WO2022167134A1 - Plant-based products fermented with b. amyloliquefaciens and p. freudenreichii having increased vit b12 content - Google Patents
Plant-based products fermented with b. amyloliquefaciens and p. freudenreichii having increased vit b12 content Download PDFInfo
- Publication number
- WO2022167134A1 WO2022167134A1 PCT/EP2021/086018 EP2021086018W WO2022167134A1 WO 2022167134 A1 WO2022167134 A1 WO 2022167134A1 EP 2021086018 W EP2021086018 W EP 2021086018W WO 2022167134 A1 WO2022167134 A1 WO 2022167134A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- plant
- cncm
- accordance
- fermentation
- bacillus amyloliquefaciens
- Prior art date
Links
- 241000186428 Propionibacterium freudenreichii Species 0.000 title claims abstract description 40
- 241000193744 Bacillus amyloliquefaciens Species 0.000 title claims abstract description 38
- 239000000203 mixture Substances 0.000 claims abstract description 69
- 238000000855 fermentation Methods 0.000 claims abstract description 52
- 230000004151 fermentation Effects 0.000 claims abstract description 49
- FDJOLVPMNUYSCM-WZHZPDAFSA-L cobalt(3+);[(2r,3s,4r,5s)-5-(5,6-dimethylbenzimidazol-1-yl)-4-hydroxy-2-(hydroxymethyl)oxolan-3-yl] [(2r)-1-[3-[(1r,2r,3r,4z,7s,9z,12s,13s,14z,17s,18s,19r)-2,13,18-tris(2-amino-2-oxoethyl)-7,12,17-tris(3-amino-3-oxopropyl)-3,5,8,8,13,15,18,19-octamethyl-2 Chemical compound [Co+3].N#[C-].N([C@@H]([C@]1(C)[N-]\C([C@H]([C@@]1(CC(N)=O)C)CCC(N)=O)=C(\C)/C1=N/C([C@H]([C@@]1(CC(N)=O)C)CCC(N)=O)=C\C1=N\C([C@H](C1(C)C)CCC(N)=O)=C/1C)[C@@H]2CC(N)=O)=C\1[C@]2(C)CCC(=O)NC[C@@H](C)OP([O-])(=O)O[C@H]1[C@@H](O)[C@@H](N2C3=CC(C)=C(C)C=C3N=C2)O[C@@H]1CO FDJOLVPMNUYSCM-WZHZPDAFSA-L 0.000 claims abstract description 39
- 229930003779 Vitamin B12 Natural products 0.000 claims abstract description 38
- 235000019163 vitamin B12 Nutrition 0.000 claims abstract description 38
- 239000011715 vitamin B12 Substances 0.000 claims abstract description 38
- LWGJTAZLEJHCPA-UHFFFAOYSA-N n-(2-chloroethyl)-n-nitrosomorpholine-4-carboxamide Chemical compound ClCCN(N=O)C(=O)N1CCOCC1 LWGJTAZLEJHCPA-UHFFFAOYSA-N 0.000 claims abstract description 34
- 230000001580 bacterial effect Effects 0.000 claims abstract description 4
- 241000196324 Embryophyta Species 0.000 claims description 61
- 108010064851 Plant Proteins Proteins 0.000 claims description 15
- 235000021118 plant-derived protein Nutrition 0.000 claims description 15
- 235000020245 plant milk Nutrition 0.000 claims description 14
- 241000208818 Helianthus Species 0.000 claims description 10
- 235000003222 Helianthus annuus Nutrition 0.000 claims description 10
- 235000013351 cheese Nutrition 0.000 claims description 6
- 239000004615 ingredient Substances 0.000 claims description 6
- 235000020275 sunflower milk Nutrition 0.000 claims description 4
- 235000012377 Salvia columbariae var. columbariae Nutrition 0.000 claims description 2
- 235000001498 Salvia hispanica Nutrition 0.000 claims description 2
- 235000014167 chia Nutrition 0.000 claims description 2
- 244000063299 Bacillus subtilis Species 0.000 claims 1
- 235000014469 Bacillus subtilis Nutrition 0.000 claims 1
- KDXKERNSBIXSRK-YFKPBYRVSA-N L-lysine Chemical compound NCCCC[C@H](N)C(O)=O KDXKERNSBIXSRK-YFKPBYRVSA-N 0.000 claims 1
- 244000292604 Salvia columbariae Species 0.000 claims 1
- 235000021135 plant-based food Nutrition 0.000 abstract description 9
- 239000000047 product Substances 0.000 description 12
- 239000000725 suspension Substances 0.000 description 11
- 244000045195 Cicer arietinum Species 0.000 description 10
- 235000010523 Cicer arietinum Nutrition 0.000 description 10
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 10
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 9
- 239000000796 flavoring agent Substances 0.000 description 8
- 235000013305 food Nutrition 0.000 description 8
- 240000004713 Pisum sativum Species 0.000 description 7
- 235000010582 Pisum sativum Nutrition 0.000 description 7
- 238000004458 analytical method Methods 0.000 description 6
- 238000000034 method Methods 0.000 description 6
- 235000019634 flavors Nutrition 0.000 description 5
- 235000013336 milk Nutrition 0.000 description 5
- 239000008267 milk Substances 0.000 description 5
- 210000004080 milk Anatomy 0.000 description 5
- 235000016709 nutrition Nutrition 0.000 description 5
- 239000000523 sample Substances 0.000 description 5
- 241000894006 Bacteria Species 0.000 description 4
- 241001465754 Metazoa Species 0.000 description 4
- 244000046052 Phaseolus vulgaris Species 0.000 description 4
- 235000010627 Phaseolus vulgaris Nutrition 0.000 description 4
- 238000010564 aerobic fermentation Methods 0.000 description 4
- 239000008367 deionised water Substances 0.000 description 4
- 229910021641 deionized water Inorganic materials 0.000 description 4
- 235000013372 meat Nutrition 0.000 description 4
- 239000002609 medium Substances 0.000 description 4
- 238000002360 preparation method Methods 0.000 description 4
- 240000006677 Vicia faba Species 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 230000000813 microbial effect Effects 0.000 description 3
- 238000002156 mixing Methods 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 239000001974 tryptic soy broth Substances 0.000 description 3
- 108010050327 trypticase-soy broth Proteins 0.000 description 3
- 235000010749 Vicia faba Nutrition 0.000 description 2
- 235000002096 Vicia faba var. equina Nutrition 0.000 description 2
- 235000002098 Vicia faba var. major Nutrition 0.000 description 2
- 244000042295 Vigna mungo Species 0.000 description 2
- 235000010721 Vigna radiata var radiata Nutrition 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- RMRCNWBMXRMIRW-BYFNXCQMSA-M cyanocobalamin Chemical compound N#C[Co+]N([C@]1([H])[C@H](CC(N)=O)[C@]\2(CCC(=O)NC[C@H](C)OP(O)(=O)OC3[C@H]([C@H](O[C@@H]3CO)N3C4=CC(C)=C(C)C=C4N=C3)O)C)C/2=C(C)\C([C@H](C/2(C)C)CCC(N)=O)=N\C\2=C\C([C@H]([C@@]/2(CC(N)=O)C)CCC(N)=O)=N\C\2=C(C)/C2=N[C@]1(C)[C@@](C)(CC(N)=O)[C@@H]2CCC(N)=O RMRCNWBMXRMIRW-BYFNXCQMSA-M 0.000 description 2
- 235000013365 dairy product Nutrition 0.000 description 2
- 235000005911 diet Nutrition 0.000 description 2
- 230000037213 diet Effects 0.000 description 2
- 235000005489 dwarf bean Nutrition 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 235000013312 flour Nutrition 0.000 description 2
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 2
- 230000000873 masking effect Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000005457 optimization Methods 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 235000018102 proteins Nutrition 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 235000021251 pulses Nutrition 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 241000894007 species Species 0.000 description 2
- 241000251468 Actinopterygii Species 0.000 description 1
- BSYNRYMUTXBXSQ-UHFFFAOYSA-N Aspirin Chemical compound CC(=O)OC1=CC=CC=C1C(O)=O BSYNRYMUTXBXSQ-UHFFFAOYSA-N 0.000 description 1
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 1
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- XFXPMWWXUTWYJX-UHFFFAOYSA-N Cyanide Chemical compound N#[C-] XFXPMWWXUTWYJX-UHFFFAOYSA-N 0.000 description 1
- 230000006820 DNA synthesis Effects 0.000 description 1
- 235000002673 Dioscorea communis Nutrition 0.000 description 1
- 241000544230 Dioscorea communis Species 0.000 description 1
- 241000186605 Lactobacillus paracasei Species 0.000 description 1
- 235000014647 Lens culinaris subsp culinaris Nutrition 0.000 description 1
- 244000043158 Lens esculenta Species 0.000 description 1
- 241001534756 Mungos Species 0.000 description 1
- 102000006386 Myelin Proteins Human genes 0.000 description 1
- 108010083674 Myelin Proteins Proteins 0.000 description 1
- 244000061176 Nicotiana tabacum Species 0.000 description 1
- 235000002637 Nicotiana tabacum Nutrition 0.000 description 1
- 208000035753 Periorbital contusion Diseases 0.000 description 1
- 235000006089 Phaseolus angularis Nutrition 0.000 description 1
- 244000045930 Phaseolus coccineus Species 0.000 description 1
- 235000010632 Phaseolus coccineus Nutrition 0.000 description 1
- 235000010617 Phaseolus lunatus Nutrition 0.000 description 1
- 244000042209 Phaseolus multiflorus Species 0.000 description 1
- 240000005481 Salvia hispanica Species 0.000 description 1
- 240000007098 Vigna angularis Species 0.000 description 1
- 235000010711 Vigna angularis Nutrition 0.000 description 1
- 235000010716 Vigna mungo Nutrition 0.000 description 1
- 235000006085 Vigna mungo var mungo Nutrition 0.000 description 1
