CN103766486A - Milk beverage containing bifidobacteria embedded beads and preparation method thereof - Google Patents
Milk beverage containing bifidobacteria embedded beads and preparation method thereof Download PDFInfo
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- CN103766486A CN103766486A CN201410006248.9A CN201410006248A CN103766486A CN 103766486 A CN103766486 A CN 103766486A CN 201410006248 A CN201410006248 A CN 201410006248A CN 103766486 A CN103766486 A CN 103766486A
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- 241000186000 Bifidobacterium Species 0.000 title claims abstract description 178
- 235000020124 milk-based beverage Nutrition 0.000 title claims abstract description 89
- 238000002360 preparation method Methods 0.000 title claims abstract description 67
- 239000011324 bead Substances 0.000 title abstract 5
- 238000003860 storage Methods 0.000 claims abstract description 22
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 79
- 239000002054 inoculum Substances 0.000 claims description 63
- 239000000243 solution Substances 0.000 claims description 59
- 230000001954 sterilising effect Effects 0.000 claims description 59
- 238000004659 sterilization and disinfection Methods 0.000 claims description 51
- 235000013336 milk Nutrition 0.000 claims description 50
- 239000008267 milk Substances 0.000 claims description 50
- 210000004080 milk Anatomy 0.000 claims description 50
- 239000000463 material Substances 0.000 claims description 43
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 claims description 42
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 claims description 42
- 239000003223 protective agent Substances 0.000 claims description 40
- 239000002994 raw material Substances 0.000 claims description 35
- 239000002253 acid Substances 0.000 claims description 31
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 30
- 229930006000 Sucrose Natural products 0.000 claims description 30
- 239000003708 ampul Substances 0.000 claims description 30
- 239000005720 sucrose Substances 0.000 claims description 30
- 238000000855 fermentation Methods 0.000 claims description 26
- 230000004151 fermentation Effects 0.000 claims description 26
- 241001474374 Blennius Species 0.000 claims description 25
- 108010010803 Gelatin Proteins 0.000 claims description 25
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 claims description 25
- 229920000159 gelatin Polymers 0.000 claims description 25
- 239000008273 gelatin Substances 0.000 claims description 25
- 235000019322 gelatine Nutrition 0.000 claims description 25
- 235000011852 gelatine desserts Nutrition 0.000 claims description 25
- 239000011734 sodium Substances 0.000 claims description 25
- 229910052708 sodium Inorganic materials 0.000 claims description 25
- 239000012153 distilled water Substances 0.000 claims description 24
- 238000009630 liquid culture Methods 0.000 claims description 24
- 239000001963 growth medium Substances 0.000 claims description 23
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical group [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims description 22
- 238000002156 mixing Methods 0.000 claims description 21
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 18
- 238000010899 nucleation Methods 0.000 claims description 18
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 claims description 16
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims description 16
- 239000003795 chemical substances by application Substances 0.000 claims description 16
- 229920002134 Carboxymethyl cellulose Polymers 0.000 claims description 14
- 239000001768 carboxy methyl cellulose Substances 0.000 claims description 14
- 235000010948 carboxy methyl cellulose Nutrition 0.000 claims description 14
- 239000008112 carboxymethyl-cellulose Substances 0.000 claims description 14
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- 239000000203 mixture Substances 0.000 claims description 14
- 239000001814 pectin Substances 0.000 claims description 14
- 235000010987 pectin Nutrition 0.000 claims description 14
- 229920001277 pectin Polymers 0.000 claims description 14
- 238000012360 testing method Methods 0.000 claims description 14
- 235000008924 yoghurt drink Nutrition 0.000 claims description 13
- 235000020183 skimmed milk Nutrition 0.000 claims description 12
- 238000003756 stirring Methods 0.000 claims description 12
- 235000020185 raw untreated milk Nutrition 0.000 claims description 11
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- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 claims description 10
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 10
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 9
- 239000012530 fluid Substances 0.000 claims description 9
- 238000005057 refrigeration Methods 0.000 claims description 9
- 241001608472 Bifidobacterium longum Species 0.000 claims description 8
- 235000010323 ascorbic acid Nutrition 0.000 claims description 8
- 229960005070 ascorbic acid Drugs 0.000 claims description 8
- 239000011668 ascorbic acid Substances 0.000 claims description 8
- 239000001110 calcium chloride Substances 0.000 claims description 8
- 229910001628 calcium chloride Inorganic materials 0.000 claims description 8
- 235000011187 glycerol Nutrition 0.000 claims description 8
- 238000005406 washing Methods 0.000 claims description 8
- 238000000034 method Methods 0.000 claims description 7
- 235000015097 nutrients Nutrition 0.000 claims description 7
- 241000186018 Bifidobacterium adolescentis Species 0.000 claims description 6
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- 229940009291 bifidobacterium longum Drugs 0.000 claims description 6
- 238000005119 centrifugation Methods 0.000 claims description 6
- 238000001556 precipitation Methods 0.000 claims description 6
- 239000010902 straw Substances 0.000 claims description 6
- 239000006228 supernatant Substances 0.000 claims description 6
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 5
- 241000186660 Lactobacillus Species 0.000 claims description 5
- 239000001888 Peptone Substances 0.000 claims description 5
- 108010080698 Peptones Proteins 0.000 claims description 5
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical class [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 claims description 5
- 235000015278 beef Nutrition 0.000 claims description 5
- 229910000019 calcium carbonate Inorganic materials 0.000 claims description 5
- 229940041514 candida albicans extract Drugs 0.000 claims description 5
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical class [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 5
- 239000000284 extract Substances 0.000 claims description 5
- 239000008103 glucose Substances 0.000 claims description 5
- 229940039696 lactobacillus Drugs 0.000 claims description 5
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 5
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 5
- 235000019319 peptone Nutrition 0.000 claims description 5
- 239000000843 powder Substances 0.000 claims description 5
- 230000008569 process Effects 0.000 claims description 5
- 239000012138 yeast extract Substances 0.000 claims description 5
- NWONKYPBYAMBJT-UHFFFAOYSA-L zinc sulfate Chemical compound [Zn+2].[O-]S([O-])(=O)=O NWONKYPBYAMBJT-UHFFFAOYSA-L 0.000 claims description 5
- 229910000368 zinc sulfate Inorganic materials 0.000 claims description 5
- 239000011686 zinc sulphate Substances 0.000 claims description 5
- 235000009529 zinc sulphate Nutrition 0.000 claims description 5
- 241001134770 Bifidobacterium animalis Species 0.000 claims description 3
- 241000186016 Bifidobacterium bifidum Species 0.000 claims description 3
- 241000186012 Bifidobacterium breve Species 0.000 claims description 3
- 241000186015 Bifidobacterium longum subsp. infantis Species 0.000 claims description 3
- 241000185999 Bifidobacterium longum subsp. longum Species 0.000 claims description 3
- 241001465754 Metazoa Species 0.000 claims description 3
- 229940118852 bifidobacterium animalis Drugs 0.000 claims description 3
