HIGH TEMPERATURE RESISTANT PROBIOTICS FOR FOOD OR BEVERAGE AND METHOD OF MAKING THE SAME

1 . A heat and acid resistant probiotics particle having a size from 20 to 250 μm, comprising:

a synbiotic core comprising:

a seed layer comprising at least one polysaccharide; and

a probiotics layer coated on the seed layer;

an acid-resistant shell layer positioned over the synbiotic core, comprising one or more pH-responsive polymers; and

a heat-resistant bilayer shell positioned over the acid-resistant shell layer, comprising:

an inner shell layer comprising a heat-resistant isoprenoid-stabilized phospholipid liposome; and

an outer layer comprising a heat-resistant disaccharide or polysaccharide.

2 . The heat and acid resistant probiotics particle of claim 1 , wherein the at least one polysaccharide of the seed layer is selected from sucrose, inulin, starch and/or cellulose.

3 . The heat and acid resistant probiotics particle of claim 1 , wherein the probiotics in the probiotics layer are selected from one or more of Bifidobacterium, Lactobacillus, Lactococcus, Leuconostoc, Streptococcus, Enterococcus, Staphylococcus, Saccharomyces and Kluyveromyces.

4 . The heat and acid resistant probiotics particle of claim 1 , wherein the probiotics layer further comprises:

one or more of polysaccharides selected from fructo-oligosaccharides, galacto-oligosaccharides, inulin and/or pectin;

one or more of binder protein selected from whey protein, soy protein, chickpea protein, rice protein, pea protein, egg protein, casein, milk protein, zein and/or bovine serum albumin; and/or

one or more of heat-sensitive vitamins selected from vitamin C, vitamin B1 and/or vitamin E.

5 . The heat and acid resistant probiotics particle of claim 1 , wherein the acid-resistant shell is a pH-responsive polymer selected from one or more of methacrylic acid-methyl methacrylate copolymer, methacrylic acid-ethyl acrylate copolymer, shellac, alginate and/or pectin.

6 . The heat and acid resistant probiotics particle of claim 1 , wherein the isoprenoid-stabilized phospholipid of the inner shell layer of the heat-resistant bilayer shell is formed by proliposomes comprising:

one or more of phospholipids selected from phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, dipalmitoyl phosphatidylcholine, dipalmitoyl phosphatidylethanolamine, dipalmitoyl phosphatidylinositol, dimyristoyl phosphatidylcholine, dimyristoyl phosphatidylethanolamine and/or dimyristoyl phosphatidylinositol;

one or more of isoprenoids selected from β-carotene, cholesterol and/or lycopene; and

one or more of water-soluble carriers selected from maltodextrin, sorbitol, mannitol, maltitol and/or xylitol.

7 . The heat and acid resistant probiotics particle of claim 1 , wherein the heat-resistant disaccharide or polysaccharide of outer shell layer of the bilayer shell is selected from one or more of sucrose, lactose, maltose, trehalose, cellobiose or chitobiose.

8 . The heat and acid resistant probiotics particle of claim 7 , wherein the outer shell layer of the bilayer shell further comprises a mineral selected from talc, kaolin, zinc oxide, titanium oxide, silicon oxide, or any combinations thereof.

9 . The heat and acid resistant probiotics microsphere of claim 1 , wherein the weight percentage of the microsphere comprises 50-79.2% seed, 0.01-0.1% live probiotics, 1.6-3% protein, 2-6.3% polymer, 4-7.9% liposome, equal to or less than 1.6% polysaccharides, equal to or less than 3.2% disaccharides, and equal to or less than 0.3% minerals.

10 . The heat and acid resistant probiotics particle of claim 1 , further comprising an additional water barrier coating layer coupled to the outer layer including shellac and dimethylaminoethyl methacrylate-copolymer.

11 . The heat and acid resistant probiotics particle of claim 1 , wherein the particle resists a temperature of up to 90° C. for a period of up to 15 minutes without releasing the probiotics core.

12 . A method for preparing a probiotics particle resistant to high temperature and gastrointestinal digestive challenges, the method comprising:

preparing a seed with size between 10-125 μm and density between 0.65 to 0.75 g/cm 3 by sieving;

preparing a homogenous live probiotics solution with prebiotics by stir-mixing;

preparing an acid resistant pH-responsive polymer by heating and stir-mixing;

preparing an isoprenoid-stabilized phospholipid liposome by stir-mixing of proliposomes;

preparing a heat resistant outer layer comprising a disaccharide by stir-mixing;

preparing the high temperature resistant probiotics particle with the seed coated with a prebiotic and a probiotic layer, followed by acid resistant layer and heat resistant bilayers by fluidized-bed coating.

13 . The method of claim 12 , wherein the probiotics particle further comprises a seed core, a probiotic with binder coating on seed surface, an acid resistant protective layer, and a plurality of heat resistant protective layers.

14 . The method of claim 12 , wherein the probiotics particle has a particle encapsulation efficiency of more than 95%.

15 . The method of claim 12 , wherein the probiotics particle has a viable count of 10 8 -10 10 CFU/g.

16 . The method of claim 12 , wherein the probiotics particle has a size of 20-250 μm.

17 . The method of claim 12 , further comprising infusing the probiotics particle in a food product, a food additive, and liquid beverages.

18 . The method of claim 17 , wherein the liquid beverages are subjected to thermal treatment including pasteurization.

19 . The method of claim 17 , wherein the food product further comprises dry food that are subjected to thermal treatment including full baking production process.