Composition and method for improving stability and extending shelf life of probiotic bacteria and food products thereof

Abstract

A composition comprising probiotic bacteria, the composition comprising: (a) a core composition containing the probiotic bacteria and a stabilizer, wherein the total amount of probiotics in the mixture is from about 10% to about 90% by weight of the core composition; (b) an innermost coating layer, layered on said core composition, comprising at least one hydrophobic solid fat or fatty acid having a melting point lower than 60° C.; (c) an intermediate coating layer layered on said innermost coating layer, which when present in an aqueous solution in the amount of 0.1% weight/weight over the weight of the solution, has a surface tension lower than 60 mN/m, when measured at 25° C.; and (d) an outer coating layer, layered on said intermediate coating layer; wherein the composition is in the form of particles; food products containing the composition and methods of preparation thereof.

1 . A composition comprising probiotic bacteria, the composition comprising: (a) a core composition containing the probiotic bacteria and a stabilizer, wherein the total amount of probiotics in the mixture is from about 10% to about 90% by weight of the core composition; (b) an innermost coating layer, layered on said core composition, comprising at least one hydrophobic solid fat, fatty acid or a wax having a melting point lower than 60° C., or a combination thereof; (c) an intermediate coating layer layered on said innermost coating layer, which when present in an aqueous solution in the amount of 0.1% weight/weight over the weight of the solution, has a surface tension lower than 60 mN/m, when measured at 25° C.; and (d) an outer coating layer, layered on said intermediate coating layer; wherein the composition is in the form of particles. 
     
     
         2 . The composition of  claim 1 , wherein said stabilizer comprises an oxygen scavenger. 
     
     
         3 . The composition of  claim 2 , wherein said oxygen scavenger comprises one or more of L-cysteine hydrochloride, L-cysteine base, 4,4 (2,3 dimethyl tetramethylene dipyrocatechol), tocopherol-rich extract (natural vitamin E), α-tocopherol (synthetic Vitamin E), β-tocopherol, γ-tocopherol, δ-tocopherol, butylhydroxinon, butyl hydroxyanisole (BHA), butyl hydroxytoluene (BHT), propyl gallate, octyl gallate, dodecyl gallate, tertiary butylhydroquinone (TBHQ), fumaric acid, malic acid, ascorbic acid (Vitamin C), sodium ascorbate, calcium ascorbate, potassium ascorbate, ascorbyl palmitate, or ascorbyl stearate. 
     
     
         4 . The composition of  claim 1 , wherein said stabilizer comprises a material selected from the group consisting of dipotassium edetate, disodium edetate, edetate calcium disodium, edetic acid, fumaric acid, malic acid, maltol, sodium edetate, and trisodium edetate. 
     
     
         5 . The composition of  claim 1 , wherein said core composition further comprises at least one sugar compound. 
     
     
         6 . The composition of  claim 5 , wherein said at least one sugar compound comprises one or more of a monosaccharide, a disaccharide, a trisaccharide, a tetrasaccharide, a higher oligosaccharide, or a polysaccharide, or a combination thereof. 
     
     
         7 . The composition of  claim 6 , wherein said monosaccharide comprises one or more of a triose, a tetrose, a pentose, a hexose, a heptose, an octose, or a nonose; or any deoxy forms thereof; or a combination thereof. 
     
     
         8 . The composition of  claim 7 , wherein said monosaccharide comprises one or more of ketotriose (dihydroxyacetone), aldotriose (glyceraldehyde), ketotetrose (erythrulose), erythrose, threose, ribulose, xylulose, ribose, arabinose, xylose, lyxose, deoxyribose, psicose, fructose, sorbose, tagatose, allose, altrose, glucose, mannose, gulose, idose, galactose, talose, fucose, fuculose, rhamnose, sedoheptulose, or neuraminic acid, or a combination thereof. 
     
     
         9 . The composition of  claim 1 , wherein said disaccharide comprises one or more of sucrose, lactose, maltose, trehalose, turanose, or cellobiose, or a combination thereof. 
     
     
         10 . The composition of  claim 1 , wherein said trisaccharide comprises one or more of raffinose, melezitose or maltotriose, or a combination thereof. 
     
     
         11 . The composition of  claim 6 , wherein said tetrasaccharide comprises one or more of acarbose or stachyose, or a combination thereof. 
     
     
         12 . The composition of  claim 6 , wherein said higher oligosaccharide comprises one or more of fructooligosaccharide (FOS), galactooligosaccharides (GOS) or mannan-oligosaccharides (MOS), or a combination thereof. 
     
     
         13 . The composition of  claim 6 , wherein said polysaccharide comprises one or more of glucose-based polysaccharides/glucan cellulose, dextrin, dextran, beta-glucan, maltodextrin, fructose-based polysaccharides/fructan, levan beta 2-6, mannose-based polysaccharides, galactose-based polysaccharides, N-acetylglucosamine-based polysaccharides, or a gum, or a combination thereof. 
     
     
         14 . The composition of  claim 13 , wherein said polysaccharide comprises one or more of amylose, amylopectin, zymosan, lentinan, sizofiran, inulin, mannan, galactan, chitin, arabic gum, or gum acacia, or a combination thereof. 
     
