CN106472817A - A kind of bird's nest polypeptide and preparation method thereof - Google Patents
A kind of bird's nest polypeptide and preparation method thereof Download PDFInfo
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- CN106472817A CN106472817A CN201610792966.2A CN201610792966A CN106472817A CN 106472817 A CN106472817 A CN 106472817A CN 201610792966 A CN201610792966 A CN 201610792966A CN 106472817 A CN106472817 A CN 106472817A
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23J3/00—Working-up of proteins for foodstuffs
- A23J3/30—Working-up of proteins for foodstuffs by hydrolysis
- A23J3/32—Working-up of proteins for foodstuffs by hydrolysis using chemical agents
- A23J3/34—Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes
- A23J3/348—Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes of proteins obtained from waste materials
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23J1/00—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites
- A23J1/10—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from hair, feathers, horn, skins, leather, bones, or the like
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/01—Hydrolysed proteins; Derivatives thereof
- A61K38/012—Hydrolysed proteins; Derivatives thereof from animals
Abstract
The invention provides a kind of bird's nest polypeptide, its weight average molecular weight is less than 2000Da；Sialic acid weight percentage is more than 7%.Present invention also offers the preparation method of described bird's nest polypeptide and purposes.The bird's nest polypeptide that the present invention provides, easily digests in human body and absorbs, can quickly beauty care, improve subhealth symptom, and preparation technology is easy, low production cost, and prospects for commercial application is good.
Description
Technical field
The invention belongs to Nidus collocaliae processing technique field is and in particular to a kind of bird's nest polypeptide and preparation method thereof.
Background technology
Nidus collocaliae, number delicacy made from bird's nests, Yan Gen, swallow vegetable, refer to the part swift of Apodiformes Apodidae and the several of aerodramus again
Esculent swift mixes built up nest with saliva and down etc..Nidus collocaliae is mild-natured, sweet in the mouth, tonifying the lung yin nourishing, there is nourishing YIN to relieve dryness, QI invigorating
Effect of invigorating middle warmer, can alleviate the symptoms such as cough, hemoptysis, be just described as nourishing precious sample from the Tang Dynasty, be a kind of and Cornu Cervi Pantotrichum, Radix Ginseng etc.
Build up one's health by taking tonic sample in number together.
In Nidus collocaliae, nutrient substance enriches, including protein, polysaccharide, lipid, trace element and sialic acid etc..Wherein protein
For main component, in Nidus collocaliae, content is up to more than 50%, more than the protein in Nidus collocaliae presented in glycoprotein, molecular weight
It is mainly distributed on 20～150kDa.Sialic acid is an important active component in Nidus collocaliae, the physiology to human body and biochemical function
There is important adjustment effect.Sialic acid content highest contained by Nidus collocaliae in nature, content accounts for more than 10%.Sialic acid is in swallow
Existed with sialoglycoprotein form in nest.
At present in nutrition and cosmetic sample industry, the conventional process of Nidus collocaliae is mainly in the roughing mode of steaming and decocting, this
Substantially original macromolecular protein structures are kept, because protein molecular weight is too big, no in the Nidus collocaliae sample preparation that processing mode obtains
Method is fully absorbed by human body so that the absorption efficiency of Nidus collocaliae is relatively low, therefore passes through suitably protein-modified method for improvement
In Nidus collocaliae protein absorb very necessary.
Presently disclosed protein-modified number carried out to Nidus collocaliae offer, the patent of such as publication number CN1858225A discloses one
Plant the method with ficin enzymatic degradation Nidus collocaliae, obtain the bird's nest polypeptide in 200-1000D for the molecular weight distribution, but do not mention
Sialic content ratio in extract；The patent of publication number CN104498572A discloses a kind of Nidus collocaliae complex enzyme hydrolysis processing work
Skill, enzymolysis is realized by being simultaneously introduced animal protease, neutral protease and food flavor enzyme, and the bird's nest polypeptide obtaining produces sample
Middle sialic acid content is 10.5%, but the number-average molecular weight of product is 5.83*103, weight average molecular weight is 1.89*104, this product
Absorption in human body and availability are still not ideal enough.
