CN106432445A - Preparation method and applications of chickpea defensins - Google Patents
Preparation method and applications of chickpea defensins Download PDFInfo
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- CN106432445A CN106432445A CN201610900310.8A CN201610900310A CN106432445A CN 106432445 A CN106432445 A CN 106432445A CN 201610900310 A CN201610900310 A CN 201610900310A CN 106432445 A CN106432445 A CN 106432445A
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/415—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from plants
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
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- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
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Abstract
The invention discloses a preparation method and applications of chickpea defensins. According to the method, chickpeas are ground and degreased with n-hexane; a product is extracted with a phosphate buffered solution; a product is precipitated with ammonium sulfate; a product is separated by anion exchange resin and cation exchange resin; a product is subjected to ultrafiltration; a product is subjected to gel column chromatography separation and purification by HW-55F gel resin, the chickpea defensins with the molecular weight being 5.483 kDa are obtained, and the N-terminal amino acid sequence is H2N-Ala-Cys-Cys-Glu-Asn-Lue-Ala-AspThr-Tyr-Arg-Gly- Pro-Cys-Phe. The chickpea defensins belong to plant defensins. Bacteriostatic and antitumor activity experiments performed on the chickpea defensins prepared with the method prove that the chickpea defensins have a stronger inhibition effect on staphylococcus aureus and escherichia coli and has a strong inhibition effect on human colonic adenocarcinoma cells Caco-2 (the C50 value is 1.74 mu g/mL). The chickpea defensins can be further studied to be applied to the pharmaceutical industry.
Description
Technical field
The present invention relates to a kind of alexinic preparation method and applications of chickpea, it is to be raw material preparation using chickpea
There is the alexin of antibacterial, tumor-suppression activity, in pharmaceutical sanitary field, there is good application prospect.
Background technology
The discovery of antibiotic was once a milestone in physianthropy history, and due to the abuse of antibiotic bring many new
Problem, the use of conventional antibiotic is faced with stern challenge, and research and application are nontoxic, nuisanceless, no drug resistance is new
Antibiotic preparation substitute antibiotics have become development trend.Antibacterial peptide is unsurpassed in excellence with it as the possible substitute of conventional antibiotic
Characteristic increasingly becomes focus of concern.
Antibacterial peptide (antibacterial peptide, ABP) is also known as antimicrobial peptide (antimicrobial
Peptide, AMP) or peptide antibiotic (peptide antibiotics), it is to be widely present in one of nature biotechnology tool
There is the active polypeptide of the biological functions such as anti-mattress, antiviral, parasiticide, raising immunity of organisms, it is biological non-specific
The immunne response product of system of defense, has broad spectrum antibiotic activity, various bacteria or funguses not only can be suppressed it might even be possible to
Suppress the tumor cell in animal body in the case of not injuring normal cell.Conventional antibiotic be by eliminate growth of microorganism or
Survive necessary function, the synthesis such as obstructing bacterioprotein or the activity that changes enzyme are reaching sterilization purpose, and antibacterial is led to
Cross and change a kind of gene just enough to tackle this attack of antibiotic.Antibacterial peptide then acts on bacterial cell membrane, leads to the logical of film
Permeability increases, and is penetrated with this, kills antibacterial.Antibacterial must change the structure of film, that is, changing substantial portion of gene could defend
The attack of antibacterial peptide, and this is nearly impossible.Therefore, antibacterial peptide considerably reduces the possibility producing drug resistance, this
It is the big advantage compared with antibiotic.Antibacterial peptide is the small molecule small peptide of animal body secretion, is also one of body composition, because
This has the biological characteristicses of safe without toxic side effect, thus avoiding the damage to body immune system for the conventional antibiotic class medicine
Evil.Antibacterial peptide remains to maintain vigour under the conditions of 100 DEG C of heating 10min, pH value is had compared with strong adaptability, good water solubility.Antibacterial
Peptide does not almost have lethal effect to normal eukaryotic cell, the eukaryotic cell acting only on prokaryotic cell and pathological changes occurring.
