CN106350561A - Tartary buckwheat active peptide and extraction method thereof - Google Patents
Tartary buckwheat active peptide and extraction method thereof Download PDFInfo
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Abstract
The invention discloses a method for extracting a tartary buckwheat active peptide, which comprises the following steps: preparing tartary buckwheat flour; degreasing the buckwheat flour; enzymatically extracting to obtain tartary buckwheat protein extractive solution; carrying out post-treatment; and separating to obtain the tartary buckwheat active peptide. The application also discloses a tartary buckwheat active peptide. According to the method disclosed by the invention, tartary buckwheat is used as the main raw material, neutral protease, alkaline protease, trypsin, plant hydrolyzed protease, pepsin and papain are screened, and orthogonal test is carried out on the enzymolysis conditions to obtain the preparation process conditions for the tartary buckwheat active peptide; and separation and freeze-drying are carried out on the prepared tartary buckwheat active peptide to finally obtain the tartary buckwheat active peptide. Also, the method lays the foundation for the development of other related products of tartary buckwheat.
Description
Technical field
The application belongs to food processing field, specifically, is related to a kind of tartary buckwheat bioactive peptide and its extracting method.
Background technology
Tartary buckwheat bioactive peptide is the mixture of small peptide that duck wheat protein powder generates after protease hydrolysiss and aminoacid, it
More easily digested.In recent years scientific research personnel constantly finds the hydrolyzate being obtained by Radix Et Rhizoma Fagopyri Tatarici powder hydrolysis both at home and abroad, has
Fall blood, hepatic cholesterol, suppression body fat accumulation, promote excretion, improve constipation, suppression tumor, slow down aging, suppression has
It is abroad considered as " high-grade nutrient health care product " by the effects such as the absorption of evil thing.Therefore, the depth with people, Radix Et Rhizoma Fagopyri Tatarici researched and developed
Enter, its nutritive value and health care will be appreciated that more and more clearer, also will increasingly cause the attention of people.
The research of duck wheat protein hydrolysis and its hydrolyzate is later, yet not comprehensively deep discussion now.At present,
Active peptide product has the bioactive peptide that can reduce hypertension and suppression angiotensin (ace), and this product is with a series of protease water
Solution Buckwheat protein, obtains a kind of structure tripeptides quite similar with crotoxin, and this tripeptides suppression angiotensin turns
Move enzyme, so that blood pressure drops.At present, China's research Radix Et Rhizoma Fagopyri Tatarici is proteoclastic more.(Cui Xia, Zhou Yanming, the Niu Sen such as Cui Xia
Deng, enzymatic hydrolysises Radix Et Rhizoma Fagopyri Tatarici albumen is prepared bioactive peptides optimum condition and is studied, food science and technology, and 2006,13 (1): 39-
41) have studied two kinds of protease (neutral protease, alkaline protease) hydrolysis duck wheat protein and prepare bioactive peptide, find alkaline egg
The hydrolysis effect of white enzyme preferably, and determines the process conditions of hydrolysis by novo Radix Et Rhizoma Fagopyri Tatarici albumen.Li Hongmin etc. (Li Hongmin,
Zhou little Li, the preparation of Buckwheat polypeptide and its antioxidant activity research, Food Science, 2006,27 (10): 302-306) utilize Fructus Chaenomeliss
Protease, protamex enzyme, alcalase enzyme, flavourzyme protease, neutrase neutral protease hydrolyze Radix Et Rhizoma Fagopyri Tatarici respectively
Albumen composition, is optimized to the hydrolysising condition of various enzymes, and research finds, the suitable enzyme preparing Semen Fagopyri Esculenti bioactive peptide is
Alcalase enzyme.Zhou little Li etc. (Zhou little Li, Li Hongmin, Monday ring etc., the research " Food Science " of Radix Et Rhizoma Fagopyri Tatarici peptide nutritious beverage,
2005,26 (11): 128-132) determine and prepare the process conditions of Radix Et Rhizoma Fagopyri Tatarici bioactive peptide liquid and joining of Radix Et Rhizoma Fagopyri Tatarici bioactive peptide nutritious drink
Side.Current research is concentrated mainly on the optimization of hydrolysis process condition, to the separation of duck wheat protein hydrolysate bioactive peptide,
Purification and exploring of Related product are gone back seldom it is necessary to carry out comprehensively deep discussion to it.
