CN107853396A - A kind of method that Lactobacillus rhamnosus protease prepares sheep breast active beverage - Google Patents
A kind of method that Lactobacillus rhamnosus protease prepares sheep breast active beverage Download PDFInfo
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- CN107853396A CN107853396A CN201710963850.5A CN201710963850A CN107853396A CN 107853396 A CN107853396 A CN 107853396A CN 201710963850 A CN201710963850 A CN 201710963850A CN 107853396 A CN107853396 A CN 107853396A
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- 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
- A23C9/00—Milk preparations; Milk powder or milk powder preparations
- A23C9/152—Milk preparations; Milk powder or milk powder preparations containing additives
- A23C9/156—Flavoured milk preparations ; Addition of fruits, vegetables, sugars, sugar alcohols or sweeteners
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- 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
- A23C9/00—Milk preparations; Milk powder or milk powder preparations
- A23C9/12—Fermented milk preparations; Treatment using microorganisms or enzymes
- A23C9/1203—Addition of, or treatment with, enzymes or microorganisms other than lactobacteriaceae
- A23C9/1209—Proteolytic or milk coagulating enzymes, e.g. trypsine
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- 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
- A23C2220/00—Biochemical treatment
- A23C2220/10—Enzymatic treatment
- A23C2220/104—Enzymatic treatment with immobilised enzymes
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Abstract
A kind of method that Lactobacillus rhamnosus protease prepares sheep breast active beverage, Lactobacillus rhamnosus inoculum concentration is accessed in Lactobacillus rhamnosus culture medium after cultivating and bacterial sediment is collected by centrifugation, washed with sterile saline centrifuge to obtain Lactobacillus rhamnosus bacterium mud afterwards；Add phosphate buffer into Lactobacillus rhamnosus bacterium mud, centrifuge protein in cell wall enzyme enzyme liquid, freeze-drying obtain Lactobacillus rhamnosus protein in cell wall enzyme bacterium powder；Lactobacillus rhamnosus protein in cell wall enzyme bacterium powder and CaCl are added into the fresh goat milk after degreasing2Add beta cyclodextrin after enzymolysis again, hovenia polysaccharides, antierythrite, xylitol, L malic acid, citric acid, after sodium citrate is well mixed the sheep breast active beverage containing Antihypertensive Peptides and anti-oxidation peptide is obtained after homogeneous, sterilization, its ACE inhibitory activity is 70 90%, DPPH free radical scavenging activities are 60 80%, and Scavenging action to hydroxyl free radical is 75 95%.For the present invention using sheep breast as raw material, prepared sheep breast active beverage can be used as aided blood pressure-lowering, anti-oxidant, hypoglycemic and anti-caries food.
Description
Technical field
The present invention relates to a kind of preparation method of feature goat milk beverage, more particularly to a kind of Lactobacillus rhamnosus protease
The method for preparing sheep breast active beverage.
Background technology
Probiotics has proteolytic system, and the system includes CEP, aminopeptidase, endopeptidase, Pro- specificity peptases
Deng, protein can be hydrolyzed and discharge peptide of different sizes and some free amino acid are to meet thalli growth by it,
Some peptides caused by wherein hydrolyzing have bioactivity, such as the function such as anti-oxidant, hypotensive, immunological regulation.In whole water
In solution preocess, protein in cell wall enzyme is most important of which enzyme, studies have found that the independent can lactoalbumin hydrolysates of some CEP simultaneously
Produce ace inhibitory peptide.
The cell surface of Bacillus acidi lactici includes many enzymes and protein, mainly there is ribosomal protein, cell surface permeability
Enzyme, the protein of superficial layer (S layers), cell embrane-associated protein enzyme (CEPs), and other molecules based on protein[10].Carefully
The mechanism of protein in cell wall enzyme (CEP) lactoalbumin hydrolysate is：Lactoprotein is hydrolyzed to a series of small peptide by CEP first, afterwards oligopeptides,
Dipeptides and tripeptides movement system these small peptides are transported it is intracellular, finally by intracellular peptase (endopeptidase, pro- specificity
Peptase, aminopeptidase etc.) it is further hydrolyzed to the biologically active peptide such as ace inhibitory peptide and anti-oxidation peptide peptide
Honey raisin tree is called papaw, scientific name trifoliate orange Dulcis, and its polysaccharide has hypoglycemic function, but hovenia polysaccharides prepare presence at present
The problem of purity is low, major impurity are monose and disaccharide, therefore it is that to prepare high-purity honey raisin tree more to remove mono-and di-saccharides and retain polysaccharide
The key of sugar and exploitation new product.
