CN102391361B - Chickpea peptide with antioxidation effect and separation and purification method and application - Google Patents
Chickpea peptide with antioxidation effect and separation and purification method and application Download PDFInfo
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Abstract
The invention discloses a chickpea peptide with antioxidation effect and a separation and purification method and an application; the chickpea peptide is expressed as CPe-III, and has a molecular weight of 1155 Da; the amino acid composition is Arg-Gln-Ser-His-Phe-Ala-Asn-Ala-Gln-Pro; and the secondary structure comprises 66% of alpha-helix. The method of the invention has a mild separation condition, a high sample recovery rate, high reproducibility of experimental results, and simple and economical equipment. The method of the invention has a simple process, and low energy consumption; the product has antioxidation effect; and basis is provided for artificial synthesis of antioxidant peptides.
Description
Technical field
The invention belongs to the preparing technical field of biologically active peptides, relate to a kind of there is antioxygenation chickpea peptides and separation purification method and purposes.
Background technology
Chickpea peptides is hydrolyzed by Chickpea Protein the product obtained, and its amino acid composition is identical with Chickpea Protein, and Coordinate specific of essential amino acid rationally, is easily absorbed by the body utilization, has the special and functional performance of the processing more superior than its albumen simultaneously.Chickpea peptides molecular weight is little, be easy to digest and assimilate, without protein denaturation, without fishy smell, soluble in water, in acid condition not coagulation, viscosity little, be heated and the processing characteristics such as do not solidify, but also have reduce serum cholesterol content, promote metabolism of fat, the functional performance such as hypotensive, resistance to reduction, rapidly alleviating fatigue.From garbanzo, antibacterial peptide, low irritability protein zymolyte, angiotensin inhibiting peptide etc. are isolated at present.
Anti-oxidation peptide is the one of biologically active peptides, the people such as Yoshinori Mine think that the oxidation-resistance of peptide at least can other biological activity of some effects, large quantity research has shown the oxidation-resistance of peptide and immunomodulatory, hypotensive, decreasing cholesterol, opioid antagonist activity, antitumor etc. has had certain contacting, and may be the basis of other physiological function.It by reducing oxyradical, hydroxy radical qiao, thus reaches antidotal function.The polypeptide with biological function is all ordered structure, is all made up of certain amino acid per-cent, puts in order and special higher structure.Therefore, from garbanzo, isolate the polypeptide with oxidation-resistance, and structure, its activity is resolved, both to the applications expanding road of Natural Antioxidant Peptides, also for synthetic anti-oxidation peptide establishes solid basis, the advantage that it is applied in fields such as medicine, food, feeds is demonstrated.
Gel filtration chromatography utilizes the molecular sieve effect with cancellated gel, and the molecular size according to separated object is separated.Separation condition is gentle, sample recovery rate is high, experimental result circulation ratio is high, simple equipments economy.Mass spectroscopy is considered to measure the accurately the sensitiveest method of small molecules molecular weight at present.In recent years, along with every technical development, the molecular weight ranges that mass spectrum can measure improves greatly.The laser desorption ionisation flight time mass spectrum of Matrix-assisted becomes the effective tool measuring biomacromolecule especially protein, polypeptide molecular weight and primary structure.The infrared spectra of material has the characteristic of height, absorption peak in spectrogram is corresponding with the vibration mode of group each in molecule, can be used for structure and the chemical bond of studying molecule, differentiating the secondary structure of polypeptide, is the novel analysis and testing technology of one that developed recently gets up.
Summary of the invention
The object of this invention is to provide a kind of chickpea peptides with antioxygenation.
Second object of the present invention is to provide a kind of separation purification method with the chickpea peptides of antioxygenation.
3rd object of the present invention is to provide a kind of purposes with the chickpea peptides of antioxygenation.
Have a chickpea peptides for antioxygenation, represent with CPe-III, the molecular weight of described CPe-III is 1155Da, and amino acid consists of Arg-Gln-Ser-His-Phe-Ala-Asn-Ala-Gln-Pro, and secondary structure accounts for 66% with alpha-helix.