Content of the invention
In view of this, the application is directed to above-mentioned problem, there is provided a kind of tartary buckwheat bioactive peptide and its extracting method.
In order to solve above-mentioned technical problem, this application discloses a kind of extracting method of tartary buckwheat bioactive peptide, including following
Step:
1) prepare Radix Et Rhizoma Fagopyri Tatarici powder；
2) ungrease treatment is carried out to Radix Et Rhizoma Fagopyri Tatarici powder；
3) Enzymatic Extraction duck wheat protein extracting solution；
4) post processing；
5) the separation preparation of bioactive peptide, prepares tartary buckwheat bioactive peptide.
Further, step 1) in prepare Radix Et Rhizoma Fagopyri Tatarici powder particularly as follows: taking the Radix Et Rhizoma Fagopyri Tatarici Wheat Seeds of drying to grind in pulverizer
Powder, crosses 40-80 mesh sieve, standby.
Further, step 2) in ungrease treatment is carried out to Radix Et Rhizoma Fagopyri Tatarici powder particularly as follows: in step 1) hardship for preparing
Petroleum ether is added, in room temperature environment defat 8-16h, period stirring simultaneously repeatedly changes petroleum ether in Fagopyrum esculentum Moench powder；By the hardship after defat
Semen Fagopyri Esculenti is placed in fume hood, and air-dried 42-54h is standby, prepares the Radix Et Rhizoma Fagopyri Tatarici powder after defat.
Further, step 3) in Enzymatic Extraction duck wheat protein extracting solution particularly as follows: adjustment defat after Radix Et Rhizoma Fagopyri Tatarici
Ph value be 6-8, temperature is 45-65 DEG C, adds papain enzymolysis, and enzyme concentration is the Radix Et Rhizoma Fagopyri Tatarici weight after defat
0.01%-0.05%, enzymolysis time is 3h-7h.
Further, step 4) in post processing particularly as follows: in 4000r/min centrifuge be centrifuged 10min, take supernatant
Liquid；Add the s-8 macroporous adsorbent resin of 1/2 volume, upper shaking table decolours 4 hours；Stay supernatant standby.
Further, step 5) in bioactive peptide separation preparation particularly as follows:
5.1) by step 4) supernatant for preparing is molten clear with the acetonitrile solution of 2ml, filters, upper high performance liquid chromatography
It is analyzed；
5.2) supernatant having filtered is carried out high performance liquid chromatography separation: preparative high-performance liquid chromatographic balance 10min, rise
Beginning gradient water 95%, acetonitrile 5%, terminates gradient water 25%, acetonitrile 75% gradient timetable 40min；Collect from detector out
Sample；
5.3) by the solution lyophilizing after separation, both obtained finished product.
The invention also discloses a kind of tartary buckwheat bioactive peptide being obtained by above-mentioned extracting method extraction.
Compared with prior art, the application can obtain including following technique effect:
The present invention with Radix Et Rhizoma Fagopyri Tatarici as main material, by centering protease (vigor 200,000 u/g), alkaline protease (vigor
200000 u/g), trypsin, plant protein hydrolysate enzyme, pepsin, papain screening, and enzymatic hydrolysis condition is carried out orthogonal
Test, obtains the process conditions of tartary buckwheat bioactive peptide preparation, and the tartary buckwheat bioactive peptide prepared is carried out separate, and lyophilizing is
Obtain tartary buckwheat bioactive peptide powder eventually, lay the foundation for the exploitation of Radix Et Rhizoma Fagopyri Tatarici other Related products of peptide simultaneously.
Certainly, the arbitrary product implementing the application must be not necessarily required to reach all the above technique effect simultaneously.
Specific embodiment
To describe presently filed embodiment below in conjunction with embodiment in detail, thereby to the application how application technology handss
Section is solving technical problem and to reach realizing process and fully understanding and implement according to this of technology effect.