Shaanxi Province is the traditional milch goat main producing region in China, and Guanzhong area is national maximum milch goat breeding production base
Ground.2015 end of the year the whole province milch goat livestock on hands more than 1,800,000, the ton of sheep milk production more than 500,000, the goat milk powder market share up to more than 95%,
Milch goat quantity, sheep milk production and goat milk powder yield are national first, but the less varieties of domestic sheep milk product, predominantly goat milk
Powder and liquid goat milk, wherein most goat milk powder as raw material of industry exportation abroad, exist the technological element of a product is low, added value not
The problems such as high and homogeneity is serious, external sheep milk product species is more, main to include fermentation sheep milk product (such as cheese, degreasing buttermilk
Or acidophilus goat milk), frozen product (such as ice cream or freezing acidophilus goat milk), liquid milk, butter, feature goat milk piece.Therefore, develop
Feature sheep breast new product is necessary.
The content of the invention
It is an object of the invention to provide a kind of sandlwood with functions such as hypotensive, anti-oxidant, hypoglycemic and anti-caries
The method that sugared lactobacillus protease prepares sheep breast active beverage.
To reach above-mentioned purpose, the technical solution adopted by the present invention is：
1) prepared by Lactobacillus rhamnosus culture medium：
By lactose 15-25g, beef extract 15-25g, magnesium sulfate 10-20mg, Tween 80 0.5-1.0mL, galactooligosaccharide 2-
5g, peptone 3-6g, disodium hydrogen phosphate 4-6g, alanine 5-10mg are added in pure water, are settled to 1L, regulation pH value to
5.5-7.0, sterilize Lactobacillus rhamnosus culture medium in 121 DEG C；
2) Lactobacillus rhamnosus culture and microorganism collection：
By Lactobacillus rhamnosus by 2%-5% inoculum concentration access Lactobacillus rhamnosus culture medium, 35-41 DEG C of constant temperature is trained
18-24h is supported, is then centrifuged for collecting bacterial sediment, is washed with sterile saline centrifuge to obtain Lactobacillus rhamnosus bacterium mud afterwards；
3) prepared by Lactobacillus rhamnosus protein in cell wall enzyme：
By the phosphate buffer that 20-30mL is added in every g Lactobacillus rhamnosus bacterium mud, centrifuged after 35-45 DEG C of insulation 1-3h
The enzyme liquid that supernatant is protein in cell wall enzyme is obtained, small-molecule substance is removed using 1-3KDa milipore filters, then added in liquid is retained
Add CaCl2, control CaCl2Concentration be 10-20mM, freeze-drying obtain Lactobacillus rhamnosus protein in cell wall enzyme bacterium powder；
4) preparation of sheep breast active beverage：
Fresh goat milk is adjusted to the rhamnose breast that pH value to 6.0-8.0 mass volume ratios add 1-3% thereto after degreasing
Bacillus protein in cell wall enzyme bacterium powder and 0.01-0.04%CaCl2, 2-4h is digested at 35-45 DEG C, is then added again by mass volume ratio
Add 0.5-1.0% beta-schardinger dextrin, 0.2-0.4% hovenia polysaccharides, 4-6% antierythrite, 3-5% xylitol, 0.1-
0.3% L MALIC ACID, 0.10-0.25% citric acid, after 0.10-0.25% sodium citrate is well mixed through homogeneous, kill
The sheep breast active beverage containing Antihypertensive Peptides and anti-oxidation peptide is obtained after bacterium, its ACE inhibitory activity is 70-90%, and DPPH free radicals are clear
Except rate is 60-80%, Scavenging action to hydroxyl free radical 75-95%.
The centrifugation of the step 2) is to centrifuge 15-25min under 4-10 DEG C, 6000-8000r/min.
The pH=7.0 of the phosphate buffer of the step 3), glycine containing 15-30g and 0.5- in every liter of phosphate buffer
1.0g lysozyme.
The centrifugation of the step 3) is to centrifuge 20-30min under 4-10 DEG C, 6000-8000r/min.
The homogeneous of the step 4) is that temperature is 15-30 DEG C.
