CN105928750A - Processing method of small red bean isolated protein with high solubility - Google Patents
Processing method of small red bean isolated protein with high solubility Download PDFInfo
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- CN105928750A CN105928750A CN201610224641.4A CN201610224641A CN105928750A CN 105928750 A CN105928750 A CN 105928750A CN 201610224641 A CN201610224641 A CN 201610224641A CN 105928750 A CN105928750 A CN 105928750A
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- semen ormosiae
- ormosiae hosiei
- protein
- microwave treatment
- separation
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- 108090000623 proteins and genes Proteins 0.000 title claims abstract description 169
- 102000004169 proteins and genes Human genes 0.000 title claims abstract description 169
- 238000003672 processing method Methods 0.000 title claims abstract description 18
- 240000001417 Vigna umbellata Species 0.000 title abstract description 9
- 235000011453 Vigna umbellata Nutrition 0.000 title abstract description 9
- 239000000843 powder Substances 0.000 claims abstract description 83
- 238000000034 method Methods 0.000 claims abstract description 40
- 238000001506 fluorescence spectroscopy Methods 0.000 claims abstract description 20
- 238000012545 processing Methods 0.000 claims abstract description 19
- 210000000582 semen Anatomy 0.000 claims description 190
- 238000000926 separation method Methods 0.000 claims description 89
- 239000000523 sample Substances 0.000 claims description 74
- 239000007788 liquid Substances 0.000 claims description 58
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 26
- 239000008367 deionised water Substances 0.000 claims description 21
- 229910021641 deionized water Inorganic materials 0.000 claims description 21
- 238000003556 assay Methods 0.000 claims description 20
- 239000002245 particle Substances 0.000 claims description 20
- 230000002209 hydrophobic effect Effects 0.000 claims description 19
- 238000002360 preparation method Methods 0.000 claims description 16
- 239000000203 mixture Substances 0.000 claims description 15
- 230000020978 protein processing Effects 0.000 claims description 15
- 239000000243 solution Substances 0.000 claims description 15
- 239000008363 phosphate buffer Substances 0.000 claims description 14
- 238000003756 stirring Methods 0.000 claims description 11
- 239000006228 supernatant Substances 0.000 claims description 9
- 238000005259 measurement Methods 0.000 claims description 7
- 229910019142 PO4 Inorganic materials 0.000 claims description 6
- 230000005284 excitation Effects 0.000 claims description 6
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 claims description 6
- 239000010452 phosphate Substances 0.000 claims description 6
- 238000001228 spectrum Methods 0.000 claims description 6
- QCVGEOXPDFCNHA-UHFFFAOYSA-N 5,5-dimethyl-2,4-dioxo-1,3-oxazolidine-3-carboxamide Chemical compound CC1(C)OC(=O)N(C(N)=O)C1=O QCVGEOXPDFCNHA-UHFFFAOYSA-N 0.000 claims description 5
- 102000002322 Egg Proteins Human genes 0.000 claims description 5
- 108010000912 Egg Proteins Proteins 0.000 claims description 5
- 238000001157 Fourier transform infrared spectrum Methods 0.000 claims description 5
- 238000004140 cleaning Methods 0.000 claims description 5
- 238000010790 dilution Methods 0.000 claims description 5
- 239000012895 dilution Substances 0.000 claims description 5
- 235000014103 egg white Nutrition 0.000 claims description 5
- 210000000969 egg white Anatomy 0.000 claims description 5
- 238000004108 freeze drying Methods 0.000 claims description 5
- MOMKYJPSVWEWPM-UHFFFAOYSA-N 4-(chloromethyl)-2-(4-methylphenyl)-1,3-thiazole Chemical compound C1=CC(C)=CC=C1C1=NC(CCl)=CS1 MOMKYJPSVWEWPM-UHFFFAOYSA-N 0.000 claims description 4
- 238000009826 distribution Methods 0.000 claims description 4
- 235000019983 sodium metaphosphate Nutrition 0.000 claims description 4
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 claims description 3
- 238000004458 analytical method Methods 0.000 claims description 3
- 239000008366 buffered solution Substances 0.000 claims description 3
- 229920002301 cellulose acetate Polymers 0.000 claims description 3
- 230000009514 concussion Effects 0.000 claims description 3
- 238000002189 fluorescence spectrum Methods 0.000 claims description 3
- 239000005457 ice water Substances 0.000 claims description 3
- 230000004845 protein aggregation Effects 0.000 claims description 3
- 239000012488 sample solution Substances 0.000 claims description 3
- PXIPVTKHYLBLMZ-UHFFFAOYSA-N sodium azide Substances [Na+].[N-]=[N+]=[N-] PXIPVTKHYLBLMZ-UHFFFAOYSA-N 0.000 claims description 3
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 claims description 3
- 229910021642 ultra pure water Inorganic materials 0.000 claims description 3
- 239000012498 ultrapure water Substances 0.000 claims description 3
- 238000005406 washing Methods 0.000 claims description 3
- 239000000872 buffer Substances 0.000 claims description 2
- 238000000295 emission spectrum Methods 0.000 claims description 2
- 238000003760 magnetic stirring Methods 0.000 claims description 2
- 238000002156 mixing Methods 0.000 claims description 2
- DWNBOPVKNPVNQG-LURJTMIESA-N (2s)-4-hydroxy-2-(propylamino)butanoic acid Chemical compound CCCN[C@H](C(O)=O)CCO DWNBOPVKNPVNQG-LURJTMIESA-N 0.000 claims 1
- 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 1
- UEZVMMHDMIWARA-UHFFFAOYSA-N Metaphosphoric acid Chemical compound OP(=O)=O UEZVMMHDMIWARA-UHFFFAOYSA-N 0.000 claims 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 claims 1
- 238000010521 absorption reaction Methods 0.000 claims 1
- 230000031709 bromination Effects 0.000 claims 1
- 238000005893 bromination reaction Methods 0.000 claims 1
- 238000001816 cooling Methods 0.000 claims 1
- 238000013507 mapping Methods 0.000 claims 1
- 229910052700 potassium Inorganic materials 0.000 claims 1
- 239000011591 potassium Substances 0.000 claims 1
- 229910052708 sodium Inorganic materials 0.000 claims 1
- 239000011734 sodium Substances 0.000 claims 1
- 238000002798 spectrophotometry method Methods 0.000 claims 1
- 238000003860 storage Methods 0.000 claims 1
- 235000018102 proteins Nutrition 0.000 description 126
- 244000046052 Phaseolus vulgaris Species 0.000 description 4
- 235000010627 Phaseolus vulgaris Nutrition 0.000 description 4
- 238000005516 engineering process Methods 0.000 description 4
- IOLCXVTUBQKXJR-UHFFFAOYSA-M potassium bromide Chemical compound [K+].[Br-] IOLCXVTUBQKXJR-UHFFFAOYSA-M 0.000 description 4