Embodiment 1
A kind of extracting method of tartary buckwheat bioactive peptide, comprises the following steps:
1) prepare Radix Et Rhizoma Fagopyri Tatarici powder: take the Radix Et Rhizoma Fagopyri Tatarici Wheat Seeds of drying pulverizing in pulverizer, cross 60 mesh sieves, standby.
2) ungrease treatment is carried out to Radix Et Rhizoma Fagopyri Tatarici powder: in step 1) add petroleum ether, in room in the Radix Et Rhizoma Fagopyri Tatarici powder for preparing
Warm environment defat 12h, period stirring simultaneously repeatedly changes petroleum ether；Radix Et Rhizoma Fagopyri Tatarici after defat is placed in fume hood, air-dried 48h is standby
With preparing the Radix Et Rhizoma Fagopyri Tatarici powder after defat.
3) Enzymatic Extraction duck wheat protein extracting solution: the ph value of the Radix Et Rhizoma Fagopyri Tatarici after adjustment defat is 7, and temperature is 60 DEG C, adds
Plus papain enzymolysis, enzyme concentration is 0.04% of the Radix Et Rhizoma Fagopyri Tatarici weight after defat, and enzymolysis time is 5h.
4) post processing: be centrifuged 10min in 4000r/min centrifuge, take supernatant；The s-8 macropore adding 1/2 volume is inhaled
Attached resin, upper shaking table decolours 4 hours；Stay supernatant standby.
5) the separation preparation of bioactive peptide: by step 4) supernatant for preparing is molten clear with the acetonitrile solution of 2ml, mistake
Filter, upper high performance liquid chromatography is analyzed；Preparative high-performance liquid chromatographic balances 10min, start gradient water 95%, acetonitrile 5%, knot
Bundle gradient water 25%, acetonitrile 75% gradient timetable 40min；Collect from detector sample out；By the solution lyophilizing after separation,
Both obtained finished product.
Embodiment 2
A kind of extracting method of tartary buckwheat bioactive peptide, comprises the following steps:
1) prepare Radix Et Rhizoma Fagopyri Tatarici powder: take the Radix Et Rhizoma Fagopyri Tatarici Wheat Seeds of drying pulverizing in pulverizer, cross 40 mesh sieves, standby.
2) ungrease treatment is carried out to Radix Et Rhizoma Fagopyri Tatarici powder: in step 1) add petroleum ether, in room in the Radix Et Rhizoma Fagopyri Tatarici powder for preparing
Warm environment defat 16h, period stirring simultaneously repeatedly changes petroleum ether；Radix Et Rhizoma Fagopyri Tatarici after defat is placed in fume hood, air-dried 42h is standby
With preparing the Radix Et Rhizoma Fagopyri Tatarici powder after defat.
3) Enzymatic Extraction duck wheat protein extracting solution: the ph value of the Radix Et Rhizoma Fagopyri Tatarici after adjustment defat is 6, and temperature is 65 DEG C, adds
Plus papain enzymolysis, enzyme concentration is 0.01% of the Radix Et Rhizoma Fagopyri Tatarici weight after defat, and enzymolysis time is 7h.
4) post processing: be centrifuged 10min in 4000r/min centrifuge, take supernatant；The s-8 macropore adding 1/2 volume is inhaled
Attached resin, upper shaking table decolours 4 hours；Stay supernatant standby.
5) the separation preparation of bioactive peptide: by step 4) supernatant for preparing is molten clear with the acetonitrile solution of 2ml, mistake
Filter, upper high performance liquid chromatography is analyzed；Preparative high-performance liquid chromatographic balances 10min, start gradient water 95%, acetonitrile 5%, knot
Bundle gradient water 25%, acetonitrile 75% gradient timetable 40min；Collect from detector sample out；By the solution lyophilizing after separation,
Both obtained finished product.
Embodiment 3
A kind of extracting method of tartary buckwheat bioactive peptide, comprises the following steps:
1) prepare Radix Et Rhizoma Fagopyri Tatarici powder: take the Radix Et Rhizoma Fagopyri Tatarici Wheat Seeds of drying pulverizing in pulverizer, cross 80 mesh sieves, standby.