Different according to source, antibacterial peptide can be divided into 6 classes：Insect antimicrobial peptide, Mammalian Antimicrobial Peptides, Amphibian antibacterial
Peptide, Fish, Mollusca, the antibacterial peptide in Crustacean source, Antimicrobial Peptides From Plants, antibacterial antibacterial peptide.Wherein, Antimicrobial Peptides From Plants
It is referred to as plant alexin.Most plants antibacterial peptide has excellent activity to pathogenic, and Activities of Some Plants antibacterial peptide is blue to leather
Family name's positive bacteria, negative bacterium, funguses, yeast and mammalian cell are all toxic.
At present, chickpea planting area in Xinjiang, more than 200,000 mu, is produced up to ten thousand tons of chickpea products, aboundresources per year, is incited somebody to action
This utilization of resources is got up, and will be greatly facilitated chickpea science and technology added value.Therefore, extract chickpea alexin, and develop correlation
Antibacterial, suppression cancer medical product, there is very strong realistic meaning.
Content of the invention
Present invention aim at, provide a kind of chickpea alexinic preparation method and applications, the method is by chickpea
After pulverizing and adopt normal hexane defat, using phosphate buffered solution extraction, ammonium sulfate precipitation, and exchange tree through anions and canons
Fat separates, and ultrafiltration purification, HW-55F gel resin gel filtration chromatography isolate and purify, and obtains the chickpea that molecular weight is 5.483kDa
Alexin, its n terminal amino acid sequence is：N- alanine-cysteine-cysteine-glutamic acid-agedoite-leucine-
Ala-Asp-threonine-tyrosine-arginine-glycine-proline-Cys-Phe, i.e. H2N-
Ala-Cys-Cys-Glu-Asn-Lue-Ala-Asp-Thr-Tyr-Arg-Gly-Pro-Cys-Phe.Belong to plant alexin one
Race, the chickpea alexin through obtaining to the method for the invention carries out the experiment of antibacterial and active anticancer and shows：This chickpea is prevented
Imperial element has stronger inhibitory action to staphylococcus aureuses or escherichia coli, human colon adenocarcinoma cell Caco-2 is had very strong
Inhibitory action (C50It is worth for 1.74 μ g/mL).
A kind of alexinic preparation method of chickpea of the present invention, the raw material in the method is chickpea, concrete grasps
It follow these steps to carry out：
A, chickpea is pulverized, sieve, and with normal hexane or petroleum ether soak degreasing；
B, by the olecranon Semen Glycines powder of step a gained defat with phosphate buffer dissolve, 4 DEG C of temperature, ultrasonic 30min, centrifugation,
Discard precipitation, obtain chickpea polypeptide extracting solution；
C, will in the chickpea polypeptide extracting solution of step b gained add ammonium sulfate so as to saturation be 30%, stirring and dissolving
And after standing 24h, precipitation, 4 DEG C of temperature, rotating speed 12000rpm is centrifuged 15min, takes out precipitation desalination and is dried, obtains thick polypeptide；
After d, 8-10h that diethylaminoethyl cellulose is soaked in water, successively with NaOH the and HCl process of 0.5N,
Then it is saved in water, the diethylaminoethyl cellulose handled well is loaded in 25 × 150mm pillar, first rinsed with water, then
Use ammonium acetate buffer solution 0.05mol/L, the thick polypeptide of pH=9.0 dissolving step c gained, 4 DEG C of temperature, rotating speed 10000rpm is centrifuged
5min, removes precipitation, the diethylaminoethyl cellulose absorption after supernatant is processed, and takes diethylaminoethyl cellulose not
Absorbed component；
After e, 8-10h that cation exchange resin CM-TSK-650M is soaked in water, entered with NaOH and HCl of 0.5N successively
Row is processed, and is then saved in water, and the cation exchange resin handled well CM-TSK-650M is loaded in 15 × 100mm pillar,
First rinsed with water, with 0.05mol/L, the ammonium acetate buffer solution balance of pH=5.0, then by step d gained diethyllaminoethyl
PH adjusted by cellulose unadsorbed component acetic acid is 5.0, is carried out point with the cation exchange resin CM-TSK-650M pillar after processing
From, take CM binding site dialysis desalting, dry；
F, the sample tri-distilled water obtaining step e dissolve so as to concentration is 1mg/mL, in 4 DEG C of temperature, rotating speed
10000rpm is centrifuged 5min, removes precipitation, supernatant is added in the ultra-filtration centrifuge tube of molecular cut off 10kDa, temperature 4
DEG C, rotating speed 3000rpm is centrifuged 3min, collects and is concentrated to give infiltration component；
G, by the infiltration component of step f gained using HW-55F gel resin carry out separate, purification, obtain with SEQ ID
NO：Aminoacid sequence shown in 1, Ala Cys Cys Glu Asn Lue Ala Asp Thr Tyr Arg Gly Pro Cys
The chickpea alexin of Phe.