Health preserving consciousness with people strengthens, and the consumption demand to Nidus collocaliae is also increasingly vigorous, for meeting people to Nidus collocaliae
Nutrition and functional requirements, find more suitably enzyme solution, obtain micromolecule active polypeptide, to give full play to the nutrition of Nidus collocaliae
It is worth very necessary.
Content of the invention
It is an object of the invention to provide a kind of bird's nest polypeptide containing micromolecule active polypeptide and preparation method thereof.
The invention provides a kind of bird's nest polypeptide, the weight average molecular weight of bird's nest polypeptide is less than 2000Da；Sialic acid weight hundred
Content is divided to be more than 7%.
Wherein, bird's nest polypeptide is prepared by Nidus collocaliae；
And/or, the weight average molecular weight of bird's nest polypeptide is 400～1500Da, and sialic acid weight percentage is 7～12%.
Present invention also offers the preparation method of above-mentioned bird's nest polypeptide, it comprises the steps：
A () takes Nidus collocaliae, extracting in water, obtain Nidus collocaliae protein stock；
B () Nidus collocaliae protein stock to step (a), adjustment temperature, to 30～65 DEG C, adjusts pH value to 8～9, adds 1
～8 weight portion alkaline proteases, react 0.5～12 hour；
C () temperature 30-65 DEG C, adds 1～8 weight portion neutral protease, react 0.5～12 hour；
D 40～55 DEG C of () temperature, adds 1～8 weight portion carbohydrase, react 0.5～12 hour；
(e) enzyme-deactivating；
F () filters, filtrate is concentrated, you can.
Described Nidus collocaliae derives from the nest of several birds of aerodramus：Nidus collocaliae.
Described Nidus collocaliae derives from swift Nidus collocaliae and the domestic Nidus collocaliae that Malaysia produces.
Wherein, in step (a), in described Nidus collocaliae, the weight percentage of protein is not less than 50%, and nitrite contains
Amount is less than 2ppm.
Wherein, in step (a), the bar number of extracting in water is：Take 100 weight portion Nidus collocaliaes, add 800～1200 parts by volume
Water, adjustment temperature, to 70～85 DEG C, is reacted 20～60 minutes；
Further, the bar number of extracting in water is：Take 100 weight portion Nidus collocaliaes, add 800 parts by volume water, adjustment temperature is extremely
70 DEG C, react 20 minutes.
Weight portion and the corresponding relation of parts by volume are " 1g corresponds to 1mL ".
Wherein, in step (c) and/or (d), it is stirred during reaction；
In step (d), before adding carbohydrase, regulation pH value to 5.5～6；
In step (e), enzyme-deactivating adopts：Adjustment temperature to 75～90 DEG C, is incubated 15～30 minutes；Preferably, enzyme-deactivating
Using：Adjustment temperature to 80 DEG C, is incubated 15 minutes；
In step (f), described concentration adopts vacuum spray drying or lyophilisation.
Wherein, after step (f), sterilization steps are also included.
Wherein, described alkaline protease is at least one in alcalase or pancreatin；Described neutral protease is
At least one in A1398, Neutrase enzyme, Flavourzyme；Described carbohydrase is lysozyme, cellulase, chitosanase
At least one.
Present invention also offers above-mentioned bird's nest polypeptide is in preparation food sample, food sample additive, health care food sample and/or medicine sample
Purposes.
Wherein, sample is eaten in described health care, medicine sample is beauty care, improves subhealth state, enhance immunity, promotes repair in trauma
Health care food sample, medicine sample.
The bird's nest polypeptide of present invention preparation, can play the biological efficiency of protein and polysaccharide, to greatest extent in people
Easily digest in body and absorb, can quickly beauty care, improve subhealth symptom, effect is better than commercially available product sample；And the present invention
Bird's nest polypeptide preparation technology is easy, low production cost, can be used for preparation food sample, food sample additive, health care food sample and medicine sample,
It is suitable for use in cosmetic sample industry, prospects for commercial application is good.
