Probiotic compositions for the treatment of malnutrition - Patent Application 20070122999

Abstract

The present invention relates to the probiotic strains Lactobacillus plantarum (Lactiprantibacillus plantarum) DSM 33363, Lactobacillus plantarum (Lactiprantibacillus plantarum) DSM 33364, Lactobacillus paracasei (Lacticasebacillus paracasei) DSM 33373, Lactobacillus reuteri (LimociLactobacillus reuteri) DSM 33374, Bacillus megaterium (Priestia megaterium) DSM 33300, Bacillus pumilus DSM 33297, Bacillus pumilus DSM 33355 for treating and preventing malnutrition in humans and animals by increasing the bioavailability of essential micronutrients and amino acids contained in the food, thereby increasing the nutritional value of cereal-based foods, particularly wheat-derived foods. ("Pronutrient" Consortium).

Description

[Technical Field]

[0001] The present invention relates to the probiotic strains Lactobacillus plantarum (Lactiplantibacillus plantarum) DSM 33363, Lactobacillus plantarum (Lactiplantibacillus plantarum) DSM 33364, Lactobacillus paracasei (Lacticaseibacillus paracasei) DSM 33373, Lactobacillus reuteri (Lactobacillus reuteri) for the treatment and prevention of malnutrition in humans and animals by increasing the nutritional value of cereal-based foods, particularly wheat-derived foods, through an increase in the bioavailability of essential micronutrients and amino acids contained in said foods. The present invention relates to a probiotic composition comprising Limosilactobacillus reuteri DSM 33374, Bacillus megaterium (Priestia megaterium) DSM 33300, Bacillus pumilus DSM 33297, Bacillus pumilus DSM 33355 ("Pro-nutrient" Consortium).

[0002] Malnutrition in the context of the present invention refers to a deficiency or imbalance in a person's energy and / or nutrient intake, but does not refer to overnutrition. Approximately 700 million people worldwide are undernourished, and given the scale of this problem, the World Health Organization has set ambitious global nutrition goals, for example, targeting stunting, anemia, and low birth weight. To achieve this goal, food availability and quality need to be improved, accompanied by technological solutions that can maximize the nutritional value of food, as disclosed in the present invention.

[0003] While malnutrition is primarily a phenomenon in low- and middle-income countries, it is also prevalent among certain groups in developed countries, such as the elderly, individuals with functional gastrointestinal disorders, vegetarians, vegans, and individuals following elimination or unbalanced diets. In particular, vegetarian, vegan, and wheat-based diets limit the intake and bioavailability of the essential amino acid L-lysine and the essential micronutrients iron (Fe), zinc (Zn), and magnesium (Mg). Wheat-based diets also limit the bioavailability of Fe, Zn, and Mg due to the presence of the antinutritional factor phytic acid, which chelates divalent cations and thereby prevents their absorption from the intestinal tract. Despite extensive food fortification efforts, such as with table salt, soil, and common crops, micronutrient deficiencies remain highly prevalent and pose a significant global health problem. [1]

[0004] The recommended intake to meet lysine requirements ranges from 64 to 30 mg / kg body weight per day [2]. The recommended intakes for Fe, Zn, and Mg range from 10 to 30 mg / day (Fe), 7 to 16 mg / day (Zn), and 300 to 350 mg / day (https: / / www.dge.de / wissenschaft / referenzwerte / ). Interestingly, the recommended amount for Zn depends on phytate intake.

[0005] The most common strategy for addressing existing or suspected micronutrient deficiencies is the use of dietary supplements or functional foods containing these micronutrients [3, 4]. However, because the bioavailability of selected nutrients is affected by numerous endogenous and exogenous factors, such as antinutritional factors, gut bacterial factors, individual health status, and food matrix effects, supplementation does not always result in satisfactory improvement of nutritional status; iron deficiency is a typical example [5]. On the other hand, excessive use of dietary supplements can lead to oversupplementation and the risk of adverse health effects, as has been shown for beta-carotene, vitamin A, and vitamin E [6]. In conclusion, potential limitations and risks of food supplementation include the failure to improve nutritional status of certain nutrients and the risk of exceeding the target range, which may result in adverse health effects.