The hovenia polysaccharides preparation process of the step 4) is：Take fresh honey raisin tree to be beaten after cleaning, then add 20
The water of times honey raisin tree weight, stirs, 2h is incubated under the conditions of 90 DEG C, then filters and obtains hovenia polysaccharides extract solution, is cooled to
By the Paula enlightening yeast activated liquid of mass volume ratio access 2% after 38 DEG C, 37 DEG C of incubated 24h remove monose in honey raisin trees and
Disaccharide, is collected by centrifugation Paula enlightening yeast thalline, and supernatant is concentrated into 1/20 with Rotary Evaporators, then by 1:3 volume ratio adds
Enter absolute ethyl alcohol, then filtered with Buchner funnel and obtain honey raisin tree Thick many candies, then honey raisin tree Thick many candies rehydration is added into chloroform and positive fourth
Alcohol removing protein, then water intaking mutually add chloroform and n-butanol again, obtain hovenia polysaccharides by being freeze-dried after de- albumen three times.
For the present invention using sheep breast as raw material, the protein in cell wall enzyme of GRAS microorganisms --- Lactobacillus rhamnosus is living things catalysis
Agent, sheep protein of milk is hydrolyzed into the small peptide with ACE inhibitory activity and inoxidizability, consumer's intake can aid in dropping
Blood pressure and anti-oxidant, the hovenia polysaccharides of addition had both had the function that stabilizer and had reduced blood glucose, can increase sheep breast active beverage
Feature；The antierythrite and xylitol of addition are functional Sugar Alcohol, its have low sugariness, it is low in calories, without carious tooth, to blood glucose
The features such as small is influenceed, sheep breast active beverage prepared by the present invention can be used as aided blood pressure-lowering, anti-oxidant, hypoglycemic and anti-caries food
Product.
Embodiment
It is that the present invention is described in further details in conjunction with specific embodiments below.
Embodiment 1：
1) prepared by Lactobacillus rhamnosus culture medium：
By lactose 20g, beef extract 15g, magnesium sulfate 13mg, Tween 80 0.7mL, galactooligosaccharide 3g, peptone 3g, phosphorus
Sour disodium hydrogen 6g, alanine 5mg are added in pure water, are settled to 1L, regulation pH value is to 6.0, in 121 DEG C of sandlwoods that sterilize
Sugared lactobacillus culture medium；
2) Lactobacillus rhamnosus culture and microorganism collection：
Lactobacillus rhamnosus is accessed in Lactobacillus rhamnosus culture medium by 3% inoculum concentration, 37 DEG C of incubated 22h,
It is then centrifuged for collecting bacterial sediment, is washed with sterile saline centrifuge to obtain Lactobacillus rhamnosus bacterium mud afterwards；
Described centrifugation is to centrifuge 20min under 8 DEG C, 7000r/min；
3) prepared by Lactobacillus rhamnosus protein in cell wall enzyme：
By the phosphate buffer of addition 23mL, pH=7.0 in every g Lactobacillus rhamnosus bacterium mud (in every liter of phosphate buffer
Glycine containing 20g and 1.0g lysozymes), under 4 DEG C, 8000r/min centrifuging 20min after 35 DEG C of insulation 3h, to obtain supernatant be born of the same parents
The enzyme liquid of wall-held protein enzyme, small-molecule substance is removed using 1-3KDa milipore filters, then adds CaCl in liquid is retained2, control
CaCl2Concentration be 17mM, freeze-drying obtain Lactobacillus rhamnosus protein in cell wall enzyme bacterium powder；
4) preparation of sheep breast active beverage：
Fresh goat milk is adjusted to the Lactobacillus rhamnosus that pH value adds 1.5% to 6.5 mass volume ratios thereto after degreasing
Protein in cell wall enzyme bacterium powder and 0.03%CaCl2, 3h is digested at 40 DEG C, then again by β-ring of mass volume ratio addition 0.8%
Dextrin, 0.2% hovenia polysaccharides, 5% antierythrite, 5% xylitol, 0.3% L MALIC ACID, 0.2% citric acid,
Obtained after 0.15% sodium citrate is well mixed after 25 DEG C of homogeneous, sterilization containing the sheep of Antihypertensive Peptides and anti-oxidation peptide breast activity drink
Material, its ACE inhibitory activity is that 80%, DPPH free radical scavenging activities are 60%, Scavenging action to hydroxyl free radical 75%.