- 230000037354 amino acid metabolism Effects 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 235000021120 animal protein Nutrition 0.000 description 1
- 230000000433 anti-nutritional effect Effects 0.000 description 1
- 239000002518 antifoaming agent Substances 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 235000015241 bacon Nutrition 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 210000001185 bone marrow Anatomy 0.000 description 1
- 229940041514 candida albicans extract Drugs 0.000 description 1
- 235000013339 cereals Nutrition 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 229940112822 chewing gum Drugs 0.000 description 1
- 235000015218 chewing gum Nutrition 0.000 description 1
- 239000002537 cosmetic Substances 0.000 description 1
- 235000014048 cultured milk product Nutrition 0.000 description 1
- 235000000639 cyanocobalamin Nutrition 0.000 description 1
- 239000011666 cyanocobalamin Substances 0.000 description 1
- 229960002104 cyanocobalamin Drugs 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 235000015872 dietary supplement Nutrition 0.000 description 1
- 229910001873 dinitrogen Inorganic materials 0.000 description 1
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 1
- 229910000396 dipotassium phosphate Inorganic materials 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 244000013123 dwarf bean Species 0.000 description 1
- 235000013601 eggs Nutrition 0.000 description 1
- 210000003743 erythrocyte Anatomy 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 239000011536 extraction buffer Substances 0.000 description 1
- 230000004129 fatty acid metabolism Effects 0.000 description 1
- 235000014106 fortified food Nutrition 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 244000080020 horsebean Species 0.000 description 1
- 238000011065 in-situ storage Methods 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 235000014655 lactic acid Nutrition 0.000 description 1
- 239000004310 lactic acid Substances 0.000 description 1
- 230000003050 macronutrient Effects 0.000 description 1
- 235000021073 macronutrients Nutrition 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000035800 maturation Effects 0.000 description 1
- 235000012054 meals Nutrition 0.000 description 1
- 238000011177 media preparation Methods 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- GBMDVOWEEQVZKZ-UHFFFAOYSA-N methanol;hydrate Chemical compound O.OC GBMDVOWEEQVZKZ-UHFFFAOYSA-N 0.000 description 1
- 238000009629 microbiological culture Methods 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 239000011785 micronutrient Substances 0.000 description 1
- 235000013369 micronutrients Nutrition 0.000 description 1
- 210000005012 myelin Anatomy 0.000 description 1
- 210000000653 nervous system Anatomy 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 238000009928 pasteurization Methods 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 235000020266 pea milk Nutrition 0.000 description 1
- 229920001296 polysiloxane Polymers 0.000 description 1
- 239000006041 probiotic Substances 0.000 description 1
- 235000018291 probiotics Nutrition 0.000 description 1
- 108010009004 proteose-peptone Proteins 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 235000014102 seafood Nutrition 0.000 description 1
- 230000001953 sensory effect Effects 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 230000001502 supplementing effect Effects 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 238000011282 treatment Methods 0.000 description 1
- 239000012137 tryptone Substances 0.000 description 1
- 235000020806 vegan diet Nutrition 0.000 description 1
- 235000003563 vegetarian diet Nutrition 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 235000019156 vitamin B Nutrition 0.000 description 1
- 239000011720 vitamin B Substances 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C11/00—Milk substitutes, e.g. coffee whitener compositions
- A23C11/02—Milk substitutes, e.g. coffee whitener compositions containing at least one non-milk component as source of fats or proteins
- A23C11/10—Milk substitutes, e.g. coffee whitener compositions containing at least one non-milk component as source of fats or proteins containing or not lactose but no other milk components as source of fats, carbohydrates or proteins
- A23C11/103—Milk substitutes, e.g. coffee whitener compositions containing at least one non-milk component as source of fats or proteins containing or not lactose but no other milk components as source of fats, carbohydrates or proteins containing only proteins from pulses, oilseeds or nuts, e.g. nut milk
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C11/00—Milk substitutes, e.g. coffee whitener compositions
- A23C11/02—Milk substitutes, e.g. coffee whitener compositions containing at least one non-milk component as source of fats or proteins
- A23C11/10—Milk substitutes, e.g. coffee whitener compositions containing at least one non-milk component as source of fats or proteins containing or not lactose but no other milk components as source of fats, carbohydrates or proteins
- A23C11/103—Milk substitutes, e.g. coffee whitener compositions containing at least one non-milk component as source of fats or proteins containing or not lactose but no other milk components as source of fats, carbohydrates or proteins containing only proteins from pulses, oilseeds or nuts, e.g. nut milk
- A23C11/106—Addition of, or treatment with, microorganisms
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C20/00—Cheese substitutes
- A23C20/02—Cheese substitutes containing neither milk components, nor caseinate, nor lactose, as sources of fats, proteins or carbohydrates
- A23C20/025—Cheese substitutes containing neither milk components, nor caseinate, nor lactose, as sources of fats, proteins or carbohydrates mainly containing proteins from pulses or oilseeds
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L11/00—Pulses, i.e. fruits of leguminous plants, for production of food; Products from legumes; Preparation or treatment thereof
- A23L11/50—Fermented pulses or legumes; Fermentation of pulses or legumes based on the addition of microorganisms
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/15—Vitamins
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/17—Amino acids, peptides or proteins
- A23L33/175—Amino acids
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/26—Preparation of nitrogen-containing carbohydrates
- C12P19/28—N-glycosides
- C12P19/42—Cobalamins, i.e. vitamin B12, LLD factor
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P39/00—Processes involving microorganisms of different genera in the same process, simultaneously
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2250/00—Food ingredients
- A23V2250/54—Proteins
- A23V2250/548—Vegetable protein
Definitions
- the present invention relates generally to the field of plant-based food.
- the present invention relates to improving the vitamin B12 content of plant-based food products.
- One embodiment of the present invention relates to the use of a combination of Bacillus amyloliquefaciens and Propionibacterium freudenreichii to enrich the vitamin B12 content in plant-based compositions through fermentation.
- the present invention further relates to a bacterial strain selected from the group consisting of Bacillus amyloliquefaciens CNCM I-5624, Propionibacterium freudenreichii CNCM I-5623, and Propionibacterium freudenreichii CNCM I-5639.
- plant-based milk alternatives do not yet achieve the nutritional profile of milk-based products, in particular if the plant-based milk alternative is based on a single-source plant-based material.
- Today the problem is usually overcome by combining several plant-based sources, or by supplementing the plant-based composition with specific micro- and/or macro nutrients.
- Another way to further increase the demand of plant-based foods is to further increase their quality and taste profile to match the products of animal origin (e.g., milk or meat).
- the inventors propose to use fermentation to improve the taste, texture and nutritional profile of plant-based products.
- fermentation is known to be technology that can reduce off-flavours, liberate new flavours, optimize textural aspects, reduce anti-nutritional factors, and - also - to synthesize nutritional compounds such as vitamins and amino acids.
- EP1625794 discloses a fermented milk product containing an increased content of in situ produced vitamin B12, wherein at least bacteria capable of producing vitamin B12 are present.