[0006] Deficiencies in macronutrients, such as protein, are often addressed using concentrated protein compositions, protein hydrolysates, peptides, or amino acid compositions, with or without additional nutrients. For example, International Publication No. 2012 / 052463 discloses the use of cysteine ​​and its derivatives for the treatment and prevention of malnutrition. International Publication No. 2019 / 230849 discloses a composition of minerals, vitamins, and grains, along with lysine and methionine, for treating malnutrition. Herbal and botanical compositions have also been described. For example, Chinese Patent Application Publication No. 104623224 and Chinese Patent Application Publication No. 105663996 describe compositions of traditional Chinese medicinal plants for treating malnutrition in infants.

[0007] The present invention utilizes a microbiota-targeting strategy as a technological solution to improve the nutritional value of various diets while simultaneously overcoming the limitations of single supplementation strategies. The gastrointestinal microbiota determines the fate of ingested materials (food, medicines, etc.) through, for example, microbial metabolism, interactions with host physiological functions such as barrier function, nutrient and water absorption, and gastrointestinal motility, and in this sense, is a crucial regulator of health in humans and animals. Microbiota-targeting strategies include the application of prebiotics, probiotics, synbiotics, and sometimes even fecal transplants, with the aim of altering the composition and activity of the microbiota. Probiotics are live microorganisms that, when administered in appropriate amounts, confer health benefits to the host. [7] The most studied and commercially available probiotics are primarily derived from species of the Lactobacillus and Bifidobacterium genera. Additionally, several other bacteria, such as Propionibacterium, Streptococcus, Bacillus, Enterococcus, Escherichia coli, and yeast, are also used. Different strains of bacteria from the same genus and species may have different effects on the host. A meta-analysis of clinical trials conducted by Barkhidarian et al. demonstrated a possible association between the intake of probiotics from Lactobacillus, Bifidobacterium, and Streptococcus thermophilus and the status of micronutrients vitamin B12, calcium, folate, iron, and zinc [8]. In general, the gut microbiome has been described as a confounding factor in the results of nutritional intervention studies [9]. Lactic acid bacteria (LAB), including some Lactobacillus species, express phytase. The use of these LAB in the production of (fermented) foods such as bread, soy milk, fruit juice, beer, and fermented vegetables has been described. A prerequisite for phytase function is sufficient survival of the LAB probiotics under gastrointestinal conditions.Although there are few reports evaluating the phytase activity of LAB under such conditions, screening of LAB isolates, including Lactobacillus species, revealed phytase activities ranging from approximately 0.5 to a maximum of 1.77 U / ml for Weissella kimchii strains

[10] .

[0008] In recent years, the taxonomic classification of some species of the genera Lactobacillus and Bacillus has been updated [11-13]. Of relevance in the context of the present invention are the following species: "Old" Name Updated Name (2020 and later) Lactobacillus paracasei Lacticaseibacillus paracasei Lactobacillus plantarum Lactiprancibacillus plantarum Lactobacillus reuteri Limocilactobacillus reuteri Bacillus megaterium Priestia megaterium Bacillus pumilus Bacillus pumilus For convenience, the former name is used in the Examples section, but both names are used in the General Description and Claims sections.

[0009] In WO 2021 / 129998 and under

[14] , the Applicant previously disclosed the combination of Lactobacillus plantarum (Lactiprantibacillus plantarum) DSM 33363, Lactobacillus plantarum (Lactiprantibacillus plantarum) DSM 33364, Lactobacillus paracasei (Lacticasibacillus paracasei) DSM 33373, Lactobacillus reuteri (Limosilactobacillus reuteri) DSM 33374, Bacillus megaterium (Priestia megaterium) DSM 33300, Bacillus pumilus DSM 33297, Bacillus pumilus DSM 33355 (= Pronutrient Consortium), among other combinations

[15] , and its ability to completely digest gluten.