Embodiment 2：
1) prepared by Lactobacillus rhamnosus culture medium：
By lactose 15g, beef extract 20g, magnesium sulfate 18mg, Tween 80 0.5mL, galactooligosaccharide 4g, peptone 5g, phosphorus
Sour disodium hydrogen 4g, alanine 8mg are added in pure water, are settled to 1L, regulation pH value is to 5.5, in 121 DEG C of sandlwoods that sterilize
Sugared lactobacillus culture medium；
2) Lactobacillus rhamnosus culture and microorganism collection：
Lactobacillus rhamnosus is accessed in Lactobacillus rhamnosus culture medium by 5% inoculum concentration, 35 DEG C of incubated 24h,
It is then centrifuged for collecting bacterial sediment, is washed with sterile saline centrifuge to obtain Lactobacillus rhamnosus bacterium mud afterwards；
Described centrifugation is to centrifuge 15min under 4 DEG C, 8000r/min；
3) prepared by Lactobacillus rhamnosus protein in cell wall enzyme：
By the phosphate buffer of addition 30mL, pH=7.0 in every g Lactobacillus rhamnosus bacterium mud (in every liter of phosphate buffer
Glycine containing 15g and 0.8g lysozymes), under 8 DEG C, 7000r/min centrifuging 5min after 40 DEG C of insulation 2h, to obtain supernatant be born of the same parents
The enzyme liquid of wall-held protein enzyme, small-molecule substance is removed using 1-3KDa milipore filters, then adds CaCl in liquid is retained2, control
CaCl2Concentration be 17mM, freeze-drying obtain Lactobacillus rhamnosus protein in cell wall enzyme bacterium powder；
4) preparation of sheep breast active beverage：
Fresh goat milk is adjusted to the Lactobacillus rhamnosus born of the same parents that pH value to 6.0 mass volume ratios add 3% thereto after degreasing
Wall-held protein enzyme bacterium powder and 0.01%CaCl2, 4h is digested at 35 DEG C, then again by β-ring paste of mass volume ratio addition 0.5%
Essence, 0.4% hovenia polysaccharides, 4% antierythrite, 3% xylitol, 0.1% L MALIC ACID, 0.15% citric acid,
Obtained after 0.10% sodium citrate is well mixed after 15 DEG C of homogeneous, sterilization containing the sheep of Antihypertensive Peptides and anti-oxidation peptide breast activity drink
Material, its ACE inhibitory activity is that 70%, DPPH free radical scavenging activities are 80%, Scavenging action to hydroxyl free radical 85%.
Embodiment 3：
1) prepared by Lactobacillus rhamnosus culture medium：
By lactose 25g, beef extract 20g, magnesium sulfate 20mg, Tween 80 1.0mL, galactooligosaccharide 2g, peptone 6g, phosphorus
Sour disodium hydrogen 5g, alanine 10mg are added in pure water, are settled to 1L, regulation pH value is to 6.5, in 121 DEG C of mouse that sterilize
Lee's sugar lactobacillus culture medium；
2) Lactobacillus rhamnosus culture and microorganism collection：
Lactobacillus rhamnosus is accessed in Lactobacillus rhamnosus culture medium by 2% inoculum concentration, 39 DEG C of incubated 20h,
It is then centrifuged for collecting bacterial sediment, is washed with sterile saline centrifuge to obtain Lactobacillus rhamnosus bacterium mud afterwards；
Described centrifugation is to centrifuge 25min under 10 DEG C, 6000r/min；
3) prepared by Lactobacillus rhamnosus protein in cell wall enzyme：
By the phosphate buffer of addition 28mL, pH=7.0 in every g Lactobacillus rhamnosus bacterium mud (in every liter of phosphate buffer
Glycine containing 30g and 0.5g lysozymes), under 6 DEG C, 6000r/min centrifuging 30min after 45 DEG C of insulation 1h, to obtain supernatant be born of the same parents
The enzyme liquid of wall-held protein enzyme, small-molecule substance is removed using 1-3KDa milipore filters, then adds CaCl in liquid is retained2, control
CaCl2Concentration be 20mM, freeze-drying obtain Lactobacillus rhamnosus protein in cell wall enzyme bacterium powder；
4) preparation of sheep breast active beverage：
Fresh goat milk is adjusted to the Lactobacillus rhamnosus born of the same parents that pH value to 8.0 mass volume ratios add 1% thereto after degreasing
Wall-held protein enzyme bacterium powder and 0.04%CaCl2, 2h is digested at 45 DEG C, then again by β-ring paste of mass volume ratio addition 1.0%
Essence, 0.3% hovenia polysaccharides, 6% antierythrite, 5% xylitol, 0.2% L MALIC ACID, 0.10% citric acid,
Obtained after 0.25% sodium citrate is well mixed after 30 DEG C of homogeneous, sterilization containing the sheep of Antihypertensive Peptides and anti-oxidation peptide breast activity drink
Material, its ACE inhibitory activity is that 90%, DPPH free radical scavenging activities are 75%, Scavenging action to hydroxyl free radical 95%.