- 229940002008 bifidobacterium bifidum Drugs 0.000 claims description 3
- -1 0.1-1 part Chemical compound 0.000 claims description 2
- 235000013960 Lactobacillus bulgaricus Nutrition 0.000 claims description 2
- 241000186672 Lactobacillus delbrueckii subsp. bulgaricus Species 0.000 claims description 2
- 241000194020 Streptococcus thermophilus Species 0.000 claims description 2
- 238000001816 cooling Methods 0.000 claims description 2
- YXVFQADLFFNVDS-UHFFFAOYSA-N diammonium citrate Chemical class [NH4+].[NH4+].[O-]C(=O)CC(O)(C(=O)O)CC([O-])=O YXVFQADLFFNVDS-UHFFFAOYSA-N 0.000 claims description 2
- AIUDWMLXCFRVDR-UHFFFAOYSA-N dimethyl 2-(3-ethyl-3-methylpentyl)propanedioate Chemical class CCC(C)(CC)CCC(C(=O)OC)C(=O)OC AIUDWMLXCFRVDR-UHFFFAOYSA-N 0.000 claims description 2
- 238000001914 filtration Methods 0.000 claims description 2
- 238000002844 melting Methods 0.000 claims description 2
- 230000008018 melting Effects 0.000 claims description 2
- 229940066779 peptones Drugs 0.000 claims description 2
- 239000012047 saturated solution Substances 0.000 claims description 2
- 235000011091 sodium acetates Nutrition 0.000 claims description 2
- 108010046845 tryptones Proteins 0.000 claims description 2
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- 210000001035 gastrointestinal tract Anatomy 0.000 abstract 1
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- 240000001624 Espostoa lanata Species 0.000 description 4
- 235000009161 Espostoa lanata Nutrition 0.000 description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 4
- 230000036541 health Effects 0.000 description 4
- 230000000968 intestinal effect Effects 0.000 description 4
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 3
- 230000001580 bacterial effect Effects 0.000 description 3
- KLOIYEQEVSIOOO-UHFFFAOYSA-N carbocromen Chemical compound CC1=C(CCN(CC)CC)C(=O)OC2=CC(OCC(=O)OCC)=CC=C21 KLOIYEQEVSIOOO-UHFFFAOYSA-N 0.000 description 3
- 230000015556 catabolic process Effects 0.000 description 3
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- 238000011081 inoculation Methods 0.000 description 3
- 239000011159 matrix material Substances 0.000 description 3
- 244000005700 microbiome Species 0.000 description 3
- 229910052760 oxygen Inorganic materials 0.000 description 3
- 239000001301 oxygen Substances 0.000 description 3
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 description 3
- 229920000053 polysorbate 80 Polymers 0.000 description 3
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- BHQCQFFYRZLCQQ-OELDTZBJSA-N cholic acid Chemical compound C([C@H]1C[C@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(O)=O)C)[C@@]2(C)[C@@H](O)C1 BHQCQFFYRZLCQQ-OELDTZBJSA-N 0.000 description 2
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Dairy Products (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The invention discloses a milk beverage containing bifidobacteria embedded beads and a preparation method of the milk beverage to solve the problems of low activity, short storage time and difficulty in storage of bifidobacteria in a bifidobacteria milk beverage product. The milk beverage contains the bifidobacteria embedded beads. The preparation method comprises the following two steps: firstly, preparing the bifidobacteria embedded beads; secondly preparing the milk beverage containing the bifidobacteria embedded beads. According to the milk beverage, the problems of low viable count, short storage time, difficulty in reaching the intestinal tract and the like of the bifidobacteria in the milk beverage are effectively solved, the viable count, the activity and the storage time of the bifidobacteria are effectively ensured, and the probiotic effect of the bifidobacteria on a human body is achieved. The preparation method is mild in preparation conditions, is simple and convenient, and is suitable for large-scale industrialization production.
Description
Technical field
The present invention relates to milk beverage of a kind of Bifidobacterium embedding pearl and preparation method thereof.
Background technology
Bifidobacterium is the normal physiological microorganism in human body, surely grows in enteron aisle, is the dominant microflora of enteron aisle, its quantity number and health closely related, be the class of generally acknowledging at present has facilitation representative beneficial bacterium to body health.The main Physiological Function of Bifidobacterium has: 1. biological barrier and Ant agonism; 2. strengthen immunity and antitumor action; 3. trophism; 4. suppress endotoxic generation; 5. the slow body aging in the court of a feudal ruler.With advancing age, the impact such as environmental pollution and antibiotic use, the quantity of Bifidobacterium in human body intestinal canal declines gradually, thereby causes microecological balance in enteron aisle to destroy, and causes the generation of various diseases aging.
Adopt exactly the oral edible way of external source to supplementing the most direct approach of Bifidobacterium in enteron aisle, increase the quantity of Bifidobacterium at human body intestinal canal by the oral dietary supplementation of external source, adjust human body intestinal canal microecological balance, improve immunity, build up health.Bifidobacterium has to nutritional requirement harshness, obligate anaerobic, to special biological characters such as the extraneous poor environment sensitivities such as oxygen, low pH, make Bifidobacterium product in production, storage and transportation, be difficult to keep high activity, Bifidobacterium is at the oral environment such as hydrochloric acid in gastric juice, cholate that cannot tolerate low pH value when edible, be easy to lose activity, be difficult to guarantee that a large amount of viable bacterias arrive enteron aisle and surely grow on intestinal mucosa.The quantity of Bifidobacterium and the active important indicator that is used as the product qualities such as functional food, health products or medicine that check contains Bifidobacterium thereof, Bifidobacterium product inactivation very easily in preservation and in oral edible process, keeping Bifidobacterium high viability is a great problem of this series products.Both at home and abroad all improve the resistivity of bacterial classification to acid and oxygen attempting screening by genetic modification or acidproof oxytolerant bacterial strain etc., in technology, make bacterium and oxygen and sour isolation by microencapsulation, to improve the survival rate of Bifidobacterium.
Summary of the invention
The object of this invention is to provide the milk beverage of a kind of Bifidobacterium embedding pearl, to solve, in Bifidobacterium milk beverage, Bifidobacterium vigor is low, storage life is short, difficult problem of storing.
Another object of the present invention is to provide the preparation method of the milk beverage of a kind of Bifidobacterium embedding pearl.
Technical solution of the present invention is as follows: a kind of milk beverage that contains Bifidobacterium embedding pearl, it contains Bifidobacterium embedding pearl, Bifidobacterium embedding pearl comprise the long subspecies of Bifidobacterium bifidobacterium longum (
bifidobacterium longum subsp. longum) CGMCC numbering 1.2186, bifidobacterium longum baby subspecies (
bifidobacterium longum subsp. infantis) CGMCC numbering 1.2202, bifidobacterium adolescentis (
bifidobacterium adolescentis) CGMCC numbering 1.2190, bifidobacterium breve (
bifidobacterium brevis) CGMCC numbering 1.2213, bifidobacterium (
bifidobacterium bifidum) CGMCC numbering 1.2212 or animal bifidobacteria (
bifidobacterium animalis) one in CGMCC numbering 1.1852, the number of active bifid bacteria in Bifidobacterium embedding pearl is not less than 1 × 10
9cFU/g.Above-mentioned Bifidobacterium is bought in Chinese microorganism strain administration committee common micro-organisms center, Pekinese (CGMCC).Preferably, Bifidobacterium embedding pearl is solid-state shape spherical in shape, and diameter is 3mm-12mm.
