certain embodiments, NaCI may be added to the fermentation broth after the cells and / or cell debris has been removed.

[0107] The fermentation product is typically a liquid fermentation product, which can be directly utilized in various applications. However, for enhanced versatility and extended shelf life, the liquid fermentation product may undergo drying processes known in the art such as spray drying, vacuum drying, or belt drying to produce a stable powder form. This powder can then be further processed and converted into any desired physical form.

[0108] The obtained fermentation product may be subsequently mixed with a natural hydrolysate, such as the natural hydrolysate disclosed in US 6 / 838,100, which is fully incorporated herein by reference.

[0109] Cell extracts as the source of glutamate

[0110] In certain embodiments, the source of glutamate may be a cell extract. The cell extract can be derived from various microbial, plant, or animal cells known to produce or contain high levels of glutamate. Examples of suitable sources include bacterial cultures such as Corynebacterium glutamicum, yeast extracts, algal extracts, or plant cell cultures. The cell extract may be obtained through processes such as fermentation, lysis, and, optionally, subsequent purification to concentrate the glutamate content. Additionally, the cell extract may be further processed to remove unwanted components, ensuring that the final product is rich in glutamate and suitable for use in nutritional products. The use of cell extracts as a source of glutamate can provide a natural and efficient means of incorporating this amino acid into various dietary formulations. Accordingly, in a particular embodiment, the invention relates to the nutritional composition or product according to the invention, wherein the source of glutamate is a cell extract, in particular a yeast extract.

[0111] In the context of the present invention, the term "yeast extract" refers to a complex mixture derived from the autolysis, hydrolysis, or enzymatic breakdown of yeast cells. Yeast extract is rich in proteins, peptides, amino acids, vitamins, minerals, and other bioactive compounds. It is typically produced by subjecting yeast cells, such as those from Saccharomyces cerevisiae, to processes that disrupt the cell walls and release the intracellular contents. The resulting extract can be further refined and concentrated to enhance specific components, depending on the intended application. Yeast extract is commonly used as a flavor enhancer, nutrient supplement, and growth medium in various food, pharmaceutical, and biotechnological applications due to its high nutritional value and functional properties.

[0112] In certain embodiments, the cell extract, in particular the yeast extract is dried as known in the art, for example by spray drying, vacuum drying, or belt drying, to produce a stable powder form.

[0113] The cell extract comprised in the nutritional composition or product according to the invention, in particular the yeast extract, preferably comprises at least 5%, at least 10%, at least 15%, at least 20%, at least 25% glutamate, at least 30% glutamate, at least 35% glutamate or at least 40% glutamate.

[0114] Additional ingredients of the composition according to the invention

[0115] The composition according to the invention, in particular the nutritional composition or product according to the invention, may comprise one or more of the ingredients described herein below in addition to the Bifidobacterium adolescentis strain NCC275 and, optionally, the source of glutamate. The ingredients described herein below may be an endogenous constituent of a (prokaryotic) fermentation product and / or a (yeast) cell extract or may be externally added to the composition according to the invention. The composition according to the invention, and, in particular, the source of glutamate comprised therein, may comprise one or more nucleotides, in particular nucleotide monophosphates.

[0116] In certain embodiments, the composition according to the invention, and, in particular, the source of glutamate comprised therein, may comprise one or more of adenosine monophosphate (AMP), cytidine monophosphate (CMP), guanosine monophosphate (GMP), uridine monophosphate (UMP) or inosine monophosphate (IMP).

[0117] In certain embodiments, the source of glutamate comprised in the nutritional composition or product according to the invention, in particular the (prokaryotic) fermentation product, may comprise between 0.01 to 70 % of nucleotides, between 0.01 to 60 % of nucleotides, between 0.01 to 50 % of nucleotides, between 0.01 to 40 % of nucleotides, between 0.01 to 30 % of nucleotides, between 0.01 to 20 % of nucleotides, between 0.05 to 20 % of nucleotides, between 0.05 to 10 % of nucleotides, between 0.05 to 5 % of nucleotides, or between 0.1 to 2.5 % of nucleotides.

[0118] In certain embodiments, the composition according to the invention, and, in particular, the source of glutamate comprised therein, may comprise guanosine monophosphate (GMP) and / or inosine monophosphate (IMP).

[0119] In certain embodiments, the source of glutamate comprised in the nutritional composition or product according to the invention, in particular the (prokaryotic) fermentation product, may comprise between 0.01 to 70 % of GMP and / or IMP, between 0.01 to 60 % of GMP and / or IMP, between 0.01 to 50 % of GMP and / or IMP, between 0.01 to 40 % of GMP and / or IMP, between 0.01 to 30 % of GMP and / or IMP, between 0.01 to 20 % of GMP and / or IMP, between 0.01 to 10 % of GMP and / or IMP, between 0.01 to 5 % of GMP and / or IMP, between 0.05 to 2.5 % of GMP and / or IMP, or between 0.05 to 1 % of GMP and / or IMP.

[0120] The composition according to the invention, and, in particular, the source of glutamate comprised therein, may comprise further amino acids in addition to glutamate. In certain embodiments, the composition according to the invention, and, in particular, the source of glutamate comprised therein, may comprise one or more proteinogenic amino acids in addition to glutamate.

[0121] In certain embodiments, the composition according to the invention, and, in particular, the source of glutamate comprised therein, may comprise one or more of: alanine, aspartic acid, glutamine, glycine, leucine, lysine, methionine, tryptophan and valine. Accordingly, in a particular embodiment, the invention relates to the nutritional composition or product according to the invention, wherein the source of glutamate comprises further amino acids, in particular wherein the further amino acids are one or more of: alanine, aspartic acid, glutamine, glycine, leucine, lysine, methionine, tryptophan and valine.

[0122] The composition according to the invention, and, in particular, the source of glutamate comprised therein, may further comprise polysaccharides and / or carbohydrates.

[0123] In certain embodiments, the source of glutamate comprised in the nutritional composition or product according to the invention, in particular the (prokaryotic) fermentation product or the (yeast) cell extract, may comprise between 0 and 30 % of polysaccharides. These polysaccharides may be, or may be derived from, cellulose, pectin, starch, maltodextrins, or a combination thereof.

[0124] Accordingly, in a particular embodiment, the invention relates to the nutritional composition or product according to the invention, wherein the source of glutamate further comprises polysaccharides, in particular wherein the polysaccharides are present between 0 and 30 %, in particular, wherein the polysaccharides are one or more of: cellulose, pectin, starch and maltodextrins.

[0125] In certain embodiments, the composition according to the invention, and, in particular, the source of glutamate comprised therein, may further comprise one or more carbohydrates or sugars selected from glucose, fructose, mannose, sorbitol, and / or glycerol.

[0126] In a particular embodiment, the source of glutamate is a (yeast) cell extract that comprises maltodextrin. The composition according to the invention, and, in particular, the source of glutamate comprised therein, may further comprise one or more organic acids, or salts thereof. Organic acids are a class of organic compounds characterized by the presence of one or more carboxyl groups (-COOH). These acids are typically weak acids and are commonly found in various natural sources, including plants, animals, and microorganisms. Organic acids play crucial roles in metabolic processes and are widely used in the food, pharmaceutical, and chemical industries due to their preservative, flavoring, and pH-regulating properties. In certain embodiments, the organic acid is lactic acid (or a lactate salt), citric acid (or a citrate salt), acetic acid (or an acetate salt) or malic acid (or a malate salt).

[0127] Accordingly, in a particular embodiment, the invention relates to the nutritional composition or product according to the invention, wherein the source of glutamate further comprises organic acids, or salts thereof, in particular wherein the organic acids are one or more of: lactic acid (lactate), citric acid (citrate), acetic acid (acetate) and malic acid (malate). The amount of lactate in the source of glutamate, in particular the (prokaryotic) fermentation product, may be between 0.01 and 8 %, that of acetate between 0.01 and 6 % and that of citrate between 0.01 and 8 %.