[0010] These strains were deposited in 2019 under the provisions of the Budapest Treaty on the International Recognition of the Deposit of Microorganisms for the Purposes of Patent Procedure at the Leibniz-Institut DSMZ Deutsche Sammlung von Mikroorganismen and Zellkulturen GmbH, Inhoffenstr. 7B, 38124 Braunschweig, Germany, under the aforementioned accession numbers, in the name of Evonik Nutrition and Care GmbH (legal predecessor of Evonik Operations GmbH).

[0011] Applicant unexpectedly discovered that the pro-nutrient consortium has the potential to be an effective treatment for malnutrition by improving the nutritional value of various foods and diets. Applicant discovered that the consortium enabled the release of essential nutrients L-lysine, Fe, Zn, and Mg from various food matrices (whole wheat bread, white bread, and wheat flour) in simulated gastrointestinal digestion. This release was significantly stronger than under control conditions and compared to the effects of other added digestive aids, such as proteases and other probiotic consortia. Applicant discovered that the strains of the pro-nutrient consortium had higher phytase activity than other probiotic strains and exhibited very good survival and storage stability in simulated gastric and small intestinal conditions. These combined unique features form a novel technological solution for improving the nutritional value of various diets and, consequently, for treating and / or preventing pathologies resulting from L-lysine, Fe, Zn, Mg deficiency, including anemia, fatigue, dizziness, nausea, dermatitis, low birth weight, growth retardation, stunting, and improving growth performance in animals.

[0012] Preparations of the pronutrient consortium can be used, for example, as dietary supplements, feed additives, in the preparation of functional foods and feeds, and in the manufacture of food and livestock feed.

[0013] One advantage of the composition of the present invention is that it can be incorporated into a person's regular diet, compared to nutritional foods that (partially) replace regular meals, such as prescription diets, medical nutrition, or functional foods. Meal replacements can be inconvenient, expensive, tasteless, and can be disadvantageous in social situations. These disadvantages limit compliance and therefore the effectiveness of such treatments.

[0014] The present invention therefore relates to a probiotic composition for use in the treatment and prevention of malnutrition, the probiotic composition comprising one or more of the following strains: Lactobacillus plantarum (Lactiplantibacillus plantarum) DSM 33363, Lactobacillus plantarum (Lactiplantibacillus plantarum) DSM 33363, Lactobacillus plantarum (Lactiplantibacillus plantarum) DSM 33364, Lactobacillus paracasei (Lacticasei Bacillus paracasei) DSM 33373, Lactobacillus reuteri (Limosilactobacillus reuteri) DSM 33374, Bacillus megaterium (Priestia megaterium) DSM 33300, Bacillus pumilus DSM 33297, and Bacillus pumilus DSM 33355. More specifically, the preparation is suitable for improving a person's nutritional status, as determined by appropriate biomarkers, in particular the status of total protein, L-lysine, L-asparagine, L-glycine, L-ornithine, Fe, Zn, and Mg. Furthermore, the preparation is suitable for the treatment and prevention of deficiencies or suboptimal states of any of the aforementioned nutrients, as well as for the treatment or prevention of any health condition or disease resulting from a chronic deficiency or suboptimal state of any of the aforementioned nutrients.

[0015] In a preferred configuration of the present invention, the probiotic composition comprises all strains of Lactobacillus paracasei (Lacticaseibacillus paracasei) DSM 33373, Bacillus megaterium (Priestia megaterium) DSM 33300, Bacillus pumilus DSM 33297, and Bacillus pumilus DSM 33355.

[0016] In another preferred configuration, the probiotic composition further comprises Lactobacillus plantarum (Lactiplantibacillus plantarum) DSM 33363, Lactobacillus plantarum (Lactiplantibacillus plantarum) DSM 33364, Lactobacillus reuteri (Limosilactobacillus reuteri) DSM 33374.