- Vitamin B12 is also known as cobalamin. Vitamin B12 is one of the B-vitamins and is soluble in water. It plays a key role in human metabolism. For example, it is involved in the maturation of developing red blood cells in the bone marrow, in fatty acid metabolism, in amino acid metabolism, in DNA synthesis, and in the myelin synthesis and - consequently - in the normal functioning of the nervous system.
- Vitamin B12 is usually obtained in the daily diet from consuming animal-sourced foods, including meat, fish, fowl, milk and eggs. Very few non-animal-based food sources are rich in vitamin B12. As a consequence, many grain-based products are fortified with vitamin B12. Also, consumers electing not to consume animal-based food are frequently advised to consume a dietary supplement comprising vitamin B12 or vitamin B12 fortified foods.
- the objective of the present invention was it to provide a particular effective method to increase the vitamin B12 content in plant-based compositions, in particular singlesource plant-based compositions, for example plant-based milk alternatives while improving their taste and/or texture profile, or to at least provide a useful alternative to existing solutions available in the art.
- the present invention provides a use of a combination of Bacillus amyloliquefaciens and Propionibacterium freudenreichii to enrich the vitamin B12 content in plant-based compositions through fermentation.
- the present invention further provides Bacillus amyloliquefaciens CNCM 1-5624, Propionibacterium freudenreichii CNCM 1-5623, and Propionibacterium freudenreichii CNCM 1-5639. These strains can be used, for example, to ferment sunflower-based plant-based ingredients to increase their vitamin B12 content.
- the present inventors have shown that the vitamin B12 content of a plant-based composition can be improved through fermentation of the plant-based composition with a combination of Bacillus amyloliquefaciens and Propionibacterium freudenreichii.
- the micronutrient profile of the plant-based composition is improved by increasing the vitamin B12 content through fermentation. This could be achieved while - at the same time - removing and/or masking undesired off-flavours of the plant-based composition. Fermentation further allowed it to produce a pleasant texture in the plant-based composition.
- the inventors could further show that Bacillus amyloliquefaciens and Propionibacterium freudenreichii act synergistically to achieve the objective of the present invention. Finally, the inventors have identified certain specific Bacillus amyloliquefaciens and Propionibacterium freudenreichii strains that allowed to produced particularly promising results. The inventors could further show that the use of a combination of Bacillus amyloliquefaciens, Propionibacterium freudenreichii and Lacticaseibacillus paracasei is particularly effective in achieving the objective of the present invention, for example in enriching the vitamin B12 content in plant-based compositions through fermentation.
- Figure 1 shows Vitamin B12 content in three kinds of plant milk before and after fermentation.
- Single means NCC1177
- double means NCC1177+NCC2511
- triple means NCC1177+NCC2511+NCC156.
- Figure 2 shows that mixed-culture fermentation yielded a more balanced volatile compounds composition.
- the present invention relates in part to the use of a combination of Bacillus amyloliquefaciens and Propionibacterium freudenreichii to enrich the vitamin B12 content in plant-based compositions through fermentation.
- the inventors were surprised to see that Bacillus amyloliquefaciens and Propionibacterium freudenreichii acted synergistically in improving the vitamin B12 content in a plant-based composition, for example in a sunflower-based plant-based composition.
- the plant-based compositions may be a single-source plant-based composition.
- a plant-based composition may be considered a single source plant-based composition, if at least at least 75 weight-%, at least 90 weight-%, at least 95 weight-%, at least 99 weight-% or 100 weight-% of its protein content is derived from one plant origin, for example from chickpea.
- Bacillus amyloliquefaciens is a well-studied species of bacterium in the genus Bacillus, frequently used in agriculture to strengthen the resistance of roots to pathogens or high salt conditions.
- Propionibacterium freudenreichii is another well-studied gram-positive bacterium frequently used in food manufacturing. It has a particularly long usage history in cheese making, in particular in the making of the famous Emmental cheese.
- a composition shall be considered as plantbased, if at least 75 weight-%, at least 90 weight-%, at least 95 weight-%, at least 99 weight-% or 100 weight-% of its protein content is not from animal origin.
- a composition may be considered as plant-based, if at least 75 weight-%, at least 90 weight-%, at least 95 weight-%, at least 99 weight-% or 100 weight-% of its protein content is from plant origin.
- the plant-based composition may be a plant-based food composition.
- the term "food” shall mean in accordance with Codex Alimentarius any substance, whether processed, semi-processed or raw, which is intended for human consumption, and includes drink, chewing gum and any substance which has been used in the manufacture, preparation or treatment of "food” but does not include cosmetics or tobacco or substances used only as drugs.
- the inventors were able to demonstrate, that by fermentation with a combination of Bacillus amyloliquefaciens and Propionibacterium freudenreichii the vitamin B12 content in plant-based compositions could be increased by at least 200% compared to unfermented plant milk.
- the inventors have carried out further studies to even further improve the vitamin B12 generation and were surprised that the addition of Lacticaseibacillus paracasei allowed to even further increase the vitamin B12 generation.
- the inventors were surprised to see that Bacillus amyloliquefaciens, Propionibacterium freudenreichii and Lacticaseibacillus paracasei acted synergistically in improving the vitamin B12 content in a plant-based composition, for example in a sunflower-based plant-based composition.
- Lacticaseibacillus paracasei (L. paracasei) is a well-studied gram-positive, facultatively heterofermentative species of lactic acid bacteria. They are frequently used in food technology, for example in dairy product fermentation and as probiotics.
- composition may further comprise Lacticaseibacillus paracasei.
- Bacillus amyloliquefaciens is Bacillus amyloliquefaciens CNCM 1-5624; the Propionibacterium freudenreichii is selected from the group consisting of CNCM 1-5639, CNCM 1-5623, or combinations thereof; and Lacticaseibacillus paracasei is ATCC 25302.
- Bacillus amyloliquefaciens may be Bacillus amyloliquefaciens CNCM 1-5624; Propionibacterium freudenreichii may be selected from the group consisting of CNCM 1-5639, CNCM 1-5623, or combinations thereof; and Lacticaseibacillus paracasei is ATCC 25302.
- Propionibacterium freudenreichii NCC 1177 was deposited under the Budapest treaty with the Institut Pasteur (28 rue du Dondel Roux 75024 Paris Cedex 15) and was designated CNCM 1-5623.
- Propionibacterium freudenreichii NCC 1186 was deposited under the Budapest treaty with the Institut Pasteur (28 rue du Dondel Roux 75024 Paris Cedex 15) and was designated CNCM 1-5639.
- Lacticaseibacillus paracasei NCC 2511 was deposited as ATCC 25302 (American Type Culture Collection) / DSM 5622 (Deutsche Sammlung von Mikroorganismen un Zellkulturen GmbH. This strain is commercially available from many sources, for example from JCM RIKEN, Japan.
- Lacticaseibacillus rhamnosus NCC 4007 is described, for example in W02010091992, was deposited under the Budapest Treaty as Lacticaseibacillus rhamnosus CGMCC 1 .3724 with the China General Microbiological Culture Collection Center (CGMCC).
- CGMCC China General Microbiological Culture Collection Center
- the subject matter of the present invention comprises Bacillus amyloliquefaciens CNCM 1-5624 and Propionibacterium freudenreichii CNCM 1-5623.
- the inventors have tested different combinations of Bacillus amyloliquefaciens and Propionibacterium freudenreichii. While they all achieved the objective of the present invention and were surprisingly more effective than one of the strains alone, a combination of Bacillus amyloliquefaciens is NCC156 and Propionibacterium freudenreichii is NCC 1177 stood out in terms of vitamin B12 generation. Hence, in one embodiment of the use of the present invention the Bacillus amyloliquefaciens is CNCM 1-5624 and the Propionibacterium freudenreichii is CNCM I- 5623.
- the inventors have also tested different combinations of Bacillus amyloliquefaciens, Propionibacterium freudenreichii and Lacticaseibacillus paracasei. While they all achieved the objective of the present invention and were surprisingly more effective than one or two of the strains alone, a combination of Bacillus amyloliquefaciens is NCC156, the Propionibacterium freudenreichii is NCC1177 and the Lacticaseibacillus paracasei is NCC2511 stood out in terms of vitamin B12 generation.
- Bacillus amyloliquefaciens is CNCM 1-5624
- the Propionibacterium freudenreichii is CNCM I- 5623
- the Lacticaseibacillus paracasei is ATCC 25302.
- the fermentation may be carried out in an anaerobic state or in an aerobic state.
