CN110183516A - A kind of preparation method of blood pressure lowering ginkgo nut protein peptides - Google Patents

A kind of preparation method of blood pressure lowering ginkgo nut protein peptides Download PDF

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CN110183516A
CN110183516A CN201910434381.7A CN201910434381A CN110183516A CN 110183516 A CN110183516 A CN 110183516A CN 201910434381 A CN201910434381 A CN 201910434381A CN 110183516 A CN110183516 A CN 110183516A
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ginkgo
blood pressure
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CN110183516B (en
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魏兆军
江力
章建国
马菲菲
胡飞
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Hefei University of Technology
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    • C07K7/00Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
    • C07K7/04Linear peptides containing only normal peptide links
    • C07K7/06Linear peptides containing only normal peptide links having 5 to 11 amino acids
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    • C12P21/00Preparation of peptides or proteins
    • C12P21/06Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products

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Abstract

The invention discloses a kind of preparation methods of blood pressure lowering ginkgo nut protein peptides.Specific step is as follows: will extract crude protein after the shelling of fresh ginkgo nut, pulverization process;Crude protein is through protease hydrolyzed;Enzymolysis liquid ultrafiltration;Ultrafiltration is through object through sephadex chromatography;Chromatographic solution is separated through liquid chromatogram, obtains the gingko protein peptide with blood pressure lowering potentiality, amino acid sequence are as follows: threonine-asparagine-leucine-aspartic acid-tryptophan-tyrosine (TNLDWY).Small peptide prepared by the present invention, angiotensin converting enzyme (ACE) inhibitory activity is high, and cysteine residues are not present in amino acid sequence, does not need the additive of addition protection activity peptide, and preparation process is simple, at low cost, and product quality is high, is suitble to industrialized production.