There is a separation purification method for the chickpea peptides of antioxygenation, comprise the steps:
(1) ratio being 1: 8-15 in mass ratio by the olecranon bean powder after degreasing and water mixes, pH=8.3 is adjusted with the NaOH aqueous solution, stir and extract 0.5-1.5h, the centrifugal 15min-25min of 3500r/min-5000r/min, the ratio that precipitation is 1: 3-7 in solid-liquid mass ratio is again extracted 2-3 time, merge supernatant liquor, pH=5.0 protein precipitation is adjusted with the HCl aqueous solution, isolate supernatant liquor, again pH=3.7 is adjusted to carry out secondary protein precipitation with the HCl aqueous solution supernatant liquor, abandon supernatant, merge twice precipitation, precipitate 2-3 time with deionized water wash, pH=7.0 is adjusted with the NaOH aqueous solution, stirring makes precipitation redissolve, lyophilize is garbanzo white protein,
(2) described garbanzo white protein is dissolved in distilled water makes the garbanzo white protein aqueous solution that mass concentration is 3-5%; Be 0.3-0.4AU/g in Sumizyme MP Alcalase (NOVO company of Denmark) and the albuminised ratio of garbanzo, Alcalase enzyme is put into the described garbanzo white protein aqueous solution, at pH=8.0, under the condition of temperature of reaction 45-55 DEG C, hydrolysis 50-70min; Be 50LAPU/g in Flavourzyme composite flavor enzyme (NOVO company of Denmark) and the albuminised ratio of garbanzo, then add Flavourzyme enzyme, pH=7.0, under the condition that temperature of reaction is 50 DEG C, hydrolysis 90-120min; Enzymolysis solution is heated to boiling, go out enzyme 10min, and after cooling, namely the lyophilize of centrifuging and taking supernatant liquor obtains chickpea peptides powder;
(3) the dextrane gel Sephdex G-25 will handled well, dress post 1.6 × 50cm, by registering instrument recorded stream fluid absorbancy situation, water elution to be distilled is steady to baseline, get chickpea peptides powder dissolution in distilled water after join in described dextrane gel Sephdex G-25 post, with distilled water wash-out, flow velocity is 0.6mL/min, elutriant absorbancy is detected at 280nm place, there are 3 elution peaks altogether at times, collect the 3rd elution peak liquid, after lyophilize, namely obtain the chickpea peptides represented with CPe-III with antioxygenation.
Above-mentioned chickpea peptides is preparing the purposes of antioxidant health-care product.
The invention has the advantages that: utilize Sephdex G-25 the peptides separation of different molecular weight in crude product chickpea peptides can be opened, obtain the CPe-III with antioxidant effect, separation condition is gentle, sample recovery rate is high, experimental result circulation ratio is high, simple equipments economy.Utilize LC/MS, to CPe-III separation and purification further, and the information such as relative molecular mass and amino acid composition can be obtained.Finally by infrared spectra, primary structure is verified, and draw the information of secondary structure.Can resolve from the Antioxidation Mechanism of structure level to CPe-III, to the application blaze the trail of Natural Antioxidant Peptides, also for synthetic anti-oxidation peptide establishes solid basis.
Separation and purification anti-oxidation peptide from chickpea peptides also measures the method for its structure.The method process is simple, and energy consumption is low, and product has oxidation resistant effect, and provides foundation to the synthetic of anti-oxidation peptide.
Accompanying drawing explanation
Fig. 1 is chickpea peptides chromatography collection of illustrative plates.
Fig. 2 is CPe-III chromatograms.
Fig. 3 is CPe-III first mass spectrometric collection of illustrative plates.
Fig. 4 is CPe-III second order ms collection of illustrative plates.
Fig. 5 is the infrared spectrogram of CPe-III.
Fig. 6 is the mensuration of each component reducing power of chickpea peptides.
Fig. 7 is the ability that each component of chickpea peptides removes DPPH free radical.
Fig. 8 is the ability that each component of chickpea peptides removes 0H free radical.
Embodiment
Embodiments of the invention understand the present invention better to enable those skilled in the art to, and can not impose any restrictions the present invention.
Below in conjunction with specific embodiment, the present invention is further illustrated.
