CN107853394A - A kind of method that lactobacillus bulgaricus protein in cell wall enzyme prepares sheep breast active beverage - Google Patents
A kind of method that lactobacillus bulgaricus protein in cell wall enzyme prepares sheep breast active beverage Download PDFInfo
- Publication number
- CN107853394A CN107853394A CN201710965217.XA CN201710965217A CN107853394A CN 107853394 A CN107853394 A CN 107853394A CN 201710965217 A CN201710965217 A CN 201710965217A CN 107853394 A CN107853394 A CN 107853394A
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- China
- Prior art keywords
- lactobacillus bulgaricus
- protein
- cell wall
- sheep breast
- wall enzyme
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
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- 229940004208 lactobacillus bulgaricus Drugs 0.000 title claims abstract description 68
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- 108090000623 proteins and genes Proteins 0.000 title claims abstract description 46
- 210000002421 cell wall Anatomy 0.000 title claims abstract description 41
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- 238000000034 method Methods 0.000 title claims abstract description 12
- 241000186672 Lactobacillus delbrueckii subsp. bulgaricus Species 0.000 title abstract 3
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Classifications
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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
- A23C9/13—Fermented milk preparations; Treatment using microorganisms or enzymes using additives
- A23C9/1307—Milk products or derivatives; Fruit or vegetable juices; Sugars, sugar alcohols, sweeteners; Oligosaccharides; Organic acids or salts thereof or acidifying agents; Flavours, dyes or pigments; Inert or aerosol gases; Carbonation methods
-
- 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
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
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- C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
- C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
- C12N9/52—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from bacteria or Archaea
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/123—Bulgaricus
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Wood Science & Technology (AREA)
- Organic Chemistry (AREA)
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- Bioinformatics & Cheminformatics (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Polymers & Plastics (AREA)
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- Biochemistry (AREA)
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- General Health & Medical Sciences (AREA)
- Food Science & Technology (AREA)
- Molecular Biology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Virology (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
- Non-Alcoholic Beverages (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
A kind of method that lactobacillus bulgaricus protein in cell wall enzyme prepares sheep breast active beverage, using sheep breast as raw material, the protein in cell wall enzyme of GRAS microorganisms --- lactobacillus bulgaricus is biocatalyst, sheep protein of milk is hydrolyzed into the oligopeptides with ACE inhibitory activity, consumer's intake can be with aided blood pressure-lowering, the hovenia polysaccharides of addition had both had the function that stabilizer and had reduced blood glucose, can increase the feature of sheep breast active beverage；The synanthrin of addition is prebiotics, the propagation of beneficial bacteria of intestinal tract and the absorption of nutriment can be promoted, antierythrite and xylitol are functional Sugar Alcohol, its have the characteristics that low sugariness, it is low in calories, without carious tooth, it is small to blood sugar influence, Antihypertensive Peptides sheep breast active beverage prepared by the present invention can be used as aided blood pressure-lowering, hypoglycemic and anti-caries food, have multiple healthcare function.
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 bulgaricus cell wall
The method that protease prepares sheep breast active beverage.
Background technology
Hypertension is the most common chronic disease in the world, is one of maximum epidemic disease, and cardiovascular and cerebrovascular diseases are most important
Hazards.Conventional antihypertensive drugs includes calcium channel blocker, angiotensin converting enzyme inhibitor (ACEI), angiotensins
ARBs (ARB), diuretics, α ARBs and the class of beta-blocker six, wherein Angiotensin-Converting suppress
Agent (ACEI) has good target-organ protection and cardiovascular endpoints event prevention effect, and antihypertensive effect for hyperpietic
Clearly, glycolipid metabolism is had no adverse effects, is one kind with fastest developing speed in current decompression class medicine.Conventional ACEI class medicine bags
Captopril, enalapril, benazepil etc. are included, but the ACEI of these synthesis can cause many adverse reactions, most commonly
Dry persistent coughs.Research shows that the ace inhibitory peptide of natural origin is that protein is caused a kind of with ACE suppressions through enzyme decomposition
The polypeptides matter of activity is made, generally contains 3-10 amino acid, molecular weight, can be quickly through digestion between 300-10000
Mucous membrane enters blood circulation, with the potent combinations of ACE so as to suppress its activity, plays a part of decompression, therefore be called Antihypertensive Peptides.Portion
The protein in cell wall enzyme energy lactoalbumin hydrolysate of probiotics is divided to discharge peptides, some of which small peptide has bioactivity, such as drops blood
Pressure, but there is presently no the goat milk beverage of hypotensive.