- the fermentation is carried out in two steps, wherein one step is carried out in an anaerobic state and the other step is carried out in an aerobic state.
- the inventors have obtained best results when the first step of the fermentation was carried out in an anaerobic state and the second step of the fermentation was carried out in an aerobic state.
- the fermentation may be carried out usingthe steps of preparing a suspension of the plant protein with water, preparation of a preculture of the microorganisms, and the fermentation of the plant protein suspension with the preculture.
- the inventors have obtained very good results when the fermentation of the plant protein suspension comprised an anaerobic and an aerobic fermentation step.
- the fermentation of the plant protein suspension may be carried out at room temperature or at temperature in the range of about 25°C to 35°C, for example 28°C to 32°C.
- the inventors have obtained good results when the fermentation of the plant protein suspension was carried out for about 12 - 72 hours, for example 24 - 48 hours.
- the fermentation of the plant protein suspension comprises a first anaerobic fermentation step for 12-48 hours, for example 20-28 hours and a second aerobic fermentation step for 24-60 hours, for example 44-52 hours.
- the fermentation of the plant protein suspension comprises a first anaerobic fermentation step for 24-60 hours, for example 44-52 hours and a second aerobic fermentation step for 12-48 hours, for example 20-28 hours.
- the fermentation may be carried out as described in the Examples.
- the plant-based compositions with an enriched vitamin B12 content may be used as a natural source of vitamin B12 for vegan products such as meat alternatives, dairy alternatives, cheese alternatives, milk alternatives, bacon alternatives, or seafood alternatives, for example, while improving taste by flavor generation and/or the removal or masking of undesired off-flavors.
- the fermented plant-based compositions may be used as a final fermented product (e.g. as cheese alternative, or as a plant-based milk alternative) or as an ingredient to be added to other products (e.g. as ingredient for a meat alternative products).
- a final fermented product e.g. as cheese alternative, or as a plant-based milk alternative
- an ingredient to be added to other products e.g. as ingredient for a meat alternative products.
- the plant-based composition may be selected from the group consisting of ingredients for plant-based products and plant- based milk product alternatives, for example, plant-based milk alternatives or plantbased cheese alternatives.
- the plant-based composition may be sunflower seed milk.
- Sunflower seed milk can be prepared, for example, by mixing 7 % (w/w) of sunflower protein Heliaflor 55 (All Organic Treasures GmbH) with deionized water, followed by pasteurization at about 95 °C for about 6 h. Prior to the microbial fermentation, the suspension can then be homogenized by shaking.
- the plant-based composition may be based on a single plant protein source or on a combination of plant protein sources.
- the plant-based composition may comprise plant proteins derived from sources selected from the group consisting of the Fabacea family, the Leguminosa family, nuts, seeds, sasha inshi, sunflower, chia or combinations thereof.
- the plant proteins may be derived from pulses.
- the pulses may be selected from the group consisting of peas, chickpeas, lentils, beans, or combinations thereof.
- the peas may be selected from the group consisting of garden peas, cow peas, blackeye peas, field peas or combinations thereof.
- the beans may be selected from the group consisting of dry beans, kidney deans, haricot beans, butter beans, adzuki beans, mungo beans, green gram, black gram, scarlet runner beans, broad beans, field beans, horse beans, faba beans, or combinations thereof.
- the plant-based composition may be based on chick-pea.
- a composition shall be considered as based on chick-pea, if at least 75 weight-%, at least 90 weight- %, at least 95 weight-%, at least 99 weight-% or 100 weight-% of its protein content is from chick-pea origin.
- the plant-based composition may be based on sunflower.
- a composition shall be considered as based on sunflower, if at least 75 weight-%, at least 90 weight-%, at least 95 weight-%, at least 99 weight-% or 100 weight-% of its protein content is from chick-pea origin.
- the plant protein in the plant-based composition may contribute at least 75 weight-% of the total protein content of the composition, at least 85 weight-% of the total protein content of the composition, or at least 95 weight-% of the total protein content of the composition. In one embodiment of the present invention, the plant protein in the plant-based composition may contribute 100 weight-% of the total protein content of the composition.
- the framework of the present invention comprises the use of the present invention to improve the smell of plant-based compositions.
- Lacticaseibacillus paracasei improves the freshness and sweetness of the smell of the plant-based compositions.
- the present inventors have also tasted the plant-based composition fermented in accordance with the present invention and have found that the fermentation leads to an improved taste and an improved mouthfeel of the tested composition.
- the framework of the present invention further comprises the use of the present invention to improve the taste and/or the texture of plant-based compositions.
- Plant milk media preparation o A chickpea suspension (in the following named chickpea milk due to its milk-like appearance) was prepared by mixing 10 % (w/w) chickpea flour with deionized water. A two-step heat treatment was applied for sterilization. First, the chickpea suspension was stirred (250 rpm) for 2 h at 75 °C, followed by autoclaving (121 °C, 15 min). o Pea milk was prepared by dissolving 10 % (w/w) pea flour in deionized water, and was autoclaved (121 °C, 15 min). o Sunflower seed milk was prepared by mixing 7 % (w/w) of 55 % sunflower protein with deionized water, then pasteurized under 95 °C for 6 h. Prior to microbial fermentation, the suspension was manually homogenized.
- Preculture condition (Pl, from glycerol stock):
- Preculture condition (Pl, from glycerol stock):
- Preculture condition (P2, 2 % from pl): MRS, incubator (30 degree, no humidity control, no shaking) overnight o Propionibacterium freudenreichii NCC 1177:
- Preculture condition (Pl, from glycerol stock):
- Fermentation stage 1 Anaerobic jar in incubator (30 degree, no humidity control, no shaking), 48 h;
- Fermentation stage 2 shaker (30 degree, 80 % humidity, 130 rpm), 24 h
- Vitamin B12 was analysed following AOAC 2014.02 with small modifications. Briefly, 6 mL of sample were mixed with an extraction buffer (2.5 mL) and cyanide solution (0.1 mL). The solutions were heated at 107 5 C for 20 min. After making up to 10 mL with water, the extracts were centrifuged, and 8.5 mL of supernatant was subjected to automated immunoaffinity clean-up on a Gilson GX-271 ASPEC system. The final eluate was evaporated to dryness and reconstituted in 0.2 mL of water. Analysis was performed by UHPLC-UV, the chromatographic conditions were modified to allow separation of cyanocobalamin and tentatively identified pseudocobalamin. Separation took place on a Waters Atlantis T3 column, 3.0 pm, 4.6 x 150 mm on isocratic mode. Mobile phase was methanol-water (25:75), flow rate 0.3 mL/min.
- Preculture preparation of strains are the same as example 1. Precultures were grown in the respective medium under anaerobic or aerobic conditions in flasks.
- the main fermentation was conducted in the 1.8L DASGIP Bioreactor system (DASGIP AG, Germany).
- the plant milk was transferred into glass bioreactor, incubated on the bench top control system for a minimum of 6 hours with the vessel temperature at 30°C.
- the DO probe was calibrated between 0% and 100%.
- precultures of Propionibacterium freudenreichii NCC 1177 and Bacillus amyloliquefaciens NCC 156 are inoculated into plant milk to start the fermentation.
- the pH was monitored using an on-line pH sensor.
- the culture temperature was controlled at 30°C.
- pO2 sensor 02 Sen lnPro6800G was utilized the DO control.
- stage 1 filter-sterilized nitrogen gas was sparged through the fermentation broth to maintain an anaerobic environment, whereas air was used to maintain an aerobic condition (stage 2).
- the DO concentration or ORP was controlled by adjusting the volume of air pumped in and the rotation speed of the stirrer for the aerobic fermentation.
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Polymers & Plastics (AREA)
- Food Science & Technology (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Nutrition Science (AREA)
- Zoology (AREA)
- Microbiology (AREA)
- Wood Science & Technology (AREA)
- Mycology (AREA)
- Biotechnology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- General Engineering & Computer Science (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Biochemistry (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Molecular Biology (AREA)
- Agronomy & Crop Science (AREA)
- Botany (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The present invention relates generally to the field of plant-based food. In particular, the present invention relates to improving the vitamin B12 content of plant-based food products. One embodiment of the present invention relates to the use of a combination of Bacillus amyloliquefaciens and Propionibacterium freudenreichii to enrich the vitamin B12 content in plant-based compositions through fermentation. The present invention further relates to a bacterial strain selected from the group consisting of Bacillus amyloliquefaciens CNCM I-5624, Propionibacterium freudenreichii CNCM I-5623, and Propionibacterium freudenreichii CNCM I-5639.