[0017] In certain configurations, the malnutrition is a macronutrient and micronutrient deficiency, preferably a protein and mineral deficiency. More specifically, the probiotic composition reduces the phytic acid content and increases the bioavailability of micronutrients selected from iron, copper, zinc, and magnesium.

[0018] In another particular configuration, the probiotic composition has a phytase activity of at least 5 phytase activity units, preferably at least 10 phytase activity units, more preferably at least 15 phytase activity units.

[0019] Specifically, the probiotic composition is used to increase the nutritional value and / or bioavailability of macro- and micronutrients, preferably protein, amino acids selected from L-lysine, L-aspartic acid, L-glycine and L-ornithine, or minerals selected from Mg, Zn, Fe in food products and diets.

[0020] The cells of the strains of the present invention may be present in the compositions of the present invention as spores (dormant), as vegetative cells (growing), as transition state cells (transitioning from vegetative cells to spores or vice versa), as cell extracts, or as a combination of at least two of these types of cells. In a preferred embodiment, the probiotic strains are present in a dormant form or as vegetative cells. In alternative embodiments, cytoplasmic extracts or cell-free supernatants or heat-killed biomass of the probiotic strains are used.

[0021] In an alternative embodiment, the preparation further comprises one or more probiotic strains.

[0022] In a further preferred embodiment, the preparation further comprises one or more of the following: microbial proteases purified from Aspergillus niger, Aspergillus oryzae, Bacillus, Lactobacillus, Pediococcus, Weissella, Rothia mucilaginosa, Rothia aeria, subtilisin, and nattokinase.

[0023] In an alternative embodiment, the preparation further comprises enzymes that facilitate digestion of carbohydrates, proteins, peptides, and lipids.

[0024] In a preferred embodiment, the preparation for use further comprises a substance, preferably alginate, that acts as a permeabilizer for microbial cell membranes of members of the genera Bacillus, Lactobacillus, Pediococcus and Weissella.

[0025] In an alternative embodiment, one or more probiotic strains selected from the genera Bacillus and Lactobacillus are immobilized individually or as a consortium. Immobilization can be achieved on a solid surface, for example, on cellulose and chitosan, in a porous matrix, for example, as encapsulation in polysaccharide gels such as alginate, k-carrageenan, agar, chitosan and polygalacturonic acid, or in other polymeric matrices such as gelatin, collagen and polyvinyl alcohol, or by aggregation and microencapsulation or electrospray techniques.

[0026] One subject of the present invention is the use of the preparation according to the invention as a food supplement or in food products. Preferred food products according to the present invention are cereals, bread, chocolate products, gummies, muesli, muesli bars, health bars, biscuits, spreads and dairy products.

[0027] A further subject of the present invention is the use of the preparation according to the invention as a synbiotic ingredient in food products.

[0028] One subject of the present invention is the use of the preparation according to the invention as a food or feed supplement or functional food or food product or pharmaceutical. Preferred food products according to the present invention are cereals, bread, chocolate products, gummies, muesli, muesli bars, health bars, biscuits, spreads and dairy products.

[0029] Thus, in a preferred embodiment, the preparation is formulated for oral use, preferably as a pill, capsule, tablet, granular powder, opercula, soluble granules, bag, pill or drinkable vial, or as a syrup or drink, or added to food, preferably cereals, gummies, bread, muesli, muesli bars, health bars, biscuits, chocolate, yogurts or spreads.

[0030] A further subject of the present invention is the use of the preparation according to the invention as a synbiotic ingredient in food products.

[0031] A further subject of the present invention is a food composition comprising the preparation according to the invention and at least one further food ingredient, preferably selected from proteins, carbohydrates, fats, further probiotics, prebiotics, enzymes, vitamins, immunomodulators, milk substitutes, minerals, amino acids, anticoccidial agents, acid-based products, medicines, and combinations thereof. Food compositions according to the invention also include dietary supplements, for example in the form of pills, capsules, tablets, powders, sachets, operculas, soluble granules, bags or drinkable vials, syrups, drinks, or other liquids.