- 235000013305 food Nutrition 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- QIVBCDIJIAJPQS-VIFPVBQESA-N L-tryptophane Chemical compound C1=CC=C2C(C[C@H](N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-VIFPVBQESA-N 0.000 description 2
- 108010073771 Soybean Proteins Proteins 0.000 description 2
- QIVBCDIJIAJPQS-UHFFFAOYSA-N Tryptophan Natural products C1=CC=C2C(CC(N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-UHFFFAOYSA-N 0.000 description 2
- 238000000862 absorption spectrum Methods 0.000 description 2
- 230000007547 defect Effects 0.000 description 2
- 238000005187 foaming Methods 0.000 description 2
- 238000002329 infrared spectrum Methods 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 230000026731 phosphorylation Effects 0.000 description 2
- 238000006366 phosphorylation reaction Methods 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 238000001179 sorption measurement Methods 0.000 description 2
- 235000019710 soybean protein Nutrition 0.000 description 2
- 244000068988 Glycine max Species 0.000 description 1
- 235000010469 Glycine max Nutrition 0.000 description 1
- 208000002720 Malnutrition Diseases 0.000 description 1
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 1
- 229910000004 White lead Inorganic materials 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 235000013361 beverage Nutrition 0.000 description 1
- 210000000481 breast Anatomy 0.000 description 1
- 230000003139 buffering effect Effects 0.000 description 1
- 238000010411 cooking Methods 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 230000001934 delay Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 235000013601 eggs Nutrition 0.000 description 1
- 239000000839 emulsion Substances 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 239000006260 foam Substances 0.000 description 1
- 238000001879 gelation Methods 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 230000001071 malnutrition Effects 0.000 description 1
- 235000000824 malnutrition Nutrition 0.000 description 1
- 238000000691 measurement method Methods 0.000 description 1
- 235000020124 milk-based beverage Nutrition 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 235000012149 noodles Nutrition 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 208000015380 nutritional deficiency disease Diseases 0.000 description 1
- 239000011574 phosphorus Substances 0.000 description 1
- 229910052698 phosphorus Inorganic materials 0.000 description 1
- XNGIFLGASWRNHJ-UHFFFAOYSA-L phthalate(2-) Chemical compound [O-]C(=O)C1=CC=CC=C1C([O-])=O XNGIFLGASWRNHJ-UHFFFAOYSA-L 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
- 230000000630 rising effect Effects 0.000 description 1
- 230000014616 translation Effects 0.000 description 1
- 235000013618 yogurt Nutrition 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume, or surface-area of porous materials
- G01N15/02—Investigating particle size or size distribution
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
- G01N21/35—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using infrared light
- G01N21/3563—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using infrared light for analysing solids; Preparation of samples therefor
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
- G01N21/35—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using infrared light
- G01N21/3563—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using infrared light for analysing solids; Preparation of samples therefor
- G01N2021/3572—Preparation of samples, e.g. salt matrices
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
- G01N21/35—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using infrared light
- G01N2021/3595—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using infrared light using FTIR
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N2021/6417—Spectrofluorimetric devices
Abstract
The invention provides a processing method of small red bean isolated protein with high solubility, and mainly overcomes the problems that the small red bean isolated protein prepared by the present processing method of small red bean isolated protein has low solubility, the microwave processing condition is not easy to control and the like. The method includes preparing the original sample, preparing the microwave-processed small red bean isolated protein powder by microwave processing, and determining the secondary structures, intrinsic fluorescence spectroscopy, grain diameters, surface hydrophobicity and solubility of the original sample and the microwave-processed small red bean isolated protein powder to represent the solubility of the small red bean isolated protein. The method has wide application and simple operation, and can save the processing time, and the prepared small red bean isolated protein has high solubility. The method overcomes the problem of low solubility of the original protein.
Description
Technical field
The present invention relates to a kind of highly dissoluble Semen Ormosiae Hosiei separation Protein processing method, belong to raw-food material application technology neck
Territory.
Background technology
Along with progress and the raising of people's living standard of science and technology, the requirement also increasingly stringent to food material, tradition
Bean product, owing to nutrient composition content is relatively low, is therefore eaten for a long time and can cause malnutrition, thus cause serious health to ask
Topic.In recent years, owing to the protein content of Semen Ormosiae Hosiei is higher, and it has the features such as certain dissolubility and foaming characteristic, therefore
One of Semen Ormosiae Hosiei albumen study hotspot becoming home and abroad the most gradually.
In productive life, the fabricated product of Semen Ormosiae Hosiei is the abundantest, and it can be directly used as cooking congee, and cooks Semen Ormosiae Hosiei mud, it is possible to
To be processed into sweetened red bean paste, Semen Ormosiae Hosiei Yoghourt, Semen Ormosiae Hosiei fibre drink, Semen Ormosiae Hosiei milk drink, the most also from Semen Ormosiae Hosiei
Extract natural pigment and utilize Semen Ormosiae Hosiei separation protein production health product etc..The Chinese patent of Publication No. CN103155998A
Disclose a kind of with Semen Ormosiae Hosiei as raw material, the preparation method of processing edibility Semen Ormosiae Hosiei steamed cold noodles.