Description

A kind of preparation method of blood pressure lowering ginkgo nut protein peptides
Technical field
The present invention relates to a kind of preparation methods of blood pressure lowering ginkgo nut protein peptides, belong to agricultural product intensive processing technology neck Domain.
Background technique
Estimate there are 17,500,000 people to die of cardiovascular disease every year according to the World Health Organization, and hypertension is cardiovascular disease The emphasis of prevention and treatment.Blood-pressure drug used at present such as captopril and enalapril are bad due to cough, fash and headache etc. It reacts and is restricted.However, the disease incidence of cardiovascular disease is continuously increased, some new, safe substitutes are needed, are such as eaten Product derive active peptide.Biologically active peptide is the general name for constituting a series of sequences not homopolypeptide of natural amino acid.They have more Kind biological function, such as anti-oxidant, Immune enhancement, hormone control, antibacterial, antithrombotic, antiviral, anti-hypertension.And blood pressure lowering Peptide, also known as angiotensin converting enzyme-inhibiting peptide are that the one kind isolated from food proteins has significant blood pressure reduction effect The short chain substance of polypeptide.The external antihypertensive activity of polypeptide is mainly the inhibitory activity for passing through measurement angiotensin converting enzyme (ACE) Come what is determined.Angiotensin converting enzyme (ACE) is a kind of dipeptidylcarboxypeptidase on cell membrane, is the pass for adjusting blood pressure Key molecule.Blood pressure lowering peptide is development functionality peptide medicament and function due to foodsafety with higher and bioavilability The potential selection of energy property food additives.
Summary of the invention
The purpose of the present invention is to provide a kind of preparation methods of blood pressure lowering ginkgo nut protein peptides.
In order to achieve the above objects and other related objects, present invention provide the technical scheme that a kind of blood pressure lowering ginkgo nut Protein peptides are made of following amino acid residue: threonine-asparagine-leucine-aspartic acid-tryptophan-tyrosine (TNLDWY).
In order to achieve the above objects and other related objects, present invention provide the technical scheme that a kind of blood pressure lowering ginkgo nut The preparation method of protein peptides, including the following steps:
Step 1;Soak degreasing is carried out to ginkgo fruit powder, obtains degreasing ginkgo fruit powder;
Step 2: the protein in degreasing ginkgo fruit powder being extracted using alkali extraction-acid precipitation, is lyophilized, obtains after obtained extracting solution dialysis To ginkgo albumen;
Step 3: albumen being configured to aqueous solution, alkali protease is added and is digested, enzyme deactivation after enzymatic hydrolysis is then centrifuged for taking Supernatant obtains ginkgo protein enzymatic hydrolyzate;
Step 4: hyperfiltration treatment being carried out to ginkgo protein enzymatic hydrolyzate, ultrafiltration permeate is collected and is lyophilized, obtain ginkgo proteolysis Object;
Step 5: ginkgo protolysate being configured to ginkgo protolysate aqueous solution, using sephadex as chromatography media pair Ginkgo protolysate solution is chromatographed, and the sample that retention time is 87.36-102.23min is collected;
Step 6: then the sample elder generation desalination that step 5 is obtained is lyophilized, then be redissolved in trifluoroacetic acid aqueous solution, Then C18 reverse-phase chromatographic column is used, mobile phase a is that volume fraction is 0.1% aqueous formic acid, and mobile phase b is to be containing mass fraction The acetonitrile solution of 0.1% formic acid, wherein the volume fraction of acetonitrile is 84%;After chromatographic column is with a solution equilibria, sample by automatically into Sample device loading, Detection wavelength 228nm;The eluent of 44.23 min of appearance time is collected, obtaining has blood pressure lowering ginkgo nut albumen Peptide solution;Mass spectrometry results show the amino acid sequence of the gingko protein peptide are as follows: threonine-asparagine-leucine-day Aspartic acid-tryptophan-tyrosine.
Preferred technical solution are as follows: in step 1, ginkgo fruit powder is soaked in ethyl acetate and carries out degreasing, degreasing terminates It filters afterwards and removes solvent, obtain degreasing ginkgo fruit powder after dry;The partial size of the ginkgo fruit powder is less than or equal to 40-80 mesh.
Preferred technical solution are as follows: in step 2, degreasing ginkgo fruit powder is dissolved in distilled water, and adjusts pH value to 10, stirred Night takes supernatant after centrifugation, then adjust the pH value of supernatant to 4.62, be centrifuged again, abandon supernatant and take precipitating, sediment is steaming It is lyophilized after dialysing in distilled water, obtains degreasing ginkgo albumen.
Preferred technical solution are as follows: in step 3, degreasing ginkgo albumen is configured to protein solution, adjusts pH value to 10, so After alkali protease, 48-52 DEG C of heat preservation 3.5-4.5 h is added.