[0128] The composition according to the invention, and, in particular, the source of glutamate comprised therein, may further comprise minerals. Minerals are essential inorganic elements required by the human body in small quantities to support various physiological functions and maintain overall health. These elements, which include, without limitation calcium, magnesium, potassium, iron, zinc, and selenium, are vital for processes such as bone development, nerve transmission, and metabolic regulation. As the body cannot synthesize these minerals, they must be obtained through dietary intake or supplementation to prevent deficiencies and ensure proper bodily function.

[0129] In certain embodiments, the invention relates to the nutritional composition or product according to the invention, wherein the source of glutamate further comprises minerals, in particular wherein the minerals are one or more of: sodium, potassium, chlorine, calcium, phosphorus, magnesium, iron, copper, zinc, iodine, manganese, selenium, chromium, molybdenum and fluorine.

[0130] The composition according to the invention, and, in particular, the source of glutamate comprised therein, may further comprise fatty acids.

[0131] A fatty acid is an organic compound composed of a hydrocarbon chain with a terminal carboxyl group (-COOH). Fatty acids are fundamental building blocks of lipids, which are essential components of cell membranes and energy storage molecules. They can vary in chain length, degree of saturation (presence of double bonds), and configuration (cis or trans). Fatty acids are classified into saturated fatty acids, which have no double bonds, and unsaturated fatty acids, which contain one or more double bonds.

[0132] In certain embodiments, the composition according to the invention, and, in particular, the source of glutamate comprised therein, comprises saturated and / or unsaturated fatty acids. In certain embodiments, the composition according to the invention, and, in particular, the source of glutamate comprised therein, is free or essentially free of saturated and / or trans fatty acids.

[0133] The composition according to the invention, and in particular the nutritional composition or product according to the invention, may comprise further ingredients in addition to the Bifidobacterium adolescentis strain NCC275 and, optionally, the source of glutamate.

[0134] For example, the composition may further include prebiotics such as inulin, fructooligosaccharides (FOS), and galactooligosaccharides (GOS) to promote the growth and activity of the probiotics. Additionally, the composition may comprise vitamins and minerals, including but not limited to, vitamin D, vitamin C, vitamin E, calcium, and zinc, to support overall health. Fiber sources such as soluble fiber and insoluble fiber may be included to aid in digestive health. The composition may also incorporate amino acids, antioxidants, and omega-3 fatty acids to provide comprehensive nutritional benefits. Flavoring agents, natural sweeteners, and essential oils may be added to improve palatability. Stabilizers and preservatives may be used to enhance the shelf life of the product and prevent caking. In certain embodiments, the nutritional composition or product according to the invention comprises inulin. Inulin is a naturally occurring polysaccharide and prebiotic fiber composed primarily of fructose units, commonly extracted from plants such as chicory root. It is utilized in food supplements and functional foods for its ability to promote digestive health by stimulating the growth of beneficial gut bacteria. Accordingly, in a particular embodiment, the invention relates to the nutritional composition or product according to the invention, wherein the nutritional composition or product further comprises inulin.

[0135] In certain embodiments, the nutritional composition or product according to the invention comprises maltodextrin. Maltodextrin is a polysaccharide derived from the partial hydrolysis of starch, typically sourced from corn, rice, potato, or wheat. It is composed of glucose units linked primarily by a-1,4 glycosidic bonds and is characterized by its varying degree of polymerization, which affects its sweetness and solubility. In nutritional products, maltodextrin is commonly used as a filler, binder, or carrier due to its neutral taste, high solubility, and ease of digestion. It serves to bulk up the volume of products, stabilize active ingredients, improve texture, and enhance the consistency of powdered and liquid formulations.

[0136] The nutritional composition or product according to the invention may further be supplemented with exogenous amino acids and / or vitamins.

[0137] Amino acids are organic compounds that serve as the building blocks of proteins and play critical roles in various physiological functions. Amino acids are essential nutrients that support muscle growth, repair tissues, and contribute to the synthesis of enzymes and hormones. They are categorized into essential amino acids, which must be obtained through the diet as the body cannot synthesize them, and non-essential amino acids, which the body can produce, nutritional products often include amino acids to enhance nutritional value, support athletic performance, promote recovery, and maintain overall health. These products may contain individual amino acids or a blend. Vitamins are organic compounds that are essential for normal physiological functions and overall health. In the context of nutritional products, vitamins are included to supplement the diet and ensure adequate intake of these crucial nutrients, which the body either cannot produce at all or cannot produce in sufficient quantities. They play vital roles in a wide range of bodily processes, including metabolism, immune function, and cell and tissue maintenance. The nutritional composition and product according to the invention may comprise include Vitamin A (Retinol), Vitamin Bl (Thiamine), Vitamin B2 (Riboflavin), Vitamin B3 (Niacin), Vitamin B5 (Pantothenic Acid), Vitamin B6 (Pyridoxine), Vitamin B7 (Biotin), Vitamin B9 (Folate / Folic Acid), Vitamin B12 (Cobalamin), Vitamin C (Ascorbic Acid), Vitamin D, Vitamin E (Tocopherol), and / or Vitamin K.

[0138] In certain embodiments, the nutritional composition or product according to the invention may comprise one or more probiotic strains in addition to Bifidobacterium adolescentis strain NCC275. Exemplary strains include, without limitation, Bifidobacterium, Lactobacillus, Limosilactobacillus, Lacticaseibacillus, Lactiplantibacillus, Saccharomyces, Enterococcus, Streptococcus, Pediococcus, Leuconostoc and Bacillus. In further , strain include, without limitation, Bifidobacterium longum, Bifidobacterium lactis subsp. lactis, Limosilactobacillus reuteri, Lacticaseibacillus paracasei, Lactobacillus johnsonii, Lacticaseibacillus rhamnosus, Lactobacillus acidophilus, Lactiplantibacillus plantarum, Streptococcus thermophilus, Bifidobacterium bifidum, Lactobacillus salivarius, Lactobacillus brevis, Lactobacillus fermentum, Bifidobacterium breve, Lactobacillus helveticus, Bacillus subtilis, Bacillus coagulans, Lactococcus lactis, and Bacillus clausii.

[0139] The Bifidobacterium adolescentis strain NCC275 may be administered in combination with one or more probiotic strains different from Bifidobacterium adolescentis strain NCC275 as described above. The Bifidobacterium adolescentis strain NCC275 and one or more probiotic strains different from Bifidobacterium adolescentis strain NCC275 as described above may be administered separately, simultaneously or sequentially. In preferred embodiments, The Bifidobacterium adolescentis strain NCC275 and one or more probiotic strains different from Bifidobacterium adolescentis strain NCC275 as described above are administered simultaneously. In the context of the present invention, all percentage values are to be understood as weight / weight (wt / wt) percentages for dry compositions and volume / volume (v / v) percentages for liquid compositions, unless otherwise specified.

[0140] In a preferred embodiment of the invention, the nutritional composition or product is a powder and the percentage values are to be understood as weight / weight (wt / wt) percentages.

[0141] Dosage forms of the composition according to the invention

[0142] The composition according to the invention, particularly the nutritional composition or product, may be formulated in any physical dosage form, provided that the probiotic microorganisms remain stable and active. Suitable physical dosage forms include, but are not limited to, powders, solids (including capsules, tablets, and granules), liquids, gels, and chewables. The choice of dosage form will depend on factors such as the intended use, ease of administration, and consumer preference. The composition may be designed to protect the probiotics from environmental factors such as moisture, heat, and oxygen, which could otherwise compromise their stability and activity.

[0143] In a particular embodiment, the invention relates to the nutritional composition or product according to the invention, wherein the nutritional composition or product is or comprises a powder, a solid, a semi-solid, a liquid, or a semi-liquid.

[0144] Within the present invention, the term "powder" refers to a dry, bulk solid composed of fine particles that can flow freely when shaken or tilted. In the context of the nutritional composition or product, a powder may include finely ground ingredients such as dried probiotics, and other dietary additives, which can be easily mixed with liquids or other carriers for consumption.