The simple Semen Ormosiae Hosiei separation albumen extracted has a higher protein content, but modified separates albumen phase with other
Ratio, its dissolubility is poor.And microwave modification is the most popular and easily operated method of modifying, microwave is as a kind of frequency range
At the electromagnetic wave of 300MHz～300GHz, affecting notable on the functional character of protein, this has been done greatly by domestic and international research worker
Quantity research.Cai Jianrong etc. are with soybean protein isolate as raw material, and research microwave treatment time is to its foaming characteristic, foam stability, breast
The property changed and the impact of emulsion stability, show that 1000 watts of microwave processes 40s, can improve the functional characteristic of soybean protein isolate.But
Be existing microwave technology defect be that temperature control is poor, Semen Ormosiae Hosiei separation protein solubility that temperature is wayward to be caused making, steady
The problem of qualitative difference.
Summary of the invention
The present invention proposes a kind of highly dissoluble Semen Ormosiae Hosiei separation Protein processing method, and the method is widely used, easy and simple to handle
And save process time, prepared Semen Ormosiae Hosiei separation albumen has relatively highly dissoluble, overcomes the defect of former protein solubility difference.
For achieving the above object, the present invention uses following technical proposals:
A kind of highly dissoluble Semen Ormosiae Hosiei separation Protein processing method, comprises the steps: first to prepare raw sample, then makes
Semen Ormosiae Hosiei separation protein liquid after standby microwave treatment: Semen Ormosiae Hosiei separated protein powder end lyophilizing obtained adds deionized water,
Both mixing of the ratio with mass volume ratio as 1:10, then it is added thereto under Polymeric sodium metaphosphate. room temperature stirring, obtain Semen Ormosiae Hosiei and divide
From mixed liquid of protein, then Semen Ormosiae Hosiei separation mixed liquid of protein deionized water is constantly cleaned, will clean after described red little
Bean separation mixed liquid of protein and deionized water equal-volume mix and blend, obtain Semen Ormosiae Hosiei separation protein liquid, by described Semen Ormosiae Hosiei
Separating protein liquid and be respectively put in microwave oven with 160 watts, 320 watts or the power of 480 and time were at 5 minutes or the bar of 10 minutes
Processing under part, during microwave treatment, temperature controls at 60-65 DEG C, obtains the Semen Ormosiae Hosiei separation protein liquid after microwave treatment；
Then the Semen Ormosiae Hosiei separation mixed liquid of protein after microwave treatment is carried out lyophilization, be placed in 4 DEG C of refrigerators storing, obtain micro-
Semen Ormosiae Hosiei separated protein powder after ripple processes is last；Respectively to the Semen Ormosiae Hosiei separated protein powder end after raw sample and microwave treatment finally
Carry out the mensuration of secondary structure, intrinsic fluorescence spectroscopy mensuration, particle diameter measuring, surface hydrophobic and deliquescent mensuration,
To characterize Semen Ormosiae Hosiei separation protein solubility.
A kind of highly dissoluble Semen Ormosiae Hosiei separation Protein processing method, is specifically realized by following steps:
A, prepare raw sample
Semen Ormosiae Hosiei separation protein liquid after b, preparation microwave treatment
(1) Semen Ormosiae Hosiei separated protein powder end lyophilizing obtained adds deionized water, described Semen Ormosiae Hosiei separated protein powder end
Mix with the deionized water ratio with mass volume ratio as 1:10, manufacture in beaker；
(2) it is added thereto to 0.1g Polymeric sodium metaphosphate. again, is stirred at room temperature 24h with stirring paddle, obtain Semen Ormosiae Hosiei separation egg
White mixed liquor；
(3) then Semen Ormosiae Hosiei separation mixed liquid of protein deionized water is constantly cleaned, until described Semen Ormosiae Hosiei separation egg
White mixed liquor color is similar with deionized water color；
(4) the Semen Ormosiae Hosiei separation mixed liquid of protein after then cleaning and deionized water equal-volume mix and blend, obtain red
Semen Phaseoli separation protein liquid；
(5) pour described Semen Ormosiae Hosiei separation protein liquid into flat board, put into 160 watts in microwave oven, 320 watts or 480
Power and time process under conditions of 5 minutes or 10 minutes, it is ensured that the sample volume every time processed is equal, and microwave
In processing procedure, needs taking-up in every 5 minutes is put in ice-water bath and is cooled down, it is ensured that temperature is at 60-65 DEG C, after obtaining microwave treatment
Semen Ormosiae Hosiei separation protein liquid；
Semen Ormosiae Hosiei separation protein liquid after c, the microwave treatment described step b obtained carries out lyophilization, is placed in 4
DEG C refrigerator stores, obtains the end of the Semen Ormosiae Hosiei separated protein powder after microwave treatment；
D, the Semen Ormosiae Hosiei separated protein powder end after the microwave treatment of preparation and raw sample are carried out the mensuration of secondary structure, interior
Source property fluorescence spectrometry, particle diameter measuring, surface hydrophobic and deliquescent mensuration；
Described secondary structure assay method is: carry out processing at the microwave obtained by raw sample and different microwave treatment conditions
P is used in being respectively placed in exsiccator in Semen Ormosiae Hosiei separated protein powder end after reason2O5Fully it is dried, respectively weighs 1mg, the most again with 100mg
Potassium bromide is ground tabletting, measures FTIR (fourier transform infrared spectrum), during data acquisition, steams to reduce water
The gas interference to Infrared spectra adsorption, persistently with the N being dried2Drip washing measuring chamber, measuring in wave-number range is 4000-400cm-1Respectively
The absorption spectrum of sample, resolution 4cm during measurement-1, wave number precision 0.01cm-1, scanning times 64 times, ambient temperature 25 DEG C；
Described intrinsic fluorescence spectroscopy assay method is: carries out processing by raw sample and different microwave treatment conditions and obtains
Semen Ormosiae Hosiei separated protein powder end sample after microwave treatment mixes with phosphate buffer same volume ratio respectively, and described phosphate delays