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 issue of sugar.
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
Sheep breast new product is necessary.
The content of the invention
It is an object of the invention to provide a kind of Bulgaria with hypotensive, hypoglycemic and anti-caries health-care efficacy
The method that lactobacillus protein in cell wall enzyme prepares sheep breast active beverage.
To reach above-mentioned purpose, the technical solution adopted by the present invention is：
1) prepared by lactobacillus bulgaricus culture medium:
By lactose 15-25g, dusty yeast 15-25g, manganese sulfate 10-20mg, Tween 80 0.5-1.0mL, stachyose 1.5-
2.5g, peptone 4-6g, disodium hydrogen phosphate 4-6g, glutamic acid 5-10mg, alanine 5-10mg are added in pure water, are settled to
1L, regulation pH value is to 5.5-7.0, and sterilize to obtain lactobacillus bulgaricus culture medium in 121 DEG C；
2) lactobacillus bulgaricus culture and microorganism collection：
Lactobacillus bulgaricus is accessed in lactobacillus bulgaricus culture medium by 2%-5% inoculum concentration, 35-41 DEG C of perseverance
Temperature culture 18-24h, is then centrifuged for collecting bacterial sediment, is washed with sterile saline centrifuge to obtain lactobacillus bulgaricus afterwards
Bacterium mud；
3) prepared by lactobacillus bulgaricus protein in cell wall enzyme：
By in every g lactobacillus bulgaricus bacterium mud add 20-30mL phosphate buffer, 35-45 DEG C insulation 1-3h after from
Supernatant obtained by the heart is the enzyme liquid of protein in cell wall enzyme, small-molecule substance is removed using 1-3KDa ultrafiltration, then in liquid is retained
Add CaCl2, control CaCl2Concentration be 8-15mM, be freeze-dried to obtain lactobacillus bulgaricus protein in cell wall enzyme bacterium powder；
4) preparation of sheep breast active beverage：
Fresh goat milk is adjusted to the Bao Jiali that pH value is added 1-3% to 6.0-8.0 thereto by mass volume ratio after degreasing
The CaCl of sub- lactobacillus protein in cell wall enzyme bacterium powder and 0.01-0.04%2, 2-4h is digested at 35-45 DEG C, then presses mass body again
Beta-schardinger dextrin of the product than adding 0.5-1.0%, 0.2-0.4% hovenia polysaccharides, 0.3-0.5% synanthrin, 4-6% erythrose
Alcohol, 3-5% xylitol, 0.1-0.3% L MALIC ACID, 0.10-0.25% citric acid, 0.10-0.25% citric acid
After sodium is well mixed the sheep breast active beverage of the enzyme of protein in cell wall containing lactobacillus bulgaricus is obtained after homogeneous, sterilization.
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 2000- in every liter of phosphate buffer
4000U mutanolysin.
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 bulgaricus is living things catalysis
Agent, sheep protein of milk is hydrolyzed into the oligopeptides with ACE inhibitory activity, consumer's intake can be added with aided blood pressure-lowering
Hovenia polysaccharides both there is stabilizer and reduced blood glucose, the feature of sheep breast active beverage can be increased；The synanthrin of addition
For prebiotics, the propagation of beneficial bacteria of intestinal tract and the absorption of nutriment can be promoted, antierythrite and xylitol are functional Sugar Alcohol,
It has the characteristics that low sugariness, it is low in calories, without carious tooth, it is small to blood sugar influence, the present invention prepare Antihypertensive Peptides sheep breast activity drink
Material can be used as aided blood pressure-lowering, hypoglycemic and anti-caries food, have multiple healthcare function.
Embodiment
It is that the present invention is described in further details in conjunction with specific embodiments below.