[0145] The term "solid" as used herein refers to a state of matter characterized by structural rigidity and resistance to changes in shape or volume. For the nutritional composition or product, a solid dosage form may include tablets, capsules, granules, or bars, where the ingredients are compressed or molded into a firm, stable shape.

[0146] The term "semi-solid" as used herein refers to a state of matter that exhibits properties between those of a solid and a liquid, having a definite shape but being deformable under pressure. In the context of the nutritional composition or product, a semi-solid may include gels, pastes, or soft chews, where the ingredients are combined to create a thick, cohesive, yet pliable form.

[0147] The term "liquid" as used herein refers to a state of matter that flows freely and conforms to the shape of its container while maintaining a constant volume. For the nutritional composition or product, a liquid dosage form may include solutions, suspensions, or emulsions, where the ingredients are dissolved or dispersed in a fluid medium for easy ingestion.

[0148] The term "semi-liquid" as used herein refers to a state of matter that exhibits properties between those of a liquid and a semi-solid, having a thicker consistency than a liquid but still able to flow. In the context of the nutritional composition or product, a semi-liquid may include syrups, thickened beverages, or viscous suspensions, where the ingredients are combined to create a dense, pourable form.

[0149] It is particularly preferred herein that the nutritional composition or product according to the invention is a powder that can be reconstituted in a liquid. Powders offer several advantages, including ease of storage, transportation, and extended shelf life due to its low moisture content. The reconstitutable powder may be conveniently mixed with water, milk, juice, or other suitable liquids to create a ready-to-consume nutritional product. The powder dosage form allows for precise dosing and flexibility in usage, making it suitable for a wide range of applications, from daily dietary supplements to specialized nutritional needs. Additionally, the powder may be easily fortified with various other bioactive compounds, ensuring a comprehensive nutritional profile. The reconstitutable nature of the powder ensures that the probiotics and other sensitive ingredients remain stable until the point of consumption, thereby maximizing their efficacy and health benefits. This form is particularly advantageous for consumers seeking a portable, customizable, and user-friendly nutritional product.

[0150] The nutritional composition or product in powder dosage form is preferably characterized by its low water activity, which is a critical factor in ensuring the stability and shelf life of the product. Water activity (aw) is a measure of the free moisture available in the powder that can support microbial growth and chemical reactions. By maintaining a low water activity, typically below 0.6, the powder inhibits the growth of bacteria, yeast, and molds, thereby enhancing its microbiological safety and extending its shelf life. Additionally, low water activity helps to preserve the potency and effectiveness of sensitive ingredients such as probiotics, vitamins, and enzymes, which can degrade in the presence of moisture.

[0151] Accordingly, in a particular product, the invention relates to the nutritional composition or product according to the invention, wherein the nutritional composition or product is a powder, in particular a reconstitutable powder.

[0152] The term "reconstitutable powder" refers to a dry, finely divided substance that is designed to be mixed with a liquid, such as water, milk, or juice, to form a solution, suspension, or emulsion that is ready for consumption. This type of powder is formulated to dissolve or disperse easily and uniformly when combined with the liquid, ensuring that the active ingredients are effectively delivered in the intended dosage. The formulation of a reconstitutable powder typically involves the inclusion of stabilizers, emulsifiers, and anticaking agents to maintain its free-flowing properties and ensure consistent reconstitution.

[0153] In the context of this invention, the term "dosage form" for a reconstitutable powder refers to the specific amount of powder that is intended to be mixed with a liquid to create a single serving of the final product. This dosage form ensures the precise and consistent delivery of active ingredients, such as probiotic bacteria, when reconstituted according to the recommended instructions. It is tailored to provide the intended therapeutic or nutritional benefits in each serving. The probiotic bacteria according to the invention may be incorporated into the composition in an amount ranging from 10e3 to 10el2 cfu / g of powder, with a more preferred range being from 10e7 to 10el2 cfu / g of powder, with an even more preferred range being from 10e7 to lOell cfu / g of powder. When the composition is an infant formula, the amount of probiotic in the infant formula may be between 105and 108cfu / g, preferably between 106and 5xl07cfu / g, of infant formula powder.

[0154] In certain embodiments, the nutritional composition or product according to the invention may be a liquid composition, such as, without limitation, a chilled dairy product. This includes, but is not limited to, products like milk, yogurt, kefir, and drinkable dairy-based smoothies.

[0155] Uses of the composition according to the invention

[0156] The composition according to the invention, in particular the nutritional composition or product according to the invention, may comprise Bifidobacterium adolescentis strain NCC275 as the main ingredient. In such embodiments, the composition is specifically formulated to deliver Bifidobacterium adolescentis strain NCC275 to the gastrointestinal tract, where it can enhance the production of GABA and thereby promote associated nutritional, health and / or cosmetic benefits.

[0157] Alternatively, the composition according to the invention, in particular the nutritional composition or product according to the invention, may comprise additional functional ingredients other than Bifidobacterium adolescentis strain NCC275 that can promote nutritional, health and / or cosmetic benefits.

[0158] In a preferred embodiment, the nutritional composition or product according to the invention is a supplement. The term "supplement" refers to a product intended to enhance the diet by providing additional nutrients, bioactive compounds, or other beneficial substances that may not be consumed in sufficient quantities through regular food intake. These products are designed to support overall health, address specific nutritional deficiencies, promote physiological functions, and / or achieve targeted health outcomes. In certain embodiments, the nutritional composition or product according to the invention may be a dietary supplement. A "dietary supplement" is a product intended to supplement the diet and provide nutrients or other bioactive compounds that may not be consumed in sufficient quantities through food alone. Dietary supplements according to the invention comprise Bifidobacterium adolescentis strain NCC275 and, optionally, a source of glutamate, to increase the production of GABA in the gastrointestinal tract and thereby promote associated dietary, health and / or cosmetic benefits.

[0159] In certain embodiments, the nutritional composition or product according to the invention may be a nutritional supplement. A "nutritional supplement" refers to a product intended to provide essential nutrients that may be missing or insufficient in a subject's diet. These nutrients may include vitamins, minerals, amino acids, fatty acids, and other essential nutrients. Nutritional supplements are designed to support overall health and well-being by ensuring that individuals receive the necessary nutrients to maintain optimal bodily functions. Nutritional supplements according to the invention may be supplemented with Bifidobacterium adolescentis strain NCC275 and, optionally, a source of glutamate, to increase the production of GABA in the gastrointestinal tract and thereby promote associated nutritional, health and / or cosmetic benefits.

[0160] In one embodiment the composition is an infant formula. The term "infant formula" may refer to a foodstuff intended for particular nutritional use by infants during the first year of life and satisfying by itself the nutritional requirements of this category of person, as defined in European Commission Directive 2006 / 141 / EC of 22 December 2006. Infants can be fed solely with infant formulas or the infant formula can be used as a complement of human milk.

[0161] The term "infant formula" includes hypoallergenic infant formulas. A hypoallergenic composition is a composition which is unlikely to cause allergic reactions.

[0162] In some embodiment, the expression "infant formula" encompasses both "starter infant formula" and "follow-up formula" or "follow-on formula". As used herein, a "follow-up formula" or "follow-on formula" is given from the 6th month onwards and thus may also be given to toddlers. It constitutes the principal liquid element in the progressively diversified diet of this category of person.

[0163] The infant formula of the invention may be in the form of a powder or liquid. The liquid may be, for example, a concentrated liquid infant formula or a ready-to-feed formula. The infant formula may be in the form of a reconstituted infant formula (i.e. a liquid infant formula that has been reconstituted from the powdered form). Preferably, the infant formula is in the form of a powder.

[0164] The powder is preferably capable of being reconstituted into a liquid composition suitable for feeding an infant, for example by the addition of water. Similarly, the concentrated liquid infant formula is preferably capable of being diluted into a liquid composition suitable for feeding an infant, for example by the addition of water.

[0165] Accordingly, in a particular embodiment, the invention relates to the nutritional composition or product according to the invention, wherein the nutritional composition or product is a dietary supplement or a nutritional supplement.

