CN102626206A - Application of mung beans to preparation of ACE (Angiotensin Converting Enzyme) inhibitor - Google Patents
Application of mung beans to preparation of ACE (Angiotensin Converting Enzyme) inhibitor Download PDFInfo
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- CN102626206A CN102626206A CN2012101377148A CN201210137714A CN102626206A CN 102626206 A CN102626206 A CN 102626206A CN 2012101377148 A CN2012101377148 A CN 2012101377148A CN 201210137714 A CN201210137714 A CN 201210137714A CN 102626206 A CN102626206 A CN 102626206A
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Abstract
The invention discloses application of mung beans to preparation of an ACE (Angiotensin Converting Enzyme) inhibitor. Preferably, the mung beans are products obtained by hydrolyzing mung beans through protease. The invention develops a brand-new approach for development and utilization of mung beans and has wider application prospect.
Description
Technical field
The invention belongs to field of food, specifically, is to be used to prepare ACE (Angiotensin Converting Enzyme, ACE) application of inhibitor about mung bean.
Background technology
Mung bean (formal name used at school: Vigna radiata) be a kind of pulse family, Papillionoideae Vigna plant, originate in India, Burma area.Generally plant the various countries, East Asia now, and also there are a small amount of plantation in Africa, Europe, the U.S., and states such as China, Burma are main mung bean exported countrys.Seed and stem are edible by extensively.
Mung bean is a kind of high protein, middle starch, low-fat integration of drinking and medicinal herbs crop, and every 100g dry contains 22 ~ 26g protein, is 2 times of cereal class, and about starch-containing 50g, about dietary fiber 6.4g, fat content is low, only accounts for 1%.In addition, also contain abundant vitamin and trace element in the mung bean, vitamin B1 is 17.5 times of chicken; Vitamin B2 is 2 ~ 4 times of cereal class; Phosphorus is cereal class and pork, chicken, fish, egg 2 times, and calcium is 4 times of cereal class, also contains trace elements such as iron, selenium, magnesium, zinc.The mung bean nutritional labeling is comprehensive, and not only protein content is high, and the protein effect occupies first of the various beans than high, and amino acid A wide selection of colours and designs and composition are better in the protein, especially with lysine content than horn of plenty.
ACE (ACE; Angiotensin Converting Enzyme) effect in human body is that angiotensin I (Angiotensin I Converting Enzyme) is changed into Angiotensin II (Angiotensin II Converting Enzyme); And Angiotensin II is strong vasoconstrictor substance; Can make vessel retraction, blood pressure raises in the body thereby make.In addition, ACE can also lose the bradykinin of in vivo regulating blood pressure, and blood pressure raises in the body thereby also make.
ACE is a kind of metallopeptidase, contains 2 and combines Zn
2+The site, promptly so-called " must binding site (Obligatory Binding Site) ".Zn
2+Binding site is the position, active group place of ACE catalytic reaction.The acting in conjunction of various ACE mortifiers is the Zn with the ACE active site
2+In conjunction with, make it inactivation.
The peptide that blood pressure lowering peptide is made up of several amino acid; Be to the stronger competitive inhibitor of ACE active region affinity; They are stronger than ACE I (AngI) or bradykinin with the affinity of ACE, and are not easy to discharge from the ACE land, reduce the ACE activity or it is lost activity; Thereby hinder ACE catalysis angiotensin and change into Angiotensin II; Blood vessel is not shunk, and the catalyzing hydrolysis bradykinin becomes two kinds of biochemical reaction processes of inactivation fragment, play hypotensive effect.
Up to the present, do not find relevant for utilizing mung bean to prepare the report of ACE mortifier or blood-pressure drug.
Summary of the invention
The present inventor finds that in the process of the purposes of the mung bean that studies for a long period of time the hydrolysate of mung bean has tangible ACE (ACE) and suppresses active.
Therefore, the object of the present invention is to provide a kind of mung bean to be used to prepare the application of ACEI.
According to the present invention, said mung bean is the product of mung bean behind protease hydrolytic.
According to a preferred embodiment of the invention, said protease is selected from: pepsin, trypsase, perhaps their combination.
Have the effect that suppresses hypertensin conversion enzyme activity based on mung bean, mung bean can be further used for developing food or the health products with antihypertensive function.