A preparation method for the milk beverage that contains Bifidobacterium embedding pearl, comprises the following steps:
(1) preparation of Bifidobacterium embedding pearl
A, actication of culture test tube are cultivated
Before actication of culture, at desinfection chamber, the Bifidobacterium ampoule bottle of preserving at subzero 18 ℃ of-4 ℃ of temperature is at room temperature placed to 2 h-3 h, then by ampoule bottle sterilization, open ampoule bottle, the culture medium of drawing 0.2 mL-0.5 mL with aseptic straw adds in ampoule bottle, by fungus in ampoule bottle, the inoculum concentration that by volume percentage is 2%-5% is inoculated in culture medium, and test tube is cultivated, under anaerobic, at the temperature of 30 ℃-42 ℃, cultivate after 20 h-48 h, stop cultivating; Activate continuously again 2 times with identical inoculum concentration, condition of culture, obtain three generations's liquid culture;
B, triangular flask first class inoculum are cultivated
Get test tube and cultivate three generations's liquid culture, the inoculum concentration that by volume percentage is 2%-5%, access triangular flask carries out first class inoculum Liquid Culture, under anaerobic, at the temperature of 30 ℃-42 ℃, cultivates after 20h-48h, stops cultivating;
C, seeding tank second class inoculum are cultivated
Get triangular flask first class inoculum liquid culture, the inoculum concentration that by volume percentage is 2%-5%, access seeding tank carries out second class inoculum Liquid Culture, under non-anaerobic condition, at the temperature of 30 ℃-42 ℃, cultivates after 20h-48h, stops cultivating;
D, fermentation tank culture
By the liquid culture of cultivating through seeding tank second class inoculum, the inoculum concentration that by volume percentage is 2%-5%, access fermentation tank, at the temperature of 30 ℃-42 ℃, cultivate, when zymotic fluid pH value is lower than 5.5 time, between NaOH solution adjusting zymotic fluid pH to 6.0-6.5, after cultivation 20 h-48 h, stop cultivating;
E, the centrifugal collection of thalline
It is 0 ℃-8 ℃ that centrifuge temperature is set, and centrifugal rotational speed is that 3000 r/min-9000 r/min, centrifugation time are 10 min-30 min, after centrifugal end, removes supernatant, and the Bifidobacterium thalline of collecting precipitation is for subsequent use;
F, interpolation protective agent
The thalline of centrifugal collection is suspended in the protective agent of centrifugal prefermentor nutrient solution original volume 1/100-1/10; At the temperature of subzero 20 ℃-10 ℃, store for subsequent use;
The embedding of G, Bifidobacterium
To be suspended in Bifidobacterium thalline in protective agent and mix with sterilizing embedded material solution, and after stirring, clamp-on curing agent, low rate mixing 5min-30min, filters, and with sterilized water washing, obtains Bifidobacterium embedding pearl;
(2) preparation of milk beverage
H, raw material milk pretreatment: raw material milk is preheated to the temperature of 65 ℃-70 ℃, opens mixer, sucrose is added in hot milk, while all melting, filter with 200 order filter cloths; Homogeneous after raw material milk preheating, homogenization pressure is 18Mpa-20Mpa; Then sterilization, sterilization temperature is 95 ℃, 5 minutes time, is cooled to 37 ℃-42 ℃;
I, prepare milk beverage: be divided into two kinds of preparation preparation shape milk beverage and yogurt drinks,
1) prepare Bifidobacterium embedding pearl blending type milk beverage: be 1-6:6-1 in pretreated feedstock milk and water ratio, carry out after proportioning, add sucrose, citric acid, edible pectin, carboxymethyl cellulose modulation, then homogeneous, sterilization, cooling, Bifidobacterium embedding pearl adds in milk beverage by the amount of 10-30%, then aseptic subpackaged;
2) prepare Bifidobacterium embedding pearl yogurt drink: after raw material milk pretreatment, the inoculum concentration that by volume percentage is 2%-5%, leavening is inoculated in pretreated milk, at the temperature of 37 ℃-43 ℃, cultivate, acidity reaches 70-80 clean Er Nieerdu, stop fermentation, then add water, milk beverage accounts for 40-50% and water accounts for 50-60%, add sucrose, citric acid, edible pectin, carboxymethyl cellulose, after homogeneous, the Bifidobacterium embedding pearl by mass percentage amount of 15-20% adds in milk beverage again, then aseptic subpackaged
J, refrigeration: will contain the milk beverage that has Bifidobacterium embedding pearl, be cooled to after 20 ℃-25 ℃, enter refrigerated storage temperature and be the freezer of 2 ℃-10, final finished.
Preferably; in described steps A, the composition of culture medium and weight proportion are: 3-15 parts of peptones; 1-10 parts of beef extracts; 3-10 parts of tryptones; 1-10 parts of yeast extract powders; 3-15 parts of glucose; 0.5-5 parts of dipotassium hydrogen phosphates, 0.5-5 parts of sodium acetates, 0.5-5 parts of diammonium hydrogen citrates; 0.1-0.5 part of white vitriol; 0.05-0.2 part of epsom salt, 0.1-1 part of Cys hydrochloride, 0.5-5 parts of FOSs; 0.5-2 parts, calcium carbonate, 1000 parts, 1-5 parts of Tween-80s and water; The pH value of culture medium is 6.0-7.5, after 0.05Mpa-0.14Mpa moist heat sterilization 15 min-30 min, uses.
Preferably, in described step F, protectant composition and weight proportion are: 5-30 parts of skim milk or skimmed milk powers, 0.5-5 parts of glycerine, 0.1-1 part, ascorbic acid, 100 parts, 3-10 parts of sucrose and water; By skim milk or skimmed milk power, glycerine and sucrose and water moist heat sterilization 15min-30min under 0.05 Mpa-0.14 Mpa pressure, ascorbic acid adopts filtration sterilization.
Preferably, in described step G, embedded material solution is formulated by the edible seaweed acid sodium of 2-10 weight portions and the water of edible gelatin and 90-98 weight portions, the weight proportion of edible seaweed acid sodium and edible gelatin is 1-5:1, preparation process is for first taking edible gelatin, add in distilled water, the temperature of distilled water is 75 ℃-95 ℃, be stirred to completely and dissolve, add again edible seaweed acid sodium, continuing to be stirred to becomes clear solution, moist heat sterilization 15 min-30min under 0.05 Mp-0.14Mpa pressure.
Preferably, in described step G, embedded material solution is that embedded material solution accounts for 50-90 parts, is suspended in Bifidobacterium thalline in protective agent and accounts for 10-50 parts with the weight proportion that is suspended in Bifidobacterium thalline in protective agent.
Preferably, in described step G, curing agent is calcium chloride solution, and process for preparation is, with distilled water, calcium chloride is mixed with to the saturated solution that concentration is 0.1mol/L, after 0.05Mpa-0.14Mpa moist heat sterilization 15min-30min, uses.
Preferably, milk beverage leavening in described step I is that number of viable is the mixture of having a liking for acid streptococci (Streptococcus thermophilus) CGMCC numbering 1.1864 and lactobacillus delbruockii subspecies bulgaricus (Lactobacillus delbrueckii subsp. bulgaricus) CGMCC numbering 1.1863 of 1:1, and bacterial classification is bought in Chinese microorganism strain administration committee common micro-organisms center, Pekinese (CGMCC).
Preferably, in described step I, the addition of sucrose, citric acid, edible pectin, carboxymethyl cellulose is, adds 8-10% sucrose by raw milk and water inventory, 0.1-0.2% citric acid, 0.1-0.3% edible pectin, 0.15-0.2% carboxymethyl cellulose.
It is bifidobacterium species that the present invention contains Bifidobacterium embedding pearl milk beverage, cultivate through fluid nutrient medium propagation, concentrating and separating, by natural high molecular substance edible seaweed acid sodium, gelatin embedding, then joins in milk beverage and the step such as refrigeration is made the milk beverage that contains Bifidobacterium embedding pearl.The milk beverage that contains Bifidobacterium embedding pearl has solved the problem that Bifidobacterium milk beverage goods low pH is unfavorable for maintaining for a long time the easy inactivation of Bifidobacterium in the number of active bifid bacteria amount problem and edible process.
Bifidobacterium embedding pearl has advantages of stomach juice-resistant, bile tolerance and enteric solubility, avoid the lethal effect of hydrochloric acid in gastric juice and the cholate Bifidobacterium to absorption human body, improve Bifidobacterium and surely grown the number of viable at human body intestinal canal, realize the prebiotic effect of Bifidobacterium to human body, guaranteed that probio determining in enteron aisle grow.The present invention makes Bifidobacterium embedding pearl milk beverage reasonably optimizing Bifidobacterium embedding pearl milk beverage technique by the Bifidobacterium embedding pearl preparing is joined in milk beverage.In milk beverage matrix, store survival rate after 28 days months for about 4 ℃ and can remain on more than 10%, storage at normal temperature after 28 days survival rate can remain on more than 1%.Bifidobacterium embedding pearl milk beverage preparation condition gentleness, method is easy, is convenient to industrialized production.