2) ungrease treatment is carried out to Radix Et Rhizoma Fagopyri Tatarici powder: in step 1) add petroleum ether, in room in the Radix Et Rhizoma Fagopyri Tatarici powder for preparing
Warm environment defat 8h, period stirring simultaneously repeatedly changes petroleum ether；Radix Et Rhizoma Fagopyri Tatarici after defat is placed in fume hood, air-dried 54h is standby
With preparing the Radix Et Rhizoma Fagopyri Tatarici powder after defat.
3) Enzymatic Extraction duck wheat protein extracting solution: the ph value of the Radix Et Rhizoma Fagopyri Tatarici after adjustment defat is 8, and temperature is 45 DEG C, adds
Plus papain enzymolysis, enzyme concentration is 0.05% of the Radix Et Rhizoma Fagopyri Tatarici weight after defat, and enzymolysis time is 3h.
4) post processing: be centrifuged 10min in 4000r/min centrifuge, take supernatant；The s-8 macropore adding 1/2 volume is inhaled
Attached resin, upper shaking table decolours 4 hours；Stay supernatant standby.
5) the separation preparation of bioactive peptide: by step 4) supernatant for preparing is molten clear with the acetonitrile solution of 2ml, mistake
Filter, upper high performance liquid chromatography is analyzed；Preparative high-performance liquid chromatographic balances 10min, start gradient water 95%, acetonitrile 5%, knot
Bundle gradient water 25%, acetonitrile 75% gradient timetable 40min；Collect from detector sample out；By the solution lyophilizing after separation,
Both obtained finished product.
The technique effect of the present invention to be described with reference to specific experimentation and experimental result:
1 experiment material and instrument
1.1 experiment material
Radix Et Rhizoma Fagopyri Tatarici (six hardships four), is provided by six department of agricultural engineering of Pan Shui Vocationl Technical College Radix Et Rhizoma Fagopyri Tatarici breeding bases.
Take the Radix Et Rhizoma Fagopyri Tatarici Wheat Seeds of drying pulverizing in pulverizer, cross 60 mesh sieves.Oil is added in the Radix Et Rhizoma Fagopyri Tatarici powder obtaining
Ether, in room temperature environment defat 12h, period stirring simultaneously repeatedly changes petroleum ether.Radix Et Rhizoma Fagopyri Tatarici powder after defat is placed in fume hood,
Air-dried 48h is standby.
1.2 experiment enzymes
Neutral protease (vigor 200,000 u/g), alkaline protease (vigor 200,000 u/g), trypsin, plant protein hydrolysate
Enzyme, pepsin, papain
1.3 experiment reagent
The complete buffer agent (Tianjin Kermel Chemical Reagent Co., Ltd.) of ph=6.864 and ph=9.182, hydroxide
Sodium, hydrochloric acid, formaldehyde, phenolphthalein reagent.
1.4 experimental apparatus
Tu-1901 dual-beam ultraviolet-uisible spectrophotometer (Beijing Puxi General Instrument Co., Ltd)；tdl-60b
Low speed desk centrifuge (Anting Scientific Instrument Factory, Shanghai)；Standard inspection sieve (mesh 60,0.3 millimeter of aperture) (Zhejiang Shangyu city
Road ruins Zhang Xingshashai factory)；Phs-3c laboratory ph meter (upper marine rainbow benefit instrument and meter company limited)；101-2 type electric heating air blast is done
Dry case (Tianjin Stettlen Instrument Ltd.)；Dk-98-1 type electric-heated thermostatic water bath (the limited public affairs of Tianjin Stettlen instrument
Department)；Al204 type electronic balance (prunus mume (sieb.) sieb.et zucc. Teller-support benefit (Shanghai) Co., Ltd.)；Dft-200 type high speed Universal pulverizer (Wenling
Lin great Machinery Co., Ltd. of city)；Freezer dryer；Lc3000 high performance liquid chromatograph (the logical perseverance of Beijing innovation).