The present invention has opened up a brand-brand-new way for the development and use of mung bean, has very application prospects.
The specific embodiment
Below in conjunction with specific embodiment, the preparation method of blood pressure lowering polypeptide of the present invention is done further explain.Should be understood that following examples only are used for embodiment of the present invention but not are used to limit scope of the present invention.
In following examples, the measuring principle and the method step of ACE inhibition activity are following:
The ACE of the protolysate of mung bean (polypeptide) suppresses determination of activity and adopts in-vitro simulated mensuration; External ACE inhibiting rate test philosophy is: substrate hippuroyl histidyl-leucine (HHL) is under the catalytic action of ACE enzyme; Can produce hippuric acid (Hip) and dipeptides; Method through liquid phase is measured the content of hippuric acid, and then enzyme that can indirect determination ACE enzyme is lived; And the existence of ACE inhibitor can suppress the activity of ACE enzyme, causes the generation of Hip to reduce, thereby has reflected the activity of ace inhibitory peptide indirectly.
The method step of test ACE inhibiting rate is following: choose the EP pipe as reactor, numbering 0,1,2,3 etc.No. 0 pipe adds 15 μ L deionized waters and is contrast, and pipes such as 1,2,3 add the ACE inhibitor of 15 μ L.All add the substrate HHL of 15 μ L then in each pipe, after 37 ℃ of temperature were bathed 5min, the ACE enzyme that adds 5 μ L started reaction, in 37 ℃ of water-baths, reacts 40min.Reaction adds 300 μ L 1M HCl cessation reactions after finishing.Reactant liquor carries out high speed centrifugation, crosses the filter of 4.5 μ m, carries out high performance liquid chromatography (HPLC) then and measures.The HPLC condition is: 10% acetonitrile: 90% water (0.1%TFA), detect wavelength 228nm, 30 ℃ of column temperatures, applied sample amount 20 μ L;
ACE inhibiting rate (X) is calculated as follows:
X=(the A contrast-A1)/A contrast * 100%
In the formula: X--ACE inhibiting rate (%)
The A contrast--control group generates the peak area of Hip
The A1--experimental group generates the peak area of Hip
Embodiment 1-8, the Semen phaseoli radiati polypeptide hydrolysate preparation
Get mung bean, clean, smash to pieces and grind the back collection, get a certain amount of Green Gram Seed, carry out drying, get dry mung bean flour.
Press shown in the table 1, the dry mung bean flour that obtains is handled with the protease of various combination respectively, specific as follows:
Take by weighing the 2g powder, be mixed with the solution of 20mg/ml, according to the protease hydrolytic condition of table 1, adjust pH adds corresponding enzyme 0.1% (w/v) to corresponding enzyme reaction pH condition, and water enzyme digestion is 0.17 ~ 10 hour under corresponding reaction temperature, stirs.When using two kinds of different Protease Treatment, after first kind of protease hydrolyzed finishes, boil the 10min enzyme that goes out and live, carry out the treatment step of second kind of protease again.After the cessation reaction, regulate pH to 7, hydrolysate is centrifugal, and (12000r/min 10min), gets supernatant ,-40 ℃ of freeze dryings, collects the hydrolysate powder.
Table 1, different protease hydrolytic condition
| Embodiment | The combination of enzyme | Temperature (℃) | pH | Time (hr) |
| 1 | Pepsin | 37 | 1.5 | 0.17 |
| 2 | Pepsin | 37 | 1.5 | 2 |
| 3 | Pepsin | 37 | 1.5 | 4 |
| 4 | Pepsin | 37 | 1.5 | 6 |
| 5 | Pepsin+trypsase | 37 | 1.5+8 | 0.085+0.085 |
| 6 | Pepsin+trypsase | 37 | 1.5+8 | 2+2 |
| 7 | Pepsin+trypsase | 37 | 1.5+8 | 3+4 |
| 8 | Pepsin+trypsase | 37 | 1.5+8 | 4+6 |
Embodiment 9-15, the Semen phaseoli radiati polypeptide hydrolysate preparation
Get mung bean, smash grinding after the cleaning to pieces, collect, get a certain amount of Green Gram Seed, carry out drying, get dry mung bean flour.