The concentration rushing liquid is 0.01mol/L, pH7.0, and the Semen Ormosiae Hosiei separation protein concentration of preparation is 0.15mg/mL, uses F-4500
The intrinsic fluorescence spectroscopy of fluorescent spectrophotometer assay Semen Ormosiae Hosiei separation albumen；
Described Particle Size Determination Method is: use ZetaPlus Particle Size Analyzer to measure raw sample and different microwave treatment conditions
Carry out hydrodynamic radius and the distribution thereof of the Semen Ormosiae Hosiei separated protein powder end sample after processing the microwave treatment obtained, with dense
Each sample is all diluted to, for the phosphate buffer of 50mmol/L, pH=7.0, the solution that protein concentration is 0.2% by degree, will
0.45 μm cellulose acetate film (water system) crossed by sample after dilution, is at room temperature measured, and takes the meansigma methods measured three times for surveying
Amount result；
Described surface hydrophobic assay method is: weigh respectively at 0.025g raw sample and different microwave treatment conditions
Semen Ormosiae Hosiei separated protein powder end sample after the microwave treatment that reason obtains, is dissolved in 50mL phosphate buffer, described phosphorus respectively
Phthalate buffer concentration is 0.01mol/L, pH=7.0, stirs 1h at ambient temperature, is then centrifuged 30 under 10000r/min
Minute, take supernatant Lowry method and measure protein concentration, and with described phosphate buffer, each sample is diluted the most successively,
Diluted concentration and increases between 0.005-0.5mol/mL in gradient, takes the sample solution 4mL of variable concentrations after dilution, respectively
Add the ANS solution (phosphate buffered solution of described ANS solution employing 0.01mol/L, pH7.0 that 40 μ L concentration are 8mmol/L
Preparation), after concussion, stand 3 minutes, then use the fluorescence intensity of F-4500 fluorescent spectrophotometer assay sample；
Described dissolubility assay method is: weigh 500mg raw sample respectively and different microwave treatment conditions carries out processing
Semen Ormosiae Hosiei separated protein powder end sample after the microwave treatment arrived, is added thereto to 10ml ultra-pure water, respectively at magnetic stirring apparatus
Upper stirring 1h so that it is fully dissolve, subsequently, was centrifuged 30 minutes under the conditions of 4000rpm/ minute, takes 2mL supernatant, add respectively
Enter 2mLNaN3Solution, described supernatant and NaN3The volume ratio of solution is 0.05, preserves as under the conditions of 4 DEG C of refrigerator subsequently, adopts
Solubility and the Semen Ormosiae Hosiei separation protein aggregation of insolubility is measured by Lowry method.
Further, the microwave treatment conditions respectively power in described step b (5) is 160 watts, and the time is 5 minutes；Merit
Rate is 160 watts, and the time is 10 minutes；Power is 320 watts, and the time is 5 minutes；Power is 320 watts, and the time is 10 minutes；Power
Being 480 watts, the time is 5 minutes；Power is 480 watts, and the time is 10 minutes.
Further, the flat board in described step b (5) is use for laboratory 15mm*15mm flat board.
Further, the microwave oven that described step b (5) uses is Panasonic's controllable temperature microwave oven, and optimum temperature controls at 60-
62℃。
Further, the Peakfit Version 4.12 of the collection of illustrative plates Systat in described secondary structure assay method is soft
Part processes, and after process, each sub-peak with secondary structure corresponding relation is: 1610-1640cm-1 is beta sheet；1640-
1650cm-1 is random coil；1650-1660cm-1 is alpha-helix；1660-1700cm-1 is β-corner.
Further, in order to reduce the contribution of tyrosine in described intrinsic fluorescence spectroscopy assay method, fluorescent emission light
Analysis of spectrum is with the Tryptophan fluorescence group within protein molecule as probe, and fluorescence spectrum excitation wavelength is 290nm, dissipates spectrum
Sweep limits is 300-400nm, excites slit and divergent slit width to be 5nm.
Further, excitation wavelength lambda ex=370nm in described surface hydrophobic assay method, emission wavelength lambda em=
490nm, crack is 5nm, maps protein concentration with fluorescence intensity, and initial segment slope is the surface hydrophobic of protein.
The invention has the beneficial effects as follows:
(1) major ingredient that the present invention uses is Semen Ormosiae Hosiei separation albumen, compensate for traditional soybean separation albumen in nutritional labeling
The deficiency of aspect.
(2) present invention use microwave phosphorylation (physics and chemistry combination method of modifying) to be processed, red by extract
Semen Phaseoli separation albumen carries out microwave treatment, thus reaches to improve the purpose of Semen Ormosiae Hosiei separation albumen solubility, it is simple to it is in health care
Application in terms of food, beverage.
(3) the Semen Ormosiae Hosiei separation albumen after microwave phosphorylation is processed, its dissolubility significantly improves, and pollution-free, tool
There are preferable thermodynamics, dynamic performance and gelation, overcome original Semen Ormosiae Hosiei separation albumen and answering in performances such as dissolubilities
By the limitation of aspect, widen it in health care of food, the development prospect of manufacture field.
Accompanying drawing illustrates:
Fig. 1 is the infrared spectrogram of the Semen Ormosiae Hosiei albumen under difference microwave condition of the present invention processes；
Fig. 2 is the Semen Ormosiae Hosiei protein fluorescence spectrogram under difference microwave treatment conditions of the present invention；
Semen Ormosiae Hosiei albumen particle size distribution measuring figure under Fig. 3 difference of the present invention microwave treatment conditions；
Fig. 4 is Semen Ormosiae Hosiei protein surface hydrophobicity mensuration figure under difference microwave treatment conditions of the present invention；
Fig. 5 is Semen Ormosiae Hosiei protein solubility mensuration figure under difference microwave treatment conditions of the present invention.
In Fig. 1-Fig. 5, each sample sequence number represents: 1-raw sample；Semen Ormosiae Hosiei separation protein liquid after 2-microwave treatment
A；Semen Ormosiae Hosiei separation protein liquid B after 3-microwave treatment；Semen Ormosiae Hosiei separation protein liquid C after 4-microwave treatment；5-microwave
Semen Ormosiae Hosiei separation protein liquid D after process；Semen Ormosiae Hosiei separation protein liquid E after 6-microwave treatment；After 7-microwave treatment
Semen Ormosiae Hosiei separation protein liquid F.