Embodiment 1
There is a separation purification method for the chickpea peptides of antioxygenation, comprise the steps:
(1) ratio being 1: 10 in mass ratio by the olecranon bean powder after degreasing and water mixes, pH=8.3 is adjusted with the NaOH aqueous solution, stir and extract 1h, the centrifugal 20min of 4000r/min, the ratio that precipitation is 1: 5 in solid-liquid mass ratio again extracts 2 times, merge supernatant liquor, pH=5.0 protein precipitation is adjusted with the HCl aqueous solution, isolate supernatant liquor, again pH=3.7 is adjusted to carry out secondary protein precipitation with the HCl aqueous solution supernatant liquor, abandon supernatant liquor, merge twice precipitation, 3 times are precipitated with deionized water wash, pH=7.0 is adjusted with the NaOH aqueous solution, stirring makes precipitation redissolve, lyophilize 24h is garbanzo white protein,
(2) described garbanzo white protein is dissolved in distilled water makes the garbanzo white protein aqueous solution that mass concentration is 4.87%; Be 0.38AU/g in Alcalase enzyme and the albuminised ratio of garbanzo, Alcalase enzyme put into the described garbanzo white protein aqueous solution, at pH=8.0, under the condition that temperature of reaction is 50 DEG C, hydrolysis 70min; Be 50LAPU/g in Flavourzyme enzyme and the albuminised ratio of garbanzo, then add Flavourzyme enzyme, at pH=7.0, temperature of reaction 50 DEG C, condition under, hydrolysis 100min; Enzymolysis solution is heated to boiling, go out enzyme 10min, and after cooling, namely the lyophilize of centrifuging and taking supernatant liquor obtains chickpea peptides powder;
(3) the dextrane gel Sephdex G-25 will handled well, dress post 1.6 × 50cm, by registering instrument recorded stream fluid absorbancy situation, water elution to be distilled is steady to baseline, get chickpea peptides powder dissolution in distilled water after join in described dextrane gel Sephdex G-25 post, with distilled water wash-out, flow velocity is 0.6mL/min, elutriant absorbancy is detected at 280nm place, there are 3 elution peaks altogether at times, use CPe-I respectively, CPe-II and CPe-III represents, collect the 3rd elution peak liquid, namely the chickpea peptides represented with CPe-III with antioxygenation is obtained after lyophilize, see Fig. 1 chromatography collection of illustrative plates.
LC-MS (HLPC-MS) determines the primary structure of CPe-III:
Chromatographic condition: sample size: 5 μ L; Moving phase: A liquid: water (0.1%TFA); B liquid: acetonitrile (0.1%TFA); Column temperature: room temperature; Flow velocity: 0.7mL/min.
Mass Spectrometry Conditions: adopt electron spray ionisation (ESI), spray voltage 4.5kv, atomization gas (N
2) flow 35 arbitrary value units, assisted gas (N
2) flow 5 arbitrary value units, capillary temperature 300 DEG C; Full scan scope: 150m/z-2000m/z; Carry out positive ion detecting pattern; Carry out online MS/MS, intensity threshold 10
5, ion width 2, collision normalized energy 30%.Application BioWorks3.3 software analysis patch information, thus obtain the primary structural information of CPe-III, molecular weight: 1155Da, amino acid forms: Arg-Gln-Ser-His-Phe-Ala-Asn-Ala-Gln-Pro.See Fig. 2 main component of CPe-III (retention time to be peak corresponding to 3.55 places be).See Fig. 3 (relative molecular mass that first mass spectrometric obtains CPe-III is 1155Da).See that (second order ms determines the amino acid composition of CPe-III to Fig. 4: Arg-Gln-Ser-His-Phe-Ala-Asn-Ala-Gln-Pro)
Infrared spectra detects primary structure, and infers the secondary structure of oxidation-resistance CPe-III:
CPe-III and KBr mixed pressuring plate, adopt total reflection spectrum assay method (ATR), European sampler samples 32 times.At 4000cm
-1-400cm
-1scan in scope.See Fig. 5.(Fig. 5-1 and Fig. 5-2 pairs of acid amides I district bands of a spectrum carry out spectrum Fourier from decurl with ask second derivative, Spectrum Analysis software OMNIC is used to carry out Gauss/Lorentz fit to CPe-III original spectrum band, calculate peak area, learn that CPe-III secondary structure is based on alpha-helix, accounts for 66%.