The present invention is Bifidobacterium milk beverage producing, store, transportation and while selling in a long time activity do not weaken, during for consumption, having enough number of viable provides guarantee, also provides assurance for a large amount of viable bacterias in oral edible process arrive enteron aisles.The milk beverage main feature that contains Bifidobacterium embedding pearl is: bifidobacteria viable bacteria content is high, product preservation term is longer, every batch of constant product quality, has improved Bifidobacterium milk beverage quality, the Bifidobacterium embedding pearl eating mouth feel resilient enough that adds, smooth good to eat.Bifidobacterium embedding pearl in milk beverage of the present invention, in the milk beverage matrix of acidity 75 clean Er Nieerdu, 2 ℃ of-8 ℃ of preservations 28 days, the number of active bifid bacteria is greater than 10
8cFU/g.In the clean Er Nieerdu milk beverage of acidity 75 matrix, room temperature (25 ℃ of left and right) storage 28 days, the number of active bifid bacteria is greater than 10
7cFU/g.
The specific embodiment
Following examples are used for illustrating the present invention, can make the present invention of those skilled in the art comprehend, are not used for limit value scope of the present invention.
Embodiment 1
(1) Bifidobacterium embedding pearl preparation
A, actication of culture test tube are cultivated
Culture medium consists of, peptone 15g, beef extract 10g, tryptone 3g, yeast extract powder 1g, glucose 15g, dipotassium hydrogen phosphate 5g, sodium acetate 0.5g, diammonium hydrogen citrate 0.5g, white vitriol 0.1 g, epsom salt 0.05g, Cys hydrochloride 0.1g, FOS 0.5g, calcium carbonate 0.5g, Tween-80 5g and water 1000g.The pH value of culture medium is 7.0, after 0.05MpaMpa moist heat sterilization 30 min, uses;
Before actication of culture, by long the bifidobacterium longum of 4 ℃ of preservations subspecies (
bifidobacterium longum subsp. longum) ampoule bottle at room temperature places 2h, then the cotton ball soaked in alcohol wipe surfaces sterilization by 75% concentration by ampoule bottle at desinfection chamber, open ampoule bottle, the culture medium of drawing 0.2 mL with aseptic straw adds in ampoule bottle, by fungus in ampoule bottle, be inoculated in culture medium by the inoculum concentration of 5% (percent by volume), test tube is cultivated, under anaerobic cultivate after 36h for 37.5 ℃, stop cultivating.Activate continuously again 2 times with identical inoculum concentration, condition of culture, obtain three generations's liquid culture;
B, triangular flask first class inoculum are cultivated
Third generation culture is carried out to first class inoculum cultivation by the inoculum concentration access triangular flask of 5% (percent by volume), under anaerobic cultivate after 24h for 37.5 ℃, stop cultivating;
C, seeding tank second class inoculum are cultivated
Get triangular flask first class inoculum liquid culture, by the inoculum concentration of 5% (percent by volume), access seeding tank carries out second class inoculum cultivation, under non-anaerobic condition, cultivates after 24h for 37.5 ℃, stops cultivating;
D, fermentation tank culture
Fermentation tank culture: by the inoculum concentration of 5% (percent by volume); access seeding tank second class inoculum liquid culture; under non-anaerobic condition, under 37.5 ℃ of conditions, cultivate, when pH regulates zymotic fluid pH to 6.0 with NaOH solution lower than 5.5 time, after 24h, stop cultivating;
E, the centrifugal collection of thalline
4 ℃ of centrifuging temperatures are set, centrifugal rotational speed 6000 r/min, centrifugation time 15 min; After centrifugal end, remove supernatant, the Bifidobacterium thalline of collecting precipitation is for subsequent use;
F, interpolation protective agent
Protective agent consists of: skim milk 15g, glycerine 2g, ascorbic acid 0.5g, sucrose 10g and water 100g; The thalline of centrifugal collection is suspended in the protective agent of centrifugal prefermentor nutrient solution original volume 1/100;
The embedding of G, Bifidobacterium
1) preparation of embedded material solution: embedded material solution is formulated by the edible seaweed acid sodium of 10 weight portions and the water of edible gelatin and 90 weight portions, edible seaweed acid sodium and edible gelatin are pressed the preparation of 2:1 part by weight, preparation process is for first taking edible gelatin, add in distilled water, the temperature of distilled water is 80 ℃, is stirred to completely and dissolves, then add edible seaweed acid sodium, continuing to be stirred to becomes clear solution, for subsequent use after moist heat sterilization 20min under 0.05 Mpa;
2) preparation of curing agent: calcium chloride is mixed with to 0.5mol/L concentration calcium chloride solution with distilled water, for subsequent use after 0.05 Mpa moist heat sterilization 20min;
3) embedding: the embedded material solution after sterilizing be suspended in protective agent in Bifidobacterium thalline mix, wherein embedded material solution with the weight proportion that is suspended in Bifidobacterium thalline in protective agent is, embedded material solution accounts for 50 parts, is suspended in Bifidobacterium thalline in protective agent and accounts for 50 parts; After stirring, clamp-on curing agent 0.5mol/L calcium chloride solution by extruder, form the Bifidobacterium embedding pearl of 12 mm diameters, low rate mixing 30min, filters, and with sterilized water washing Bifidobacterium embedding pearl 5 min, subzero 20 ℃ of low tempertaure storages are for subsequent use;
(2) prepare Bifidobacterium embedding pearl yogurt drink
H, raw material milk pretreatment: by 70 ℃ of raw milk raw material milk preheatings, open mixer, the sucrose of material quality 5% is added in hot milk, after all dissolving, filter with 200 order filter cloths.Homogeneous after raw material milk preheating, homogenization pressure is 18 Mpa, then sterilization, sterilization temperature is 95 ℃, 5 minutes time, is cooled to 42 ℃;
I, inoculation, fermentation: milk beverage leavening is for having a liking for acid streptococci and lactobacillus delbruockii subspecies bulgaricus is prepared according to 1:1, press 5%(percent by volume) inoculum concentration, milk beverage leavening is inoculated in after pretreatment in raw material milk, 43 ℃ of fermentations, milk beverage titratable acidity arrives 70 clean Er Nieerdu, stops fermentation.Stir breakdown of emulsion and be cooled to 25 ℃, be 1:1 in the ratio of milk beverage and water, add pure water, add 8% sucrose by milk beverage and water inventory, 0.1% citric acid, 0.1% edible pectin, 0.2% carboxymethyl cellulose, under homogenization pressure 20MPa, after homogeneous, Bifidobacterium embedding pearl by mass percentage 20% amount adds milk beverage, then aseptic subpackaged, will add Bifidobacterium embedding pearl yogurt drink;
J, refrigeration, by stopping the Bifidobacterium embedding pearl yogurt drink of fermentation, be cooled to after 25 ℃ of temperature, enters refrigerated storage temperature and be the freezer final finished of 4 ℃.