Description
PLANT-BASED PRODUCTS FERMENTED WITH B. AMYLOLIQUEFACIENS AND P. FREUDENREICHII HAVING INCREASED VIT B12 CONTENT
The present invention relates generally to the field of plant-based food. In particular, the present invention relates to improving the vitamin B12 content of plant-based food products. One embodiment of the present invention relates to the use of a combination of Bacillus amyloliquefaciens and Propionibacterium freudenreichii to enrich the vitamin B12 content in plant-based compositions through fermentation. The present invention further relates to a bacterial strain selected from the group consisting of Bacillus amyloliquefaciens CNCM I-5624, Propionibacterium freudenreichii CNCM I-5623, and Propionibacterium freudenreichii CNCM I-5639.
A growing number of consumers are choosing to enjoy food based on animal protein sources less often, be it as part of a flexitarian diet or as part of a vegetarian or vegan diet. A 2017 study found that 69% of Germans, for example, and 38% of Americans eat meatless meals once a week or more. Global plant-based foods sales are set to hit USD 5bn this year. Plant-based foods can have a positive impact on the environment as well, as they may use less water and CO2 in production.
One way to further increase the demand of plant-based foods is to further increase their nutritional profile. Oftentimes, today, for example plant-based milk alternatives do not yet achieve the nutritional profile of milk-based products, in particular if the plant-based milk alternative is based on a single-source plant-based material. Today the problem is usually overcome by combining several plant-based sources, or by supplementing the plant-based composition with specific micro- and/or macro nutrients.
Another way to further increase the demand of plant-based foods is to further increase their quality and taste profile to match the products of animal origin (e.g., milk or meat).
Here the inventors propose to use fermentation to improve the taste, texture and nutritional profile of plant-based products.
In general, fermentation is known to be technology that can reduce off-flavours, liberate new flavours, optimize textural aspects, reduce anti-nutritional factors, and - also - to synthesize nutritional compounds such as vitamins and amino acids.
For example, EP1625794 discloses a fermented milk product containing an increased content of in situ produced vitamin B12, wherein at least bacteria capable of producing vitamin B12 are present.
Vitamin B12 is also known as cobalamin. Vitamin B12 is one of the B-vitamins and is soluble in water. It plays a key role in human metabolism. For example, it is involved in the maturation of developing red blood cells in the bone marrow, in fatty acid metabolism, in amino acid metabolism, in DNA synthesis, and in the myelin synthesis and - consequently - in the normal functioning of the nervous system.
Vitamin B12 is usually obtained in the daily diet from consuming animal-sourced foods, including meat, fish, fowl, milk and eggs. Very few non-animal-based food sources are rich in vitamin B12. As a consequence, many grain-based products are fortified with vitamin B12. Also, consumers electing not to consume animal-based food are frequently advised to consume a dietary supplement comprising vitamin B12 or vitamin B12 fortified foods.
It would hence be desirable to have available a process that allows it to improve taste and texture of plant-based compositions, in particular, single-source plant-based compositions, while - at the same time - increasing their vitamin B12 content.
To achieve these benefits via fermentation, the right microbial strains have to be selected. Further improvements can be achieved by optimizing process conditions, especially to combine the optimization of flavour and nutritional aspects.
It would therefore be desirable to have available a method to treat plant-based compositions, in particular, single-source plant-based compositions, for example, plant-based milk alternatives, that allows it to increase their vitamin B12 content while also having a positive impact on taste and flavor. As different bacterial strains deliver different taste and textures in fermentation, it would be desirable to have available an alternative method to what is described in EP1625794. It would also be desirable to have available a fermentation method that delivers particularly high vitamin B12 contents.
Any reference to prior art documents in this specification is not to be considered an admission that such prior art is widely known or forms part of the common general knowledge in the field.
The objective of the present invention was it to provide a particular effective method to increase the vitamin B12 content in plant-based compositions, in particular singlesource plant-based compositions, for example plant-based milk alternatives while improving their taste and/or texture profile, or to at least provide a useful alternative to existing solutions available in the art.
The inventors were surprised to see that the objective of the present invention could be achieved by the subject matter of the independent claims. The dependent claims further develop the idea of the present invention.
Accordingly, the present invention provides a use of a combination of Bacillus amyloliquefaciens and Propionibacterium freudenreichii to enrich the vitamin B12 content in plant-based compositions through fermentation.
The present invention further provides Bacillus amyloliquefaciens CNCM 1-5624, Propionibacterium freudenreichii CNCM 1-5623, and Propionibacterium freudenreichii CNCM 1-5639. These strains can be used, for example, to ferment sunflower-based plant-based ingredients to increase their vitamin B12 content.
As used in this specification, the words "comprises", "comprising", and similar words, are not to be interpreted in an exclusive or exhaustive sense. In other words, they are intended to mean "including, but not limited to".
The present inventors have shown that the vitamin B12 content of a plant-based composition can be improved through fermentation of the plant-based composition with a combination of Bacillus amyloliquefaciens and Propionibacterium freudenreichii. The micronutrient profile of the plant-based composition is improved by increasing the vitamin B12 content through fermentation. This could be achieved while - at the same time - removing and/or masking undesired off-flavours of the plant-based composition. Fermentation further allowed it to produce a pleasant texture in the plant-based composition.
The inventors could further show that Bacillus amyloliquefaciens and Propionibacterium freudenreichii act synergistically to achieve the objective of the present invention. Finally, the inventors have identified certain specific Bacillus amyloliquefaciens and Propionibacterium freudenreichii strains that allowed to produced particularly promising results.
The inventors could further show that the use of a combination of Bacillus amyloliquefaciens, Propionibacterium freudenreichii and Lacticaseibacillus paracasei is particularly effective in achieving the objective of the present invention, for example in enriching the vitamin B12 content in plant-based compositions through fermentation.
Figure 1 shows Vitamin B12 content in three kinds of plant milk before and after fermentation. Single means NCC1177, double means NCC1177+NCC2511, triple means NCC1177+NCC2511+NCC156.
Figure 2 shows that mixed-culture fermentation yielded a more balanced volatile compounds composition.
Consequently, the present invention relates in part to the use of a combination of Bacillus amyloliquefaciens and Propionibacterium freudenreichii to enrich the vitamin B12 content in plant-based compositions through fermentation.
The inventors were surprised to see that Bacillus amyloliquefaciens and Propionibacterium freudenreichii acted synergistically in improving the vitamin B12 content in a plant-based composition, for example in a sunflower-based plant-based composition.
The plant-based compositions may be a single-source plant-based composition. A plant-based composition may be considered a single source plant-based composition, if at least at least 75 weight-%, at least 90 weight-%, at least 95 weight-%, at least 99 weight-% or 100 weight-% of its protein content is derived from one plant origin, for example from chickpea.
Bacillus amyloliquefaciens is a well-studied species of bacterium in the genus Bacillus, frequently used in agriculture to strengthen the resistance of roots to pathogens or high salt conditions.
Propionibacterium freudenreichii is another well-studied gram-positive bacterium frequently used in food manufacturing. It has a particularly long usage history in cheese making, in particular in the making of the famous Emmental cheese.
For the purpose of the present invention, a composition shall be considered as plantbased, if at least 75 weight-%, at least 90 weight-%, at least 95 weight-%, at least 99 weight-% or 100 weight-% of its protein content is not from animal origin. For example, a composition may be considered as plant-based, if at least 75 weight-%, at least 90 weight-%, at least 95 weight-%, at least 99 weight-% or 100 weight-% of its protein content is from plant origin.
The plant-based composition may be a plant-based food composition. For the purpose of the present invention, the term "food" shall mean in accordance with Codex Alimentarius any substance, whether processed, semi-processed or raw, which is intended for human consumption, and includes drink, chewing gum and any substance which has been used in the manufacture, preparation or treatment of "food" but does not include cosmetics or tobacco or substances used only as drugs.
The inventors were able to demonstrate, that by fermentation with a combination of Bacillus amyloliquefaciens and Propionibacterium freudenreichii the vitamin B12 content in plant-based compositions could be increased by at least 200% compared to unfermented plant milk.
The inventors have carried out further studies to even further improve the vitamin B12 generation and were surprised that the addition of Lacticaseibacillus paracasei allowed to even further increase the vitamin B12 generation.
The inventors were surprised to see that Bacillus amyloliquefaciens, Propionibacterium freudenreichii and Lacticaseibacillus paracasei acted synergistically in improving the vitamin B12 content in a plant-based composition, for example in a sunflower-based plant-based composition.