[0166] Therapeutic uses

[0167] The Bifidobacterium adolescentis strain NCC275 may be comprised in a food for special medical purpose (FSMP). As a food for special medical purpose (FSMP), the composition may be specifically designed to meet the nutritional needs of individuals with particular medical conditions or dietary restrictions. This may include formulations for individuals with malabsorption disorders, metabolic conditions, or those requiring specialized nutrition during recovery from illness or surgery. The FSMP can be tailored to provide precise nutrient profiles, including macronutrients, micronutrients, and therapeutic agents, to support the management of specific health conditions. Accordingly, in a particular embodiment, the invention relates to the nutritional composition or product according to the invention, wherein the nutritional composition or product is a food for special medical purpose (FSMP).

[0168] The probiotic bacteria according to the invention, or the composition comprising the probiotic bacteria according to the invention and, optionally, a source of glutamate, can be used in therapy. Thus, the invention also provides for such probiotic bacteria and such compositions for use in therapy.

[0169] For example, the probiotic bacteria or the composition according to the invention comprising the probiotic bacteria may be used in the treatment or prevention of a disease or medical condition. The treatment or prevention of a disease or condition includes managing, alleviating, mitigating, or curing the symptoms or underlying causes of a disease or medical condition. The term "treatment" encompasses actions taken to improve the health status of an individual already affected by a disease or condition, including reducing symptoms, slowing disease progression, and achieving remission or cure. The term "prevention" involves measures taken to avert the onset of a disease or condition in individuals who are at risk but have not yet manifested symptoms. This includes prophylactic interventions that reduce the likelihood of developing the disease or condition. The terms "delay" or "delaying" refer to interventions that postpone the onset or progression of a disease or condition, thereby extending the period of time before symptoms appear or worsen.

[0170] Accordingly, in certain embodiments, the invention provides the probiotic bacteria or the composition as described above for use in the following specific therapeutic methods: a) for use in reducing one or more symptoms of stress in a subject; b) for use in reducing one or more symptoms of premenstrual syndrome in a subject; c) for use in reducing pain in a subject; d) for use in improving sleep in a subject; e) for use in the prevention or delay of age-related decline in physiological and / or health functions in a subject; and f) for use in the prevention or delay of conditions associated with aging in a subject. The probiotic bacteria or the composition according to the invention comprising the probiotic bacteria may also be used in the dietary management of a disease or condition. The term "manage," "managing," and "management" as used herein, including in the context of "dietary management", refers to the prevention or reduction in severity or frequency of one or more symptoms of a disease or condition, including ameliorating one or more existing symptoms of a disease or condition, preventing one or more existing symptoms of disease or condition, preventing one or more underlying causes of a disease or condition, ameliorating one or more underlying cause of a disease or condition, reducing the prevalence of one or more symptoms of a disease or condition, and / or reducing the occurrence of one or more symptoms of a disease or condition. It will be understood to include stabilizing a disease or condition and preventing progression of a disease or condition. In the context of the nutritional compositions or products disclosed herein, it is understood that such nutritional compositions or products can be used in the dietary management of a disease or condition.

[0171] Accordingly, in certain embodiments, the invention provides the probiotic bacteria or the composition as described above for use in the following specific therapeutic methods: a) for use in the dietary management of stress in a subject; b) for use in the dietary management of premenstrual syndrome in a subject; c) for use in the dietary management of pain in a subject; d) for use in the dietary management of sleep in a subject; e) for use in the dietary management of age-related decline in physiological and / or health functions in a subject; and f) for use in the dietary management of conditions associated with aging in a subject.

[0172] Also encompassed herein are methods of treatment utilizing the probiotic bacteria according to the invention or the compositions described herein. Accordingly, in certain embodiments, the invention relates to a method for: a) reducing one or more symptoms of stress in a subject; b) reducing one or more symptoms of premenstrual syndrome in a subject; c) reducing pain in a subject; d) improving sleep in a subject; e) preventing or delaying age-related decline in physiological and / or health functions in a subject; or f) preventing or delaying conditions associated with aging in a subject; comprising administering to the subject the probiotic bacteria or the composition or product according to the invention.

[0173] In further embodiments, the invention relates to a method for: a) the dietary management of stress in a subject; b) the dietary management of premenstrual syndrome in a subject; c) the dietary management of pain in a subject; d) the dietary management of sleep in a subject; e) the dietary management of age-related decline in physiological and / or health functions in a subject; or f) the dietary management of conditions associated with aging in a subject; comprising administering to the subject the probiotic bacteria or the composition or product according to the invention.

[0174] GABA has been reported in the literature to positively impact various symptoms of stress. Specifically, the probiotic bacteria or the composition according to the invention may be used to increase relaxation and / or calmness in a subject, to reduce morning drowsiness in a subject, to restore mental calmness in a subject, and / or to support mental relaxation in a subject.

[0175] Such stress-reducing effects of GABA have been reported, inter alia, by Abdou et al. (Biofactors (2006) 26(3):201-8), Yoto et al. (Amino Acids (2012) 43(3):1331-7), Nobile et al. (Neuropsychobiology (2022) 81(2) :116-126), Patterson et al. (Sci Rep (2019) 9(1) :16323) and Ko et al. (Process Biochemistry (2013) 48(4) :559-568). In certain embodiments, the symptom of stress may be depression. Accordingly, in certain embodiment, the invention relates to the bacteria or composition according to the invention for use in treating or preventing depression in a subject. In certain embodiment, the invention relates to the bacteria or composition according to the invention for use in the dietary management of depression in a subject.

[0176] GABA has also been reported in the literature to exhibit sleep-promoting effects. For example, it was reported by Yamatsu et al. (Journal of Nutritional Science and Vitaminology (2015) 61(2):182-187; and Food Sci Biotechnol (2016) 25(2):547-551) that GABA can shorten sleep latency. Okada et al. (Nippon Shokuhin Kagaku Kogaku Kaishi (2000) 47(8) :596-603) reported the use of GABA for improving sleeplessness in post-menopausal women. Yamatsu et al. (Japanese Pharmacology and Therapeutics (2013) 41( 10) :985-988) reported the use of GABA for improving sleep quality and for decreasing night urination in elderly people.

[0177] Accordingly, the improvement of sleep and / or the dietary management of sleep may be characterized, inter alia, by a shorter sleep latency, reduced sleeplessness / insomnia, improved sleep quality and / or reduced night urination.

[0178] The probiotic bacteria or composition according to the invention may be used for reducing the symptoms of premenstrual syndrome and / or for the dietary management of premenstrual syndrome. The term "premenstrual syndrome" (also referred to as 'PMS') as used herein encompasses conditions associated with the female menstrual cycle involving a variety of physical and mental symptoms, which symptoms typically peak during the late luteal phase (i.e. after ovulation) of a woman's cycle and abate with the beginning of menstrual blood loss. The severity of symptoms associated with PMS range from mild to incapacitating and it has been estimated that up to 90% of women who menstruate suffer from some degree of PMS, while 20-40% suffer symptoms severe enough to lead to physical or mental incapacitation. The severity of symptoms can vary from month to month or year to year for a particular woman. Symptoms associated with PMS typically include depression, anger, irritability, anxiety, bloating, weight gain, abdominal pain, muscles and / or joint pain, breast pain and swelling, headache, fatigue, difficulty sleeping, difficulty concentrating, confusion, food cravings, lack of sexual interest and / or decreased efficiency. The probiotic bacteria or composition according to the invention may also be used for reducing pain and / or the dietary management of pain. The term "pain" as used herein is given its broadest sense and includes an unpleasant sensory and emotional experience associated with actual or potential tissue damage, or described in terms of such damage and includes the more or less localized sensation of discomfort, distress, or agony, resulting from the stimulation of specialized nerve endings.

[0179] The pain may be of any kind, including, without limitation, acute pain, chronic pain, nociceptive pain (including visceral and somatic pain), neuropathic pain, and inflammatory pain.