Embodiment 2:
(1) preparation of Bifidobacterium embedded block
A, actication of culture test tube are cultivated
Culture medium consists of, peptone 15g, beef extract 10g, tryptone 3g, yeast extract powder 1g, glucose 15g, dipotassium hydrogen phosphate 5g, sodium acetate 0.5g, diammonium hydrogen citrate 0.5g, white vitriol 0.1g, epsom salt 0.05g, Cys hydrochloride 0.1g, FOS 0.5g, calcium carbonate 0.5g, Tween-80 5g and water 1000g; The pH value of culture medium is 7.5, after 0.14Mpa moist heat sterilization 15 min, uses;
Before actication of culture, by the bifidobacterium longum baby subspecies of 4 ℃ of preservations (
bifidobacterium longum subsp. infantis) ampoule bottle at room temperature places 3h, then the cotton ball soaked in alcohol wipe surfaces sterilization by 75% concentration by ampoule bottle at desinfection chamber, open ampoule bottle, the culture medium of drawing 0.25mL with aseptic straw adds in ampoule bottle, by fungus in ampoule bottle, be inoculated in culture medium by the inoculum concentration of 3% (percent by volume), test tube is cultivated, under anaerobic cultivate after 20h for 42 ℃, stop cultivating.Activate continuously again 2 times with identical inoculum concentration, condition of culture, obtain three generations's liquid culture;
B, triangular flask first class inoculum are cultivated
Third generation culture is carried out to first class inoculum cultivation by the inoculum concentration access triangular flask of 3% (percent by volume), under anaerobic cultivate after 20 h for 42 ℃, stop cultivating;
C, seeding tank second class inoculum are cultivated
Get triangular flask first class inoculum liquid culture, by the inoculum concentration of 3% (percent by volume), access seeding tank carries out second class inoculum cultivation, under non-anaerobic condition, cultivates after 20h for 42 ℃, stops cultivating;
D, fermentation tank culture
Fermentation tank culture: by the inoculum concentration of 3% (percent by volume), access seeding tank second class inoculum liquid culture, under non-anaerobic condition, under 42 ℃ of conditions, cultivate, when pH regulates between zymotic fluid pH to 6.5 with NaOH solution lower than 5.5 time, after 20 h, stop cultivating.
E, the centrifugal collection of thalline
8 ℃ of centrifuging temperatures are set, centrifugal rotational speed 3000 r/min, centrifugation time 30 min; After centrifugal end, remove supernatant, the Bifidobacterium thalline of collecting precipitation is for subsequent use;
F, interpolation protective agent
Protective agent consists of, skimmed milk power 30g, glycerine 2g, ascorbic acid 0.1g, sucrose 3g and water 100g; The thalline of centrifugal collection is suspended in the protective agent of centrifugal prefermentor nutrient solution original volume 1/10;
The embedding of G, Bifidobacterium
1) preparation of embedded material solution: embedded material solution is formulated by the edible seaweed acid sodium of 2 weight portions and the water of edible gelatin and 98 weight portions, and edible seaweed acid sodium and edible gelatin are pressed the preparation of 5:1 part by weight,
Preparation process, for first taking edible gelatin, adds in distilled water, and the temperature of distilled water is 95 ℃, is stirred to completely and dissolves, then add edible seaweed acid sodium, and continuing to be stirred to becomes clear solution, for subsequent use after moist heat sterilization 30 min under 0.14 Mpa;
2) preparation of curing agent: calcium chloride is mixed with to 0.5mol/L concentration calcium chloride solution with distilled water, for subsequent use after 0.05 Mpa moist heat sterilization 20min;
3) embedding: the embedded material solution after sterilizing be suspended in protective agent in Bifidobacterium thalline mix, wherein embedded material solution with the weight proportion that is suspended in Bifidobacterium thalline in protective agent is, embedded material solution accounts for 90 parts, is suspended in Bifidobacterium thalline in protective agent and accounts for 10 parts; After stirring, clamp-on curing agent 0.5mol/L calcium chloride solution by extruder, form 12 mm wide, 12 mm are thick, Bifidobacterium embedding bar, low rate mixing 15min, filter, be cut into long 12 mm Bifidobacterium embedded blocks, with sterilized water washing Bifidobacterium embedded block 1 min, 10 ℃ of low tempertaure storages are for subsequent use;
(2) prepare Bifidobacterium embedded block blending type milk beverage
H, raw material milk pretreatment: by 65 ℃ of raw milk raw material milk preheatings, open mixer, the sucrose of material quality 5% is added in hot milk, after all dissolving, filter with 200 order filter cloths.Homogeneous after raw material milk preheating, homogenization pressure is 20Mpa, then sterilization, sterilization temperature is 95 ℃, 5 minutes time, is cooled to 37 ℃;
I, preparation: take raw milk as raw material, after raw material milk pretreatment, adding water, is 6:4 in pretreated feedstock milk and water ratio, carries out after proportioning, add 10% sucrose by raw milk and water inventory, 0.2% citric acid, 0.3% edible pectin, 0.2% carboxymethyl cellulose carries out the preparation of blending type milk beverage, under homogenization pressure 18MPa after homogeneous, 95 ℃ of sterilization temperatures, 5 minutes time, are cooled to 25 ℃.Bifidobacterium embedding pearl adds milk beverage by 30% amount, then aseptic subpackaged, will add Bifidobacterium embedding pearl blending type milk beverage;
J, refrigeration: be cooled to after 20 ℃ of temperature, enter 2 ℃ of freezers of refrigerated storage temperature, final finished.
Embodiment 3:
(1) preparation of Bifidobacterium embedding pearl
A, actication of culture test tube are cultivated
Culture medium consists of, peptone 15g, beef extract 10g, tryptone 3g, yeast extract powder 1g, glucose 15g, dipotassium hydrogen phosphate 5g, sodium acetate 0.5g, diammonium hydrogen citrate 0.5g, white vitriol 0.1 g, epsom salt 0.05g, Cys hydrochloride 0.1g, FOS 0.5g, calcium carbonate 0.5g, Tween-80 5g and water 1000g.The pH value of culture medium is 6.0, after 0.10Mpa moist heat sterilization 20min, uses;
Before actication of culture, by the bifidobacterium adolescentis of subzero 18 ℃ of preservations (
bifidobacterium adolescentis) ampoule bottle at room temperature places 2.5 h, then the cotton ball soaked in alcohol wipe surfaces sterilization by 75% concentration by ampoule bottle at desinfection chamber, open ampoule bottle, the culture medium of drawing 0.3 mL with aseptic straw adds in ampoule bottle, by fungus in ampoule bottle, be inoculated in culture medium by the inoculum concentration of 2% (percent by volume), test tube is cultivated, under anaerobic cultivate after 48 h for 30 ℃, stop cultivating.Activate continuously again 2 times with identical inoculum concentration, condition of culture, obtain three generations's liquid culture.
B, triangular flask first class inoculum are cultivated
Third generation culture is carried out to first class inoculum cultivation by the inoculum concentration access triangular flask of 2% (percent by volume), under anaerobic cultivate after 48h for 30 ℃, stop cultivating.
C, seeding tank second class inoculum are cultivated
Get triangular flask first class inoculum liquid culture, by the inoculum concentration of 2% (percent by volume), access seeding tank carries out second class inoculum cultivation, under non-anaerobic condition, cultivates after 48 h for 30 ℃, stops cultivating.
D, fermentation tank culture
Fermentation tank culture: by the inoculum concentration of 2% (percent by volume), access seeding tank second class inoculum liquid culture, under non-anaerobic condition, under 30 ℃ of conditions, cultivate, when pH regulates zymotic fluid pH to 6.2 with NaOH solution lower than 5.5 time, after 48 h, stop cultivating.