[0032] A further subject of the present invention is a pharmaceutical composition comprising a preparation according to the invention and a pharmaceutically acceptable carrier.

[0033] Another subject of the present invention is the use as a feed additive to increase the feed conversion ratio and reduce the luminal content of phytic acid, thereby increasing the bioavailability of one or more of the micronutrients iron, copper, zinc and magnesium and one or more of the amino acids L-lysine, L-aspartic acid, L-glycine and L-ornithine. [Brief explanation of the drawings]

[0034] [Figure 1] This figure shows that the probiotic composition of the present invention significantly increases the release of L-lysine from foods during simulated gastric and small intestinal digestion. The composition increases L-lysine release from gluten extracted from wheat flour, white bread, and whole wheat bread by 33-400% compared to the control. Treatment with proteases (Protease 1: proline-specific oligopeptidase, Protease 2: casein protease) and alternative probiotic compositions had only a slight or even negative effect on L-lysine release under the same conditions. The black-bordered rectangle highlights the proportion of L-lysine in the bar graph. MC16: Microbial Consortium 16 = Pronutrient Consortium. [Figure 2]This figure shows that the probiotic composition of the present invention significantly increases the release of asparagine, lysine, glycine, and ornithine from food during simulated gastric and small intestinal digestion. The heat map shows the clustering of control-treated and enzyme-treated samples compared to probiotic-treated samples MC12 and MC16. The color scale reflects the Euclidean distance between samples based on high (dark brown) or low (blue) score values ​​for compound concentrations formed during digestion of gluten extracted from wheat flour, white bread, and whole wheat bread. The b1 cluster contains high scores for asparagine, lysine, glycine, and ornithine, clearly separating MC16 from the other treatments. [Figure 3] This figure shows that the pronutrient consortium removes phytic acid from wheat-based foods through its high phytase activity. Panel A: Phytase activity and phytic acid assays from aqueous extracts of digested control (dough containing 10 grams of gluten (CG) or 100 grams of white wheat bread and whole wheat bread (CB and CWB), respectively) tested with or without two commercially available enzymes, Tolerase® G and Promod™, and digested dough containing the test microbial consortiums MC12 and MC16. Panel B: Phytase activity and phytic acid assays from aqueous extracts of pure cultures of the strains included in MC12 (gray circles) and MC16 (black circles), as well as the activity of both MC12 and MC16. Different superscript capital letters in A-F indicate significantly different phytase activity values ​​(two-way ANOVA test). Different superscript lowercase letters in af indicate significantly different phytic acid values ​​(two-way ANOVA test). "*" means that the p-value between MC12 and MC16 is <0.05 (two-tailed, Student's t-test).

[0035] Example Example 1. Probiotic compositions significantly increase the release of L-lysine from foods Digests of three foods (whole wheat bread, white bread, and wheat flour protein), each containing 10 g of gluten protein, were prepared under simulated gastrointestinal conditions with and without the addition of a microbial consortium or proteases, as described in

[14] . Protease 1 is a proline-specific oligopeptidase, and protease 2 is a casein protease.

[0036] L-Lysine Quantification: The digests were assayed for the content of individual free amino acids (FAA) in the pH 4.6 soluble nitrogen fraction using a Biochrom 30 Series Amino Acid Analyzer (Biochrom Ltd., Cambridge Science Park, England) equipped with a sodium cation exchange column (20 × 0.46 cm internal diameter). A mixture of amino acids of known concentrations (Sigma Chemical Co., Milan, Italy) spiked with tryptophan, ornithine, asparagine, and GABA served as a standard. Proteins and peptides in the samples were precipitated by adding 5% (vol / vol) cold solid sulfosalicylic acid, holding the samples at 4°C for 1 hour, and centrifuging them at 15,000 × g for 15 minutes. The supernatant was filtered through a 0.22 μm pore size filter and diluted, if necessary, with sodium citrate (0.2 M, pH 2.2) loading buffer. Amino acids were post-column derivatized with ninhydrin reagent and detected by absorbance at 440 nm (proline and hydroxyproline) or 570 nm (all other amino acids).