Obviously, the above of the radical present invention, according to ordinary technical knowledge and the customary means of this area, without departing from
Under the premise of the present invention above-mentioned basic fundamental thought, modification, replacement or the change of other various ways can also be made.
The specific embodiment of form by the following examples, remakes further specifically to the above of the present invention
Bright.But this scope being interpreted as the above-mentioned theme of the present invention should not be only limitted to Examples below.All based on the above of the present invention
The technology realized belongs to the scope of the present invention.
Brief description
Fig. 1 is the Malaysian bird's nest polypeptide collection of illustrative plates of the present invention；
Fig. 2 is sialic HPLC-UV detection in Nidus collocaliae；
Fig. 3 is the domestic bird's nest polypeptide chromatograms of the present invention.
Specific embodiment
It is described further with embodiment below, but the present invention is not limited to these embodiments.
Present invention process flow process：
Nidus collocaliae study → cleaning → enzymolysis → inactivator → filtration → spray drying (or lyophilizing) → sterilizing of raw material shape；
Or, the Nidus collocaliae study → cleaning → enzymolysis → inactivator → filtration → spray drying (or lyophilizing) of raw material shape, that is,
Can.
Experiment reagent used by the present invention is as follows：
Amylase, cellulase, chitosanase：It is purchased from German AB company；
Alcalase, pancreatin, Neutrase enzyme, Flavourzyme：It is purchased from Novi's letter；
A1398：Purchased from Beijing Fang County enzyme preparation head factory.
The detection method of each composition is as follows：
First, sialic acid detection method：
(1) instrument inspection equipment：
Shimadzu CBM-10A VP PLUS chromatograph of liquid (UV-detector：SPD-10A VP PLUS, infusion pump：LC-
10AVP PLUS), KQ-300DE Ultrasound Instrument (city of Kunshan's Ultrasound Instrument examines company limited), FA2004N electronic balance (Shanghai cyanines Hai Yi
Inspection company limited, d=0.1mg), HH-6 thermostat water bath (Guo Hua electric-examination company limited), 10mL headspace sampling bottle, C18 chromatograph
Post.
(2) reagent：Standard sample sialic acid (N-acetyl-neuraminate)：SIGMA-ALDRICH, 100mg, o-phenylenediamine salt
Hydrochlorate, analyzes pure, sodium bisulfate, and analysis is pure.
(3) sample prepares：Take Nidus collocaliae zymolyte sample and standard sample 10mg, add the sodium bisulfate of 0.5 molar concentration molten
Liquid 1ml, heating in water bath for reaction 30 minutes at 80 DEG C, take out cooling, obtain hydrolyzation sample solution.Take this solution, add 10mg/ml
O-phenylenediamine solution (with the sodium bisulfate dissolving of 0.25 molar concentration) 1ml, then it is placed in 80 DEG C of heating in water bath 40 minutes, take
Go out cooling, be settled to 10ml with water, filter and both obtained.
(4) operational approach：Chromatostrip number：4.6mm×250mm；With 1% tetrahydrofuran aqueous solution (containing 0.2% phosphoric acid)-
Acetonitrile (92:8) it is mobile phase；Detection wavelength is 230nm, and flow velocity is 1.0ml/min, and sampling volume is 20 microlitres.
The preparation of reference standard sample solution：Precision weighs sialic acid (N-acetyl-neuraminate) standard sample 2mg, and supplies sample
Product are equally processed, and the control sample solution of about 0.2mg/ml is obtained.
Calibrating：Draw above-mentioned control sample and 20 microlitres of zymolyte sample solution respectively, injection high performance liquid chromatograph is carried out
Calibrating.