Preferred technical solution are as follows: in step 4, using molecular cut off is the ultrafiltration membrane of 3000 Da to ginkgo protease It solves liquid and carries out hyperfiltration treatment.
Preferred technical solution are as follows: in step 5, using G-15 sephadex column to ginkgo protolysate solution into Row chromatography collects the sample that retention time is 87.36-102.23min, and ultrapure water is eluent, flow velocity 1mL/min, loading Concentration is 80 mg/mL.
Since above-mentioned technical proposal is used, the present invention has the advantage, that compared with prior art
Gingko protein peptide prepared by the present invention, angiotensin converting enzyme (ACE) inhibitory activity is high, does not deposit in amino acid sequence In cysteine residues, the additive of addition protection activity peptide is not needed, preparation process is simple, and at low cost, product quality is high, fits Close industrialized production.
Detailed description of the invention
Fig. 1 is ginkgo nut protein zymolyte through sephadex chromatography figure.
Specific embodiment
Embodiments of the present invention are illustrated by particular specific embodiment below, those skilled in the art can be by this explanation Content disclosed by book is understood other advantages and efficacy of the present invention easily.
Please refer to Fig. 1.It should be clear that this specification structure depicted in this specification institute accompanying drawings, ratio, size etc., only to cooperate The revealed content of specification is not intended to limit the invention enforceable so that those skilled in the art understands and reads Qualifications, therefore do not have technical essential meaning, the modification of any structure, the change of proportionate relationship or the adjustment of size, It does not influence still fall in disclosed technology contents under the effect of present invention can be generated and the purpose that can reach It obtains in the range of capable of covering.Meanwhile cited such as "upper", "lower", "left", "right", " centre " and " one " in this specification Term, be merely convenient to being illustrated for narration, rather than to limit the scope of the invention, the change of relativeness or Adjustment, under the content of no substantial changes in technology, when being also considered as the enforceable scope of the present invention.
Blood pressure lowering is investigated with the inhibitory activity to ACE, sample ACE inhibitory activity detection method:
Substrate hippuroyl-histidine-Leucine (HHL, 5 mM) and sample are dissolved in 0.1M borate buffer solution, and (pH8.3 contains Have 0.3M NaCl) in.Then, the substrate of the sample and 150 μ L that take 50 μ L is added in centrifuge tube, in 37 DEG C of water after mixing 5 min of middle preheating.The ACE solution of 0.1U, 37 DEG C of 60 min of incubation are added later.It is whole that 250 μ L of 1M hydrochloric acid is added at the end of reaction Only react, solution is after 0.45 μm of membrane filtration, in intsil ODS-3(25 × 4.6mm, 5 μm of partial sizes) on carry out reversed phase high efficiency Liquid-phase chromatographic analysis.Mobile phase is distilled water: acetonitrile=75:25(v/v, 0.1% trifluoroacetic acid), 0.5 mL/min of flow velocity, detection Wavelength is 228 nm.Using captopril as positive control, distilled water is blank control, and it is as follows to measure its inhibitory activity (%):
ACE inhibitory activity (%)=[(A-B)/A] × 100;
The peak area of hippuric acid when A is blank control, B are the peak area of the hippuric acid of when containing sample.
A kind of embodiment 1: preparation method of blood pressure lowering ginkgo nut protein peptides
(1) pretreatment of raw material: the fresh ginkgo nut of 500 g is shelled, and takes off kind of a skin processing, and chopping is placed in 45 DEG C of freeze-day with constant temperature 24 h are dried in case, cross 60 meshes after pulverization process.
(2) degreasing: taking 220 g ginkgo fruit powders, and 400mL ethyl acetate is added and is impregnated, and every 10min stirring is primary, leaching 2h is steeped, filters and removes solvent, it is dry, obtain degreasing ginkgo powder;
(3) ginkgo protein extraction: taking 200g degreasing ginkgo powder to be dissolved in distilled water, solid-liquid ratio 1:15, and pH is adjusted to 10.0, stirred Night, 8000 g are centrifuged 20 min, take supernatant, then adjust pH to 4.62, and 8000 g are centrifuged 20 min, abandon supernatant and take precipitating, sediment It using distilled water as dialyzate, is lyophilized after dialysis, obtains ginkgo albumen.
(4) it digests: preparing the ginkgo protein solution that 200mL mass fraction is 3%, be placed in 50 DEG C of water-bath dissolutions, pH tune It is 10.0, waits 5 min, be added alkali protease (enzyme activity 4000U), digest 4h, NaOH solution is during which added, keeps pH value of solution It is constant for 10.0, after enzymatic hydrolysis, 5min enzyme deactivation is boiled, pH recalls to neutrality after cooling, and is centrifuged.
(5) ultrafiltration: taking 150 mL enzymolysis liquids to carry out ultrafiltration, and interception 3kDa constantly collects ultrafiltration component, and freezing is dry It is dry.