[0180] Numerous reports exist linking GABA to the treatment of different kinds of pain.

[0181] For example, it has been reported by Pokusaeva et al. (Neurogastroenterol Motil (2017) 29(l):el2904) that GABA-producing probiotics can reduce visceral hypersensitivity and modulate sensory neuron activity to treat abdominal pain. The use of GABA-producing probiotics for inhibiting visceral hypersensitivity triggered by partial restraint stress has been disclosed in WO2020 / 157297 and Laroute et al. (Elife, 2022. 11: p. e77100). It is demonstrated in Fig.3 of Laroute et al. that GABA-producing probiotics can reduce the frequency of abdominal contractions in a stress-induced visceral hypersensitivity rat model.

[0182] Thus, in a particular embodiment, the pain is visceral pain. Further evidence linking GABA signaling with the treatment of visceral pain is provided, for example, by Loeza-Alcocer et al. (J Physiol, 2019, 597(13):p.3425-3439; e.g., last sentence of Abstract).

[0183] Visceral pain is a type of nociceptive pain that originates from the internal organs located within the thoracic, abdominal, or pelvic cavities. Unlike somatic pain, which arises from the skin, muscles, and joints, visceral pain is often diffuse, poorly localized, and can be described as deep, aching, or cramping. It is frequently associated with autonomic symptoms such as nausea, vomiting, changes in heart rate, and blood pressure fluctuations. Visceral pain is commonly linked to conditions affecting the internal organs, such as irritable bowel syndrome (IBS), inflammatory bowel disease (IBD), endometriosis, and other gastrointestinal or gynecological disorders. The complexity of visceral pain arises from the intricate network of sensory nerves that transmit pain signals from the internal organs to the central nervous system, often making it challenging to diagnose and treat effectively.

[0184] In certain embodiments, the pain, in particular the visceral pain, may be associated with menstruation and / or endometriosis. For example, it has been reported by Sandstrom et al. (Acta Obstet Gynecol Scand, 2023, 102(10): 1316-1322) that enhancing GABAergic function may play a role managing pain associated with endometriosis.

[0185] Another report by Mills (Journal of Nutrition and Metabolism, 2021:6655064) links pain associated with old age to a decline in GABAergic function. Mills further links GABA with the amelioration of neuropathic pain and inflammatory pain. Thus, in a particular embodiment, the pain is pain associated with aging, neuropathic pain and / or inflammatory pain.

[0186] A report by Benke (Current Topics in Behavioral Neurosciences, vol 52. Springer, Cham. https: / / doi.org / 10.1007 / 7854_2020_130) proposes the GABAB receptor as a target for the treatment of chronic pain. Therefore, in certain embodiments, the pain may be chronic pain.

[0187] Zeevenhooven et al. (Nature Reviews, 2018, 15: p.479-496) link pain associated with colic in infants to increased visceral hypersensitivity and colonic contractions. In light of the findings by Laroute et al. (Elife, 2022, 11: p. e77100) that GABA-producing probiotics can inhibit visceral hypersensitivity, it is plausible that GABA-producing probiotics can be used to prevent or treat pain in infants or toddlers with colic, potentially preventing or at least reducing the frequency of colic episodes. Therefore, in certain embodiments, the bacteria or composition according to the invention may be used in the prevention, treatment or dietary management of pain associated with colic in infants or toddlers. In certain embodiments, the pain associated with colic in infants or toddlers is visceral pain. In certain embodiments, the bacteria or composition according to the invention may be used in the prevention, treatment or dietary management of colic in infants or toddlers. In such embodiments, the composition according to the invention is preferably an infant formula. Colic in infants is a condition characterized by frequent, prolonged, and intense episodes of crying or fussiness in an otherwise healthy baby, typically occurring in the first months of life without an apparent cause. Colic may be diagnosed as known in the art, for example as described in Zeevenhooven et al. (Pediatr Gastroenterol Hepatol Nutr, 2017, 20(1): p.1-13). While colic is most commonly associated with infants, toddlers can also be affected by colic, experiencing similar symptoms of gastrointestinal discomfort and distress.

[0188] In certain embodiments, the bacteria or composition according to the invention may be used in the treatment, prevention or dietary management of symptoms of irritable bowel syndrome (IBS) or inflammatory bowel diseases (IBD), in particular wherein the symptoms are visceral pain and / or anxiety.

[0189] A decline in GABAergic functions has been associated with various aspects of aging, as summarized by Mills (Journal of Nutrition and Metabolism, 2021:6655064). For instance, reduced GABAergic activity has been linked to the decline of several physiological and health functions in aging individuals. "Age-related decline in physiological and / or health functions" refers to the deterioration of essential biological processes and regulatory mechanisms over time, which contributes to the overall decline in health and vitality.

[0190] These include:

[0191] - decreased cognitive function,

[0192] - decreased neuronal activity,

[0193] - increased secretion of inflammatory cytokines and / or histamine,

[0194] - immune system dysregulation,

[0195] - decreased autophagy,

[0196] - elevated blood pressure,

[0197] - lowered blood cell levels,

[0198] - decreased thyroid hormone levels, and / or

[0199] - imbalances in glucagon and / or insulin levels.

[0200] Moreover, reduced GABAergic function has been associated with several conditions commonly linked to aging. A "condition associated with aging" may be understood as a specific disease or health issue that becomes more prevalent or severe as individuals grow older, often resulting from the cumulative decline in various physiological and / or health functions.

[0201] These conditions include:

[0202] - neurodegenerative diseases, in particular Alzheimer's disease,

[0203] - neuropathic pain,

[0204] - depression,

[0205] - inflammation, in particular low-grade chronic systemic inflammation and inflammatory pain,

[0206] - diabetes, in particular type 1 diabetes (T1D) and type 2 diabetes (T2D),

[0207] - hypertension,

[0208] - heart rhythm disorders,

[0209] - vision decline,

[0210] - bone and / or cartilage degeneration,

[0211] - respiratory dysfunction,

[0212] - renal dysfunction,

[0213] - anemia,

[0214] - thyroid dysfunction,

[0215] - muscle atrophy,

[0216] - hearing loss,

[0217] - cancer,

[0218] - metabolic syndrome,

[0219] - glucose intolerance,

[0220] - hyperglycemia,

[0221] - insulin resistance,

[0222] - obesity and obesity-related inflammation,

[0223] - pancreatic dysregulation, and

[0224] - liver dysfunction.

[0225] That is, the GABA-producing probiotic strain according to the invention may be used to mitigate the effects of aging and promote healthy aging. This probiotic strain may be beneficial for a broad range of individuals, including both healthy individuals and those who are experiencing age-related declines in physiological and / or health functions. Specifically, the target patient group may include healthy individuals who are looking to maintain their health and prevent the onset of age-related conditions.

[0226] The probiotic strain of the invention, or compositions comprising the same, is particularly advantageous for aged individuals over 50 years old, preferably over 55 years old, more preferably over 60 years old, even more preferably over 63 years old, more preferably over 65 years old, and most preferably over 70 years old. These individuals are more likely to experience declines in GABAergic function, which can lead to various age-related conditions.

[0227] Cosmetic / non-therapeutic uses

[0228] The probiotic bacteria or composition according to the invention may also be used for non- therapeutic or cosmetic applications.

[0229] For example, it was reported by Zhao et al. (Biomol Ther (Seoul) (2023) 31(6): 640-647) that GABA treatment enhanced the migration ability of skin fibroblasts, inhibited UVB-induced MMP-1 production, and increased filaggrin and aquaporin 3 levels in keratinocytes in an in vitro setting. In vivo findings further demonstrated that oral administration of GABA significantly improved skin wrinkles, epidermal thickness, and restored skin moisture and elasticity in UVB-irradiated hairless mice.