(5) computational methods：
Computing formula：
In formula：
SFor：Main peak peak area in need testing solution chromatogram；SRight：Main peak peak area in reference substance solution chromatogram；
mRight：The sample weighting amount (mg) of reference substance；PRight：The percentage composition of reference substance；mFor：The sample weighting amount (mg) of test sample；
VRight：Reference substance extension rate；VFor：Test sample extension rate (VRight=VFor=10))
2nd, molecular weight detection method：Ninhydrin method inspection degree of hydrolysis and molecular weight
(1) instrument inspection ＆ reagent
Beijing general analysis TU-1900 type ultraviolet-uisible spectrophotometer or the inspection of similar instrument
Pipet (2ml)
Liquid-transfering gun
Volumetric flask (100ml, 50mL)
Color-comparison tube (10ml)
1,2,3-indantrione monohydrate, glycine, Fructose, disodium hydrogen phosphate, potassium dihydrogen phosphate (analysis is pure)
Ethanol
(2) experimental procedure
1st, 20 μ g/mL glycine standard solution：2mg glycine constant volume is in 100mL volumetric flask.
2nd, 1,2,3-indantrione monohydrate developer is prepared：Ninhydrin 0.5g, Fructose 0.3g, disodium hydrogen phosphate 11.2g, potassium dihydrogen phosphate
6g, constant volume to 100mL, slightly heat of solution, low temperature can preserve 1 week.
3rd, the drafting of standard curve：Take 9 brace plug test tubes, the accurate standard solution of drawing of according to the form below has plug test tube to 10mL
In, respectively plus pure water is to 2.0mL, add 1,2,3-indantrione monohydrate developer 1.0mL, use in boiling water bath after mixing and heat 15min, then use rapidly
Cold water is cooled to room temperature, adds 40% ethanol 5mL to shake up, and under 570nm wavelength, inspection absorbance (A) is vertical after placing 15min
Coordinate, content makes standard curve for abscissa.
4th, sample calibrating
4.1 sample solution：Hydrolyzed solution 0.50mL is taken to be settled to 50mL.
4.2 sample calibratings：Pipette samples solution 0.5mL, puts into color-comparison tube (each sample is repeated 2 times), plus pure water
To 2.0mL, add 1,2,3-indantrione monohydrate developer 1.0mL, use in boiling water bath after mixing and heat 15min, be then cooled to cold water rapidly
Room temperature, adds 40% ethanol 5mL to shake up, and places after 15min colorimetric under 570nm, makees reference, calibrating suction with " 0 " number test tube
Luminosity, and content is checked in by standard curve.
Total peptide bond mM number=each peptide bond mM number sum
4.3 results calculate：
Hydrolyzed solution-NH2Content (μm ol/mL)=[C/ (75*V)] * 100
In formula, N hydrolyzed solution protein content (kjeldahl determination)
htotThe peptide bond mM number of every gram of material protein
The content (μ g) that C standard curve checks in
75 glycine molecule amounts
A V sample solution institute corpse or other object for laboratory examination and chemical testing amasss
100 diluted sample multiples
4.4 average peptide chain lengths (PCL) and the calibrating of mean molecule quantity (Mw)：
PCL=1/DH
Mw=110*PCL+18
The preparation of embodiment 1 bird's nest polypeptide of the present invention
1st, raw material：Import Malaysia Nidus collocaliae.
2nd, preparation method：
By 100 grams of Nidus collocaliae studies, wash, plus 1000 milliliters of deionized water be warming up to 30 minutes at 85 DEG C, after be cooled to 45
DEG C, adjust pH value to 8, add 3 grams of alcalase enzymes, degrade 2 hours, with adding 4 grams of A1398 enzyme under batten number, reaction 2 is little
When, Nidus collocaliae dissolving becomes white translucent reactant liquor, adjusts pH value to be 6 afterwards, adds lysozyme, 40 DEG C are reacted 2 hours, Ran Housheng
Temperature, to 80 DEG C, is incubated 15 minutes, so that enzyme is inactivated, filtered while hot.Supernatant is in vacuum 0.06～0.10Mpa, temperature 40～65
DEG C vacuum distillation, last lyophilizing, obtain final product bird's nest polypeptide of the present invention.
3rd, testing result：
(1) polypeptide weight average molecular weight：860Da.