Embodiment 1：
1) prepared by lactobacillus bulgaricus culture medium:
By lactose 15g, dusty yeast 15g, manganese sulfate 10mg, Tween 80 0.5mL, stachyose 1.5g, peptone 4g, phosphoric acid
Disodium hydrogen 4g, glutamic acid 5mg, alanine 5mg are added in pure water, are settled to 1L, regulation pH value to 6.0, in 121 DEG C of sterilizings
Obtain lactobacillus bulgaricus culture medium；
2) lactobacillus bulgaricus culture and microorganism collection：
By lactobacillus bulgaricus by 2% inoculum concentration access lactobacillus bulgaricus culture medium, 37 DEG C incubated
18h, it is then centrifuged for collecting bacterial sediment, centrifuges to obtain lactobacillus bulgaricus bacterium mud after being washed afterwards with sterile saline again；
Described centrifugation is to centrifuge 15min under 4 DEG C, 6000r/min；
3) prepared by lactobacillus bulgaricus protein in cell wall enzyme：
Phosphate buffer (every liter of phosphate buffer that pH by addition 20mL in every g lactobacillus bulgaricus bacterium mud is 7.0
In glycine containing 15g and 2000U mutanolysin), 35 DEG C insulation 1h after under 4 DEG C, 6000r/min centrifuge 20min obtain supernatant
The as enzyme liquid of protein in cell wall enzyme, small-molecule substance is removed using 1-3KDa ultrafiltration, then adds CaCl in liquid is retained2, control
CaCl processed2Concentration be 8mM, be freeze-dried to obtain lactobacillus bulgaricus protein in cell wall enzyme bacterium powder；
4) preparation of sheep breast active beverage：
Fresh goat milk is adjusted after degreasing pH value to 6.0 add thereto by mass volume ratio 1% bulgarian milk bar
Mycetocyte wall-held protein enzyme bacterium powder and 0.01% CaCl2, 2h is digested at 37 DEG C, then again by mass volume ratio addition 0.5%
Beta-schardinger dextrin, 0.2% hovenia polysaccharides, 0.3% synanthrin, 4% antierythrite, 3% xylitol, 0.1% L- apples
Acid, 0.10% citric acid, obtained containing Bulgaria after 20 DEG C of homogeneous, sterilization after 0.10% sodium citrate is well mixed
The sheep breast active beverage of lactobacillus protein in cell wall enzyme, its ACE inhibitory activity are 72%.
Embodiment 2：
1) prepared by lactobacillus bulgaricus culture medium:
By lactose 20g, dusty yeast 20g, manganese sulfate 15mg, Tween 80 0.8mL, stachyose 2g, peptone 5g, phosphoric acid hydrogen
Disodium 5g, glutamic acid 8mg, alanine 7mg are added in pure water, are settled to 1L, and regulation pH value sterilizes to 6.5 in 121 DEG C
Lactobacillus bulgaricus culture medium；
2) lactobacillus bulgaricus culture and microorganism collection：
By lactobacillus bulgaricus by 4% inoculum concentration access lactobacillus bulgaricus culture medium, 37 DEG C incubated
20h, it is then centrifuged for collecting bacterial sediment, centrifuges to obtain lactobacillus bulgaricus bacterium mud after being washed afterwards with sterile saline again；
Described centrifugation is to centrifuge 25min under 4 DEG C, 7000r/min；
3) prepared by lactobacillus bulgaricus protein in cell wall enzyme：
Phosphate buffer (every liter of phosphate buffer that pH by addition 25mL in every g lactobacillus bulgaricus bacterium mud is 7.0
In glycine containing 20g and 3000U mutanolysin), 40 DEG C insulation 2h after under 4 DEG C, 7000r/min centrifuge 25min obtain supernatant
The as enzyme liquid of protein in cell wall enzyme, small-molecule substance is removed using 1-3KDa ultrafiltration, then adds CaCl in liquid is retained2, control
CaCl processed2Concentration be 12mM, be freeze-dried to obtain lactobacillus bulgaricus protein in cell wall enzyme bacterium powder；
4) preparation of sheep breast active beverage：
Fresh goat milk is adjusted after degreasing pH value to 7.0 add thereto by mass volume ratio 2% bulgarian milk bar
Mycetocyte wall-held protein enzyme bacterium powder and 0.02% CaCl2, 2.5h is digested at 40 DEG C, then again by mass volume ratio addition 0.8%
Beta-schardinger dextrin, 0.3% hovenia polysaccharides, 0.4% synanthrin, 5% antierythrite, 4% xylitol, 0.2% L- apples
Tartaric acid, 0.15% citric acid, obtain containing Bao Jiali after 20 DEG C of homogeneous, sterilization after 0.15% sodium citrate is well mixed
The sheep breast active beverage of sub- lactobacillus protein in cell wall enzyme, its ACE inhibitory activity are 80%.