[0037] Figure 1 shows that the probiotic composition significantly increases the release of L-lysine from foods during simulated gastric and small intestinal digestion. The composition increases L-lysine release from gluten extracted from wheat flour, white bread, and whole wheat bread by 33-400% compared to the control. Treatment with proteases (Protease 1: proline-specific oligopeptidase, Protease 2: casein protease) and alternative probiotic compositions had only a slight or even negative effect on L-lysine release under the same conditions. Black-bordered rectangles highlight the proportion of L-lysine in the bar graph. MC16: Microbial Consortium 16 = Pronutrient Consortium.

[0038] Figure 2 shows that the probiotic composition significantly increases the release of asparagine, lysine, glycine, and ornithine from food during simulated gastric and small intestinal digestion. The heat map shows the clustering of control- and enzyme-treated samples compared to probiotic-treated samples MC12 and MC16. The color scale reflects the Euclidean distance between samples based on high (dark brown) or low (blue) score values ​​for compound concentrations formed during digestion of gluten extracted from wheat flour, white bread, and whole wheat bread. Cluster b1 contains high scores for asparagine, lysine, glycine, and ornithine, clearly separating MC16 from the other treatments.

[0039] Figures 1 and 2 show that the pronutrient consortium (MC16) releases significant amounts of L-asparagine, L-lysine, L-glycine, and L-ornithine from different foods during simulated gastric and small intestinal digestion. Importantly, this release is much more pronounced compared to the control treatment and compared to other microbial consortia and two proteases.

[0040] Example 2. Probiotic strains with phytase activity Figure 3 shows that the pronutrient consortium removes phytic acid from wheat-based foods through enhanced phytase activity. Panel A: Phytase activity and phytic acid assays from aqueous extracts of digested control (dough containing 10 grams of gluten (CG) or 100 grams of white wheat bread and whole wheat bread (CB and CWB), respectively), tested with or without two commercially available enzymes, Tolerase® G and Promod™, and digested dough containing the test microbial consortia MC12 and MC16. Panel B: Phytase activity and phytic acid assays from aqueous extracts of pure cultures of the strains included in MC12 (gray circles) and MC16 (black circles), as well as the activity of both MC12 and MC16. Different superscript capital letters (A-F) indicate significantly different phytase activity values ​​(two-way ANOVA test). Different superscript lowercase letters (a-f) indicate significantly different phytic acid values ​​(two-way ANOVA test). "*" means that the p-value between MC12 and MC16 is <0.05 (two-tailed, Student's t-test).

[0041] Figure 3B displays the phytase activity of individual strains from both consortia. Importantly, when the strains were separated into the two consortia, the phytase activity was much higher compared to other wild-type probiotics mentioned in the literature

[16] , with the pro-nutrient consortium (M16) having the highest activity of ~24 U / ml.

[0042] Both consortia were used in food digestion experiments under simulated gastrointestinal conditions to evaluate their ability to reduce the phytic acid (PA) content of these wheat-based foods. As shown in Figure 3, the pronutrient consortium reduced PA in all tested foods by more than 84%; for wheat-derived gluten, white wheat bread, and whole wheat bread, the reductions were 84%, 86.1%, and 87.1%, respectively. These reductions were significantly stronger than those reported in other literature for other wild-type probiotic strains, such as Lactobacillus species [16, 17].