(6) gel chromatography: the product of step (5) is configured to aqueous solution, using G-15 sephadex column to ultrafiltration object It is further purified, ultrapure water is eluent, and flow velocity 1mL/min, applied sample amount 350mg, loading volume is 4 mL, loading Concentration is 80 mg/mL, collects the chromatographic solution of retention time 87.36-102.23min.
(7) sample is first crossed into the desalination of DEAE cellulose chromatography, be then lyophilized, being redissolved in volume fraction is 0.1% In trifluoroacetic acid solution.Then C18 reverse-phase chromatographic column is used, mobile phase a is 0.1% aqueous formic acid, and mobile phase b is containing 0.1% The acetonitrile solution (acetonitrile 84%) of formic acid.After chromatographic column is with 95% a solution equilibria, sample is by autosampler loading, inspection Survey 228 nm of wavelength, 1 mL/min of flow velocity.The eluent for collecting 44.23 min of appearance time, obtaining blood pressure lowering (has ACE suppression Make active) ginkgo nut albumen peptide solution.A solution is " mobile phase a " -0.1% aqueous formic acid, takes 0.1 mL formic acid and 99.9 The mixing of mL distilled water;The acetonitrile solution of mobile phase b-0.1% formic acid, wherein the volume fraction of acetonitrile is 84%.First by acetonitrile with Water is hybridly prepared into acetonitrile solution according to the volume ratio of 84:16, then takes 0.1 mL formic acid and 99.9 mL acetonitrile solutions mixed It closes.
(8) ACE inhibitory activity of eluent, and the amino acid sequence of LC-MS detection protein peptides are measured.
ACE inhibitory activity testing result is 54.75%(1mg/mL), LC-MS tests and analyzes 44.23 peak min substances Composition sequence are as follows: threonine-asparagine-leucine-aspartic acid-tryptophan-tyrosine (TNLDWY).
A kind of embodiment 2: preparation method of blood pressure lowering ginkgo nut protein peptides
(1) pretreatment of raw material: the fresh ginkgo nut of 700g is shelled, and is taken off kind of a skin processing, is placed in thermostatic drying chamber and is done Dry 30h crosses 80 meshes after pulverization process.
(2) degreasing: taking 350 g ginkgo fruit powders, and 100 mL ethyl acetate are added and are impregnated, and every 10 min stirring is primary, 2h is impregnated, filters and removes solvent, it is dry, obtain degreasing ginkgo powder;
(3) leach protein: taking 300 g degreasing ginkgo powders to be dissolved in distilled water, and solid-liquid ratio 1:18, pH are adjusted to 10, are placed in magnetic agitation 6 h are stirred on device, is centrifuged, takes supernatant, then carries out acid and sinks, and centrifugation, freeze-drying obtains ginkgo albumen.
(4) it digests: preparing 250 mL, 3 % protein solution and be placed in 45 DEG C and dissolve, pH is adjusted to 10.0, and basic protein is added 3500 U of enzyme digests 5 h, during which, NaOH solution is added, and keeping pH value of solution is 10.0 constant, after enzymatic hydrolysis, boils 10 min Enzyme deactivation, pH recalls to 7 after cooling, and is centrifuged, freeze-drying.
(5) ultrafiltration: taking 200 mL enzymolysis liquids to carry out ultrafiltration, and interception 3kDa constantly collects ultrafiltration component, and freezing is dry It is dry.
(6) gel chromatography: applied sample amount 800mg, loading volume 10mL collect retention time 87.36-102.23min Chromatographic solution.
(7) it by after sample desalination and freeze-drying, is redissolved in 0.1% trifluoroacetic acid solution.Using C18 reverse-phase chromatographic column, Mobile phase a is 0.1% aqueous formic acid, and mobile phase b is the acetonitrile solution (acetonitrile 84%) containing 0.1% formic acid.Chromatographic column with After 95% a solution equilibria, sample is by autosampler loading, 228 nm of Detection wavelength, 1 mL/min of flow velocity.When collecting appearance Between 44.23 min eluent;
(8) ACE inhibitory activity of eluent, and the amino acid sequence of LC-MS detection protein peptides are measured.
ACE inhibitory activity testing result is 52.27%(1mg/mL), LC-MS tests and analyzes 44.23 peak min substances Composition sequence are as follows: threonine-asparagine-leucine-aspartic acid-tryptophan-tyrosine (TNLDWY).
A kind of embodiment 3: preparation method of blood pressure lowering ginkgo nut protein peptides
A kind of blood pressure lowering ginkgo nut protein peptides, are made of following amino acid residue: threonine-asparagine-leucine-asparagus fern ammonia Acid-tryptophan-tyrosine (TNLDWY).
Preparation method includes the following steps:
Step 1;Soak degreasing is carried out to ginkgo fruit powder, obtains degreasing ginkgo fruit powder;
Step 2: the protein in degreasing ginkgo fruit powder being extracted using alkali extraction-acid precipitation, is lyophilized, obtains after obtained extracting solution dialysis To ginkgo albumen;
Step 3: albumen being configured to aqueous solution, alkali protease is added and is digested, enzyme deactivation after enzymatic hydrolysis is then centrifuged for taking Supernatant obtains ginkgo protein enzymatic hydrolyzate;
Step 4: hyperfiltration treatment being carried out to ginkgo protein enzymatic hydrolyzate, ultrafiltration permeate is collected and is lyophilized, obtain ginkgo proteolysis Object;