Embodiment 3：
1) prepared by lactobacillus bulgaricus culture medium:
By lactose 25g, dusty yeast 23g, manganese sulfate 13mg, Tween 80 1.0mL, stachyose 2.5g, peptone 4.5g, phosphorus
Sour disodium hydrogen 5.5g, glutamic acid 10mg, alanine 10mg are added in pure water, are settled to 1L, regulation pH value is to 7.0, in 121
DEG C sterilize to obtain lactobacillus bulgaricus culture medium；
2) lactobacillus bulgaricus culture and microorganism collection：
By lactobacillus bulgaricus by 3% inoculum concentration access lactobacillus bulgaricus culture medium, 41 DEG C incubated
22h, it is then centrifuged for collecting bacterial sediment, centrifuges to obtain lactobacillus bulgaricus bacterium mud after being washed afterwards with sterile saline again；
Described centrifugation is to centrifuge 18min under 8 DEG C, 8000r/min；
3) prepared by lactobacillus bulgaricus protein in cell wall enzyme：
Phosphate buffer (every liter of phosphate buffer that pH by addition 28mL in every g lactobacillus bulgaricus bacterium mud is 7.0
In glycine containing 30g and 4000U mutanolysin), 45 DEG C insulation 1h after under 10 DEG C, 6000r/min centrifuge 30min obtain supernatant
The as enzyme liquid of protein in cell wall enzyme, small-molecule substance is removed using 1-3KDa ultrafiltration, then adds CaCl in liquid is retained2, control
CaCl processed2Concentration be 15mM, be freeze-dried to obtain lactobacillus bulgaricus protein in cell wall enzyme bacterium powder；
4) preparation of sheep breast active beverage：
Fresh goat milk is adjusted after degreasing pH value to 8.0 add thereto by mass volume ratio 3% bulgarian milk bar
Mycetocyte wall-held protein enzyme bacterium powder and 0.03% CaCl2, 4h is digested at 35 DEG C, then again by mass volume ratio addition 1.0%
Beta-schardinger dextrin, 0.35% hovenia polysaccharides, 0.5% synanthrin, 5% antierythrite, 5% xylitol, 0.15% L- apples
Tartaric acid, 0.2% citric acid, obtain containing Bao Jiali after 15 DEG C of homogeneous, sterilization after 0.25% sodium citrate is well mixed
The sheep breast active beverage of sub- lactobacillus protein in cell wall enzyme, its ACE inhibitory activity are 90%.
Embodiment 4：
1) prepared by lactobacillus bulgaricus culture medium:
By lactose 18g, dusty yeast 25g, manganese sulfate 20mg, Tween 80 0.6mL, stachyose 2.3g, peptone 6g, phosphoric acid
Disodium hydrogen 6g, glutamic acid 9mg, alanine 6mg are added in pure water, are settled to 1L, regulation pH value to 6.8, in 121 DEG C of sterilizings
Obtain lactobacillus bulgaricus culture medium；
2) lactobacillus bulgaricus culture and microorganism collection：
By lactobacillus bulgaricus by 5% inoculum concentration access lactobacillus bulgaricus culture medium, 35 DEG C incubated
24h, it is then centrifuged for collecting bacterial sediment, centrifuges to obtain lactobacillus bulgaricus bacterium mud after being washed afterwards with sterile saline again；
Described centrifugation is to centrifuge 23min under 10 DEG C, 6000r/min；
3) prepared by lactobacillus bulgaricus protein in cell wall enzyme：
Phosphate buffer (every liter of phosphate buffer that pH by addition 30mL in every g lactobacillus bulgaricus bacterium mud is 7.0
In glycine containing 25g and 3000U mutanolysin), 42 DEG C insulation 3h after under 6 DEG C, 8000r/min centrifuge 28min obtain supernatant
The as enzyme liquid of protein in cell wall enzyme, small-molecule substance is removed using 1-3KDa ultrafiltration, then adds CaCl in liquid is retained2, control
CaCl processed2Concentration be 10mM, be freeze-dried to obtain lactobacillus bulgaricus protein in cell wall enzyme bacterium powder；
4) preparation of sheep breast active beverage：
Fresh goat milk is adjusted after degreasing pH value to 7.0 add thereto by mass volume ratio 2.5% bulgarian milk
Bacillus protein in cell wall enzyme bacterium powder and 0.04% CaCl2, 3h is digested at 45 DEG C, then again by mass volume ratio addition 0.9%
Beta-schardinger dextrin, 0.4% hovenia polysaccharides, 0.35% synanthrin, 6% antierythrite, 4% xylitol, 0.3% L- apples
Tartaric acid, 0.25% citric acid, obtain containing Bao Jiali after 30 DEG C of homogeneous, sterilization after 0.20% sodium citrate is well mixed
The sheep breast active beverage of sub- lactobacillus protein in cell wall enzyme, its ACE inhibitory activity are 85%.