Detailed description of the invention
Embodiment 1
A kind of highly dissoluble Semen Ormosiae Hosiei separation Protein processing method, is specifically realized by following steps:
A, prepare raw sample
Semen Ormosiae Hosiei separation protein liquid after b, preparation microwave treatment
(1) Semen Ormosiae Hosiei separated protein powder end lyophilizing obtained adds deionized water, described Semen Ormosiae Hosiei separated protein powder end
Mix with the deionized water ratio with mass volume ratio as 1:10, manufacture in beaker；
(2) it is added thereto to 0.1g Polymeric sodium metaphosphate. again, is stirred at room temperature 24h with stirring paddle, obtain Semen Ormosiae Hosiei separation egg
White mixed liquor；
(3) then Semen Ormosiae Hosiei separation mixed liquid of protein deionized water is constantly cleaned, until described Semen Ormosiae Hosiei separation egg
White mixed liquor color is similar with deionized water color；
(4) the Semen Ormosiae Hosiei separation mixed liquid of protein after then cleaning and deionized water equal-volume mix and blend, obtain red
Semen Phaseoli separation protein liquid；
(5) pour described Semen Ormosiae Hosiei separation protein liquid into use for laboratory 15mm*15mm flat board, be respectively put in microwave oven
Being 160 watts with power, the time is to process under conditions of 5 minutes, and in microwave processing process, needs taking-up in every 5 minutes is put
Enter under ice-water bath cools down, it is ensured that temperature, at 60 DEG C, obtains the Semen Ormosiae Hosiei separation protein liquid after microwave treatment, is labeled as red little
Bean separation protein liquid A；The microwave oven used is Panasonic's controllable temperature microwave oven；
Semen Ormosiae Hosiei separation protein liquid A after c, the microwave treatment described step b obtained carries out lyophilization, is placed in
4 DEG C of refrigerators store, obtains the Semen Ormosiae Hosiei after the Semen Ormosiae Hosiei separated protein powder end after microwave treatment is labeled as microwave treatment and separate
Protein powder A；
D, Semen Ormosiae Hosiei separated protein powder end A and the raw sample after the microwave treatment of preparation is carried out secondary structure mensuration,
Intrinsic fluorescence spectroscopy mensuration, particle diameter measuring, surface hydrophobic and deliquescent mensuration.
Embodiment 2
A kind of highly dissoluble Semen Ormosiae Hosiei separation Protein processing method, is specifically realized by following steps:
Difference from Example 1 is, the treatment conditions of described step b (5) be power be 160 watts, the time is 10 points
Clock, it is ensured that temperature, at 62 DEG C, obtains the Semen Ormosiae Hosiei separation protein liquid after microwave treatment, is labeled as the Semen Ormosiae Hosiei after microwave treatment
Separate protein liquid B.
Described step c obtain microwave treatment after Semen Ormosiae Hosiei separated protein powder end be labeled as microwave treatment after Semen Ormosiae Hosiei divide
From protein powder B.
Described step d is that the Semen Ormosiae Hosiei separated protein powder end B after the microwave treatment to preparation carries out two grades of knots with raw sample
The mensuration of structure, intrinsic fluorescence spectroscopy mensuration, particle diameter measuring, surface hydrophobic and deliquescent mensuration.
Embodiment 3
Difference from Example 1 is, the treatment conditions of described step b (5) be power be 320 watts, the time is 5 points
Clock, it is ensured that temperature, at 61 DEG C, obtains the Semen Ormosiae Hosiei separation protein liquid after microwave treatment, is labeled as the Semen Ormosiae Hosiei after microwave treatment
Separate protein liquid C.
Described step c obtain microwave treatment after Semen Ormosiae Hosiei separated protein powder end be labeled as microwave treatment after Semen Ormosiae Hosiei divide
From protein powder C.
Described step d is that the Semen Ormosiae Hosiei separated protein powder end C after the microwave treatment to preparation carries out two grades of knots with raw sample
The mensuration of structure, intrinsic fluorescence spectroscopy mensuration, particle diameter measuring, surface hydrophobic and deliquescent mensuration.
Embodiment 4
Difference from Example 1 is, the treatment conditions of described step b (5) be power be 320 watts, the time is 10 points
Clock, it is ensured that temperature, at 62 DEG C, obtains the Semen Ormosiae Hosiei separation protein liquid after microwave treatment, is labeled as the Semen Ormosiae Hosiei after microwave treatment
Separate protein liquid D.
Described step c obtain microwave treatment after Semen Ormosiae Hosiei separated protein powder end be labeled as microwave treatment after Semen Ormosiae Hosiei divide
From protein powder D.
Described step d is that the Semen Ormosiae Hosiei separated protein powder end D after the microwave treatment to preparation carries out two grades of knots with raw sample
The mensuration of structure, intrinsic fluorescence spectroscopy mensuration, particle diameter measuring, surface hydrophobic and deliquescent mensuration.
Embodiment 5
Difference from Example 1 is, the treatment conditions of described step b (5) be power be 480 watts, the time is 5 points
Clock, it is ensured that temperature, at 60 DEG C, obtains the Semen Ormosiae Hosiei separation protein liquid after microwave treatment, is labeled as the Semen Ormosiae Hosiei after microwave treatment
Separate protein liquid E.
Described step c obtain microwave treatment after Semen Ormosiae Hosiei separated protein powder end be labeled as microwave treatment after Semen Ormosiae Hosiei divide
From protein powder E.
Described step d is that the Semen Ormosiae Hosiei separated protein powder end E after the microwave treatment to preparation carries out two grades of knots with raw sample
The mensuration of structure, intrinsic fluorescence spectroscopy mensuration, particle diameter measuring, surface hydrophobic and deliquescent mensuration.
Embodiment 6
Difference from Example 1 is, the treatment conditions of described step b (5) be power be 480 watts, the time is 10 points
Clock, it is ensured that temperature, at 62 DEG C, obtains the Semen Ormosiae Hosiei separation protein liquid after microwave treatment, is labeled as the Semen Ormosiae Hosiei after microwave treatment
Separate protein liquid F.
Described step c obtain microwave treatment after Semen Ormosiae Hosiei separated protein powder end be labeled as microwave treatment after Semen Ormosiae Hosiei divide
From protein powder F.
Described step d is that the Semen Ormosiae Hosiei separated protein powder end F after the microwave treatment to preparation carries out two grades of knots with raw sample
The mensuration of structure, intrinsic fluorescence spectroscopy mensuration, particle diameter measuring, surface hydrophobic and deliquescent mensuration.