The mensuration of each component reducing power of chickpea peptides
Reducing power measuring method: get the certain density chickpea peptides solution of 1mL, add the PBS buffered soln of 2.5mL 0.2mol/L pH6.6 and the potassium ferricyanide solution of 2.5mL 1%, abundant mixing, react 20min in 50 DEG C of water-baths after, rapid cooling, add the trichoroacetic acid(TCA) of 2.5mL 10% again, in the centrifugal 10min of 3000r/min.Go precipitation, get supernatant liquor 2mL, add the FeCl of 2mL distilled water and 0.4mL 0.1% successively
3, fully mix, after leaving standstill 10min, in 700nm wavelength place colorimetric.
As can be seen from Figure 6, the reducing power of CPe-III is apparently higher than component CPe-I and II, and in the concentration range of 0.1 ~ 25mg/mL, along with concentration increases, reducing power significantly increases.
The each component of chickpea peptides removes the ability of DPPH free radical:
Remove the measuring method of DPPH free radical: the CPe solution getting 1mL different concns mixes with 1mL 0.1mmol/LDPPH reference liquid and concuss, and at room temperature lucifuge is reacted for some time (about 20min), then surveys light absorption value A with 517nm place
i.Sample blank group replaces DPPH reference liquid with equal-volume dehydrated alcohol, then surveying light absorption value with 517nm place, is designated as A
j, control group replaces sample solution with equal-volume distilled water, then surveying light absorption value with 517nm place, is designated as A
0.Clearance rate calculates according to the following formula:
Clearance rate (%)=[1-(A
i-A
j)/A
0] × 100%
Wherein A
o, A
i, A
jrepresent control group respectively, the absorbancy of sample sets and sample blank group
As shown in Figure 7, chickpea peptides respectively collects part all has significant scavenging(action) to DPPH free radical.In the scope of concentration 0.2 ~ 5mg/mL, along with the increase of concentration, the ability removing DPPH free radical also increases to some extent.Under same concentrations, the ability that chickpea peptides respectively collects the removing DPPH free radical of part is as follows: CPe-III > CPe-II > CPe-I.CPe-III is in the scope of 0.1 ~ 1mg/mL, and increase with concentration, clearance rate significantly increases, and shows obvious dose-effect relationship, and in the concentration range of 1 ~ 5mg/mL, clearance rate tends towards stability.
The each component of chickpea peptides removes the ability of 0H free radical:
Measuring method to 0H Scavenging activity: add 0.4mL 50mmol/L phosphate buffered saline buffer (pH=7.5) in the test tube of drying successively, 0.1mL certain density chickpea peptides solution (blank tube replaces chickpea peptides solution with isopyknic distilled water), the EDTA solution of 0.1mL 1.04mmol/L, 0.1mL 10mmol/L H
2o
2, 0.1mL 2mmol/L VC, 0.1mL 60mmol/L ribodesose (DR) (control group does not add DR, supplements distilled water), 0.1ml 1mmol/L FeC1
3each pipe final volume 1.0ml, take out after the water bath with thermostatic control being placed in 37 DEG C is incubated 1h, add rapidly 1mL 25% (v/v) HCI termination reaction, add 1mL1% thiobarbituricacidα-again, boiling water bath reaction 15min, cools after taking-up immediately, measure light absorption value in wavelength 532nm place, be calculated as follows clearance rate:
0H clearance rate (%)={ [A
0-(A
1-A
2)]/A
0} × 100
Wherein A
0, A
1, A
2represent the absorbancy of control group, sample sets and sample blank group respectively
As seen from Figure 8, chickpea peptides respectively collects part all has significant scavenging(action) to 0H free radical, and shows obvious dose-effect relationship, and in the concentration range of 0.1 ~ 20mg/mL, the Scavenging activity of CPe-III is all higher than component CPe-I and II.