The hovenia polysaccharides of above example are prepared via a method which：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 bulgaricus protein in cell wall enzyme prepares sheep breast active beverage, it is characterised in that：
1) prepared by lactobacillus bulgaricus culture medium:
By lactose 15-25g, dusty yeast 15-25g, manganese sulfate 10-20mg, Tween 80 0.5-1.0mL, stachyose 1.5-2.5g,
Peptone 4-6g, disodium hydrogen phosphate 4-6g, glutamic acid 5-10mg, alanine 5-10mg are added in pure water, are settled to 1L, adjust
PH value is saved to 5.5-7.0, sterilize to obtain lactobacillus bulgaricus culture medium in 121 DEG C；
2) lactobacillus bulgaricus culture and microorganism collection：
By lactobacillus bulgaricus by 2%-5% inoculum concentration access lactobacillus bulgaricus 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 bulgaricus bacterium mud afterwards；
3) prepared by lactobacillus bulgaricus protein in cell wall enzyme：
By the phosphate buffer that 20-30mL is added in every g lactobacillus bulgaricus bacterium mud, institute is 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 ultrafiltration, then added in liquid is retained
CaCl2, control CaCl2Concentration be 8-15mM, be freeze-dried to obtain lactobacillus bulgaricus protein in cell wall enzyme bacterium powder；
4) preparation of sheep breast active beverage：
Fresh goat milk is adjusted to the bulgarian milk that pH value is added 1-3% to 6.0-8.0 thereto by mass volume ratio after degreasing
The CaCl of bacillus protein in cell wall enzyme bacterium powder and 0.01-0.04%2, 2-4h is digested at 35-45 DEG C, then presses mass volume ratio again
Add 0.5-1.0% beta-schardinger dextrin, 0.2-0.4% hovenia polysaccharides, 0.3-0.5% synanthrin, 4-6% antierythrite,
3-5% xylitol, 0.1-0.3% L MALIC ACID, 0.10-0.25% citric acid, 0.10-0.25% sodium citrate mix
The sheep breast active beverage of the enzyme of protein in cell wall containing lactobacillus bulgaricus is obtained after closing uniformly after homogeneous, sterilization, its ACE suppresses
Activity is 70-90%.
2. the method that lactobacillus bulgaricus protein in cell wall enzyme according to claim 1 prepares sheep breast active beverage, it is special
Sign is：The centrifugation of the step 2) is to centrifuge 15-25min under 4-10 DEG C, 6000-8000r/min.
3. the method that lactobacillus bulgaricus protein in cell wall enzyme according to claim 1 prepares sheep breast active beverage, it is special
Sign is：The pH=7.0 of the phosphate buffer of the step 3), glycine containing 15-30g and 2000- in every liter of phosphate buffer
4000U mutanolysin.
4. the method that lactobacillus bulgaricus protein in cell wall enzyme according to claim 1 prepares sheep breast active beverage, it is special
Sign is：The centrifugation of the step 3) is to centrifuge 20-30min under 4-10 DEG C, 6000-8000r/min.
5. the method that lactobacillus bulgaricus protein in cell wall enzyme according to claim 1 prepares sheep breast active beverage, it is special
Sign is：The homogeneous of the step 4) is that temperature is 15-30 DEG C.
6. the method that lactobacillus bulgaricus protein in cell wall enzyme according to claim 1 prepares sheep breast active beverage, it is special
Sign is：The hovenia polysaccharides preparation process of the step 4) is：Take fresh honey raisin tree to be beaten after cleaning, then add 20 times
The water of honey raisin tree weight, is stirred, and 2h is incubated under the conditions of 90 DEG C, is then filtered and is obtained hovenia polysaccharides extract solution, is cooled to 38
By the Paula enlightening yeast activated liquid of mass volume ratio access 2% after DEG C, 37 DEG C of incubated 24h remove monose in honey raisin trees and double
Sugar, 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
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 n-butanol
Removing protein, then water intaking mutually add chloroform and n-butanol again, obtain hovenia polysaccharides by being freeze-dried after de- albumen three times.
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| 陈合等: "Alcalase酶解羊乳酪蛋白制备ACE抑制肽的研究", 《陕西科技大学学报(自然科学版)》 * |
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