     
         15 . The composition of  claim 1 , wherein said core further comprises one or more of a filler, a surfactant or binder, or a combination thereof. 
     
     
         16 . The composition of  claim 15 , wherein said filler comprises one or more of microcrystalline cellulose, a sugar; dicalcium phosphate; sugar alcohols, hydrogenated starch hydrolysates; corn starch; and potato starch; and/or a mixture thereof. 
     
     
         17 . The composition of  claim 16 , wherein said filler comprises one or more of lactose, glucose, galactose, fructose, sucrose, sorbitol, manitol, mantitol, lactitol, xylitol, isomalt, or erythritol, or a combination thereof. 
     
     
         18 . The composition of  claim 17 , wherein said filler is lactose. 
     
     
         19 . The composition of  claim 15 , wherein said binder comprises one or more of Povidone (PVP: polyvinyl pyrrolidone), Copovidone (copolymer of vinyl pyrrolidone and vinyl acetate), polyvinyl alcohol, low molecular weight HPC (hydroxypropyl cellulose), low molecular weight HPMC (hydroxypropyl methylcellulose), low molecular weight hydroxymethyl cellulose (MC), low molecular weight sodium carboxy methyl cellulose, low molecular weight hydroxyethylcellulose, low molecular weight hydroxymethylcellulose, cellulose acetate, gelatin, hydrolyzed gelatin, polyethylene oxide, acacia, dextrin, starch, water soluble polyacrylates and/or polymethacrylates, low molecular weight ethylcellulose, hydrophobic solid fat or fatty acid or a wax or a polyethylene glycol having a melting point lower than 60° C. or a mixture thereof. 
     
     
         20 . The composition of  claim 19 , wherein said binder is low molecular weight HPMC. 
     
     
         21 . The composition of  claim 15 , wherein said surfactant comprises tween 80 (polysorbate 80, Polyoxyethylene (20) sorbitan monooleate), tween 20 (polysorbate 20, Polyoxyethylene (20) sorbitan monolaurate), tween 85 (Polyoxyethylene sorbitan trioleate) glycereth-2-cocoate (Levenol® C-421), glycereth-6-cocoate (Levenol® F-200), glycereth-7-cocoate (Levenol® C-301), glycereth-17-cocoate (Levenol® C-201) or a mixture thereof. 
     
     
         22 . The composition of  claim 1 , wherein said probiotic bacteria comprises one or more of  Bacillus coagulans  GBI-30, 6086 , Bacillus subtilis  var  natt, Bifidobacterium  LAFTI® B94 , Bifidobacterium  sp LAFTI B94 , Bifidobacterium bifidum, Bifidobacterium bifidum  rosell-71 , Bifidobacterium breve, Bifidobacterium  breve Rosell-70 , Bifidobacterium infantis, Bifidobacterium lactis, Bifidobacterium longum, Bifidobacterium longum  Rosell-175 , Bifidobacterium  anima/is,  Bifidobacterium animalis  subsp.  lactis  BB-12 , Bifidobacterium animalis  subsp.  lactis  HN019 , Bifidobacterium infantis  35624 , Escherichia coli  M-17 , Escherichia coli  Nissle 1917 , Lactobacillus acidophilus, Lactobacillus acidophilus  LAFTI® L10 , Lactobacillus acidophilus  LAFTI L10 , Lactobacillus casei  LAFTI® L26 , Lactobacillus casei  LAFTI L26 , Lactobacillus brevis, Lactobacillus bulgaricus, Lactobacillus casei, Lactobacillus gasseri, Lactobacillus paracasei, Lactobacillus plantarum, Lactobacillus reuteri  ATTC 55730 ( Lactobacillus reuteri  SD2112),  Lactobacillus rhamnosus, Lactobacillus salivarius, Lactobacillus delbrueckii, Lactobacillus fermentum, Lactococcus lactis, Lactococcus lactis  subsp,  Lactococcus lactis  Rosell-1058 , Lactobacillus paracasei  St11 (or NCC2461 ] Lactobacillus fortis  Nestlé,  Lactobacillus johnsonii  La1 (= Lactobacillus  LC1 , Lactobacillus johnsonii  NCC533) Nestlé,  Lactobacillus rhamnosus  Rosell-11 , Lactobacillus acidophilus  Rosell-52 , Streptococcus thermophilus, Diacetylactis, Saccharomyces cerevisiae , and a mixture thereof. 
     
     
         23 . The composition of  claim 1 , wherein said innermost coating layer comprises at least one hydrophobic solid fat or fatty acid having a melting point lower than 50° C. 
     
     
         24 . The composition of  claim 23 , wherein said melting point is higher than 25° C. 
     
     
         25 . The composition of  claim 24 , wherein said melting point is lower than 45° C. and higher than 30° C. 
     
     
         26 . The composition of  claim 25 , wherein said melting point is lower than 40° C. and higher than 35° C. 
     
     
         27 . The composition of  claim 23 , wherein said innermost coating layer comprises fatty acids, fatty acid esters, fatty acid triesters; aluminum, sodium, potassium and magnesium salts of fatty acids; fatty alcohols, phospholipids, solid fats, waxes, and a combination thereof. 
     