Embodiment 4：
1) prepared by Lactobacillus rhamnosus culture medium：
By lactose 18g, beef extract 22g, magnesium sulfate 10mg, Tween 80 0.8mL, galactooligosaccharide 5g, peptone 4g, phosphorus
Sour disodium hydrogen 4g, alanine 6mg are added in pure water, are settled to 1L, regulation pH value is to 7.0, in 121 DEG C of sandlwoods that sterilize
Sugared lactobacillus culture medium；
2) Lactobacillus rhamnosus culture and microorganism collection：
Lactobacillus rhamnosus is accessed in Lactobacillus rhamnosus culture medium by 4% inoculum concentration, 41 DEG C of incubated 18h,
It is then centrifuged for collecting bacterial sediment, is washed with sterile saline centrifuge to obtain Lactobacillus rhamnosus bacterium mud afterwards；
Described centrifugation is to centrifuge 18min under 6 DEG C, 8000r/min；
3) prepared by Lactobacillus rhamnosus protein in cell wall enzyme：
By the phosphate buffer of addition 20mL, pH=7.0 in every g Lactobacillus rhamnosus bacterium mud (in every liter of phosphate buffer
Glycine containing 25g and 0.6g lysozymes), 28min, which is centrifuged, under 10 DEG C, 6000r/min after 38 DEG C of insulation 3h obtains supernatant and be
The enzyme liquid of protein in cell wall enzyme, small-molecule substance is removed using 1-3KDa milipore filters, then adds CaCl in liquid is retained2, control
CaCl2Concentration be 10mM, freeze-drying obtain Lactobacillus rhamnosus protein in cell wall enzyme bacterium powder；
4) preparation of sheep breast active beverage：
Fresh goat milk is adjusted to the Lactobacillus rhamnosus born of the same parents that pH value to 7.0 mass volume ratios add 2% thereto after degreasing
Wall-held protein enzyme bacterium powder and 0.02%CaCl2, 3h is digested at 42 DEG C, then again by β-ring paste of mass volume ratio addition 0.6%
Essence, 0.4% hovenia polysaccharides, 5% antierythrite, 4% xylitol, 0.1% L MALIC ACID, 0.25% citric acid,
Obtained after 0.20% sodium citrate is well mixed after 20 DEG C of homogeneous, sterilization containing the sheep of Antihypertensive Peptides and anti-oxidation peptide breast activity drink
Material, its ACE inhibitory activity is that 85%, DPPH free radical scavenging activities are 70%, Scavenging action to hydroxyl free radical 90%.
Hovenia polysaccharides in above example are prepared as follows：Fresh honey raisin tree is taken to be beaten after cleaning, then
The water of 20 times of honey raisin tree weight is added, stirs, 2h is incubated under the conditions of 90 DEG C, then filters and obtains hovenia polysaccharides extract solution,
It is cooled to after 38 DEG C and is removed by the Paula enlightening yeast activated liquid of mass volume ratio access 2%, 37 DEG C of incubated 24h in honey raisin tree
Monose and disaccharide, are collected by centrifugation Paula enlightening yeast thalline, and supernatant is concentrated into 1/20 with Rotary Evaporators, then by 1:3 body
Then product is filtered with Buchner funnel than adding absolute ethyl alcohol and obtains honey raisin tree Thick many candies, then honey raisin tree Thick many candies rehydration is added into chloroform
With n-butanol removing protein, then water intaking mutually adds chloroform and n-butanol, turned after taking off albumen three times by being freeze-dried again
Jujube polysaccharide.