(2) sialic acid content testing result：Sialic acid 8.4%.
(3) total amino acid content：50.63%.The content of each aminoacid is shown in Table 1.
The amino acid content of table 1 bird's nest polypeptide of the present invention
| Aminoacid | Bird's nest polypeptide (i.e. Nidus collocaliae zymolyte 201406) |
| Aspartic acid | 5.16 |
| Threonine | 3.86 |
| Serine | 4.32 |
| Glutamic acid | 4.00 |
| Glycine | 2.22 |
| Alanine | 1.57 |
| Cystine | 1.45 |
| L-Valine | 3.87 |
| Methionine | 0.44 |
| Isoleucine | 1.59 |
| Leucine | 3.99 |
| Tyrosine | 3.10 |
| Phenylalanine | 3.40 |
| Lysine | 2.06 |
| Histidine | 2.09 |
| Arginine | 4.11 |
| Proline | 3.40 |
| Total amino acid content | 50.63 |
The preparation of embodiment 2 bird's nest polypeptide of the present invention
1st, raw material：Import Malaysia Nidus collocaliae.
2nd, preparation method：
By 100 grams of Nidus collocaliae studies, wash, plus 1200 milliliters of deionized water be warming up to 60 minutes at 80 DEG C, after be cooled to 65
DEG C, adjust pH value to 9, add 8 grams of pancreatin, degrade 12 hours, with adding 4 grams of Neutrase enzyme under batten number, react 12 hours,
Nidus collocaliae dissolving becomes white translucent reactant liquor, adds lysozyme, and 40 DEG C are reacted 12 hours, are then warmed up to 90 DEG C, are incubated 30
Minute, so that enzyme is inactivated, filtered while hot.Supernatant in vacuum 0.06～0.10Mpa, 40～65 DEG C of vacuum distillations of temperature, finally
Lyophilizing, obtains final product bird's nest polypeptide of the present invention.
3rd, testing result：
(1) polypeptide weight average molecular weight：956Da.
(2) sialic acid content testing result：Sialic acid 9.2%.
(3) total amino acid content：46.46%.The content of each aminoacid is shown in Table 2.
The bird's nest polypeptide spectrogram of the present invention is shown in Fig. 1, and in Nidus collocaliae, sialic HPLC-UV detection is shown in Fig. 2.
The amino acid content of table 2 bird's nest polypeptide of the present invention
| Aminoacid | Bird's nest polypeptide |
| Aspartic acid | 4.81 |
| Threonine | 3.52 |
| Serine | 3.84 |
| Glutamic acid | 3.51 |
| Glycine | 1.99 |
| Alanine | 1.54 |
| Cystine | 1.27 |
| L-Valine | 3.76 |
| Methionine | 0.42 |
| Isoleucine | 1.58 |
| Leucine | 3.65 |
| Tyrosine | 2.63 |
| Phenylalanine | 3.14 |
| Lysine | 1.96 |
| Histidine | 1.78 |
| Arginine | 3.36 |
| Proline | 3.7 |
| Total amino acid content | 46.46 |
The preparation of embodiment 3 bird's nest polypeptide of the present invention
1st, raw material：Import Malaysia Nidus collocaliae.
2nd, preparation method：
By 100 grams of Nidus collocaliae studies, wash, plus 800 milliliters of deionized water be warming up to 20 minutes at 70 DEG C, after be cooled to 30
DEG C, adjust pH value to 8, add 1 gram of alcalase enzyme, degrade 0.5 hour, with adding 1 gram of Flavourzyme enzyme under batten number,
Reaction 0.5 hour, Nidus collocaliae dissolving becomes white translucent reactant liquor, adds lysozyme, and 40 DEG C are reacted 0.5 hour, then heat up
To 75 DEG C, it is incubated 15 minutes, so that enzyme is inactivated, filtered while hot.Supernatant is in vacuum 0.06～0.10Mpa, 40～65 DEG C of temperature
Vacuum distillation, last lyophilizing, sterilizing, obtain final product bird's nest polypeptide of the present invention.