Lacticaseibacillus paracasei (L. paracasei) is a well-studied gram-positive, facultatively heterofermentative species of lactic acid bacteria. They are frequently used in food technology, for example in dairy product fermentation and as probiotics.
Consequently, in the use of the present invention the composition may further comprise Lacticaseibacillus paracasei.
The inventors have obtained particular promising results when the Bacillus amyloliquefaciens is Bacillus amyloliquefaciens CNCM 1-5624; the Propionibacterium freudenreichii is selected from the group consisting of CNCM 1-5639, CNCM 1-5623, or combinations thereof; and Lacticaseibacillus paracasei is ATCC 25302.
For example, the Bacillus amyloliquefaciens may be Bacillus amyloliquefaciens CNCM 1-5624; Propionibacterium freudenreichii may be selected from the group consisting of CNCM 1-5639, CNCM 1-5623, or combinations thereof; and Lacticaseibacillus paracasei is ATCC 25302.
All strains were deposited under the Budapest treaty.
Bacillus amyloliquefaciens NCC 156 was deposited under the Budapest treaty with the Institut Pasteur (28 rue du Docteur Roux 75024 Paris Cedex 15) and was designated CNCM 1-5624.
Propionibacterium freudenreichii NCC 1177 was deposited under the Budapest treaty with the Institut Pasteur (28 rue du Docteur Roux 75024 Paris Cedex 15) and was designated CNCM 1-5623.
Propionibacterium freudenreichii NCC 1186 was deposited under the Budapest treaty with the Institut Pasteur (28 rue du Docteur Roux 75024 Paris Cedex 15) and was designated CNCM 1-5639.
Lacticaseibacillus paracasei NCC 2511 was deposited as ATCC 25302 (American Type Culture Collection) / DSM 5622 (Deutsche Sammlung von Mikroorganismen un Zellkulturen GmbH. This strain is commercially available from many sources, for example from JCM RIKEN, Japan.
Lacticaseibacillus rhamnosus NCC 4007 is described, for example in W02010091992, was deposited under the Budapest Treaty as Lacticaseibacillus rhamnosus CGMCC 1 .3724 with the China General Microbiological Culture Collection Center (CGMCC).
Hence, the subject matter of the present invention comprises Bacillus amyloliquefaciens CNCM 1-5624 and Propionibacterium freudenreichii CNCM 1-5623.
The inventors have tested different combinations of Bacillus amyloliquefaciens and Propionibacterium freudenreichii. While they all achieved the objective of the present invention and were surprisingly more effective than one of the strains alone, a combination of Bacillus amyloliquefaciens is NCC156 and Propionibacterium freudenreichii is NCC 1177 stood out in terms of vitamin B12 generation.
Hence, in one embodiment of the use of the present invention the Bacillus amyloliquefaciens is CNCM 1-5624 and the Propionibacterium freudenreichii is CNCM I- 5623.
The inventors have also tested different combinations of Bacillus amyloliquefaciens, Propionibacterium freudenreichii and Lacticaseibacillus paracasei. While they all achieved the objective of the present invention and were surprisingly more effective than one or two of the strains alone, a combination of Bacillus amyloliquefaciens is NCC156, the Propionibacterium freudenreichii is NCC1177 and the Lacticaseibacillus paracasei is NCC2511 stood out in terms of vitamin B12 generation.
Hence, in one embodiment of the use of the present invention Bacillus amyloliquefaciens is CNCM 1-5624, the Propionibacterium freudenreichii is CNCM I- 5623 and the Lacticaseibacillus paracasei is ATCC 25302.
The fermentation may be carried out in an anaerobic state or in an aerobic state. In a particular preferred embodiment of the present invention, the fermentation is carried out in two steps, wherein one step is carried out in an anaerobic state and the other step is carried out in an aerobic state. The inventors have obtained best results when the first step of the fermentation was carried out in an anaerobic state and the second step of the fermentation was carried out in an aerobic state.
In the use of the present invention the fermentation may be carried out usingthe steps of preparing a suspension of the plant protein with water, preparation of a preculture of the microorganisms, and the fermentation of the plant protein suspension with the preculture.
The inventors have obtained very good results when the fermentation of the plant protein suspension comprised an anaerobic and an aerobic fermentation step. For example, the fermentation of the plant protein suspension may be carried out at room temperature or at temperature in the range of about 25°C to 35°C, for example 28°C to 32°C. The inventors have obtained good results when the fermentation of the plant protein suspension was carried out for about 12 - 72 hours, for example 24 - 48 hours.
In one embodiment, the fermentation of the plant protein suspension comprises a first anaerobic fermentation step for 12-48 hours, for example 20-28 hours and a second aerobic fermentation step for 24-60 hours, for example 44-52 hours.
In another embodiment, the fermentation of the plant protein suspension comprises a first anaerobic fermentation step for 24-60 hours, for example 44-52 hours and a second aerobic fermentation step for 12-48 hours, for example 20-28 hours.
For example, the fermentation may be carried out as described in the Examples.
After fermentation the plant-based compositions with an enriched vitamin B12 content may be used as a natural source of vitamin B12 for vegan products such as meat alternatives, dairy alternatives, cheese alternatives, milk alternatives, bacon alternatives, or seafood alternatives, for example, while improving taste by flavor generation and/or the removal or masking of undesired off-flavors.
The fermented plant-based compositions may be used as a final fermented product (e.g. as cheese alternative, or as a plant-based milk alternative) or as an ingredient to be added to other products (e.g. as ingredient for a meat alternative products).
Hence, in the use of the present invention the plant-based composition may be selected from the group consisting of ingredients for plant-based products and plant-
based milk product alternatives, for example, plant-based milk alternatives or plantbased cheese alternatives.
For example, the plant-based composition may be sunflower seed milk. Sunflower seed milk can be prepared, for example, by mixing 7 % (w/w) of sunflower protein Heliaflor 55 (All Organic Treasures GmbH) with deionized water, followed by pasteurization at about 95 °C for about 6 h. Prior to the microbial fermentation, the suspension can then be homogenized by shaking.
Depending on the intended purpose of the fermented composition and its desired properties, the plant-based composition may be based on a single plant protein source or on a combination of plant protein sources.
For example, in the use of the present invention the plant-based composition may comprise plant proteins derived from sources selected from the group consisting of the Fabacea family, the Leguminosa family, nuts, seeds, sasha inshi, sunflower, chia or combinations thereof.
In one embodiment of the present invention, the plant proteins may be derived from pulses. For example, the pulses may be selected from the group consisting of peas, chickpeas, lentils, beans, or combinations thereof.
Typically, the peas may be selected from the group consisting of garden peas, cow peas, blackeye peas, field peas or combinations thereof. The beans may be selected from the group consisting of dry beans, kidney deans, haricot beans, butter beans, adzuki beans, mungo beans, green gram, black gram, scarlet runner beans, broad beans, field beans, horse beans, faba beans, or combinations thereof.
For example, the plant-based composition may be based on chick-pea. A composition shall be considered as based on chick-pea, if at least 75 weight-%, at least 90 weight- %, at least 95 weight-%, at least 99 weight-% or 100 weight-% of its protein content is from chick-pea origin.
Further for example, the plant-based composition may be based on sunflower. A composition shall be considered as based on sunflower, if at least 75 weight-%, at least 90 weight-%, at least 95 weight-%, at least 99 weight-% or 100 weight-% of its protein content is from chick-pea origin.
In the use of the present invention the plant protein in the plant-based composition may contribute at least 75 weight-% of the total protein content of the composition, at least 85 weight-% of the total protein content of the composition, or at least 95 weight-% of the total protein content of the composition. In one embodiment of the present invention, the plant protein in the plant-based composition may contribute 100 weight-% of the total protein content of the composition.
A flavor analysis and a sensory evaluation has shown the fermentation of the use of the present invention allows it to improve the taste and/or the smell of the plant-based composition. Hence, the framework of the present invention comprises the use of the present invention to improve the smell of plant-based compositions.
In particular, it was possible to impart a freshness and a sweetness to the smell of the plant-based composition. Hence, in the use of the present invention the fermentation with Bacillus amyloliquefaciens, Propionibacterium freudenreichii and/or
Lacticaseibacillus paracasei improves the freshness and sweetness of the smell of the plant-based compositions.
The present inventors have also tasted the plant-based composition fermented in accordance with the present invention and have found that the fermentation leads to an improved taste and an improved mouthfeel of the tested composition. Hence, the framework of the present invention further comprises the use of the present invention to improve the taste and/or the texture of plant-based compositions.