[0043] References 1. Keats, E.C.; Neufeld, L.M.; Garrett, G.S.; Mbuya, M.N.N.; Bhutta, Z.A. Improved micronutrient status and health outcomes in low- and middle-income countries following large-scale fortification: evidence from a systematic review and meta-analysis. Am J Clin Nutr 2019, 109, 1696-1708, doi:10.1093 / ajcn / nqz023. 2. Tome, D.; Bos, C. Lysine requirement through the human life cycle. J Nutr 2007, 137, 1642S-1645S, doi:10.1093 / jn / 137.6.1642S. 3. Qato, D.M.; Alexander, G.C.; Conti, R.M.; Johnson, M.; Schumm, P.; Lindau, S.T. Use of prescription and over-the-counter medications and dietary supplements among older adults in the United States. JAMA 2008, 300, 从2867年到2878年,doi:10.1001 / jama.2008.892. 4. Kantor, E.D.; Rehm, C.D.; Du, M.; White, E.; Giovannucci, E.L. Trends in Dietary Supplement Use Among US Adults From 1999-2012. JAMA 2016, 316, 1464-1474, doi:10.1001 / jama.2016.14403. It should be noted that there is an error in the original text of item , where "from 2867 to 2878" should be "2867-2878". The above translation has been corrected according to the correct content.5. Kumar, S.B.; Arnipalli, S.R.; Mehta, P.; Carrau, S.; Ziouzenkova, O. Iron Deficiency Anemia: Efficacy and Limitations of Nutritional and Comprehensive Mitigation Strategies. Nutrients 2022, 14, doi:10.3390 / nu14142976. 6. Bjelakovic, G.; Nikolova, D.; Gluud, L.L.; Simonetti, R.G.; Gluud, C. Mortality in randomized trials of antioxidant supplements for primary and secondary prevention: systematic review and meta-analysis. JAMA 2007, 297, 842-857, doi:10.1001 / jama.297.8.842. 7. Hill, C.; Guarner, F.; Reid, G.; Gibson, G.R.; Merenstein, D.J.; Pot, B.; Morelli, L.; Canani, R.B.; Flint, H.J.; Salminen, S., et al. Expert consensus document. The International Scientific Association for Probiotics and Prebiotics consensus statement on the scope and appropriate use of the term probiotic. Nat Rev Gastroenterol Hepatol 2014, 11, 506-514, doi:10.1038 / nrgastro.2014.66. 8. Barkhidarian, B.; Roldos, L.; Iskandar, M.M.; Saedisomeolia, A.; Kubow, S. Probiotic Supplementation and Micronutrient Status in Healthy Subjects: A Systematic Review of Clinical Trials. Nutrients 2021, 13, doi:10.3390 / nu13093001. 9. Vandeputte, D. Personalized Nutrition Through The Gut Microbiota: Current Insights And Future Perspectives. Nutr Rev 2020, 78, 66-74, doi:10.1093 / nutrit / nuaa098. 10. Andrabi, S.T.; Bhat, B.; Gupta, M.; Bajaj, B.K. Phytase-Producing Potential and Other Functional Attributes of Lactic Acid Bacteria Isolates for Prospective Probiotic Applications. Probiotics Antimicrob Proteins 2016, 8, 121-129, doi:10.1007 / s12602-016-9220-3. 11. Zheng, J.; Wittouck, S.; Salvetti, E.; Franz, C.; Harris, H.M.B.; Mattarelli, P.; O'Toole, P.W.; Pot, B.; Vandamme, P.; Walter, J., et al. A taxonomic note on the genus Lactobacillus: Description of 23 novel genera, emended description of the genus Lactobacillus Beijerinck 1901, and union of Lactobacillaceae and Leuconostocaceae. Int J Syst Evol Microbiol 2020, 70, 2782-2858, doi:10.1099 / ijsem.0.004107. 12. Patel, S.; Gupta, R.S. A phylogenomic and comparative genomic framework for resolving the polyphyly of the genus Bacillus: Proposal for six new genera of Bacillus species, Peribacillus gen. nov., Cytobacillus gen. nov., Mesobacillus gen. nov., Neobacillus gen. nov., Metabacillus gen. nov. and Alkalihalobacillus gen. nov. Int J Syst Evol Microbiol 2020, 70, 406-438, doi:10.1099 / ijsem.0.003775. 13. Gupta, R.S.; Patel, S.; Saini, N.; Chen, S. Robust demarcation of 17 distinct Bacillus species clades, proposed as novel Bacillaceae genera, by phylogenomics and comparative genomic analyses: description of Robertmurraya kyonggiensis sp. nov. and proposal for an emended genus Bacillus limiting it only to the members of the Subtilis and Cereus clades of species. Int J Syst Evol Microbiol 2020, 70, 5753-5798, doi:10.1099 / ijsem.0.004475. 14. De Angelis, M.; Siragusa, S.; Vacca, M.; Di Cagno, R.; Cristofori, F.; Schwarm, M.; Pelzer, S.; Flugel, M.; Speckmann, B.; Francavilla, R., et al. Selection of Gut-Resistant Bacteria and Construction of Microbial Consortia for Improving Gluten Digestion under Simulated Gastrointestinal Conditions. Nutrients 2021, 13, doi:10.3390 / nu13030992. 15. Lenhart, A.; Dong, T.; Joshi, S.; Jaffe, N.; Choo, C.; Liu, C.; Jacobs, J.P.; Lagishetty, V.; Shih, W.; Labus, J.S., et al. Effect of Exclusion Diets on Symptom Severity and the Gut Microbiota in Patients With Irritable Bowel Syndrome. 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Claims