Claims (6)
1. a kind of method that Lactobacillus rhamnosus protease prepares sheep breast active beverage, it is characterised in that：
1) prepared by Lactobacillus rhamnosus culture medium：
By lactose 15-25g, beef extract 15-25g, magnesium sulfate 10-20mg, Tween 80 0.5-1.0mL, galactooligosaccharide 2-5g,
Peptone 3-6g, disodium hydrogen phosphate 4-6g, alanine 5-10mg are added in pure water, are settled to 1L, regulation pH value to 5.5-
7.0, sterilize Lactobacillus rhamnosus culture medium in 121 DEG C；
2) Lactobacillus rhamnosus culture and microorganism collection：
By Lactobacillus rhamnosus by 2%-5% inoculum concentration access Lactobacillus rhamnosus culture medium, 35-41 DEG C incubated
18-24h, it is then centrifuged for collecting bacterial sediment, is washed with sterile saline centrifuge to obtain Lactobacillus rhamnosus bacterium mud afterwards；
3) prepared by Lactobacillus rhamnosus protein in cell wall enzyme：
By the phosphate buffer that 20-30mL is added in every g Lactobacillus rhamnosus bacterium mud, centrifuged after 35-45 DEG C of insulation 1-3h
Clear liquid is the enzyme liquid of protein in cell wall enzyme, removes small-molecule substance using 1-3KDa milipore filters, is then added in liquid is retained
CaCl2, control CaCl2Concentration be 10-20mM, freeze-drying obtain Lactobacillus rhamnosus protein in cell wall enzyme bacterium powder；
4) preparation of sheep breast active beverage：
Fresh goat milk is adjusted to the Lactobacillus rhamnosus that pH value adds 1-3% to 6.0-8.0 mass volume ratios thereto after degreasing
Protein in cell wall enzyme bacterium powder and 0.01-0.04%CaCl2, 2-4h is digested at 35-45 DEG C, is then added again by mass volume ratio
0.5-1.0% beta-schardinger dextrin, 0.2-0.4% hovenia polysaccharides, 4-6% antierythrite, 3-5% xylitol, 0.1-
0.3% L MALIC ACID, 0.10-0.25% citric acid, after 0.10-0.25% sodium citrate is well mixed through homogeneous, kill
The sheep breast active beverage containing Antihypertensive Peptides and anti-oxidation peptide is obtained after bacterium, its ACE inhibitory activity is 70-90%, and DPPH free radicals are clear
Except rate is 60-80%, Scavenging action to hydroxyl free radical 75-95%.
2. the method that Lactobacillus rhamnosus protease according to claim 1 prepares sheep breast active beverage, it is characterised in that：
The centrifugation of the step 2) is to centrifuge 15-25min under 4-10 DEG C, 6000-8000r/min.
3. the method that Lactobacillus rhamnosus protease according to claim 1 prepares sheep breast active beverage, it is characterised in that：
The pH=7.0 of the phosphate buffer of the step 3), glycine containing 15-30g and 0.5-1.0g bacteriolyzes in every liter of phosphate buffer
Enzyme.
4. the method that Lactobacillus rhamnosus protease according to claim 1 prepares sheep breast active beverage, it is characterised in that：
The centrifugation of the step 3) is to centrifuge 20-30min under 4-10 DEG C, 6000-8000r/min.
5. the method that Lactobacillus rhamnosus protease according to claim 1 prepares sheep breast active beverage, it is characterised in that：
The homogeneous of the step 4) is that temperature is 15-30 DEG C.
6. the method that Lactobacillus rhamnosus protease according to claim 1 prepares sheep breast active beverage, it is characterised in that：
The hovenia polysaccharides preparation process of the step 4) is：Take fresh honey raisin tree to be beaten after cleaning, then add 20 times of honey raisin tree weights
The water of amount, is stirred, and 2h is incubated under the conditions of 90 DEG C, then filter obtain hovenia polysaccharides extract solution, after being cooled to 38 DEG C by
The Paula enlightening yeast activated liquid of mass volume ratio access 2%, 37 DEG C of incubated 24h remove monose and disaccharide in honey raisin tree, from
The heart collects Paula enlightening yeast thalline, and supernatant is concentrated into 1/20 with Rotary Evaporators, then by 1:3 volume ratio adds anhydrous second
Alcohol, then filtered with Buchner funnel and obtain honey raisin tree Thick many candies, then honey raisin tree Thick many candies rehydration is added into chloroform and n-butanol removing protein,
Then water intaking mutually adds chloroform and n-butanol again, obtains hovenia polysaccharides by being freeze-dried after de- albumen three times.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109161496A (en) * | 2018-08-19 | 2019-01-08 | 东北农业大学 | Lactobacillus rhamnosus high density bacterium solution and its preparation method for embedding bacterium powder |
| CN113186176A (en) * | 2021-04-22 | 2021-07-30 | 华东理工大学 | Method for producing rhizomucor miehei lipase by total synthesis culture medium and fermentation method |
| CN113755365A (en) * | 2021-08-11 | 2021-12-07 | 天津科技大学 | Preparation method and application of lactobacillus extract |
| CN114903083A (en) * | 2022-06-09 | 2022-08-16 | 陕西科技大学 | Biological active peptide probiotic fermented sheep milk and preparation method thereof |
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