Those skilled in the art will understand that they can freely combine all features of the present invention disclosed herein. Features described for different embodiments of the present invention may be combined.
Although the invention has been described by way of example, it should be appreciated that variations and modifications may be made without departing from the scope of the invention as defined in the claims. Furthermore, where known equivalents exist to specific features, such equivalents are incorporated as if specifically referred in this specification. Further advantages and features of the present invention are apparent from the figures and non-limiting examples.
Examples:
Example 1
1. Plant milk media preparation o A chickpea suspension (in the following named chickpea milk due to its milk-like appearance) was prepared by mixing 10 % (w/w) chickpea flour with deionized water. A two-step heat treatment was applied for sterilization. First, the chickpea suspension was stirred (250 rpm) for 2 h at 75 °C, followed by autoclaving (121 °C, 15 min). o Pea milk was prepared by dissolving 10 % (w/w) pea flour in deionized water, and was autoclaved (121 °C, 15 min). o Sunflower seed milk was prepared by mixing 7 % (w/w) of 55 % sunflower protein with deionized water, then pasteurized under 95 °C for 6 h. Prior to microbial fermentation, the suspension was manually homogenized.
2. Preculture preparation o Bacillus amyloliquefaciens NCC 156:
Preculture condition (Pl, from glycerol stock):
Media: Modified tryptic soy broth, shaker (30 degree, 80 % humidity, 230 rpm), overnight
Preculture condition (P2, 2 % from pl):
Media: Modified tryptic soy broth, shaker (30 degree, 80 % humidity, 230 rpm), around 8 h
Modified tryptic soy broth containing 17.0 g of tryptone (Becton Dickinson), 5.0 g of NaCI, 3.0 g of soytone (Becton Dickinson), 2.5 g of K2HPO4, and 1.0 mL of 30% silicone antifoam (Sigma-Aldrich) per litre. o Lacticaseibacillus paracasei N CC 2511 :
Preculture condition (Pl, from glycerol stock):
MRS(De Man, Rogosa and Sharpe broth), incubator (30 degree, no humidity control, no shaking) overnight
Preculture condition (P2, 2 % from pl):
MRS, incubator (30 degree, no humidity control, no shaking) overnight o Propionibacterium freudenreichii NCC 1177:
Preculture condition (Pl, from glycerol stock):
Medium 91, anaerobic jar in incubator (30 degree, no humidity control, no shaking), 48 h
Preculture condition (P2, 2 % from pl):
Medium 91, anaerobic jar in incubator (30 degree, no humidity control, no shaking), overnight
Composition of Medium 91
Casein peptone, tryptic digest 10.0 g
Yeast extract 5.0 g
Na-lactate 10.0 g
Distilled water 1000.0 ml
Adjust pH to 7.0 - 7.2.
3. Main culture fermentation
Fermentation stage 1) Anaerobic jar in incubator (30 degree, no humidity control, no shaking), 48 h;
Fermentation stage 2) shaker (30 degree, 80 % humidity, 130 rpm), 24 h
4. Analysis of Vitamin B12
Vitamin B12 was analysed following AOAC 2014.02 with small modifications. Briefly, 6 mL of sample were mixed with an extraction buffer (2.5 mL) and cyanide solution (0.1 mL). The solutions were heated at 107 5C for 20 min. After making up to 10 mL with water, the extracts were centrifuged, and 8.5 mL of supernatant was subjected to automated immunoaffinity clean-up on a Gilson GX-271 ASPEC system. The final eluate was evaporated to dryness and reconstituted in 0.2 mL of water. Analysis was performed by UHPLC-UV, the chromatographic conditions were modified to allow separation of cyanocobalamin and tentatively identified pseudocobalamin. Separation
took place on a Waters Atlantis T3 column, 3.0 pm, 4.6 x 150 mm on isocratic mode. Mobile phase was methanol-water (25:75), flow rate 0.3 mL/min.
Example 2
Fermentation condition optimization
1. Preculture preparation of strains are the same as example 1. Precultures were grown in the respective medium under anaerobic or aerobic conditions in flasks.
2. Main fermentation
The main fermentation was conducted in the 1.8L DASGIP Bioreactor system (DASGIP AG, Germany). The plant milk was transferred into glass bioreactor, incubated on the bench top control system for a minimum of 6 hours with the vessel temperature at 30°C. During this time the DO probe was calibrated between 0% and 100%. After the calibration of the DO probe, precultures of Propionibacterium freudenreichii NCC 1177 and Bacillus amyloliquefaciens NCC 156 are inoculated into plant milk to start the fermentation. The pH was monitored using an on-line pH sensor. The culture temperature was controlled at 30°C. For the control of dissolved oxygen (DO) level, pO2 sensor (02 Sen lnPro6800G) was utilized the DO control.
All sensor signals were amplified and then transmitted to a personal computer interfaced with an A/D converter. For anaerobic fermentation (stage 1), filter-sterilized nitrogen gas was sparged through the fermentation broth to maintain an anaerobic environment, whereas air was used to maintain an aerobic condition (stage 2). The DO concentration (or ORP) was controlled by adjusting the volume of air pumped in and the rotation speed of the stirrer for the aerobic fermentation.
3. Sample analysis of vitamin B12
During and after fermentation, samples are taken for vitamin B12 analysis. The analysis method is the sample as in example 1.
(Original in Electronic Form)
(This sheet is not part of and does not count as a sheet of the international application)
(Original in Electronic Form)
Claims
1. Use of a combination of Bacillus amyloliquefaciens and Propionibacterium freudenreichii to enrich the vitamin B12 content in plant-based compositions through fermentation.
2. Use in accordance with claim 1, wherein the combination further comprises Lacticaseibacillus paracasei.
3. Use in accordance with one of the preceding claims, wherein Bacillus amyloliquefaciens is Bacillus amyloliquefaciens CNCM 1-5624;
Propionibacterium freudenreichii is selected from the group consisting of CNCM 1-5639, CNCM 1-5623, or combinations thereof; and Lacticaseibacillus paracasei is ATCC 25302.
4. Use in accordance with one of the preceding claims, wherein the Bacillus amyloliquefaciens is CNCM 1-5624 and the Propionibacterium freudenreichii is CNCM 1-5623.
5. Use in accordance with one of the preceding claims, wherein the Bacillus amyloliquefaciens is CNCM 1-5624, the Propionibacterium freudenreichii is CNCM 1-5623 and the Lacticaseibacillus paracasei is ATCC 25302.
6. Use in accordance with one of the preceding claims wherein the fermentation is carried out in two steps, wherein the first step of the fermentation is carried out in an anaerobic state and the second step of the fermentation is carried out in an aerobic state.
7. Use in accordance with one of the preceding claims, wherein the plant-based composition comprises plant proteins derived from sources selected from the
group consisting of the Fabacea family, the Leguminosa family, nuts, seeds, sasha inshi, sunflower, chia or combinations thereof. Use in accordance with one of the preceding claims, wherein the plant-based composition is a sunflower-based ingredient, for example sunflower seed milk. Use in accordance with one of the preceding claims wherein in the plant-based composition plant protein contributes at least 75 weight-% of the total protein content of the composition, at least 85 weight-% of the total protein content of the composition, or at least 95 weight-% of the total protein content of the composition. Use in accordance with one of the preceding claims, wherein the plant-based composition is selected from the group consisting of ingredients for plant-based products and plant-based milk product alternatives, for example, plant-based milk alternatives or plant-based cheese alternatives. Use in accordance with one of the preceding claims to further improve the smell of plant-based compositions. Use in accordance with claim 11, wherein the fermentation with Lacticaseibacillus paracasei and Bacillus natto improves the freshness and sweetness of the smell of the plant-based compositions. Use in accordance with one of the preceding claims to further improve the taste of plant-based compositions. Use in accordance with one of the preceding claims to enrich the l-lysine content in plant-based compositions.
Bacterial strain selected from the group consisting of Bacillus amyloliquefaciens
CNCM 1-5624, Propionibacterium freudenreichii CNCM 1-5623, and Propionibacterium freudenreichii CNCM 1-5639.