Step 5: ginkgo protolysate aqueous solution (80 mg/mL) being chromatographed using sephadex as chromatography media, is collected Retention time is the sample of 87.36-102.23min;
Step 6: the sample that step 5 is obtained first crosses the desalination of DEAE cellulose chromatography, is then lyophilized, then be redissolved in In 0.1% trifluoroacetic acid aqueous solution of volume fraction, C18 reverse-phase chromatographic column is then used, mobile phase a is that mass fraction is 0.1% first Aqueous acid, mobile phase b are the acetonitrile solution for being 0.1% formic acid containing mass fraction, and wherein the volume fraction of acetonitrile is 84%;Color After column is composed with a solution equilibria, sample is by autosampler loading;The eluent for collecting 44.23 min of appearance time, is had The ginkgo nut albumen peptide solution of ACE inhibitory activity.A solution be " mobile phase a " -0.1% aqueous formic acid, take 0.1 mL formic acid with The mixing of 99.9 mL distilled water;The acetonitrile solution of mobile phase b-0.1% formic acid, wherein the volume fraction of acetonitrile is 84%.First by second Nitrile and water are hybridly prepared into acetonitrile solution according to the volume ratio of 84:16, then take 0.1 mL formic acid and 99.9 mL acetonitrile solutions Mixing.
Preferred embodiment are as follows: in step 1, ginkgo fruit powder is soaked in ethyl acetate and carries out degreasing, degreasing terminates It filters afterwards and removes solvent, obtain degreasing ginkgo fruit powder after dry;The partial size of the ginkgo fruit powder is less than or equal to 40 mesh.
Preferred embodiment are as follows: in step 2, degreasing ginkgo fruit powder is dissolved in distilled water, and adjusts pH value to 10.0, is stirred Overnight, supernatant is taken after centrifugation, then adjust the pH value of supernatant to 4.62, be centrifuged again, abandon supernatant and take precipitating, sediment exists It is lyophilized after dialysing in distilled water, obtains degreasing ginkgo albumen.
Preferred embodiment are as follows: in step 3, degreasing ginkgo albumen is configured to protein solution, adjusts pH value to 10, so After alkali protease, 48 DEG C of 3.5 h of heat preservation are added.
In step 4, molecular cut off is used to carry out hyperfiltration treatment to ginkgo protein enzymatic hydrolyzate for the ultrafiltration membrane of 3000 Da.
Preferred embodiment are as follows: in step 5, ginkgo protolysate is carried out into one using G-15 sephadex column Step purifying, ultrapure water are eluent, and flow velocity 1mL/min, sample concentration is 80 mg/mL.
A kind of embodiment 4: preparation method of blood pressure lowering ginkgo nut protein peptides
(1) pre-process: after the shelling of fresh Semen Ginkgo, peeling, chopping is placed in 45 DEG C of baking oven drying, then is crushed, and crosses 50 Mesh;
(2) degreasing: being impregnated using ethyl acetate, and every 10 min stirring is primary, impregnates 2h, filters and removes solvent, dry, is obtained To degreasing ginkgo powder;
(3) ginkgo protein extraction: degreasing ginkgo powder is dissolved in distilled water, and adjusting pH is 10.0, is stirred overnight, is centrifuged, takes Clearly, pH is tuned into 4.62, is centrifuged again, abandons supernatant and takes precipitating, is lyophilized after sediment dialysis, obtains ginkgo albumen;
(4) it digests: ginkgo albumen is configured to 3% protein solution, adjusting pH is 10.0, and alkali protease is added and adjusts enzyme It lives to 4000U, 50 DEG C of 4 h of heat preservation;
(5) ultrafiltration: using molecular cut off to carry out ultrafiltration for the ultrafiltration membrane of 3000 Da, collects ultrafiltration permeate and is lyophilized;
(6) gel chromatography: being further purified ultrafiltration object using G-15 sephadex column, and ultrapure water is eluent, stream Speed is 1mL/min, and sample concentration is 80 mg/mL, collects the chromatographic solution of retention time 87.36-102.23min;
(7) it liquid chromatogram separation function peptide: after sample desalination and freeze-drying, is redissolved in 0.1% trifluoroacetic acid solution.It adopts With C18 reverse-phase chromatographic column, mobile phase a is 0.1% aqueous formic acid, and mobile phase b is the acetonitrile solution (acetonitrile containing 0.1% formic acid For 84%).After chromatographic column is with 95% a solution equilibria, sample is by autosampler loading, 228 nm of Detection wavelength, 1 mL/ of flow velocity min.The eluent of 44.23 min of appearance time is collected, the ginkgo nut albumen peptide solution with ACE inhibitory activity is obtained;
(8) mass spectrometry results show the amino acid sequence of the gingko protein peptide are as follows: threonine-asparagine-leucine-day Aspartic acid-tryptophan-tyrosine (TNLDWY).
As described above is only to be not intended to tool to explain the preferred embodiments of the invention to do any shape to the present invention Limitation in formula should all wrap therefore all have any modification or change for making the related present invention under identical spirit It includes in the scope that the invention is intended to protect.