[0230] Accordingly, in a particular embodiment, the invention relates to a (non-therapeutic) use of the probiotic bacteria, composition or product according to the invention for reducing one or more appearance of aging. The appearances of aging that may be reduced with the nutritional composition or product according to the invention include, without limitation, skin wrinkles and skin thickness. Moreover, the nutritional composition or product according to the invention may be used for increasing skin moisture and / or skin elasticity. Accordingly, in a particular embodiment, the invention relates to the (non-therapeutic) use according to the invention, wherein the reduction of one or more appearance of aging is characterized by: reducing skin wrinkles, decreasing skin thickness, increasing skin moisture, and / or increasing skin elasticity. In another embodiment, the invention relates to the probiotic bacteria, composition or product according to the invention for use in reducing one or more symptoms of aging.

[0231] In further embodiment, the invention relates to a method for reducing one or more symptoms of aging in a subject, comprising administering to the subject the probiotic bacteria, composition or product according to the invention.

[0232] In certain embodiments, the invention relates to the probiotic bacteria for use, the composition for use, the product for use or the method according to the invention, wherein the reduction of one or more appearance of aging is characterized by: reducing skin wrinkles, decreasing skin thickness, increasing skin moisture, and increasing skin elasticity.

[0233] The probiotic bacteria or composition according to the invention may also be used to protect the skin from UV exposure and / or to prevent UV damage. Accordingly, in a particular embodiment, the invention relates to a (non-therapeutic) use of the probiotic bacteria or the composition or product according to the invention for reducing one or more symptoms of UV exposure, sun exposure and / or sun damage on skin.

[0234] In another embodiment, the invention relates to the probiotic bacteria or the composition or product according to the invention for use in reducing one or more symptoms of UV exposure, sun exposure and / or sun damage on skin.

[0235] In a further embodiment, the invention relates to a method for reducing one or more symptoms of UV exposure, sun exposure and / or sun damage on skin in a subject, comprising administering to the subject the probiotic bacteria or the composition or product according to the invention.

[0236] The symptoms of UV exposure, sun exposure and / or sun damage on skin may be, without limitation, skin wrinkles, increased skin thickness, reduced skin moisture and reduced skin elasticity. Accordingly, in a particular embodiment, the invention relates to the (non- therapeutic) use, the probiotic bacteria, composition or product for use, or the method according to the invention, wherein reducing the one or more symptoms of UV exposure, sun exposure and / or sun damage on skin is characterized by: reducing skin wrinkles, decreasing skin thickness, increasing skin moisture, and / or increasing skin elasticity.

[0237] The effect of GABA on growth hormone (GH) concentrations was investigated by Powers et al. (Med Sci Sports Exerc (2008) 40(l):104-10) in resistance-trained men through a double-blind, placebo-controlled study. Participants ingested GABA or a placebo, followed by rest or resistance exercise. The results demonstrated that GABA significantly increased both immunoreactive and immunofunctional GH levels, particularly post-exercise, suggesting that GABA enhances GH secretion, potentially improving muscle recovery, growth, and exercise- induced adaptations, or more generally, improves exercise performance.

[0238] Accordingly, in a particular embodiment, the invention relates to a (non-therapeutic) use of the probiotic bacteria, composition or product according to the invention for improving recovery after exercise and / or for improving exercise tolerance.

[0239] In one embodiment, the invention relates to the probiotic bacteria or the composition or product according to the invention for use in improving recovery after exercise and / or for improving exercise tolerance.

[0240] In another embodiment, the invention relates to a method for improving recovery after exercise and / or for improving exercise tolerance in a subject, comprising administering to the subject the probiotic bacteria or the composition or product according to the invention.

[0241] In a preferred embodiment, improvement of recovery after exercise and / or exercise tolerance is characterized by improved muscle recovery and / or improved muscle growth. Muscle recovery and / or muscle growth are preferably improved through increased production of immunoreactive and / or immunofunctional growth hormones in response to GABA production by the probiotic bacteria according to the invention.

[0242] Immunoreactive growth hormone (irGH) refers to the total pool of growth hormone molecules detected by immunoassays, including both biologically active and inactive forms. Immunofunctional growth hormone (ifGH), on the other hand, represents the subset of irGH that is biologically active and capable of binding to growth hormone receptors to initiate signal transduction and physiological effects. For the purpose of this patent application, irGH encompasses all detectable GH molecules, while ifGH specifically denotes those GH isoforms that possess the necessary receptor-binding domains to exert functional biological activity, thereby contributing to muscle growth and recovery. Methods for detecting irGH and ifGH levels in a subject are known to the person skilled in the art and disclosed, inter alia, by Powers et al. (Med Sci Sports Exerc (2008) 40(l):104-10).

[0243] Forthe uses disclosed herein, the probiotic bacteria or composition according to the invention is to be administered to a subject. Preferably, the probiotic bacteria or composition according to the invention is administered orally, for example in the form of a nutritional composition or product.

[0244] Forthe uses disclosed herein, the probiotic bacteria or composition according to the invention is to be administered to a subject to confer the intended benefits. Preferably, the probiotic bacteria or composition is administered orally. The oral administration ensures that the probiotic bacteria reach the gut, where they can colonize and exert their beneficial effects, such as producing bioactive compounds like gamma-aminobutyric acid (GABA).

[0245] The term "subject" as used herein refers to a mammal including human and non-human animals. More particularly, the mammal is a human. In certain examples, the subject may be an adult or an infant or a toddler or a child (pediatric) subject. The infant, toddler and children subjects may be altogether designated as pediatric subjects.

[0246] As used herein, the term "infant" means a child under the age of 12 months. The expression "young child" or "toddler" means a child aged between one and less than three years. The expression "child" means a between three and seven years of age.

[0247] The probiotic bacteria or composition according to the invention is preferably administered in an effective amount to achieve the therapeutic or non-therapeutic effects disclosed herein. An "effective amount" refers to at least an amount effective, at dosages and for periods of time necessary, to achieve the desired result. An effective amount can be provided in one or more administrations. In some embodiments of the present disclosure, the term "effective amount" is used to refer to an amount necessary to effect treatment or prevention of a disease or condition as hereinbefore described. The effective amount may vary according to the disease or condition to be treated and also according to the weight, age, racial background, sex, health and / or physical condition and other factors relevant to the subject being treated. Typically, the effective amount will fall within a relatively broad range (e.g. a "dosage" range) that can be determined through routine trial and experimentation by a medical practitioner. The effective amount can be administered in a single dose or in a dose repeated once or several times over a treatment period.

[0248] All numeric ranges provided herein are inclusive of narrower ranges; delineated upper and lower range limits are interchangeable to create further ranges not explicitly delineated. The number of significant digits conveys neither limitation on the indicated amounts nor on the accuracy of the measurements.

[0249] In this document, the terms "a" or "an" are used to include one or more than one and the term "or" is used to refer to a nonexclusive "or" unless otherwise indicated.

[0250] The term "about," as used herein, means approximately, in the region of, roughly, or around. When the term "about" is used in conjunction with a numerical range, it modifies that range by extending the boundaries above and below the numerical values set forth. In general, the term "about" is used herein to modify a numerical value above and below the stated value by a variance of 10%. Therefore, about 50% means in the range of 45%-55%. Numerical ranges recited herein by endpoints include all numbers and fractions subsumed within that range (e.g. 1 to 5 includes 1, 1.5, 2, 2.75, 3, 3.90, 4, and 5). It is also to be understood that all numbers and fractions thereof are presumed to be modified by the term "about."

[0251] BRIEF DESCRIPTION OF THE DRAWINGS

[0252] FIG.l: Overview of the reactor design. SIFR® technology was used to assess the impact on GABA production in healthy adults (n = 6) upon treatment with the test products. FIG.2: Concentration of gamma amino butyric acid (GABA) measured after 24h of exposure of adult microbiota in vitro to B. adolescentis NCC275 (initial load: 5xl07cfu / mL), Glutamate Powder A (GPA) having approximately 40 wt% glutamate (2.415 g / L), and the combination of both ingredients.

[0253] FIG.3: Concentration of gamma amino butyric acid (GABA) measured after 24h of exposure of adult microbiota in vitro to B. adolescentis NCC275, three reference strains and combinations in presence of Glutamate Powder A (GPA) (2.415 g / L).