2 experimental techniques
The preparation of 2.1 protein extracts: the Radix Et Rhizoma Fagopyri Tatarici powder after extracting degreasing, enzyme concentration is 0.03%, 65 DEG C of temperature, ph=
11, solid-liquid ratio is water-bath extraction 4h under conditions of 1:20.It is centrifuged 10min in 4000r/min centrifuge, take supernatant.Add
The s-8 macroporous adsorbent resin of 1/2 volume, upper shaking table decolours 4 hours.Stay supernatant standby.
The screening of 2.2 protease
Respectively using alkaline protease, trypsin, plant protein hydrolysate enzyme, neutral protease, pepsin, Fructus Chaenomeliss egg
White enzyme carries out the hydrolysising experiment of Buckwheat protein.Carry out the screening and optimizing of enzyme with degree of hydrolysis for index, determine hydrolysis Radix Et Rhizoma Fagopyri Tatarici
Aleuronat is conveniently papain with enzyme.
To acid protease (ph=3), neutral protease (ph=7), alkaline protease (ph=10), trypsin ph
=8), plant protein hydrolysate enzyme (ph=6), pepsin (ph=2), papain (ph=7), by ratio 1:10, extracts temperature
50 DEG C of degree, enzyme concentration 0.3%, hydrolysis time 6h.100 DEG C of enzyme denaturing 10min, are centrifuged 10min in 4000r/min centrifuge, take
Supernatant.The degree of hydrolysis measuring protein extract, according to ph-stat method, is slightly changed.Only need to be after the end of the hydrolysis by reactant
The ph of system is adjusted to 7.0, and records the amount of alkali liquor used.Caseic degree of hydrolysis can be calculated as the following formula: hydrolysis
Degree dh%=b (mb) (1/t) (1/mp) (1/htot) × 100
Wherein: b is the volume (ml) of nao h；Mb is the concentration (mol/l) of nao h；In ph7.0,50 DEG C of experiment condition
Under, for casein, 1/ α is 2.26；Mp is the quality (g) of protein；Htot is the mmoles of peptide bond in every gram of material protein
That number, for casein, this value takes 8.2.
Three repetitions are done in every group of experiment, average, and calculate degree of hydrolysis, find out the best protease of hydrolysis effect effect.
2.4 Enzymatic Extraction experiment of single factor
2.4.1 enzyme concentration
Reaction condition: temperature 60 C；Time 5h；Solid-liquid ratio 1:15；Ph=7.Enzyme concentration sets 0.01%, 0.02%,
0.03%, 0.04%, 0.05% 5 gradient.100 DEG C of enzyme denaturing 10min, are centrifuged 10min in 4000r/min centrifuge, take
Clear liquid.Measure the degree of hydrolysis of protein extract.Find out optimal enzyme concentration.
2.4.2 hydrolysis time
Reaction condition: temperature 60 C；Enzyme concentration 0.03%；Ph=7.Extraction time sets 3h, five ladders of 4h, 5h, 6h, 7h
Degree.100 DEG C of enzyme denaturing 10min, are centrifuged 10min in 4000r/min centrifuge, take supernatant.Measure the hydrolysis of protein extract
Degree.Find out the optimum hydrolysis time.
2.4.3 temperature
Reaction condition: time 5h；Enzyme concentration 0.03%；Ph=7.Temperature sets, 45 DEG C, 50 DEG C, 55 DEG C, 60 DEG C, 65 DEG C
Five gradients.100 DEG C of enzyme denaturing 10min, are centrifuged 10min in 4000r/min centrifuge, take supernatant.Measure protein extract
Degree of hydrolysis.Find out the most suitable hydrolysis temperature.
2.4.4ph value
Reaction condition: temperature 60 C；Time 5h；Enzyme concentration 0.3%.Set ph=6, ph=6.5, ph=7, ph=7.5,
Five gradients of ph=8.100 DEG C of enzyme denaturing 10min, are centrifuged 10min in 4000r/min centrifuge, take supernatant.Measure albumen to carry
Take the degree of hydrolysis of liquid.Find out optimum ph.