Claims (7)

1. a kind of blood pressure lowering ginkgo nut protein peptides, it is characterised in that: be made of following amino acid residue: threonine-asparagine- Leucine-aspartic acid-tryptophan-tyrosine.
2. a kind of preparation method of blood pressure lowering ginkgo nut protein peptides, it is characterised in that: include the following steps:
Step 1;Soak degreasing is carried out to ginkgo fruit powder, obtains degreasing ginkgo fruit powder;
Step 2: the protein in degreasing ginkgo fruit powder being extracted using alkali extraction-acid precipitation, is lyophilized, obtains after obtained extracting solution dialysis To ginkgo albumen;
Step 3: albumen being configured to aqueous solution, alkali protease is added and is digested, enzyme deactivation after enzymatic hydrolysis is then centrifuged for taking Supernatant obtains ginkgo protein enzymatic hydrolyzate;
Step 4: hyperfiltration treatment being carried out to ginkgo protein enzymatic hydrolyzate, ultrafiltration permeate is collected and is lyophilized, obtain ginkgo proteolysis Object;
Step 5: ginkgo protolysate being configured to ginkgo protolysate aqueous solution, using sephadex as chromatography media pair Ginkgo protolysate solution is chromatographed, and the sample that retention time is 87.36-102.23min is collected;
Step 6: then the sample elder generation desalination that step 5 is obtained is lyophilized, then be redissolved in trifluoroacetic acid aqueous solution, Then C18 reverse-phase chromatographic column is used, mobile phase a is that volume fraction is 0.1% aqueous formic acid, and mobile phase b is to be containing mass fraction The acetonitrile solution of 0.1% formic acid, wherein the volume fraction of acetonitrile is 84%;After chromatographic column is with a solution equilibria, sample by automatically into Sample device loading, Detection wavelength 228nm;The eluent of 44.23 min of appearance time is collected, obtaining has blood pressure lowering ginkgo nut albumen Peptide solution;Mass spectrometry results show the amino acid sequence of the gingko protein peptide are as follows: threonine-asparagine-leucine-day Aspartic acid-tryptophan-tyrosine.
3. the preparation method of blood pressure lowering ginkgo nut protein peptides according to claim 2, it is characterised in that:, will be silver-colored in step 1 Apricot fruit powder, which is soaked in ethyl acetate, carries out degreasing, filters after degreasing and removes solvent, obtains degreasing ginkgo fruit powder after dry; The partial size of the ginkgo fruit powder is less than or equal to 40-80 mesh.
4. the preparation method of blood pressure lowering ginkgo nut protein peptides according to claim 2, it is characterised in that: in step 2, degreasing Ginkgo fruit powder is dissolved in distilled water, and adjusts pH value to 10, is stirred overnight, supernatant is taken after centrifugation, then adjust the pH value of supernatant To 4.62, it is centrifuged again, abandons after supernatant takes precipitating, sediment to dialyse in distilled water and be lyophilized, obtain degreasing ginkgo albumen.
5. the preparation method of blood pressure lowering ginkgo nut protein peptides according to claim 2, it is characterised in that: in step 3, will take off Rouge ginkgo albumen is configured to protein solution, adjusts pH value to 10, alkali protease, 48-52 DEG C of heat preservation 3.5-4.5 is then added h。
6. the preparation method of blood pressure lowering ginkgo nut protein peptides according to claim 2, it is characterised in that: in step 4, use The ultrafiltration membrane that molecular cut off is 3000 Da carries out hyperfiltration treatment to ginkgo protein enzymatic hydrolyzate.
7. the preparation method of blood pressure lowering ginkgo nut protein peptides according to claim 2, it is characterised in that: in step 5, use G-15 sephadex column is chromatographed to ginkgo protolysate solution, and collection retention time is 87.36-102.23min Sample, ultrapure water is eluent, flow velocity 1mL/min, and sample concentration is 80 mg/mL.
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Cited By (2)