Embodiment 2
There is a separation purification method for the chickpea peptides of antioxygenation, comprise the steps:
(1) ratio being 1: 8 in mass ratio by the olecranon bean powder after degreasing and water mixes, pH=8.3 is adjusted with the NaOH aqueous solution, stir and extract 1.5h, the centrifugal 15min of 5000r/min, the ratio that precipitation is 1: 3 in solid-liquid mass ratio again extracts 3 times, merge supernatant liquor, pH=5.0 protein precipitation is adjusted with the HCl aqueous solution, isolate supernatant liquor, again pH=3.7 is adjusted to carry out secondary protein precipitation with the HCl aqueous solution supernatant liquor, abandon supernatant liquor, merge twice precipitation, 2 times are precipitated with deionized water wash, pH=7.0 is adjusted with the NaOH aqueous solution, stirring makes precipitation redissolve, lyophilize 24h is garbanzo white protein,
(2) described garbanzo white protein is dissolved in distilled water makes the garbanzo white protein aqueous solution that mass concentration is 3%; Be 0.4AU/g in Alcalase enzyme and the albuminised ratio of garbanzo, Alcalase enzyme put into the described garbanzo white protein aqueous solution, at pH=8.0, under the condition that temperature of reaction is 45 DEG C, hydrolysis 60min; Be 50LAPU/g in Flavourzyme enzyme and the albuminised ratio of garbanzo, add Flavourzyme enzyme, at pH=7.0, temperature of reaction 50 DEG C, condition under, hydrolysis 100min; Enzymolysis solution is heated to boiling, go out enzyme 10min, and after cooling, namely the lyophilize of centrifuging and taking supernatant liquor obtains chickpea peptides powder.
(3) with embodiment 1.
Embodiment 3
There is a separation purification method for the chickpea peptides of antioxygenation, comprise the steps:
(1) ratio being 1: 15 in mass ratio by the olecranon bean powder after degreasing and water mixes, pH=8.3 is adjusted with the NaOH aqueous solution, stir and extract 0.5h, the centrifugal 25min of 3500r/min, the ratio that precipitation is 1: 7 in solid-liquid mass ratio again extracts 2 times, merge supernatant liquor, pH=5.0 protein precipitation is adjusted with the HCl aqueous solution, isolate supernatant liquor, again pH=3.7 is adjusted to carry out secondary protein precipitation with the HCl aqueous solution supernatant liquor, abandon supernatant liquor, merge twice precipitation, 3 times are precipitated with deionized water wash, pH=7.0 is adjusted with the NaOH aqueous solution, stirring makes precipitation redissolve, lyophilize 24h is garbanzo white protein,
(2) described garbanzo white protein is dissolved in distilled water makes the garbanzo white protein aqueous solution that mass concentration is 5%; Be 0.3AU/g in Alcalase enzyme and the albuminised ratio of garbanzo, Alcalase enzyme put into the described garbanzo white protein aqueous solution, at pH=8.0, under the condition that temperature of reaction is 55 DEG C, hydrolysis 50min; Be 50LAPU/g in Flavourzyme enzyme and the albuminised ratio of garbanzo, add Flavourzyme enzyme, at pH=7.0, temperature of reaction 50 DEG C, condition under, hydrolysis 90min; Enzymolysis solution is heated to boiling, go out enzyme 10min, and after cooling, namely the lyophilize of centrifuging and taking supernatant liquor obtains chickpea peptides powder.
(3) with embodiment 1.
Claims (3)
1. have a chickpea peptides for antioxygenation, represent with CPe-III, it is characterized in that the molecular weight of described CPe-III is 1155Da, amino acid consists of Arg-Gln-Ser-His-Phe-Ala-Asn-Ala-Gln-Pro, and secondary structure accounts for 66% with alpha-helix.