The orthogonal experiment of 2.5 bioactive peptide hydrolysis processs
According to single factor experiment result, select enzyme concentration (a), temperature (b), solid-liquid ratio (c), ph value (d) as detect because
Element, every 3 preferable levels (table 1) of selecting factors, by l9(34) design orthogonal test scheme (table 7).
Table 1 orthogonal test factor level table
The separation preparation of 2.6 bioactive peptide
1st, sample is taken to put in vessel, molten clear with the acetonitrile solution of 2ml, filter, upper high performance liquid chromatography is analyzed.
2nd, prepare: the sample having dissolved does sample introduction and prepares.Preparation hplc balance 10min, start gradient water 95%, second
Nitrile 5%, terminates gradient water 25%, acetonitrile 75% gradient timetable 40min.Collect from detector sample out.
3rd, identify: by the sample collected, in sampling, analytical type colleges and universities liquid phase is identified.
4th, finally by the solution lyophilizing after separation, both obtained finished product.
3 results and analysis
The screening of 3.1 protease
The different impact to Buckwheat protein degree of hydrolysis for the protease of table 2
Result shows, under the optimum reaction conditionses of this seven kinds of enzymes, preferably, degree of hydrolysis can for the hydrolysis effect of papain
Reach 48.6%.
3.2 Enzymatic Extraction experiment of single factor
3.2.1 enzyme concentration
The results are shown in Table 3.
The impact to Buckwheat protein degree of hydrolysis for table 3 enzyme concentration
As seen from Table 3, when enzyme concentration is 0.03%, the degree of hydrolysis highest of protein.
3.2.2 hydrolysis time
The impact to Buckwheat protein degree of hydrolysis for table 4 hydrolysis time
As seen from Table 4, during hydrolysis time 4 to 5 hours, protein degree improves rapidly.With hydrolysis time
Prolongation, degree of hydrolysis is held essentially constant, so time of extracting should control in 4h.
3.2.3 temperature
The impact to Buckwheat protein degree of hydrolysis for table 5 temperature
As seen from Table 5, with the rising of temperature, the degree of hydrolysis of protein gradually steps up, and reaches highest when 60 DEG C, when
When temperature raises again, enzymatic activity is suppressed, so that degree of hydrolysis declines.Therefore, should choose 60 DEG C is optimum extraction temperature.
3.2.4ph value
The impact to Buckwheat protein degree of hydrolysis for the table 6ph value
As shown in Table 6, in the case that other reaction conditions are constant, with the increase of ph value, extraction rate of protein is in bright
Aobvious ascendant trend, reaches highest in ph=7；When ph increases again, protease loses activity, and protein degree declines, and says
Bright selection and withdrawal ph=7 is more suitable.
3.3 Enzymatic Extraction orthogonal experiments
According to single factor experiment result, select enzyme concentration (a), temperature (b), hydrolysis time (c), ph value (d) as detection
Factor, every 3 preferable levels (table 15) of selecting factors, by l9(34) design orthogonal test scheme.
Table 7 orthogonal experiments and range analysiss
Orthogonal test analysis result shows, optimize Radix Et Rhizoma Fagopyri Tatarici peptide preparation process condition: enzyme dosage 0.04%, ph value=7,
60 DEG C of hydrolysis time 5h, hydrolysis temperature.
3.4 Enzymatic Extraction confirmatory experiments
Because table 7 does not have Optimum combinational scheme, therefore it is necessary to do confirmatory experiment to it, to further confirm that orthogonal test
Feasibility.Weigh 3 parts of the defat Radix Et Rhizoma Fagopyri Tatarici powder of drying, every part of 1g, be placed in 10ml centrifuge tube, by the optimal production work preferably going out
Skill condition is extracted, and measures light absorption value, and calculates extraction ratio, experimental result is shown in Table at ultraviolet spectrophotometer 680nm
8.
Table 8 optimised process checking test result table
From the result of the test of table 8, under optimal extracting factor, degree of hydrolysis is 49.69%, higher than orthogonal examination
Top level (a testing3b1c3d2), its degree of hydrolysis is 48.43%.Therefore the success of optimised process checking test, this extraction process is described
Reliable and stable.