[0254] FIG.4: Absolute concentration of gamma amino butyric acid (GABA) measured by LC-HRMS after 24h of exposure of adult microbiota in vitro to B. adolescentis NCC275, three reference strains and combinations in presence of Glutamate Powder A (GPA) (2.415 g / L).

[0255] FIG.5: Concentration of glutamate in media A before and after fermentation with B. adolescentis NCC275.

[0256] FIG.6: GABA enhancement (calculated as provided in example 3, section "Results") after fermentation with B. adolescentis NCC275 in media A with or without supplementation of glutamate in media A.

[0257] FIG.7: The absolute concentration of GABA after fermentation with B. adolescentis NCC275 in media A or fermentation with two commercially available probiotic strains in media A.

[0258] EXAMPLES

[0259] Example 1

[0260] The aim of this Example was to assess the impact of glutamate sources, probiotic strains and combinations thereof on metabolite production by the human adult gut microbiota. All products were tested in the presence of inulin to ensure strong acidification, thus facilitating GABA production (= acid stress response). The high throughput of the ex vivo SIFR® technology enabled including 6 test subjects, thus ensuring representative findings.

[0261] Methods

[0262] Test products

[0263] Glutamate source (as GABA precursor): Glutamate Powder A (GPA) (corn starch fermented with a Corynebacterium species, comprising about 40 wt% glutamate (WO 2010 / 108542));

[0264] 4 probiotic strains which include:

[0265] • Bifidobacterium adolescentis NCC275

[0266] • Lactococcus lactis NCDO2727

[0267] • Lactococcus lactis NCDO2118

[0268] • Lactobacillus acidophilus NCC2619

[0269] Two probiotic combinations, and combinations of Glutamate Powder A (GPA) with the probiotics were tested in the presence of inulin. Inulin was added at 6 g / L. Glutamate Power A (GPA) was added at 2.415 g / L (corresponding to lg / L glutamate), and the probiotic strains were each administered at 5 x 107CFU / mL.

[0270] Table 1: Study arms, test products and respective test doses

[0271] Experimental design SIFR® incubations

[0272] An ex vivo SIFR® (Van den Abbeele, P., et al., Bridging preclinical and clinical gut microbiota research using the ex vivo SIFR® technology. Frontiers in Microbiology, 2023. 14: p. 1131662.) study was implemented, simulating the colonic fermentation of test products by the gut microbiota derived from healthy human adults (n = 6; see FIG.1).

[0273] GABA measurement

[0274] GABA content was measured according to literature (Seo, M.-J., et al., y-aminobutyric acid production in skim milk co-fermented with Lactobacillus brevis 877G and Lactobacillus sakei 795. Food Science and Biotechnology, 2013. 22: p. 751-755), using a spectrophotometric assay at Oh and 24h (all test arms).

[0275] Results

[0276] Exposing adult gut microbiota to B. adolescentis NCC275 or a glutamate source (Glutamate Powder A (GPA)) did not result in a significant increase of GABA production. If, however, adult gut microbiota were exposed to a combination of B. adolescentis NCC275 and a glutamate source (Glutamate Powder A (GPA)), GABA production was significantly increased (see FIG.2).

[0277] Of note, the combination of B. adolescentis NCC275 and a glutamate source (Glutamate Powder A (GPA)) resulted in significantly higher production of GABA compared to the combination of L. lactis NCDO2118 with the same glutamate source (see FIG.3). This is surprising in that L. lactis NCDO2118 was previously reported to increase GABA production in rat (Laroute, V., et al., Lactococcus lactis NCDO2118 exerts visceral antinociceptive properties in rat via GABA production in the gastro-intestinal tract. Elife, 2022. 11: p. e77100). Combination of L. lactis NCDO2118 with B. adolescentis NCC275 did not further improve GABA production.

[0278] Conclusion

[0279] The combination of a glutamate source (for example Glutamate Powder A (GPA)) and B. adolescentis NCC275 triggered significant increase of GABA production. In contrast, the combination of Glutamate Powder A (GPA) with other strains did not show any effect. Moreover, the combination of B. adolescentis NCC275 with other probiotics did not modify its ability to produce GABA.

[0280] Example 2

[0281] The aim of this Example was to confirm the impact of glutamate sources, probiotic strains and combinations thereof on GABA production by the human adult gut microbiota. All products were tested in the presence of inulin to ensure strong acidification, thus facilitating GABA production (= acid stress response). The high throughput of the ex vivo SIFR® technology enabled including 6 test subjects, thus ensuring representative findings. Methods

[0282] Test products Glutamate source (as GABA precursor): Glutamate Powder A (GPA) (corn starch fermented with a Corynebacterium species, comprising about 40 wt% glutamate (WO 2010 / 108542));

[0283] 4 probiotic strains which include:

[0284] • Bifidobacterium adolescentis NCC275

[0285] • Lactococcus lactis NCDO2727 • Lactococcus lactis NCDO2118

[0286] • Lactobacillus acidophilus NCC2619

[0287] Two probiotic combinations, and combinations of Glutamate Powder A (GPA) with the probiotics were tested in the presence of inulin. Inulin was added at 6 g / L. Glutamate Power A (GPA) was added at 2.415 g / L (corresponding to lg / L glutamate), and the probiotic strains were each administered at 5 x 107CFU / mL.

[0288] Table 2: Study arms, test products and respective test doses

[0289] Experimental design SIFR® incubations

[0290] An ex vivo SIFR® (Van den Abbeele, P., et al., Bridging preclinical and clinical gut microbiota research using the ex vivo SIFR® technology. Frontiers in Microbiology, 2023. 14: p. 1131662.) study was implemented, simulating the colonic fermentation of test products by the gut microbiota derived from healthy human adults (n = 6; see FIG.1).

[0291] GABA measurement

[0292] GABA absolute quantification was performed by applying an analytical method based on liquid chromatography-high resolution mass spectrometry (LC-HRMS). The identification of GABA was based on the combination of observed ion, retention time and product ion spectrum. The quantification of GABA was done by applying an external calibration ranging from 0.8 to 2000 pM, using commercial standards and deuteurated internal standards.

[0293] Statistical analysis has been done by performing ordinary one-way ANOVA (analysis of variance) test followed by calculating the Fisher's LSD (least significant difference) to assess the significance of all pairwise comparison at end of fermentation.

[0294] Statistical information reported on the figures are: ns for not significant * for significance at p < 0.05

[0295] **for significance at p < 0.01

[0296] ***for significance at p < 0.001

[0297] ****forsignificance at p < 0.0001

[0298] All tests have been performed at 95% confidence level.

[0299] Results

[0300] The combination of B. adolescentis NCC275 and a glutamate source (Glutamate Powder A (GPA)) resulted in significantly higher production of GABA (p< 0.01) compared to the combination of L. lactis NCDO2118 with the same glutamate source (see FIG.4). The combination of L. lactis NCDO2118 with B. adolescentis NCC275 or L. acidophilus NCC2619 with B. adolescentis NCC275 did not further increase GABA production.

[0301] Conclusion

[0302] The combination of a glutamate source (for example Glutamate Powder A (GPA)) and B. adolescentis NCC275 triggered significant increase of GABA production, confirmed by absolute quantification of GABA in experimental study samples. It also confirmed none of the other strain test showed an effect on GABA production. In addition, the combination of B. adolescentis NCC275 with other probiotics did not modify its ability to boost GABA production.

[0303] Example 3

[0304] Methods

[0305] Fermentation media (fermentation media A with our without glutamate supplementation) was inoculated by a starter culture comprising B. adolescentis NCC275 and fermented with said starter culture under the condition provided in below media for 16 hours and under a stirring speed of 120 rpm. Fermentation was performed in 250 ml sterile Erlenmeyer flask using a laminar flow and sterile pipettes. An anaerobic incubator was employed for the fermentation step and the oxygen levels were set at 0.5%.

[0306] Table 3: Fermentation media A composition

[0307] Note: The initial concentration of free glutamate in the supernatant of media A is 1.65g / 100ml.