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Publication number Priority date Publication date Assignee Title
CN112442112A (en) * 2020-11-23 2021-03-05 东北农业大学 Ginkgo protein source ACE inhibitory peptide composition and preparation method and application thereof
CN112940079A (en) * 2021-03-19 2021-06-11 广州明创生物科技有限公司 Rice antihypertensive peptide and enzymolysis preparation method thereof

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CN103122022A (en) * 2013-01-31 2013-05-29 陕西科技大学 Angiotensin converting enzyme inhibitory peptide derived from bitter almond protein and preparation method thereof
CN105131083A (en) * 2015-07-30 2015-12-09 陕西师范大学 Flat almond peptides capable of inhibiting activity of angiotensin converting enzyme (ACE) and preparation method thereof

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103122022A (en) * 2013-01-31 2013-05-29 陕西科技大学 Angiotensin converting enzyme inhibitory peptide derived from bitter almond protein and preparation method thereof
CN105131083A (en) * 2015-07-30 2015-12-09 陕西师范大学 Flat almond peptides capable of inhibiting activity of angiotensin converting enzyme (ACE) and preparation method thereof

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112442112A (en) * 2020-11-23 2021-03-05 东北农业大学 Ginkgo protein source ACE inhibitory peptide composition and preparation method and application thereof
CN112940079A (en) * 2021-03-19 2021-06-11 广州明创生物科技有限公司 Rice antihypertensive peptide and enzymolysis preparation method thereof
CN112940079B (en) * 2021-03-19 2022-09-09 广州明创生物科技有限公司 Rice antihypertensive peptide and enzymolysis preparation method thereof

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