4 conclusions
The preparation process condition of Radix Et Rhizoma Fagopyri Tatarici peptide: enzyme dosage 0.04%, ph value=7, hydrolysis time 5h, 60 DEG C of hydrolysis temperature.
Described above illustrate and describes some preferred embodiments of invention, but as previously mentioned it should be understood that inventing not
It is confined to form disclosed herein, be not to be taken as the exclusion to other embodiment, and can be used for various other combinations, modification
And environment, and can be carried out by the technology or knowledge of above-mentioned teaching or association area in invention contemplated scope described herein
Change.And the change that those skilled in the art are carried out and change without departing from the spirit and scope of invention, then all should weighed appended by invention
In the protection domain that profit requires.
Claims (7)
1. a kind of extracting method of tartary buckwheat bioactive peptide is it is characterised in that comprise the following steps:
1) prepare Radix Et Rhizoma Fagopyri Tatarici powder；
2) ungrease treatment is carried out to Radix Et Rhizoma Fagopyri Tatarici powder；
3) Enzymatic Extraction duck wheat protein extracting solution；
4) post processing；
5) the separation preparation of bioactive peptide, prepares tartary buckwheat bioactive peptide.
2. the extracting method of tartary buckwheat bioactive peptide according to claim 1 is it is characterised in that described step 1) in preparation
Radix Et Rhizoma Fagopyri Tatarici powder, particularly as follows: taking the Radix Et Rhizoma Fagopyri Tatarici Wheat Seeds of drying pulverizing in pulverizer, crosses 40-80 mesh sieve, standby.
3. the extracting method of tartary buckwheat bioactive peptide according to claim 1 is it is characterised in that described step 2) in hardship
Fagopyrum esculentum Moench powder carries out ungrease treatment particularly as follows: in step 1) add petroleum ether in the Radix Et Rhizoma Fagopyri Tatarici powder for preparing, take off in room temperature environment
Fat 8-16h, period stirring simultaneously repeatedly changes petroleum ether；Radix Et Rhizoma Fagopyri Tatarici after defat is placed in fume hood, air-dried 42-54h is standby.
4. the extracting method of tartary buckwheat bioactive peptide according to claim 1 is it is characterised in that described step 3) in enzyme process
Extract duck wheat protein extracting solution particularly as follows: the ph value adjusting the Radix Et Rhizoma Fagopyri Tatarici after defat is 6-8, temperature is 45-65 DEG C, add wood
Melon protease hydrolyzed, enzyme concentration is the 0.01%-0.05% of the Radix Et Rhizoma Fagopyri Tatarici weight after defat, and enzymolysis time is 3h-7h.
5. the extracting method of tartary buckwheat bioactive peptide according to claim 1 is it is characterised in that described step 4) in rear place
Reason, particularly as follows: being centrifuged 10min in 4000r/min centrifuge, takes supernatant；Add the s-8 macroporous adsorbent resin of 1/2 volume,
Upper shaking table decolours 4 hours；Stay supernatant standby.
6. the extracting method of tartary buckwheat bioactive peptide according to claim 1 is it is characterised in that described step 5) in activity
Peptide separation preparation particularly as follows:
5.1) by step 4) supernatant for preparing is molten clear with the acetonitrile solution of 2ml, filters, upper high performance liquid chromatography is carried out
Analysis；
5.2) supernatant having filtered is carried out high performance liquid chromatography separation: preparative high-performance liquid chromatographic balance 10min, initiate ladder
Degree water 95%, acetonitrile 5%, terminates gradient water 25%, acetonitrile 75% gradient timetable 40min；Collect from detector sample out；
5.3) by the solution lyophilizing after separation, both obtained finished product.
7. a kind of tartary buckwheat bioactive peptide being obtained by the extracting method extraction described in claim 1.