3rd, testing result
(1) polypeptide weight average molecular weight：1012Da.
(2) sialic acid content testing result：Sialic acid 10.3%.
(3) total amino acid content：56.02%.The content of each aminoacid is shown in Table 3.
The amino acid content of table 3 bird's nest polypeptide of the present invention
| Aminoacid | Bird's nest polypeptide |
| Aspartic acid | 5.34 |
| Threonine | 3.95 |
| Serine | 4.37 |
| Glutamic acid | 3.94 |
| Glycine | 2.16 |
| Alanine | 1.77 |
| Cystine | 1.65 |
| L-Valine | 3.91 |
| Methionine | 0.47 |
| Isoleucine | 1.74 |
| Leucine | 4.22 |
| Tyrosine | 3.84 |
| Phenylalanine | 3.77 |
| Lysine | 2.21 |
| Histidine | 2.01 |
| Arginine | 3.89 |
| Proline | 4.86 |
| Tryptophan | 0.6 |
| Total amino acid content | 56.02 |
The preparation of embodiment 4 bird's nest polypeptide of the present invention
1st, raw material：Domestic Nidus collocaliae.
2nd, preparation method：
Weigh 100 grams of dry sample chunky shape Nidus collocaliaes in 1000 milliliters of beakers, be heated to constant temperature 30 minutes at 80 DEG C, swelling after
It is cooled to 50 DEG C, adjust pH value to 8, add 2.5 grams of pancreatin, react 3 hours；With adding 4 grams of A1398 enzyme under batten number, react 3
Hour, Nidus collocaliae dissolving becomes white translucent reactant liquor, adjusts ph value to be 5.5 afterwards, adds 1-8 gram of cellulase, 55 DEG C of reactions 3
Hour, then it is warmed up to 80 DEG C, is incubated 15 minutes, so that enzyme is inactivated, filtered while hot.Supernatant in vacuum 0.06～0.10Mpa,
40～65 DEG C of vacuum distillations of temperature, last lyophilizing, obtain final product bird's nest polypeptide of the present invention.
3rd, testing result：
(1) yield：66.7%
(2) weight average molecular weight：988Da.
(3) sialic acid content testing result：Sialic acid 7.12%.
(4) total amino acid content：46.46%.The content of each aminoacid is shown in Table 4.
The bird's nest polypeptide spectrogram of the present invention is shown in Fig. 3.
The amino acid content of table 4 bird's nest polypeptide of the present invention
| Aminoacid | Bird's nest polypeptide (Nidus collocaliae zymolyte 20140814) |
| Aspartic acid | 5.18 |
| Threonine | 3.79 |
| Serine | 3.95 |
| Glutamic acid | 3.78 |
| Glycine | 2.19 |
| Alanine | 1.67 |
| Cystine | 1.32 |
| L-Valine | 3.99 |
| Methionine | 0.27 |
| Isoleucine | 1.72 |
| Leucine | 4.07 |
| Tyrosine | 3.59 |
| Phenylalanine | 3.62 |
| Lysine | 2.06 |
| Histidine | 1.88 |
| Arginine | 3.49 |
| Proline | 3.81 |
| Tryptophan | 0.84 |
| Total amino acid content | 52.29 |
The preparation of embodiment 5 bird's nest polypeptide of the present invention
1st, raw material：Domestic Nidus collocaliae.
2nd, preparation method：
Weigh 100 grams of dry sample chunky shape Nidus collocaliaes in 1200 milliliters of beakers, be heated to constant temperature 20 minutes at 70 DEG C, swelling after
It is cooled to 30 DEG C, adjust pH value to 8, add 1 gram of pancreatin, react 0.5 hour；With adding 8 grams of Neutrase enzyme under batten number, instead
Answer 12 hours, Nidus collocaliae dissolving becomes white translucent reactant liquor, adjust ph value to be 6 afterwards, add 8 grams of cellulase, 40 DEG C of reactions
12 hours, then it is warmed up to 90 DEG C, is incubated 15 minutes, so that enzyme is inactivated, filtered while hot.Supernatant in vacuum 0.06～
0.10Mpa, 40～65 DEG C of vacuum distillations of temperature, last lyophilizing, obtain final product bird's nest polypeptide of the present invention.