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
EP21155253 | 2021-02-04 | ||
EP21155253.4 | 2021-02-04 |
Publications (1)
Publication Number | Publication Date |
---|---|
WO2022167134A1 true WO2022167134A1 (en) | 2022-08-11 |
Family
ID=74553682
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/EP2021/086018 WO2022167134A1 (en) | 2021-02-04 | 2021-12-15 | Plant-based products fermented with b. amyloliquefaciens and p. freudenreichii having increased vit b12 content |
Country Status (1)
Country | Link |
---|---|
WO (1) | WO2022167134A1 (en) |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP1625794A1 (en) | 2004-08-10 | 2006-02-15 | Campina Nederland Holding B.V. | Increased vitamin content in fermented milk product |
WO2010091992A1 (en) | 2009-02-10 | 2010-08-19 | Nestec S.A. | Lactobacillus rhamnosus ncc4007, a probiotic mixture and weight control |
KR20190012336A (en) * | 2017-07-27 | 2019-02-11 | 주식회사 바이오리더스 | Method for Preparing Fermented Soy Bean Milk |
-
2021
- 2021-12-15 WO PCT/EP2021/086018 patent/WO2022167134A1/en active Application Filing
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP1625794A1 (en) | 2004-08-10 | 2006-02-15 | Campina Nederland Holding B.V. | Increased vitamin content in fermented milk product |
WO2010091992A1 (en) | 2009-02-10 | 2010-08-19 | Nestec S.A. | Lactobacillus rhamnosus ncc4007, a probiotic mixture and weight control |
KR20190012336A (en) * | 2017-07-27 | 2019-02-11 | 주식회사 바이오리더스 | Method for Preparing Fermented Soy Bean Milk |
Non-Patent Citations (11)
Title |
---|
"Functional Properties of Traditional Foods", 2016, SPRINGER US, Boston, MA, ISBN: 978-1-4899-7662-8, article YONGSMITH BUSABA ET AL: "Bioenrichment of Vitamin B12 in Fermented Foods", pages: 17 - 37, XP055893044, DOI: 10.1007/978-1-4899-7662-8_3 * |
ARICI M ET AL: "Some characteristics of Lactobacillus isolates from infant faeces", FOOD MICROBIOLOGY., vol. 21, no. 1, February 2004 (2004-02-01), GB, pages 19 - 24, XP055893651, ISSN: 0740-0020, DOI: 10.1016/S0740-0020(03)00044-3 * |
BHALLA, T. C.: "Production of Metabolites, Industrial enzymes, Amino acids, Organic acids, Antibiotics, Vitamins and Single Cell Proteins", 29 May 2007 (2007-05-29), XP055220387, Retrieved from the Internet <URL:http://nsdl.niscair.res.in/jspui/bitstream/123456789/129/1/Metabolites.pdf> [retrieved on 20151013] * |
CHAMLAGAIN BHAWANI ET AL: "Effect of the lower ligand precursors on vitamin B12 production by food-grade Propionibacteria", LWT- FOOD SCIENCE AND TECHNOLOGY, ACADEMIC PRESS, UNITED KINGDOM, vol. 72, 19 April 2016 (2016-04-19), pages 117 - 124, XP029561911, ISSN: 0023-6438, DOI: 10.1016/J.LWT.2016.04.023 * |
HAJFARAJOLLAH HAMIDREZA ET AL: "Vitamin B12 biosynthesis over waste frying sunflower oil as a cost effective and renewable substrate", JOURNAL OF FOOD SCIENCE AND TECHNOLOGY, vol. 52, no. 6, May 2014 (2014-05-01), India, pages 3273 - 3282, XP055821826, ISSN: 0022-1155, DOI: 10.1007/s13197-014-1383-x * |
JANICKI J ET AL: "The biosynthesis of vitamins of the B12 group in mixed cultures of bacteria", ACTA MICROBIOLOGICA POLONICA, PANSTWOWE WYDAWNICTWO NAUKOWE, WARSZAWA, PL, vol. 15, no. 4, 1966, pages 343 - 347, XP009528444, ISSN: 0137-1320 * |
PHOKASEM PATCHARIN ET AL: "Optimization of Mixed Bacillus Cultures as An Inoculant in Northern Thai Style Fermented Soybeans (Thua-nao) by Mixture Design", CHIANG MAI J. SCI, 2017, XP055893399, Retrieved from the Internet <URL:https://epg.science.cmu.ac.th/ejournal/dl.php?journal_id=8017> [retrieved on 20220218] * |
TANGYU MUZI ET AL: "Co-cultures of Propionibacterium freudenreichii and Bacillus amyloliquefaciens cooperatively upgrade sunflower seed milk to high levels of vitamin B12 and multiple co-benefits", MICROBIAL CELL FACTORIES, vol. 21, no. 1, 26 March 2022 (2022-03-26), XP055926148, DOI: 10.1186/s12934-022-01773-w * |
TANGYU MUZI ET AL: "Fermentation of plant-based milk alternatives for improved flavour and nutritional value", APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, SPRINGER BERLIN HEIDELBERG, BERLIN/HEIDELBERG, vol. 103, no. 23-24, 4 November 2019 (2019-11-04), pages 9263 - 9275, XP036948093, ISSN: 0175-7598, [retrieved on 20191104], DOI: 10.1007/S00253-019-10175-9 * |
TANGYU MUZI ET AL: "Genome-based selection and application of food-grade microbes for chickpea milk fermentation towards increased l-lysine content, elimination of indigestible sugars, and improved favour", MICROBIAL CELL FACTORIES, vol. 20, no. 1, 28 May 2021 (2021-05-28), XP055822761, DOI: 10.1186/s12934-021-01595-2 * |
XIE CHONG ET AL: "Co-fermentation of Propionibacterium freudenreichii and Lactobacillus brevis in Wheat Bran for in situ Production of Vitamin B12", FRONTIERS IN MICROBIOLOGY, vol. 10, 5 July 2019 (2019-07-05), XP055893376, DOI: 10.3389/fmicb.2019.01541 * |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Kök-Taş et al. | Effects of different fermentation parameters on quality characteristics of kefir | |
Patel et al. | Biosynthesis of vitamins and enzymes in fermented foods by lactic acid bacteria and related genera-A promising approach | |
Drgalic et al. | Growth and survival of probiotic bacteria in reconstituted whey | |
Kesenkaş et al. | Physicochemical, microbiological and sensory characteristics of soymilk kefir | |
Kumar Mishra et al. | Biofunctional attributes and storage study of soy milk fermented by Lactobacillus rhamnosus and Lactobacillus helveticus | |
JP7272822B2 (en) | Milk-containing food and drink containing lactic acid bacteria | |
El-Kholy et al. | Research Article Tallaga Cheese as a New Functional Dairy Product | |
EP0192986B2 (en) | Acid-resistant bifidus bacteria culture | |
WO2008003782A1 (en) | Method for preparing a dairy analogue | |
KR20210014744A (en) | Soy milk fermented substance | |
CN110741075A (en) | Method for producing mutant strains of streptococcus thermophilus | |
da Silva et al. | Quantification of lactic acid bacteria and bifidobacteria in goat milk based yoghurts with added water-soluble soy extract | |
Rakib et al. | Starter cultures used in the production of probiotic dairy products and their potential applications: A Review | |
Sani et al. | Traditional beverages in different countries: Milk-based beverages | |
Mohamed et al. | Production of vegetable yoghurt like from lupin milk | |
WO2022167135A1 (en) | Plant-based products having increased lysine content fermented with l. paracasei and optionally further bacillus amyloliquefaciens | |
KR101702206B1 (en) | Soybean pudding comprising Lactic acid producing bacterial fermented solution and preparation method thereof | |
WO2022167134A1 (en) | Plant-based products fermented with b. amyloliquefaciens and p. freudenreichii having increased vit b12 content | |
WO2012036575A1 (en) | Method of production of fermented, pro-healthy fruit beverages | |
Aita et al. | Utilization of sweet whey and ultra filtration-milk permeate in manufacture of yoghurt drink | |
KR20140131024A (en) | The manufacturing method of Yogurt drink using Wasong | |
Shah et al. | Recent advances in biosynthesis of vitamin and enzyme from food grade bacteria | |
RU2312506C2 (en) | Method for producing of fermented milk product | |
KR20160036450A (en) | Processes for preparing fermented Lentil bean | |
Pratama et al. | The potential of probiotic frozen yoghurt with the addition of fruits tamarillo to increase immunity |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
121 | Ep: the epo has been informed by wipo that ep was designated in this application |
Ref document number: 21824411 Country of ref document: EP Kind code of ref document: A1 |
|
NENP | Non-entry into the national phase |
Ref country code: DE |
|
122 | Ep: pct application non-entry in european phase |
Ref document number: 21824411 Country of ref document: EP Kind code of ref document: A1 |