The chickpea alexin that methods described obtains is in preparing anti-Staphylococcus aureus or colibacillary medicine
Purposes.
Use in the medicine of preparation treatment human colon adenocarcinoma cell Caco-2 for the chickpea alexin that methods described obtains
On the way.
Chickpea alexin (YZDCM) the employing LC-MS analysis side being obtained by the method for the invention
Method and Edman edman degradation Edman are identified：
(1) using with the alexinic molecular weight of LC-MS analysis chickpea, concrete testing conditions are：Solid swashs
Light source, wavelength 355nm；Ionic type：Cation；Detection mode：Linear mode (flight pipe range 1.22m, accelerating potential 20kV)；
Substrate：CHCA；
(2) Edman edman degradation Edman is adopted to measure the alexinic aminoacid sequence of chickpea, its n terminal amino acid sequence is：For
N- alanine-cysteine-cysteine-glutamic acid-agedoite-Leu-Ala-aspartic acid-threonine-cheese ammonia
Acid-arginine-glycine-proline-Cys-Phe, i.e. H2N-Ala-Cys-Cys-Glu-Asn-Lue-Ala-
Asp-Thr-Tyr-Arg-Gly-Pro-Cys-Phe, this polypeptide is to separate first to obtain from chickpea；Amino acid sequence analysis
Figure is shown in accompanying drawing.
A kind of alexinic preparation method and applications of chickpea of the present invention, the method Binding peptide class compound
Physicochemical properties, combine the chickpea alexin powder that modern reversed phase column chromatography and ultrafiltration membrane technique technique are made.The method
The alexin obtaining is the natural polypeptide in olecranon bean material, is verified it and has anti-Staphylococcus aureus, large intestine bar
Bacterium biological activity, has very strong inhibitory action to human colon adenocarcinoma cell, can develop and prepare anti-Staphylococcus aureus, large intestine
The medicine of bacillus biological activity and the medicine of preparation suppression human colon adenocarcinoma cell.
Brief description
Fig. 1 is the polypeptide moiety collection of illustrative plates of efficient liquid phase chromatographic analysis chickpea seed of the present invention；
Fig. 2 is that the present invention extracts the diethyllaminoethyl of solution by the chickpea polypeptide of the ammonium sulfate precipitation of saturation 30%
Cellulose elution curve；
Fig. 3 is that the present invention extracts the diethyllaminoethyl of solution by the chickpea polypeptide of the ammonium sulfate precipitation of saturation 30%
The CM elution curve of the unadsorbed component of cellulose；
Fig. 4 is that the present invention extracts the precipitation position alkalescence of solution by the chickpea polypeptide of the ammonium sulfate precipitation of saturation 30%
Gel resin (the model of the infiltration component of polypeptide fractions：HW-55F) elution profile；
Fig. 5 is the alexinic liquid of chickpea of the present invention-matter figure；
Fig. 6 is the inhibitory action figure to staphylococcus aureuses for the chickpea alexin of the present invention；
Fig. 7 is chickpea alexin of the present invention to colibacillary inhibitory action figure.