2. a kind of separation purification method with the chickpea peptides of antioxygenation of claim 1, it is characterized in that comprising the steps: that the ratio that the olecranon bean powder after degreasing and water are 1:8-15 in mass ratio mixes by (1), pH=8.3 is adjusted with the NaOH aqueous solution, stir and extract 0.5-1.5h, the centrifugal 15min-25min of 3500r/min-5000r/min, the ratio that precipitation is 1:3-7 in solid-liquid mass ratio is again extracted 2-3 time, merge supernatant liquor, pH=5.0 protein precipitation is adjusted with the HCl aqueous solution, isolate supernatant liquor, again pH=3.7 is adjusted to carry out secondary protein precipitation with the HCl aqueous solution supernatant liquor, abandon supernatant, merge twice precipitation, precipitate 2-3 time with deionized water wash, pH=7.0 is adjusted with the NaOH aqueous solution, stirring makes precipitation redissolve, lyophilize is garbanzo white protein,
(2) described garbanzo white protein is dissolved in distilled water makes the garbanzo white protein aqueous solution that mass concentration is 3-5%; Be 0.3-0.4AU/g in Sumizyme MP Alcalase and the albuminised ratio of garbanzo, Alcalase enzyme put into the described garbanzo white protein aqueous solution, at pH=8.0, under the condition of temperature of reaction 45-55 DEG C, hydrolysis 50-70min; Be 50LAPU/g in Flavourzyme composite flavor enzyme and the albuminised ratio of garbanzo, then add Flavourzyme enzyme, pH=7.0, under the condition that temperature of reaction is 50 DEG C, hydrolysis 90-120min; Enzymolysis solution is heated to boiling, go out enzyme 10min, and after cooling, namely the lyophilize of centrifuging and taking supernatant liquor obtains chickpea peptides powder;
(3) the dextrane gel Sephadex G-25 will handled well, dress post 1.6 × 50cm, by registering instrument recorded stream fluid absorbancy situation, water elution to be distilled is steady to baseline, get chickpea peptides powder dissolution in distilled water after join in described dextrane gel Sephadex G-25 post, with distilled water wash-out, flow velocity is 0.6mL/min, elutriant absorbancy is detected at 280nm place, there are 3 elution peaks altogether at times, collect the 3rd elution peak liquid, after lyophilize, namely obtain the chickpea peptides represented with CPe-III with antioxygenation.
3. chickpea peptides according to claim 1 is preparing the purposes of antioxidant health-care product.
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| CN103172706B (en) * | 2013-03-15 | 2015-05-20 | 中国科学院过程工程研究所 | Preparation method of chick-pea oligopeptide with antioxidation function |
| CN103194517B (en) * | 2013-04-11 | 2016-04-06 | 中国科学院过程工程研究所 | A kind of Chickpea short-peptide mixture and preparation method thereof |
| CN104789629B (en) * | 2015-04-30 | 2018-10-26 | 中国食品发酵工业研究院 | A kind of chick-pea oligopeptide and preparation method thereof |
| CN107223985A (en) * | 2016-03-23 | 2017-10-03 | 新疆丝路弘毅农业科技有限公司 | It is a kind of with the chick-pea composition of antioxidation and application |
| CN107361369B (en) * | 2017-07-18 | 2021-02-05 | 杏辉天力（杭州）药业有限公司 | Chickpea extract and preparation method and application thereof |
| CN108191961A (en) * | 2018-01-22 | 2018-06-22 | 山西师范大学 | The albuminised method and antioxidant that there is high scavenging capacity to superoxide anion are prepared from coconut cake |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1386862A (en) * | 2002-07-09 | 2002-12-25 | 邹远东 | Process for preparing cicer protein polypeptide by enzyme method and its use |
| CN101602799A (en) * | 2009-07-21 | 2009-12-16 | 中国科学院新疆理化技术研究所 | The fast preparation method of primary antimicrobial polypeptide in a kind of garbanzo |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1386862A (en) * | 2002-07-09 | 2002-12-25 | 邹远东 | Process for preparing cicer protein polypeptide by enzyme method and its use |
| CN101602799A (en) * | 2009-07-21 | 2009-12-16 | 中国科学院新疆理化技术研究所 | The fast preparation method of primary antimicrobial polypeptide in a kind of garbanzo |
Non-Patent Citations (2)
| Title |
|---|
| 鹰嘴豆肽、大豆肽功能性质的研究;崔竹梅等;《中国油脂》;20070920;第32卷(第09期);27-31 * |
| 鹰嘴豆肽清除自由基作用的研究;刘宇等;《食品科技》;20090320;第34卷(第03期);173-176 * |
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