Semen Ormosiae Hosiei separated protein powder end A after microwave treatment embodiment 1-6 obtained, the Semen Ormosiae Hosiei after microwave treatment divide
Semen Ormosiae Hosiei separated protein powder end C after protein powder B, microwave treatment, the Semen Ormosiae Hosiei separated protein powder end D after microwave treatment,
Semen Ormosiae Hosiei separated protein powder end E after microwave treatment, the Semen Ormosiae Hosiei separated protein powder end F after microwave treatment enter respectively with raw sample
The mensuration of row secondary structure, intrinsic fluorescence spectroscopy mensuration, particle diameter measuring, surface hydrophobic and deliquescent mensuration, tool
Body measurement method is as follows:
(1) secondary structure assay method is: carry out processing at the microwave obtained by raw sample and different microwave treatment conditions
Semen Ormosiae Hosiei after Semen Ormosiae Hosiei separated protein powder end A after reason, the Semen Ormosiae Hosiei separated protein powder end B after microwave treatment, microwave treatment
Semen Ormosiae Hosiei separated protein powder end D after separated protein powder end C, microwave treatment, the Semen Ormosiae Hosiei separated protein powder after microwave treatment end
Semen Ormosiae Hosiei separated protein powder end F after E, microwave treatment uses P in being respectively placed in exsiccator2O5Fully it is dried, respectively weighs 1mg, respectively
It is ground tabletting again with 100mg potassium bromide, measures FTIR (fourier transform infrared spectrum), during data acquisition, for
Reduce the steam interference to Infrared spectra adsorption, persistently with the N being dried2Drip washing measuring chamber, mensuration in wave-number range is
4000-400cm-1The absorption spectrum of each sample, resolution 4cm during measurement-1, wave number precision 0.01cm-1, scanning times 64 times, ring
Border temperature 25 DEG C；Peakfit Version 4.12 software of collection of illustrative plates Systat processes, each Zi Feng and two grades after process
Structure corresponding relation is: 1610-1640cm-1For beta sheet；1640-1650cm-1For random coil；1650-1660cm-1For
Alpha-helix；1660-1700cm-1For β-corner.
Measurement result as it is shown in figure 1, in figure abscissa represent scanned infrared analyzer scanning wavelength scope, vertical coordinate represents
The infrared absorbency of sample, thus, it can be known that the Semen Ormosiae Hosiei after the microwave treatment obtained after different microwave treatment conditions process separates
The secondary structure of protein powder occurs substantially to understand, α spiral and β-bend all raise, and go up high 23% and 17% respectively.
(2) intrinsic fluorescence spectroscopy assay method is: carries out raw sample and different microwave treatment conditions processing and obtains
After Semen Ormosiae Hosiei separated protein powder end A after microwave treatment, the Semen Ormosiae Hosiei separated protein powder end B after microwave treatment, microwave treatment
Semen Ormosiae Hosiei separated protein powder end D after Semen Ormosiae Hosiei separated protein powder end C, microwave treatment, the Semen Ormosiae Hosiei separation egg after microwave treatment
Semen Ormosiae Hosiei separated protein powder end F sample after white lead end E, microwave treatment mixes with phosphate buffer same volume ratio respectively, institute
The concentration stating phosphate buffer is 0.01mol/L, pH 7.0, and the Semen Ormosiae Hosiei separation protein concentration of preparation is 0.15mg/mL,
Use the intrinsic fluorescence spectroscopy of F-4500 fluorescent spectrophotometer assay Semen Ormosiae Hosiei separation albumen；In order to reduce the tribute of tyrosine
Offering, fluorescent emission spectrum analysis is with the Tryptophan fluorescence group within protein molecule as probe, and fluorescence spectrum excitation wavelength is
290nm, dissipating spectrum sweep limits is 300-400nm, excites slit and divergent slit width to be 5nm.
Measurement result as in figure 2 it is shown, in figure abscissa represent fluorescent scanning wave-length coverage, vertical coordinate representative sample is in difference
Fluorescence length in wavelength, thus, it can be known that there is the rising of different condition in fluorescence intensity, when 320 in different wavelength range
Watt, the time is when being under conditions of 10 minutes, reaches maximum fluorescence intensity.
(3) Particle Size Determination Method is: use ZetaPlus Particle Size Analyzer to measure raw sample and different microwave treatment conditions
Carry out the end of the Semen Ormosiae Hosiei separated protein powder after the Semen Ormosiae Hosiei separated protein powder end A after processing the microwave treatment obtained, microwave treatment
After Semen Ormosiae Hosiei separated protein powder end C after B, microwave treatment, the Semen Ormosiae Hosiei separated protein powder end D after microwave treatment, microwave treatment
Semen Ormosiae Hosiei separated protein powder end E, the hydrodynamic radius of Semen Ormosiae Hosiei separated protein powder end F sample after microwave treatment and
Distribution, with the phosphate buffer that concentration is 50mmol/L, pH=7.0, each sample being all diluted to protein concentration is 0.2%
Solution, the sample after dilution is crossed 0.45 μm cellulose acetate film (water system), is at room temperature measured, take measure flat three times
Average is measurement result；
Measurement structure is as it is shown on figure 3, the size of abscissa representative sample molecule in figure, and vertical coordinate representative sample is at list
In the volume of position, the volume fraction of its particle diameter contained, is 320 watts at power, and the time is under 10 minutes microwave treatment conditions, and particle diameter is
Little, reach 3.2 μm, form good setting off with dissolubility, thus further illustrate its dissolubility and be improved.
(4) surface hydrophobic assay method is: weigh respectively at 0.025g raw sample and different microwave treatment conditions
Semen Ormosiae Hosiei separated protein powder end A after the microwave treatment that reason obtains, the Semen Ormosiae Hosiei separated protein powder end B after microwave treatment, microwave
Red little after Semen Ormosiae Hosiei separated protein powder end C after process, the Semen Ormosiae Hosiei separated protein powder end D after microwave treatment, microwave treatment
Semen Ormosiae Hosiei separated protein powder end F sample after bean separated protein powder end E, microwave treatment, is dissolved in 50mL phosphate buffer respectively
In (described phosphate buffering liquid concentration is 0.01mol/L, pH=7.0), stir 1h at ambient temperature, then at 10000r/
It is centrifuged 30 minutes under min, takes supernatant Lowry method and measure protein concentration, and with described phosphate buffer by each sample
Be diluted to the most successively 5 become gradients concentration, be 0.005mol/L respectively, 0.05mol/L, 0.1mol/L, 0.3mol/L and
0.5mol/L, respectively takes sample solution 4mL, and (described ANS solution is adopted to be separately added into the ANS solution that 40 μ L concentration are 8mmol/L
Prepare by the phosphate buffered solution of 0.01mol/L, pH7.0), after concussion, stand 3 minutes, then use F-4500 fluorescence to divide
The fluorescence intensity of light photometric determination sample；Excitation wavelength lambdaex=370nm, emission wavelength lambdaem=490nm, crack is 5nm, with
Protein concentration is mapped by fluorescence intensity, and initial segment slope is the surface hydrophobic of protein.