Specific embodiment
Embodiment 1
A, chickpea is pulverized, sieve, and use normal hexane soak degreasing；
B, by the olecranon Semen Glycines powder 10g of step a gained defat with phosphate buffer dissolve, 4 DEG C of temperature, ultrasonic 30min, from
The heart, discards precipitation, obtains chickpea polypeptide extracting solution；
C, will in the chickpea polypeptide extracting solution of step b gained add ammonium sulfate so as to saturation be 30%, stirring and dissolving
And after standing 24h, precipitation, low temperature, rotating speed 12000rpm is centrifuged 15min, takes out precipitation desalination and is dried, obtains thick polypeptide；
D, take 100mL diethylaminoethyl cellulose DE-52 to be soaked in water after 8-10h, use NaOH and HCl of 0.5N successively
Processed, be then saved in water, diethylaminoethyl cellulose DE-52 handled well is loaded in 25 × 150mm pillar,
First rinsed with water, then use ammonium acetate buffer solution 0.05mol/L, the thick polypeptide of pH=9.0 dissolving step c gained, 4 DEG C of temperature, turn
Fast 10000rpm is centrifuged 5min, removes precipitation, the diethylaminoethyl cellulose absorption after supernatant is processed, and takes diethylamino
The unadsorbed component of base ethyl cellulose；
E, take 100mL cation exchange resin CM-TSK-650M to be soaked in water after 8-10h, use successively 0.5N NaOH and
HCl process, is then saved in water, and the cation exchange resin handled well CM-TSK-650M is loaded 15 × 100mm post
In son, first rinsed with water, with 0.05mol/L, the ammonium acetate buffer solution balance of pH=5.0, then by step d gained lignocaine
PH adjusted by ethyl cellulose unadsorbed component acetic acid is 5.0, is entered with the cation exchange resin CM-TSK-650M pillar after processing
Row separates, and takes CM binding site dialysis desalting, is dried；
F, the sample aqueous solution obtaining step e, are dissolved so as to concentration is 1mg/mL with tri-distilled water, in 4 DEG C of temperature, turn
Fast 10000rpm is centrifuged 5min, removes precipitation, supernatant is added in the ultra-filtration centrifuge tube of molecular cut off 10kDa, temperature 4
DEG C, rotating speed 3000rpm is centrifuged 3min, collects and is concentrated to give infiltration component；
G, by the infiltration component 1mL of step f gained using HW-55F (25 × 1000mm) gel resin carry out separate, pure
Change, obtain with SEQ ID NO：Aminoacid sequence shown in 1, Ala Cys Cys Glu Asn Lue Ala Asp Thr
The chickpea alexin (YZDCM) of Tyr Arg Gly Pro Cys Phe.
Embodiment 2
A, chickpea is pulverized, sieve, and use petroleum ether soak degreasing；
B, by the olecranon Semen Glycines powder 10g of step a gained defat with phosphate buffer dissolve, 4 DEG C of temperature, ultrasonic 30min, from
The heart, discards precipitation, obtains chickpea polypeptide extracting solution；
C, will in the chickpea polypeptide extracting solution of step b gained add ammonium sulfate so as to saturation be 30%, stirring and dissolving
And after standing 24h, precipitation, low temperature, rotating speed 12000rpm is centrifuged 15min, takes out precipitation desalination and is dried, obtains thick polypeptide；
D, take 100mL diethylaminoethyl cellulose DE-52 to be soaked in water after 8-10h, use NaOH and HCl of 0.5N successively
Processed, be then saved in water, diethylaminoethyl cellulose DE-52 handled well is loaded in 25 × 150mm pillar,
First rinsed with water, then use ammonium acetate buffer solution 0.05mol/L, the thick polypeptide of pH=9.0 dissolving step c gained, 4 DEG C of temperature, turn
Fast 10000rpm is centrifuged 5min, removes precipitation, the diethylaminoethyl cellulose absorption after supernatant is processed, and takes diethylamino
The unadsorbed component of base ethyl cellulose；
E, take 100mL cation exchange resin CM-TSK-650M to be soaked in water after 8-10h, use successively 0.5N NaOH and
HCl process, is then saved in water, and the cation exchange resin handled well CM-TSK-650M is loaded 15 × 100mm post
In son, first rinsed with water, use 0.05mol/L afterwards, the ammonium acetate buffer solution balance of pH=5.0, then by step d gained diethylamino
PH adjusted by base ethyl cellulose unadsorbed component acetic acid is 5.0, with the cation exchange resin CM-TSK-650M pillar after process
Carry out separating, take CM binding site dialysis desalting, be dried；
F, the sample aqueous solution obtaining step e, are dissolved so as to concentration is 1mg/mL with tri-distilled water, in 4 DEG C of temperature, turn
Fast 10000rpm is centrifuged 5min, removes precipitation, supernatant is added in the ultra-filtration centrifuge tube of molecular cut off 10kDa, temperature 4
DEG C, rotating speed 3000rpm is centrifuged 3min, collects and is concentrated to give infiltration component；
G, by the infiltration component 1mL of step f gained using HW-55F (25 × 1000mm) gel resin carry out separate, pure
Change, obtain with SEQ ID NO：Aminoacid sequence shown in 1, Ala Cys Cys Glu Asn Lue Ala Asp Thr
The chickpea alexin (YZDCM) of Tyr Arg Gly Pro Cys Phe.