E, the centrifugal collection of thalline
0 ℃ of centrifuging temperature is set, centrifugal rotational speed 9000 r/min, centrifugation time 10 min; After centrifugal end, remove supernatant, the Bifidobacterium thalline of collecting precipitation is for subsequent use;
F, interpolation protective agent
Protective agent consists of, skim milk 10 g, glycerine 5 g, ascorbic acid 1 g, sucrose 5 g and water 100 g; The thalline of centrifugal collection is suspended in the protective agent of centrifugal prefermentor nutrient solution original volume 1/50;
The embedding of G, Bifidobacterium
1) preparation of embedded material solution: embedded material solution is formulated by the edible seaweed acid sodium of 2 weight portions and the water of edible gelatin and 98 weight portions, and edible seaweed acid sodium and edible gelatin are pressed the preparation of 1:1 part by weight,
Preparation process, for first taking edible gelatin, adds in distilled water, and the temperature of distilled water is 75 ℃, is stirred to completely and dissolves, then add edible seaweed acid sodium, and continuing to be stirred to becomes clear solution, for subsequent use after moist heat sterilization 15 min under 0.10 Mpa;
2) preparation of curing agent: calcium chloride is mixed with to 0.5mol/L concentration calcium chloride solution with distilled water, for subsequent use after 0.05 Mpa moist heat sterilization 20min;
3) embedding: the embedded material solution after sterilizing be suspended in protective agent in Bifidobacterium thalline mix, wherein embedded material solution with the weight proportion that is suspended in Bifidobacterium thalline in protective agent is, embedded material solution accounts for 60 parts, is suspended in Bifidobacterium thalline in protective agent and accounts for 40 parts; After stirring, clamp-on curing agent 0.5mol/L calcium chloride solution by extruder, form diameter 3 mm, Bifidobacterium embedding pearl, low rate mixing 5 min, filter, and with sterilized water washing Bifidobacterium embedding pearl 2min, 5 ℃ of low tempertaure storages are for subsequent use;
(2) prepare Bifidobacterium embedding pearl blending type milk beverage
H, raw material milk pretreatment: will the preheating of raw sheep dairy milk starting material milk 68 ℃, open mixer, the sucrose of material quality 5% is added in hot milk, after all dissolving, filter with 200 order filter cloths, homogeneous after raw material milk preheating, homogenization pressure is 19Mpa, then sterilization, and sterilization temperature is 95 ℃, 5 minutes time, be cooled to 40 ℃;
I, preparation: take raw milk as raw material, after raw material milk pretreatment, adding water, is 1:1 in pretreated feedstock milk and water ratio, carries out after proportioning, add 10% sucrose by raw milk and water inventory, 0.2% citric acid, 0.3% edible pectin, 0.2% carboxymethyl cellulose carries out the preparation of blending type milk beverage, under homogenization pressure 18MPa after homogeneous, 95 ℃ of sterilization temperatures, 5 minutes time, are cooled to 25 ℃.Bifidobacterium embedding pearl adds milk beverage by 30% amount, then aseptic subpackaged, will add Bifidobacterium embedding pearl blending type milk beverage;
J, refrigeration: be cooled to after 25 ℃ of temperature, enter refrigerated storage temperature and be the freezer of 10 ℃, final finished product.
Embodiment 4:
(1) preparation of Bifidobacterium embedded block
A, actication of culture test tube are cultivated
Culture medium forms with embodiment 1, before actication of culture, by the bifidobacterium breve of subzero 10 ℃ of preservations (
bifidobacterium brevis) ampoule bottle at room temperature places 2h, then the cotton ball soaked in alcohol wipe surfaces sterilization by 75% concentration by ampoule bottle at desinfection chamber, open ampoule bottle, the culture medium of drawing 0.2 mL with aseptic straw adds in ampoule bottle, by fungus in ampoule bottle, be inoculated in culture medium by the inoculum concentration of 4% (percent by volume), test tube is cultivated, under anaerobic cultivate after 36 h for 35 ℃, stop cultivating.Activate continuously again 2 times with identical inoculum concentration, condition of culture, obtain three generations's liquid culture;
B, triangular flask first class inoculum are cultivated
Third generation culture is carried out to first class inoculum cultivation by the inoculum concentration access triangular flask of 5% (percent by volume), under anaerobic cultivate after 24 h for 35 ℃, stop cultivating;
C, seeding tank second class inoculum are cultivated
Get triangular flask first class inoculum liquid culture, by the inoculum concentration of 4% (percent by volume), access seeding tank carries out second class inoculum cultivation, under non-anaerobic condition, cultivates after 24 h for 38 ℃, stops cultivating;
D, fermentation tank culture
Fermentation tank culture: by the inoculum concentration of 4% (percent by volume), access seeding tank second class inoculum liquid culture, under non-anaerobic condition, under 37.5 ℃ of conditions, cultivate, when pH regulates zymotic fluid pH to 6.5 with NaOH solution lower than 5.5 time, after 24 h, stop cultivating;
E, the centrifugal collection of thalline
3 ℃ of centrifuging temperatures are set, centrifugal rotational speed 5000 r/min, centrifugation time 20 min; After centrifugal end, remove supernatant, the Bifidobacterium thalline of collecting precipitation is for subsequent use;
F, interpolation protective agent
Protective agent consists of, skim milk 10 g, glycerine 3 g, ascorbic acid 0.3 g, sucrose 5 g and water 100 g; The thalline of centrifugal collection is suspended in the protective agent of centrifugal prefermentor nutrient solution original volume 1/20;
The embedding of G, Bifidobacterium
1) preparation of embedded material solution: embedded material solution is formulated by the edible seaweed acid sodium of 10 weight portions and the water of edible gelatin and 90 weight portions, and edible seaweed acid sodium and edible gelatin are pressed the preparation of 3:1 part by weight,
Preparation process, for first taking edible gelatin, adds in distilled water, and the temperature of distilled water is 80 ℃, is stirred to completely and dissolves, then add edible seaweed acid sodium, and continuing to be stirred to becomes clear solution, for subsequent use after moist heat sterilization 15 min under 0.08 Mpa;
2) preparation of curing agent: calcium chloride is mixed with to 2mol/L concentration calcium chloride solution with distilled water, for subsequent use after 0.08 Mpa moist heat sterilization 15min;
3) embedding: the embedded material solution after sterilizing be suspended in protective agent in Bifidobacterium thalline mix, wherein embedded material solution with the weight proportion that is suspended in Bifidobacterium thalline in protective agent is, embedded material solution accounts for 70 parts, is suspended in Bifidobacterium thalline in protective agent and accounts for 30 parts; After stirring, clamp-on curing agent 2mol/L calcium chloride solution by extruder, form 5 mm wide, the Bifidobacterium embedding bar that 5 mm are thick, low rate mixing 10 min, filter, be cut into 5 mm bifidobacterium longum embedded blocks, with sterilized water washing Bifidobacterium embedded block 2 min, 5 ℃ of low tempertaure storages are for subsequent use;
(2) prepare Bifidobacterium embedding pearl yogurt drink
H, raw material milk pretreatment: will the preheating of raw sheep dairy milk starting material milk 70 ℃, open mixer, the sucrose of material quality 5% is added in hot milk, after all dissolving, filter with 200 order filter cloths.Homogeneous after raw material milk preheating, homogenization pressure is 20 Mpa, then sterilization, sterilization temperature is 95 ℃, 5 minutes time, is cooled to 37 ℃;
I, inoculation, fermentation: milk beverage leavening is for having a liking for acid streptococci and lactobacillus delbruockii subspecies bulgaricus is prepared according to 1:1, press 5%(percent by volume) inoculum concentration, milk beverage leavening is inoculated in after pretreatment in raw material milk, 43 ℃ of fermentations, milk beverage titratable acidity arrives 70 clean Er Nieerdu, stops fermentation.Stir breakdown of emulsion and be cooled to 25 ℃, be 6:4 in the ratio of milk beverage and water, add pure water, add 9% sucrose, 0.2% citric acid, 0.1% edible pectin by milk beverage and water inventory, 0.2% carboxymethyl cellulose, under homogenization pressure 18MPa, after homogeneous, Bifidobacterium embedding pearl by mass percentage 15% amount adds milk beverage, then aseptic subpackaged, will add Bifidobacterium embedding pearl yogurt drink;
J, refrigeration, by stopping the Bifidobacterium embedding pearl yogurt drink of fermentation, be cooled to after 22 ℃ of temperature, enters refrigerated storage temperature and be the freezer final finished of 5 ℃.