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108969681A (en) * | 2018-10-15 | 2018-12-11 | 王召利 | A kind of blood pressure lowering plant active peptides and preparation method thereof |
| CN109022527A (en) * | 2018-08-28 | 2018-12-18 | 中国农业科学院作物科学研究所 | A kind of quinoa polypeptide and preparation method thereof with hypotensive activity |
| CN109485693A (en) * | 2018-12-19 | 2019-03-19 | 山西昇力元保健品有限公司 | A method of it extracting quinoa native peptides from quinoa and digests quinoa peptide processed |
| CN110256262A (en) * | 2019-07-01 | 2019-09-20 | 成都郝尔斯生物科技有限公司 | The method of 2- hydroxy benzylamine is extracted from bitter buckwheat |
| CN111067108A (en) * | 2019-12-31 | 2020-04-28 | 北京农学院 | Rose tartary buckwheat polypeptide oral liquid and preparation method thereof |
| CN113755317A (en) * | 2021-10-12 | 2021-12-07 | 六盘水师范学院 | Method for extracting tartary buckwheat active peptide and special device |
| CN115725681A (en) * | 2022-11-23 | 2023-03-03 | 龙岩嘉麒生物科技有限公司 | Enzymatic extraction process and equipment for bioactive peptide |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1526299A (en) * | 2003-09-23 | 2004-09-08 | 江苏大学 | Wheat plumule protein hydrolysate and its prepn process and use |
| CN101381759A (en) * | 2008-10-24 | 2009-03-11 | 西北农林科技大学 | Preparation method of buckwheat protein biological activity peptides |
| CN104059127A (en) * | 2014-05-29 | 2014-09-24 | 无限极（中国）有限公司 | Natural high-activity antihypertensive peptide, and preparation method and application thereof |
-
2016
- 2016-11-11 CN CN201610995983.6A patent/CN106350561A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1526299A (en) * | 2003-09-23 | 2004-09-08 | 江苏大学 | Wheat plumule protein hydrolysate and its prepn process and use |
| CN101381759A (en) * | 2008-10-24 | 2009-03-11 | 西北农林科技大学 | Preparation method of buckwheat protein biological activity peptides |
| CN104059127A (en) * | 2014-05-29 | 2014-09-24 | 无限极（中国）有限公司 | Natural high-activity antihypertensive peptide, and preparation method and application thereof |
Non-Patent Citations (3)
| Title |
|---|
| 周小理 等: "苦荞蛋白水解过程及其水解产物抗氧化活性的初探", 《食品工业科技》 * |
| 谭萍 等: "苦荞麦蛋白质的酶提工艺研究", 《食品研究与开发》 * |
| 邓修: "《中药制药工程与技术》", 31 May 2008, 华东理工大学出版社 * |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109022527A (en) * | 2018-08-28 | 2018-12-18 | 中国农业科学院作物科学研究所 | A kind of quinoa polypeptide and preparation method thereof with hypotensive activity |
| CN108969681A (en) * | 2018-10-15 | 2018-12-11 | 王召利 | A kind of blood pressure lowering plant active peptides and preparation method thereof |
| CN109485693A (en) * | 2018-12-19 | 2019-03-19 | 山西昇力元保健品有限公司 | A method of it extracting quinoa native peptides from quinoa and digests quinoa peptide processed |
| CN110256262A (en) * | 2019-07-01 | 2019-09-20 | 成都郝尔斯生物科技有限公司 | The method of 2- hydroxy benzylamine is extracted from bitter buckwheat |
| CN110256262B (en) * | 2019-07-01 | 2022-02-11 | 北京浩鼎瑞健康科技中心(有限合伙) | Method for extracting 2-hydroxybenzylamine from tartary buckwheat |
| CN111067108A (en) * | 2019-12-31 | 2020-04-28 | 北京农学院 | Rose tartary buckwheat polypeptide oral liquid and preparation method thereof |
| CN113755317A (en) * | 2021-10-12 | 2021-12-07 | 六盘水师范学院 | Method for extracting tartary buckwheat active peptide and special device |
| CN113755317B (en) * | 2021-10-12 | 2023-06-09 | 六盘水师范学院 | Tartary buckwheat active peptide extraction method and special device |
| CN115725681A (en) * | 2022-11-23 | 2023-03-03 | 龙岩嘉麒生物科技有限公司 | Enzymatic extraction process and equipment for bioactive peptide |
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