Measurement result as shown in Figure 4, abscissa representative sample label in figure, the surface hydrophobic of vertical coordinate representative sample,
Result shows, is 320 watts at power, and the time is that under 10 minutes microwave treatment conditions, hydrophobic value reaches the highest, is 125, with former state
Condition ratio improves 50.3%, thus secondary evidence microwave phosphatizing treatment is greatly improved the dissolving of Semen Ormosiae Hosiei separation albumen further
Property.
(5) dissolubility assay method is: weigh 500mg raw sample respectively and different microwave treatment conditions carries out process and obtains
Microwave treatment after Semen Ormosiae Hosiei separated protein powder end A, Semen Ormosiae Hosiei separated protein powder end B after microwave treatment, after microwave treatment
Semen Ormosiae Hosiei separated protein powder end C, the Semen Ormosiae Hosiei separated protein powder end D after microwave treatment, the Semen Ormosiae Hosiei after microwave treatment separate
Semen Ormosiae Hosiei separated protein powder end F sample after protein powder E, microwave treatment, is added thereto to 10ml ultra-pure water, respectively at magnetic force
Stir 1h on agitator so that it is fully dissolve, subsequently, be centrifuged 30 minutes under the conditions of 4000rpm/ minute, take 2mL supernatant,
It is separately added into 2mLNaN3Solution, described supernatant and NaN3The volume ratio of solution is 0.05, protects as under the conditions of 4 DEG C of refrigerator subsequently
Deposit, use Lowry method to measure solubility and the Semen Ormosiae Hosiei separation protein aggregation of insolubility.
Measurement result is as it is shown in figure 5, abscissa representative sample label in figure, and vertical coordinate represents the molten of sample in unit volume
Xie Du, thus, it can be known that when being 320 watts at power, under the conditions of the time is 10 minutes, it is 44% that dissolubility reaches optimal value, with former
Sample is compared dissolubility and is promoted 65%.
Claims (8)
1. a highly dissoluble Semen Ormosiae Hosiei separation Protein processing method, comprises the steps: first to prepare raw sample, then prepares
Semen Ormosiae Hosiei separation protein liquid after microwave treatment: Semen Ormosiae Hosiei separated protein powder end lyophilizing obtained adds deionized water, institute
State Semen Ormosiae Hosiei separated protein powder end to mix with the deionized water ratio with mass volume ratio as 1:10, then be added thereto to Metaphosphoric acid
Stir under sodium room temperature, obtain Semen Ormosiae Hosiei separation mixed liquid of protein, then by Semen Ormosiae Hosiei separation mixed liquid of protein with deionized water not
Disconnected cleaning, the described Semen Ormosiae Hosiei separation mixed liquid of protein after cleaning and deionized water equal-volume mix and blend, obtain Semen Ormosiae Hosiei
Separate protein liquid, described Semen Ormosiae Hosiei separation protein liquid is respectively put in microwave oven with 160 watts, 320 watts or the power of 480
Processing under conditions of 5 minutes or 10 minutes with the time, during microwave treatment, temperature controls, at 60-65 DEG C, to obtain at microwave
Semen Ormosiae Hosiei separation protein liquid after reason；Then the Semen Ormosiae Hosiei separation protein liquid after microwave treatment is carried out lyophilization, and
It is placed in 4 DEG C of refrigerators storage, obtains the end of the Semen Ormosiae Hosiei separated protein powder after microwave treatment；After finally to raw sample and microwave treatment
Semen Ormosiae Hosiei separated protein powder end carry out the mensuration of secondary structure, intrinsic fluorescence spectroscopy mensuration, particle diameter measuring, table respectively
Face hydrophobicity and deliquescent mensuration, to characterize Semen Ormosiae Hosiei separation protein solubility.