[0308] Two levels of glutamate are supplemented into media A 0.04% and 0.2%, to test the impact of glutamate dosage on GABA production. Quantification of GABA is based on an enzymatic microtiter plate assay using GABA aminotransferase and succinic semialdehyde dehydrogenase in the presence of NADP+ and a- ketoglutarate. Analysis is performed according to the manual of GABA enzymatic assay kit of Sigma Aldrich®, said kit being described in the following publication: Nishiyama et al., GABA enzymatic assay kit, Biosci Biotechnol Biochem, 2020 Jan; 84(1):118-125.

[0309] Results

[0310] The results are shown in Table 4, Figures 5 and 6.

[0311] Enhancement of GABA was calculated by subtracting the absolute concentration of GABA in the absolute concentration of GABA in Media A with NCC275 with glutamate supplementation (i.e. 0.04% or 0.2%) with the absolute concentration of GABA in the absolute concentration of

[0312] GABA in Media A with NCC275 without glutamate supplementation.

[0313] Table 4: Absolute concentration of Glutamate and GABA after fermentation with B. adolescentis NCC 275 with or without glutamate supplementation in media A.

[0314] Conclusion

[0315] The data shows that the supplementation of the media with glutamate boosts the production of GABA by B. adolescentis NCC275.

[0316] Example 4

[0317] Methods

[0318] Media A as shown in table 3 of example 3 (without glutamate supplementation, i.e. having a glutamate content of 1.65g / 100ml.) was inoculated with B. adolescentis or commercial strains known to be GABA producers, i.e. B. adolescentis GabaPral® or LactiplantibaciUus plantarum 815. After inoculation, the media were fermented with said strains under the condition provided in table 5 for 16 hours and under a stirring speed of 120 rpm. Fermentation was performed in 250 ml sterile Erlenmeyer flask using a laminar flow and sterile pipettes. For the genus Bifidobacterium, an anaerobic incubator was employed for the fermentation step and the oxygen levels were set at 0.5%.

[0319] Table 5: Details about strains used in the experiment

[0320] Quantification of GABA in the fermentation broth follows the same method as in Example 3.

[0321] Results

[0322] The GABA production of B. adolescentis NCC275 is much higher compared with commercial probiotics in media A.

[0323] Table 6: Absolute GABA concentration after fermentation with B. adolescentis NCC275 or after fermentation with the different commercial probiotic strains (table shows results for each duplicate per column).

[0324] Conclusion

[0325] NCC275 produces higher level of GABA compared to the two commercially available strains with the same level of glutamate in the media A.

[0326] (Original in Electronic Form) (This sheet is not part of and does not count as a sheet of the international application)

[0327] FOR RECEIVING OFFICE USE ONLY

[0328] FOR INTERNATIONAL BUREAU USE ONLY

[0329] 0-5 This form was rec international Bure

[0330] 0-5- Authorized officer

Claims

CLAIMS1. Bifidobacterium adolescentis strain NCC275 as deposited with the Collection Nationale de Cultures de Microorganismes (CNCM) at the Institut Pasteur under the Accession Number CNCM 1-6090 on June 17, 2024.

2. A composition, preferably nutritional composition comprising Bifidobacterium adolescentis strain NCC275 (CNCM 1-6090).

3. A composition comprising (i) Bifidobacterium adolescentis strain NCC275 (CNCM 1-6090) and (ii) at least one excipient, and / or wherein the Bifidobacterium adolescentis strain NCC275 is spray-dried or freeze-dried.

4. A nutritional composition or product comprising Bifidobacterium adolescentis strain NCC275 (CNCM 1-6090) and a source of glutamate.

5. The nutritional composition or product according to claim 4, wherein the source of glutamate comprises or consists of purified glutamate, or wherein the source of glutamate is a natural product, in particular a fermentation product or a cell extract.

6. The nutritional composition or product according to claim 4 or claim 5, wherein the source of glutamate is obtainable through prokaryotic fermentation of a suitable substrate with one or more of the following bacteria: Corynebacterium glutamicum, Corynebacterium ammoniagenes, Corynebacterium casei, Corynebacterium efficiens, Brevibacterium lactofermentum and Bacillus subtilis.

7. The Bifidobacterium adolescentis strain according to claim 1, the composition according to claims 2 or 3, or the nutritional composition or product according to any one of claims 4 to 6 for use as a medicament.

628. The Bifidobacterium adolescentis strain according to claim 1, the composition according to claim 2 or 3, or the nutritional composition or product according to any one of claims 4 to 6 for use in reducing one or more symptoms of stress and / or for use in the dietary management of stress in a subject, in particular wherein reducing the symptoms of stress and / or the dietary management of stress involves one or more of: increasing relaxation, increasing calmness, reducing morning drowsiness, restoring mental calmness, and / or supporting mental relaxation in said subject.

9. The Bifidobacterium adolescentis strain according to claim 1, the composition according to claim 2 or 3, or the nutritional composition or product according to any one of claims 4 to 6 for use in reducing one or more symptoms of premenstrual syndrome and / or for use in the dietary management of premenstrual syndrome in a subject.

10. The Bifidobacterium adolescentis strain according to claim 1, the composition according to claim 2 or 3, or the nutritional composition or product according to any one of claims 4 to 6 for use in reducing pain and / or for use in the dietary management of pain in a subject, in particular wherein the pain is visceral pain, chronic pain, neuropathic pain or inflammatory pain; and / or wherein the pain is associated with menstruation and / or endometriosis; and / or wherein the pain is associated with colic in infants or toddlers.

11. The Bifidobacterium adolescentis strain according to claim 1, the composition according to claim 2 or 3, or the nutritional composition or product according to any one of claims 4 to 6 for use in improving sleep and / or for use in the dietary management of sleep in a subject.

12. The Bifidobacterium adolescentis strain according to claim 1, the composition according to claim 2 or 3, or the nutritional composition or product according to any one of claims 4 to 6 for use in prevention, delay and / or dietary management of age-related decline in physiological and / or health functions or conditions associated with aging in a subject.6313. A (non-therapeutic) use of the Bifidobacterium adolescentis strain according to claim 1, the composition according to claim 2 or 3, or the nutritional composition or product according to any one of claims 4 to 6 for reducing one or more appearance of aging, in particular wherein reducing the one or more appearance of aging is characterized by: reducing skin wrinkles, decreasing skin thickness, increasing skin moisture, and / or increasing skin elasticity.

14. A (non-therapeutic) use of the Bifidobacterium adolescentis strain according to claim 1, the composition according to claim 2 or 3, or the nutritional composition or product according to any one of claims 4 to 6 for reducing one or more symptoms of UV exposure, sun exposure and / or sun damage on skin, in particular wherein reducing the one or more symptoms of UV exposure, sun exposure and / or sun damage on skin is characterized by: reducing skin wrinkles, decreasing skin thickness, increasing skin moisture, and increasing skin elasticity.

15. The Bifidobacterium adolescentis strain according to claim 1, the composition according to claim 2 or 3, or the nutritional composition or product according to any one of claims 4 to 6 for use in reducing one or more symptoms of aging or for use in reducing one or more symptoms of UV exposure, sun exposure and / or sun damage on skin, in particular wherein reducing the one or more symptoms of aging or the one or more symptoms of UV exposure, sun exposure and / or sun damage on skin is characterized by: reducing skin wrinkles, decreasing skin thickness, increasing skin moisture, and increasing skin elasticity.

16. A (non-therapeutic) use of the Bifidobacterium adolescentis strain according to embodiment 1, the composition according to embodiment 2 or 3, or the nutritional composition or product according to any one of embodiments 4 to 6 for improving recovery after exercise and / or for improving exercise tolerance.

17. The Bifidobacterium adolescentis strain according to embodiment 1, the composition according to embodiment 2 or 3, or the nutritional composition or product according to64any one of embodiments 4 to 6 for use in improving recovery after exercise and / or for improving exercise tolerance.65

Images