Embodiment 5:
The step that repeats embodiment 1, has following difference:
The bifidobacterium species that embedded block comprises be bifidobacterium (
bifidobacterium bifidum),
(1) Bifidobacterium embedding pearl preparation
The embedding of G, Bifidobacterium
1) preparation of embedded material solution: embedded material solution is formulated by the edible seaweed acid sodium of 5 weight portions and the water of edible gelatin and 95 weight portions, and edible seaweed acid sodium and edible gelatin are pressed the preparation of 4:1 part by weight;
Preparation process, for first taking edible gelatin, adds in distilled water, and the temperature of distilled water is 80 ℃, is stirred to completely and dissolves, then add edible seaweed acid sodium, and continuing to be stirred to becomes clear solution, for subsequent use after moist heat sterilization 1530 min under 0.12Mpa;
2) preparation of curing agent: calcium chloride is mixed with to 0.1mol/L concentration calcium chloride solution with distilled water, for subsequent use after 0.12 Mpa moist heat sterilization 15min;
3) embedding: the embedded material solution after sterilizing be suspended in protective agent in Bifidobacterium thalline mix, wherein embedded material solution with the weight proportion that is suspended in Bifidobacterium thalline in protective agent is, embedded material solution accounts for 80 parts, is suspended in Bifidobacterium thalline in protective agent and accounts for 20 parts; After stirring, clamp-on curing agent 0.5mol/L calcium chloride solution by extruder, form the Bifidobacterium embedding pearl of 12 mm diameters, low rate mixing 30min, filters, and forms diameter 5mm Bifidobacterium embedding pearl, low rate mixing 10 min, filter, with sterilized water washing Bifidobacterium embedding pearl 2 min, 5 ℃ of low tempertaure storages are for subsequent use;
(2) Bifidobacterium embedding pearl blending type milk beverage preparation
H, raw material milk pretreatment: raw material milk pretreatment: by 65 ℃ of raw material milk preheatings, open mixer, the sucrose of material quality 5% is added in hot milk, after all dissolving, filter with 200 order filter cloths.Homogeneous after raw material milk preheating, homogenization pressure is 18 Mpa, then sterilization, sterilization temperature is 95 ℃, 5 minutes time, is cooled to 37 ℃.
I, preparation: take raw milk as raw material, after raw material milk pretreatment, adding water, is 4:6 in pretreated feedstock milk and water ratio, carry out after proportioning, add 9% sucrose by raw milk and water inventory, 0.25% citric acid dosage, 0.15% edible pectin, 0.15% carboxymethyl cellulose, carry out the preparation of blending type milk beverage, under homogenization pressure 18MPa after homogeneous, 95 ℃ of sterilization temperatures, 5 minutes time, be cooled to 25 ℃.Bifidobacterium embedding pearl adds milk beverage by 10% amount, then aseptic subpackaged, will add Bifidobacterium embedding pearl blending type milk beverage;
J, refrigeration: be cooled to after 25 ℃ of temperature, enter refrigerated storage temperature and be the freezer of 6 ℃, final finished product.
Embodiment 6:
Repeat the step of embodiment 1, have following difference: the bifidobacterium species that embedded block comprises be bifidobacterium species be animal bifidobacteria (
bifidobacterium animalis),
(1) Bifidobacterium embedded block preparation
The embedding of G, Bifidobacterium
1) preparation of embedded material solution: embedded material solution is formulated by the edible seaweed acid sodium of 10 weight portions and the water of edible gelatin and 90 weight portions, and edible seaweed acid sodium and edible gelatin are pressed the preparation of 3:1 part by weight;
Preparation process, for first taking edible gelatin, adds in distilled water, and the temperature of distilled water is 80 ℃, is stirred to completely and dissolves, then add edible seaweed acid sodium, and continuing to be stirred to becomes clear solution, for subsequent use after moist heat sterilization 20min under 0.10 Mpa;
2) preparation of curing agent: calcium chloride is mixed with to 0.3mol/L concentration calcium chloride solution with distilled water, for subsequent use after 0.10 Mpa moist heat sterilization 20min;
3) embedding: the embedded material solution after sterilizing be suspended in protective agent in Bifidobacterium thalline mix, wherein embedded material solution with the weight proportion that is suspended in Bifidobacterium thalline in protective agent is, embedded material solution accounts for 70 parts, is suspended in Bifidobacterium thalline in protective agent and accounts for 30 parts; After stirring, clamp-on curing agent 0.5mol/L calcium chloride solution by extruder, form the Bifidobacterium embedding pearl of 12 mm diameters, low rate mixing 30min, filter, form 3mm wide, the Bifidobacterium embedding bar that 3mm is thick, low rate mixing 5min, filter, be cut into 3 mm bifidobacterium longum embedded blocks, with sterilized water washing Bifidobacterium embedded block 2 min, 10 ℃ of low tempertaure storages are for subsequent use;
(2) prepare Bifidobacterium embedded block yogurt drink
H, raw material milk processing: by 65 ℃ of raw milk raw material milk preheatings, open stirring, enter 5% sucrose by raw material dosage, add in hot milk, after all dissolving, filter with 200 order filter cloths.Homogeneous after raw material milk preheating, homogenization pressure is 18 Mpa, 95 ℃ of sterilization temperatures, are cooled to 42 ℃ at 5 minutes time;
I, inoculation, fermentation: milk beverage leavening is for having a liking for acid streptococci and lactobacillus delbruockii subspecies bulgaricus is prepared according to 1:1, press 5%(percent by volume) inoculum concentration, milk beverage leavening is inoculated in after pretreatment in raw material milk, 42 ℃ of fermentations, milk beverage titratable acidity arrives 70 clean Er Nieerdu, stops fermentation.Stir breakdown of emulsion and be cooled to 25 ℃, be 4:6 in the ratio of milk beverage and water, add pure water, add 8% sucrose by milk beverage and water inventory, 0.2% citric acid, 0.1% edible pectin, 0.2% carboxymethyl cellulose, under homogenization pressure 20MPa, after homogeneous, Bifidobacterium embedding pearl by mass percentage 20% amount adds milk beverage, then aseptic subpackaged, will add Bifidobacterium embedding pearl yogurt drink;
J, refrigeration, by stopping the Bifidobacterium embedding pearl yogurt drink of fermentation, be cooled to after 25 ℃ of temperature, enters refrigerated storage temperature and be the freezer final finished of 8 ℃.
Claims (10)
1. a milk beverage that contains Bifidobacterium embedding pearl, is characterized in that: it contains Bifidobacterium embedding pearl, Bifidobacterium embedding pearl comprise the long subspecies of Bifidobacterium bifidobacterium longum (
bifidobacterium longum subsp. longum) CGMCC numbering 1.2186, bifidobacterium longum baby subspecies (
bifidobacterium longum subsp. infantis) CGMCC numbering 1.2202, bifidobacterium adolescentis (
bifidobacterium adolescentis) CGMCC numbering 1.2190, bifidobacterium breve (
bifidobacterium brevis) CGMCC numbering 1.2213, bifidobacterium (
bifidobacterium bifidum) CGMCC numbering 1.2212 or animal bifidobacteria (
bifidobacterium animalis) one in CGMCC numbering 1.1852, the number of active bifid bacteria in Bifidobacterium embedding pearl is not less than 1 × 10
9cFU/g.
2. a kind of milk beverage that contains Bifidobacterium embedding pearl claimed in claim 1, is characterized in that: Bifidobacterium embedding pearl is solid-state shape spherical in shape, and diameter is 3 mm-12 mm.