1. 1. A probiotic composition for use in the treatment or prevention of a deficiency of total protein, L-lysine, L-asparagine, L-glycine, L-ornithine, or a mineral selected from Fe, Zn, or Mg, the probiotic composition comprising one or more of the following strains: Lactobacillus plantarum (Lactiplantibacillus plantarum) DSM 33363, Lactobacillus plantarum (Lactiplantibacillus plantarum) DSM 33364, Lactobacillus paracasei (Lacticaseibacillus paracasei) DSM 33373, Lactobacillus reuteri (Limosilactobacillus reuteri) DSM 33374, Bacillus megaterium Bacillus megaterium (Priestia megaterium) DSM 33300, Bacillus pumilus DSM 33297, and Bacillus pumilus DSM 33355.

2. 2. The composition of claim 1, wherein the probiotic composition comprises all of the strains of Lactobacillus paracasei (Lacticaseibacillus paracasei) DSM 33373, Bacillus megaterium (Priestia megaterium) DSM 33300, Bacillus pumilus DSM 33297, and Bacillus pumilus DSM 33355.

3. 3. The composition of claim 1 or 2, wherein the probiotic composition further comprises Lactobacillus plantarum (Lactiprancibacillus plantarum) DSM 33363, Lactobacillus plantarum (Lactiprancibacillus plantarum) DSM 33364, Lactobacillus reuteri (Limocilactobacillus reuteri) DSM 33374.

4. 4. The composition according to any one of claims 1 to 3, wherein the probiotic composition has a phytase activity of at least 5 phytase activity units, preferably at least 10 phytase activity units, more preferably at least 15 phytase activity units.

5. 5. The composition according to any one of claims 1 to 4, wherein the probiotic composition is a food or feed supplement or a functional food or food product or a pharmaceutical.

6. 6. The composition of claim 5, wherein the probiotic composition is formulated for oral use, preferably as a pill, capsule, tablet, granular powder, opercula, soluble granules, bag, pill or drinkable vial, or as a syrup or drink, or added to a food product, preferably cereal, gummies, bread, muesli, muesli bars, health bars, biscuits, chocolate, yogurts or spreads.

7. 6. The composition of claim 5 as a feed additive for increasing feed conversion ratio and decreasing the luminal content of phytic acid to increase the bioavailability of one or more of the micronutrients iron, copper, zinc and magnesium and one or more of the amino acids L-lysine, L-aspartic acid, L-glycine and L-ornithine.

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