2. a kind of highly dissoluble Semen Ormosiae Hosiei separation Protein processing method as claimed in claim 1, is specifically realized by following steps:
A, prepare raw sample
Semen Ormosiae Hosiei separation protein liquid after b, preparation microwave treatment
(1) the Semen Ormosiae Hosiei separated protein powder end that lyophilizing obtained adds deionized water, described Semen Ormosiae Hosiei separated protein powder end with go
The ionized water ratio mixing with mass volume ratio as 1:10, manufactures in beaker；
(2) it is added thereto to 0.1g Polymeric sodium metaphosphate. again, is stirred at room temperature 24h with stirring paddle, obtain Semen Ormosiae Hosiei separation albumen and mix
Close liquid；
(3) then Semen Ormosiae Hosiei separation mixed liquid of protein deionized water is constantly cleaned, until described Semen Ormosiae Hosiei separation albumen mixes
Close liquid color similar with deionized water color；
(4) the Semen Ormosiae Hosiei separation mixed liquid of protein after then cleaning and deionized water equal-volume mix and blend, obtain Semen Ormosiae Hosiei
Separate protein liquid；
(5) pour described Semen Ormosiae Hosiei separation protein liquid into flat board, put into 160 watts in microwave oven, 320 watts or the power of 480
Process under conditions of 5 minutes or 10 minutes with the time, it is ensured that the sample volume every time processed is equal, and microwave treatment
During needs taking-up in every 5 minutes put in ice-water bath under cooling, it is ensured that temperature is at 60-65 DEG C, and obtain after microwave treatment is red
Semen Phaseoli separation protein liquid；
Semen Ormosiae Hosiei separation protein liquid after c, the microwave treatment described step b obtained carries out lyophilization, is placed in 4 DEG C of ice
Case stores, obtains the end of the Semen Ormosiae Hosiei separated protein powder after microwave treatment；
D, to preparation microwave treatment after Semen Ormosiae Hosiei separated protein powder end with raw sample carry out the mensuration of secondary structure, endogenous
Fluorescence spectrometry, particle diameter measuring, surface hydrophobic and deliquescent mensuration；
Described secondary structure assay method is: after carrying out processing the microwave treatment obtained by raw sample and different microwave treatment conditions
Semen Ormosiae Hosiei separated protein powder end be respectively placed in exsiccator in use P2O5Fully it is dried, respectively weighs 1mg, the most again with 100mg bromination
Potassium is ground tabletting, measures fourier transform infrared spectrum, during data acquisition, in order to reduce steam to infrared light
The interference that spectrum absorbs, persistently with the N being dried2Drip washing measuring chamber, measuring in wave-number range is 4000-400cm-1The absorption of each sample
Spectrum, resolution 4cm during measurement-1, wave number precision 0.01cm-1, scanning times 64 times, ambient temperature 25 DEG C；
Described intrinsic fluorescence spectroscopy assay method is: carry out processing the microwave obtained by raw sample and different microwave treatment conditions
Semen Ormosiae Hosiei separated protein powder end sample after process mixes with phosphate buffer same volume ratio respectively, described phosphate buffer
Concentration be 0.01mol/L, pH7.0, the Semen Ormosiae Hosiei separation protein concentration of preparation is 0.15mg/mL, uses F-4500 fluorescence
The intrinsic fluorescence spectroscopy of spectrophotometric determination Semen Ormosiae Hosiei separation albumen；
Described Particle Size Determination Method is: use ZetaPlus Particle Size Analyzer to measure raw sample and different microwave treatment conditions is carried out
Process hydrodynamic radius and the distribution thereof of the Semen Ormosiae Hosiei separated protein powder end sample after the microwave treatment obtained, by concentration be
Each sample is all diluted to the solution that protein concentration is 0.2% by the phosphate buffer of 50mmol/L, pH=7.0, will dilution
After sample cross 0.45 μm cellulose acetate film, be at room temperature measured, take three times measure meansigma methods be measurement result；
Described surface hydrophobic assay method is: weigh 0.025g raw sample respectively and different microwave treatment conditions carries out processing
Semen Ormosiae Hosiei separated protein powder end sample after the microwave treatment arrived, is dissolved in 50mL phosphate buffer, described phosphate respectively
Buffer concentration is 0.01mol/L, pH=7.0, stirs 1h at ambient temperature, is then centrifuged 30 points under 10000r/min
Clock, takes supernatant Lowry method and measures protein concentration, and diluted the most successively by each sample with described phosphate buffer, dilute
Release concentration and to increase in gradient between 0.005-0.5mol/mL, take the sample solution 4mL of variable concentrations after dilution, add respectively
Entering the ANS solution that 40 μ L concentration are 8mmol/L, described ANS solution uses the phosphate buffered solution of 0.01mol/L, pH7.0 to join
System, stands 3 minutes after concussion, then uses the fluorescence intensity of F-4500 fluorescent spectrophotometer assay sample；
Described dissolubility assay method is: weigh 500mg raw sample respectively and different microwave treatment conditions carries out processing and obtains
Semen Ormosiae Hosiei separated protein powder end sample after microwave treatment, is added thereto to 10ml ultra-pure water respectively, stirs on magnetic stirring apparatus
Mix 1h so that it is fully dissolve, subsequently, be centrifuged 30 minutes under the conditions of 4000rpm/ minute, take 2mL supernatant, be separately added into
2mLNaN3Solution, described supernatant and NaN3The volume ratio of solution is 0.05, preserves as under the conditions of 4 DEG C of refrigerator subsequently, uses
Lowry method measures solubility and the Semen Ormosiae Hosiei separation protein aggregation of insolubility.
3. a kind of highly dissoluble Semen Ormosiae Hosiei separation Protein processing method as claimed in claim 2, it is characterised in that described step
Microwave treatment conditions respectively power in b (5) is 160 watts, and the time is 5 minutes；Power is 160 watts, and the time is 10 minutes；Merit
Rate is 320 watts, and the time is 5 minutes；Power is 320 watts, and the time is 10 minutes；Power is 480 watts, and the time is 5 minutes；Power is
480 watts, the time is 10 minutes.
4. a kind of highly dissoluble Semen Ormosiae Hosiei separation Protein processing method as claimed in claim 2, it is characterised in that described step
Flat board in b (5) is use for laboratory 15mm*15mm flat board.
5. a kind of highly dissoluble Semen Ormosiae Hosiei separation Protein processing method as claimed in claim 2, it is characterised in that described step
The microwave oven that b (5) uses is Panasonic's controllable temperature microwave oven, and optimum temperature controls at 60-62 DEG C.
6. a kind of highly dissoluble Semen Ormosiae Hosiei separation Protein processing method as claimed in claim 2, it is characterised in that described two grades
Peakfit Version 4.12 software of the collection of illustrative plates Systat in structure determination method processes, after process each sub-peak with
Secondary structure corresponding relation is: 1610-1640cm-1For beta sheet；1640-1650cm-1For random coil；1650-1660cm-1For alpha-helix；1660-1700cm-1For β-corner.
7. a kind of highly dissoluble Semen Ormosiae Hosiei separation Protein processing method as claimed in claim 2, it is characterised in that described endogenous
In order to reduce the contribution of tyrosine in property fluorescence spectrometry method, fluorescent emission spectrum analysis is with the color within protein molecule
Propylhomoserin fluorophor is probe, and fluorescence spectrum excitation wavelength is 290nm, and dissipating spectrum sweep limits is 300-400nm, excites narrow
Seam and divergent slit width are 5nm.
8. a kind of highly dissoluble Semen Ormosiae Hosiei separation Protein processing method as claimed in claim 2, it is characterised in that described surface
Excitation wavelength lambda in hydrophobicity assay methodex=370nm, emission wavelength lambdaem=490nm, crack is 5nm, with fluorescence intensity to egg
White concentration mapping, initial segment slope is the surface hydrophobic of protein.
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