3. the preparation method of a kind of milk beverage that contains Bifidobacterium embedding pearl claimed in claim 1, is characterized in that it comprises the following steps:
(1) preparation of Bifidobacterium embedding pearl
A, actication of culture test tube are cultivated
Before actication of culture, at desinfection chamber, the Bifidobacterium ampoule bottle of preserving at subzero 18 ℃ of-4 ℃ of temperature is at room temperature placed to 2 h-3 h, then by ampoule bottle sterilization, open ampoule bottle, the culture medium of drawing 0.2 mL-0.5 mL with aseptic straw adds in ampoule bottle, by fungus in ampoule bottle, the inoculum concentration that by volume percentage is 2%-5% is inoculated in culture medium, and test tube is cultivated, under anaerobic, at the temperature of 30 ℃-42 ℃, cultivate after 20 h-48 h, stop cultivating; Activate continuously again 2 times with identical inoculum concentration, condition of culture, obtain three generations's liquid culture;
B, triangular flask first class inoculum are cultivated
Get test tube and cultivate three generations's liquid culture, the inoculum concentration that by volume percentage is 2%-5%, access triangular flask carries out first class inoculum Liquid Culture, under anaerobic, at the temperature of 30 ℃-42 ℃, cultivates after 20h-48h, stops cultivating;
C, seeding tank second class inoculum are cultivated
Get triangular flask first class inoculum liquid culture, the inoculum concentration that by volume percentage is 2%-5%, access seeding tank carries out second class inoculum Liquid Culture, under non-anaerobic condition, at the temperature of 30 ℃-42 ℃, cultivates after 20h-48h, stops cultivating;
D, fermentation tank culture
By the liquid culture of cultivating through seeding tank second class inoculum, the inoculum concentration that by volume percentage is 2%-5%, access fermentation tank, at the temperature of 30 ℃-42 ℃, cultivate, when zymotic fluid pH value is lower than 5.5 time, between NaOH solution adjusting zymotic fluid pH to 6.0-6.5, after cultivation 20 h-48 h, stop cultivating;
E, the centrifugal collection of thalline
It is 0 ℃-8 ℃ that centrifuge temperature is set, and centrifugal rotational speed is that 3000 r/min-9000 r/min, centrifugation time are 10 min-30 min, after centrifugal end, removes supernatant, and the Bifidobacterium thalline of collecting precipitation is for subsequent use;
F, interpolation protective agent
The thalline of centrifugal collection is suspended in the protective agent of centrifugal prefermentor nutrient solution original volume 1/100-1/10; At the temperature of subzero 20 ℃-10 ℃, store for subsequent use;
The embedding of G, Bifidobacterium
To be suspended in Bifidobacterium thalline in protective agent and mix with sterilizing embedded material solution, and after stirring, clamp-on curing agent, low rate mixing 5min-30min, filters, and with sterilized water washing, obtains Bifidobacterium embedding pearl;
(2) preparation of milk beverage
H, raw material milk pretreatment: raw material milk is preheated to the temperature of 65 ℃-70 ℃, opens mixer, sucrose is added in hot milk, while all melting, filter with 200 order filter cloths; Homogeneous after raw material milk preheating, homogenization pressure is 18Mpa-20Mpa; Then sterilization, sterilization temperature is 95 ℃, 5 minutes time, is cooled to 37 ℃-42 ℃;
I, prepare milk beverage: be divided into two kinds of preparation preparation shape milk beverage and yogurt drinks,
1) prepare Bifidobacterium embedding pearl blending type milk beverage: be 1-6:6-1 in pretreated feedstock milk and water ratio, carry out after proportioning, add sucrose, citric acid, edible pectin, carboxymethyl cellulose modulation, then homogeneous, sterilization, cooling, Bifidobacterium embedding pearl adds in milk beverage by the amount of 10-30%, then aseptic subpackaged;
2) prepare Bifidobacterium embedding pearl yogurt drink: after raw material milk pretreatment, the inoculum concentration that by volume percentage is 2%-5%, leavening is inoculated in pretreated milk, at the temperature of 37 ℃-43 ℃, cultivate, acidity reaches 70-80 clean Er Nieerdu, stop fermentation, then add water, milk beverage accounts for 40-50% and water accounts for 50-60%, add sucrose, citric acid, edible pectin, carboxymethyl cellulose, after homogeneous, the Bifidobacterium embedding pearl by mass percentage amount of 15-20% adds in milk beverage again, then aseptic subpackaged
J, refrigeration: will contain the milk beverage that has Bifidobacterium embedding pearl, be cooled to after 20 ℃-25 ℃, enter refrigerated storage temperature and be the freezer of 2 ℃-10, final finished.
4. the preparation method of a kind of milk beverage that contains Bifidobacterium embedding pearl according to claim 3, it is characterized in that: in described steps A, the composition of culture medium and weight proportion are: 3-15 parts of peptones, 1-10 parts of beef extracts, 3-10 parts of tryptones, 1-10 parts of yeast extract powders, 3-15 parts of glucose, 0.5-5 parts of dipotassium hydrogen phosphates, 0.5-5 parts of sodium acetates, 0.5-5 parts of diammonium hydrogen citrates, 0.1-0.5 part of white vitriol, 0.05-0.2 part of epsom salt, 0.1-1 part of Cys hydrochloride, 0.5-5 parts of FOSs, 0.5-2 parts, calcium carbonate, 1000 parts, 1-5 parts of Tween-80s and water, the pH value of culture medium is 6.0-7.5, after 0.05Mpa-0.14Mpa moist heat sterilization 15 min-30 min, uses.
5. the preparation method of a kind of milk beverage that contains Bifidobacterium embedding pearl according to claim 4, it is characterized in that: in described step F, protectant composition and weight proportion are: 5-30 parts of skim milk or skimmed milk powers, 0.5-5 parts of glycerine, 0.1-1 part, ascorbic acid, 100 parts, 3-10 parts of sucrose and water; By skim milk or skimmed milk power, glycerine and sucrose and water moist heat sterilization 15min-30min under 0.05 Mpa-0.14 Mpa pressure, ascorbic acid adopts filtration sterilization.
6. the preparation method of a kind of milk beverage that contains Bifidobacterium embedding pearl claimed in claim 5, it is characterized in that: in described step G, embedded material solution is formulated by the edible seaweed acid sodium of 2-10 weight portions and the water of edible gelatin and 90-98 weight portions, the weight proportion of edible seaweed acid sodium and edible gelatin is 1-5:1, preparation process is for first taking edible gelatin, add in distilled water, the temperature of distilled water is 75 ℃-95 ℃, be stirred to completely and dissolve, add again edible seaweed acid sodium, continuing to be stirred to becomes clear solution, moist heat sterilization 15 min-30min under 0.05 Mp-0.14Mpa pressure.
7. the preparation method of a kind of milk beverage that contains Bifidobacterium embedding pearl claimed in claim 6; it is characterized in that: in described step G, embedded material solution with the weight proportion that is suspended in Bifidobacterium thalline in protective agent is; embedded material solution accounts for 50-90 parts, is suspended in Bifidobacterium thalline in protective agent and accounts for 10-50 parts.
8. the preparation method of a kind of milk beverage that contains Bifidobacterium embedding pearl claimed in claim 7, it is characterized in that: in described step G, curing agent is calcium chloride solution, process for preparation is, with distilled water, calcium chloride is mixed with to the saturated solution that concentration is 0.1mol/L, after 0.05Mpa-0.14Mpa moist heat sterilization 15min-30min, uses.
9. the preparation method of a kind of milk beverage that contains Bifidobacterium embedding pearl claimed in claim 8, is characterized in that: the milk beverage leavening in described step I is that number of viable is the mixture of having a liking for acid streptococci (Streptococcus thermophilus) CGMCC numbering 1.1864 and lactobacillus delbruockii subspecies bulgaricus (Lactobacillus delbrueckii subsp. bulgaricus) CGMCC numbering 1.1863 of 1:1.
10. the preparation method of a kind of milk beverage that contains Bifidobacterium embedding pearl claimed in claim 9, it is characterized in that: in described step I, the addition of sucrose, citric acid, edible pectin, carboxymethyl cellulose is, add 8-10% sucrose by raw milk and water inventory, 0.1-0.2% citric acid, 0.1-0.3% edible pectin, 0.15-0.2% carboxymethyl cellulose.
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| CN105495322A (en) * | 2015-12-08 | 2016-04-20 | 黑龙江八一农垦大学 | Pure natural lactobacillus millet enzyme |
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