Press shown in the table 2, the dry mung bean flour that obtains is mixed with the hydrolyzate of variable concentrations, handle with different pepsins and tryptic combination then, specific as follows:
Get Green Gram Seed,, be configured to the solution of 0.1 ~ 10mg/ml according to the mung bean hydrolyzate concentration of table 2; Adjust pH adds corresponding enzyme 0.1% (w/v) to corresponding enzyme reaction pH condition, water-bath digestion under corresponding reaction temperature; After first kind of protease hydrolyzed finishes, boil the 10min enzyme that goes out and live, carry out the treatment step of second kind of protease again; Two kinds of enzymes are enzymolysis 4 hours respectively, boils the 10min enzyme that goes out at last and lives cessation reaction.Regulate pH to 7, hydrolysate is centrifugal, and (12000r/min 10min), gets supernatant ,-40 ℃ of freeze dryings, collects the hydrolysate powder.
The hydrolysising condition of table 2, variable concentrations mung bean hydrolyzate
Embodiment 16-30, ACE suppresses active mensuration
Get the mung bean hydrolysate powder that embodiment 1-8 and 9-15 obtain respectively, the solution of preparation variable concentrations is taked in-vitro simulated assay method that ACE is suppressed activity and is measured, and the result is respectively shown in following table 3 and table 4:
Table 3, ACE suppress active-1
Table 4, ACE suppress active-2
Visible by the result of table 3, the product of mung bean after pepsin hydrolysis, pepsin and trypsase combined hydrolysis all has tangible ACE and suppresses active, and along with the prolongation in reaction time, the ACE inhibiting rate increases.It is the strongest that the ACE of pepsin hydrolysis product suppresses activity, reached 94.8%.
Result by table 4 is visible; The product of the mung bean hydrolysate of variable concentrations after pepsin and trypsase combined hydrolysis also all has tangible ACE and suppresses active; According to calculating, mung bean is 0.634mg/ml through the IC50 value of the ACE inhibiting rate of pepsin and trypsase combined hydrolysis product.
In sum; The hydrolyzate of protease of mung bean provided by the invention has obvious inhibiting activity to ACE (ACE); Thereby can be used to prepare the ACE inhibitor; Perhaps be further used for processing food or health products with antihypertensive function, this is conspicuous for a person skilled in the art.
Claims (4)
1. mung bean is used to prepare the application of ACEI.
2. application as claimed in claim 1 is characterized in that, said mung bean is the product of mung bean behind protease hydrolytic.
3. application as claimed in claim 2 is characterized in that, said protease is selected from: pepsin, trypsase, perhaps their combination.
4. mung bean is used to prepare the application of blood pressure reducing food or health products, it is characterized in that, said blood pressure reducing food or health products are used to suppress the activity of ACE.
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| Application Number | Priority Date | Filing Date | Title |
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| CN2012101377148A CN102626206A (en) | 2012-05-04 | 2012-05-04 | Application of mung beans to preparation of ACE (Angiotensin Converting Enzyme) inhibitor |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103290086A (en) * | 2013-06-25 | 2013-09-11 | 广州合诚实业有限公司 | A mung bean protein peptide having ACE inhibitory activity and a preparation method and applications thereof |
| CN107805652A (en) * | 2017-09-26 | 2018-03-16 | 中国农业科学院作物科学研究所 | A kind of preparation method and application of the Semen phaseoli radiati polypeptide with active anticancer |
-
2012
- 2012-05-04 CN CN2012101377148A patent/CN102626206A/en active Pending
Non-Patent Citations (1)
| Title |
|---|
| 龚琴等: "响应面法优化酶解制备绿豆蛋白ACE抑制肽的研究", 《食品工业科技》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103290086A (en) * | 2013-06-25 | 2013-09-11 | 广州合诚实业有限公司 | A mung bean protein peptide having ACE inhibitory activity and a preparation method and applications thereof |
| CN103290086B (en) * | 2013-06-25 | 2015-01-14 | 广州合诚实业有限公司 | A mung bean protein peptide having ACE inhibitory activity and a preparation method and applications thereof |
| CN107805652A (en) * | 2017-09-26 | 2018-03-16 | 中国农业科学院作物科学研究所 | A kind of preparation method and application of the Semen phaseoli radiati polypeptide with active anticancer |
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Application publication date: 20120808 |