     
         28 . The composition of  claim 27 , wherein the innermost coating layer comprises a solid fat comprising one or more of lauric acid, hydrogenated coconut oil, cacao butter or a combination thereof. 
     
     
         29 . The composition of  claim 1 , wherein said innermost coating layer forms a stable hydrophobic matrix which embeds the core composition within and/or forms a film around the probiotic core composition. 
     
     
         30 . The composition of  claim 29 , wherein said core comprises a plurality of cores and said plurality of cores is embedded in said stable hydrophobic matrix. 
     
     
         31 . The composition of  claim 1 , wherein said intermediate coating layer, when present in an aqueous solution in the amount of 0.1% weight/weight over the weight of the solution, has a surface tension lower than 50 mN/m when measured at 25° C. 
     
     
         32 . The composition of  claim 31 , wherein said surface tension is lower than 45 mN/m. 
     
     
         33 . The composition of  claim 32 , wherein said intermediate coating layer comprises a polymer, comprising one or more of Hydroxypropylmethylcellulose (HPMC), Hydroxypropylethylcellulose (HPEC), Hydroxypropylcellulose (HPC), hydroxypropylethylcellulose (HPEC), hydroxymethylpropylcellulose (HMPC), ethylhydroxyethylcellulose (EHEC) (Ethulose), hydroxyethylmethylcellulose (HEMC), hydroxymethylethylcellulose (HMEC), propylhydroxyethylcellulose (PHEC), methylhydroxyethylcellulose (M H EC), hydrophobically modified hydroxyethylcellulose (NEXTON), carboxymethyl hydroxyethylcellulose (CMHEC), Methylcellulose, Ethylcellulose, water soluble vinyl acetate copolymers, gums, polysaccharides such as alginic acid and alginates such as ammonia alginate, sodium alginate, potassium alginate, acid phthalate of carbohydrates, amylose acetate phthalate, cellulose acetate phthalate (CAP), cellulose ester phthalates, cellulose ether phthalates, hydroxypropylcellulose phthalate (HPCP), hydroxypropylethylcellulose phthalate (HPECP), hydroxyproplymethylcellulose phthalate (HPMCP), hydroxyproplymethylcellulose acetate succinate (HPMCAS), methylcellulose phthalate (MCP), polyvinyl acetate phthalate (PVAcP), polyvinyl acetate hydrogen phthalate, sodium CAP, starch acid phthalate, cellulose acetate trimellitate (CAT), styrene-maleic acid dibutyl phthalate copolymer, styrene-maleic acid/polyvinylacetate phthalate copolymer, styrene and maleic acid copolymers, or polyacrylic acid derivatives or a combination thereof. 
     
     
         34 . The composition of  claim 33  wherein said polyacrylic acid derivatives comprise one or more of acrylic acid and acrylic ester copolymers, polymethacrylic acid and esters thereof, polyacrylic and methacrylic acid copolymers, shellac, and vinyl acetate and crotonic acid copolymers. 
     
     
         35 . The composition of  claim 34 , wherein said polymer comprises one or more of shellac, phthalate derivatives, CAT, HPMCAS, polyacrylic acid derivatives, copolymers comprising acrylic acid and at least one acrylic acid ester, Eudragit™ S (poly(methacrylic acid, methyl methacrylate)1:2); Eudragit L100™ S (poly(methacrylic acid, methyl methacrylate)1:1); Eudragit L30D™, (poly(methacrylic acid, ethyl acrylate)1:1); and (Eudragit L100-55) (poly(methacrylic acid, ethyl acrylate)1:1) (Eudragit™ L is an anionic polymer synthesized from methacrylic acid and methacrylic acid methyl ester), polymethyl methacrylate blended with acrylic acid and acrylic ester copolymers, alginic acid, ammonia alginate, sodium, potassium, magnesium or calcium alginate, vinyl acetate copolymers, polyvinyl acetate 30D (30% dispersion in water), a neutral methacrylic ester comprising poly(dimethylaminoethylacrylate) (“Eudragit E™), a copolymer of methylmethacrylate and ethylacrylate with trimethylammonioethyl methacrylate chloride, a copolymer of methylmethacrylate and ethylacrylate, Zein, shellac, gums, or polysaccharides, or a combination thereof. 
     
     
         36 . The composition of  claim 1 , wherein said outer coating layer comprises a polymer having an oxygen transmission rate of less than 1000 cc/m 2 /24 hr, measured at standard test conditions. 
     
     
         37 . The composition of  claim 36 , wherein said polymer has an oxygen transmission rate less than 500 cc/m 2 /24 hr. 
     
     
         38 . The composition of  claim 37 , wherein said oxygen transmission rate is less than 100 cc/m 2 /24 hr. 
     
     
         39 . The composition of  claim 1 , wherein said polymer of said outer coating layer has a water vapor transmission rate of less than 400 g/m 2 /day. 
     
     
         40 . The composition of  claim 39 , wherein said water vapor transmission rate is less than 350 g/m 2 /day. 
     
     
         41 - 61 . (canceled)