3rd, testing result：
(1) yield：68.2%
(2) weight average molecular weight：1130Da.
(3) sialic acid content testing result：Sialic acid 9.87%.
(4) total amino acid content：52.67%.The content of each aminoacid is shown in Table 5.
The amino acid content of table 5 bird's nest polypeptide of the present invention
| Aminoacid | Bird's nest polypeptide (i.e. Nidus collocaliae zymolyte 2015) |
| Aspartic acid | 5.25 |
| Threonine | 3.89 |
| Serine | 4.31 |
| Glutamic acid | 3.94 |
| Glycine | 2.2 |
| Alanine | 1.66 |
| Cystine | 1.28 |
| L-Valine | 3.87 |
| Methionine | 0.32 |
| Isoleucine | 1.78 |
| Leucine | 4.1 |
| Tyrosine | 4.03 |
| Phenylalanine | 3.51 |
| Lysine | 2.07 |
| Histidine | 1.91 |
| Arginine | 3.7 |
| Proline | 3.96 |
| Total amino acid content | 52.67 |
With the mode of test example, beneficial effects of the present invention are described below：
The effect experimental of test example 1 bird's nest polypeptide of the present invention
The bird's nest polypeptide of Example 3 preparation, for beauty care, the effect experimental that improves subhealth state, the results are shown in Table 6：
Table 6 bird's nest polypeptide of the present invention and the Contrast on effect of commercially available Nidus collocaliae
From table 6, bird's nest polypeptide of the present invention to beauty care, improve subhealth symptom and have good effect, especially suitable
For middle aged and aged women.With the contrast of commercially available Nidus collocaliae, bird's nest polypeptide of the present invention takes effect quickly, has significant advantage.
To sum up, the bird's nest polypeptide that the present invention provides, can play the biological efficiency of protein and polysaccharide to greatest extent,
Human body easily digests and absorbs, can quickly beauty care, improve subhealth symptom, effect is better than commercially available product sample；And this
Invention bird's nest polypeptide preparation technology is easy, low production cost, and prospects for commercial application is good.
Claims (10)
1. a kind of bird's nest polypeptide it is characterised in that：Its weight average molecular weight is less than 2000Da；Sialic acid weight percentage is more than
7%.
2. the bird's nest polypeptide described in radical claim 1 it is characterised in that：It is prepared by Nidus collocaliae；
And/or, the weight average molecular weight of bird's nest polypeptide is 400～1500Da, and sialic acid weight percentage is 7～12%.
3. the preparation method of the bird's nest polypeptide described in claim 1 or 2, it comprises the steps：
A () takes Nidus collocaliae, extracting in water, obtain Nidus collocaliae protein stock；
B () Nidus collocaliae protein stock to step (a), adjustment temperature, to 30～65 DEG C, adjusts pH value to 8～9, adds 1～8 weight
Amount part alkaline protease, reacts 0.5～12 hour；
C () temperature 30-65 DEG C, adds 1～8 weight portion neutral protease, react 0.5～12 hour；
D 40～55 DEG C of () temperature, adds 1～8 weight portion carbohydrase, react 0.5～12 hour；
(e) enzyme-deactivating；
F () filters, filtrate is concentrated, you can.
4. the preparation method described in radical claim 3 it is characterised in that：In step (a), the weight of protein in described Nidus collocaliae
Amount percentage composition is not less than 50%, and content of nitrite is less than 2ppm.
5. the preparation method described in radical claim 3 it is characterised in that：In step (a), the bar number of extracting in water is：Take 100
Weight portion Nidus collocaliae, adds 800～1200 parts by volume water, and adjustment temperature, to 70～85 DEG C, is reacted 20～60 minutes；
Further, the bar number of extracting in water is：Take 100 weight portion Nidus collocaliaes, add 800 parts by volume water, adjust temperature to 70 DEG C,
Reaction 20 minutes.
6. the preparation method described in radical claim 3 it is characterised in that：In step (c) and/or (d), stirred during reaction
Mix；
In step (d), before adding carbohydrase, regulation pH value to 5.5～6；
In step (e), enzyme-deactivating adopts：Adjustment temperature to 75～90 DEG C, is incubated 15～30 minutes；Preferably, enzyme-deactivating adopts：
Adjustment temperature to 80 DEG C, is incubated 15 minutes；
In step (f), described concentration adopts vacuum spray drying or lyophilisation.
7. the preparation method described in radical claim 3 it is characterised in that：After step (f), also include sterilization steps.
8. the preparation method described in radical claim 3 it is characterised in that：Described alkaline protease is in alcalase or pancreatin
At least one；Described neutral protease is A1398, Neutrase enzyme, at least one in Flavourzyme；Described carbohydrase
It is at least one of lysozyme, cellulase, chitosanase.
9. the bird's nest polypeptide described in claim 1 or 2 eats the use in sample and/or medicine sample in preparation food sample, food sample additive, health care
On the way.
10. the purposes described in radical claim 9 is it is characterised in that described health care food sample, medicine sample are beauty care, improve Asia
Health, enhance immunity, the health care food sample promoting repair in trauma, medicine sample.
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| CN107411069A (en) * | 2017-07-27 | 2017-12-01 | 杭州玖拾伍度鲜生物科技有限公司 | The cubilose product and its preparation technology of a kind of low temperature process |
| CN107485029A (en) * | 2017-07-27 | 2017-12-19 | 杭州玖拾伍度鲜生物科技有限公司 | One kind liquefaction cubilose product and its preparation technology |
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| CN108148880A (en) * | 2018-03-02 | 2018-06-12 | 姚之龙 | A kind of method that sialic acid and oligopeptide mixture are prepared using hair bird's nest as raw material |
| CN108371324A (en) * | 2018-01-18 | 2018-08-07 | 浙江燕宝丽堂电子商务有限公司 | A kind of preparation process of bird's nest essence oral solution |
| CN110257459A (en) * | 2019-05-16 | 2019-09-20 | 广西慧宝源医药科技有限公司 | Bird's nest small-molecular peptides and its instant Gly-His-Lys |
| WO2022032803A1 (en) * | 2020-08-10 | 2022-02-17 | 广西自贸区燕握生物科技有限公司 | Extraction method for edible bird's nest, and extract and product thereof |
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| CN107411069A (en) * | 2017-07-27 | 2017-12-01 | 杭州玖拾伍度鲜生物科技有限公司 | The cubilose product and its preparation technology of a kind of low temperature process |
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| CN107668312B (en) * | 2017-11-22 | 2020-12-29 | 福建农林大学 | Integrated defoaming method for cubilose protein liquid |
| CN108371324A (en) * | 2018-01-18 | 2018-08-07 | 浙江燕宝丽堂电子商务有限公司 | A kind of preparation process of bird's nest essence oral solution |
| CN108148880A (en) * | 2018-03-02 | 2018-06-12 | 姚之龙 | A kind of method that sialic acid and oligopeptide mixture are prepared using hair bird's nest as raw material |
| CN108148880B (en) * | 2018-03-02 | 2021-10-08 | 姚之龙 | Method for preparing sialic acid and oligopeptide mixture by using cubilose as raw material |
| CN110257459A (en) * | 2019-05-16 | 2019-09-20 | 广西慧宝源医药科技有限公司 | Bird's nest small-molecular peptides and its instant Gly-His-Lys |
| CN110257459B (en) * | 2019-05-16 | 2021-04-13 | 广西慧宝源医药科技有限公司 | Bird's nest small molecular peptide and instant peptide powder thereof |
| WO2022032803A1 (en) * | 2020-08-10 | 2022-02-17 | 广西自贸区燕握生物科技有限公司 | Extraction method for edible bird's nest, and extract and product thereof |
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