Embodiment 3
The chickpea alexin that methods described obtains carries out antibacterial activity test：
Staphylococcus aureuses or coli strain are cultivated, the inhibitory action to both bacterium for the determination sample,
And the antibacterial ring size size to two kinds of bacterium for the determination sample, with the bacteriostasis of this judgement sample, concrete grammar is：Using agar plate
The bacteriostatic activity of method detection sample；Gram positive bacteria staphylococcus aureuses and gram negative bacteria escherichia coli are in LB liquid
In culture medium, 37 DEG C of 175rpm, temperature, incubated overnight to OD6002.0 about (1 × 106CFU/mL)；Take the bacterium of incubated overnight
The LB culture medium containing 0.7% agar for the liquid 10 μ L and 8mL is laid in the LB culture medium containing 25mL 1.5% agar after mixing, and works as top
Layer agar solidification after, Deca filter with 0.22 μm of microporous filter membrane after sample solution, 37 DEG C of incubated overnight of temperature, measure antibacterial ring size straight
Footpath, with ampicillin as positive control, normal saline is negative control.
Embodiment 4
Chickpea polypeptide powder suppression human colon adenocarcinoma cell's activity assay that methods described obtains：
Using the polypeptide position inhibitory action of human colon adenocarcinoma cell Caco-2 is measured in chickpea, component and pure
Change the ability of polypeptide anti-tumor activity, be measured using MTT analytic process：
Caco-2 cell is placed in Dole primary section improvement Iger (DMEM) culture medium containing 10% hyclone, temperature 37
DEG C, 5%CO2, 100% humidified incubator culture, trophophase cell of taking the logarithm tested；Take culture 4-5 days, be in index life
One bottle of long-term cell culture fluid, adds trypsin-EDTA liquid, so that attached cell is come off, and contains 5% tire with 10mL
Loews dimension (RPMI) RPMI-1640 of Ox blood serum is made into suspension, after Trypan Blue, makees cytometer on blood counting chamber
Number, non-staining living cells should be more than 97%；
With complete medium diluting cells suspension, it is made into every 100mL and contains 5000-40000 cell, take 96 hole plates, often
Hole adds cell suspension 100 μ L, and the process of refinement born of the same parents should control the 5%CO that flat board is placed in 4h 37 DEG C of temperature2Incubator 24h；
Test-compound makees 5 dilution factors, and Cmax is 10-4Mol/L, makees 1 successively：10 dilutions (10-4, 10-5, 10-6,
10-7, 10-8mol/L)；In document report, the crude extract Cmax of natural product, with 250 μ g/mL, also makees 1：10 dilutions, sample
Do not need heating disinfection or filtration, in order to avoid pollution, the complete medium of dilute sample adds 50 μ g/mL gentamycins
(Gentamicin)；
By flat board containing 5%CO2It is incubated 2-4 days in the incubator of air and 100% humidity；
Tetrazolium bromide (MTT) is made into 1mg/mL solution with serum-free RPMI RPMI-1640, and every hole adds 50 μ L, 37 DEG C of temperature
Incubate 4h, make tetrazolium bromide (MTT) be reduced to first；
Suction out supernatant, add the dimethyl sulfoxide (DMSO) of 150 μ L so that first is dissolved, with plate shaker (plate
Shaker) shake up；
With automatization's spectrophotometric plate reader (Model 312, Biotech Research Laboratories,
Inc., Rockville, Md) optical density of each aperture of mensure at 550nm；
Interpretation of result：
The survival rate of cell：The OD value of each instrument connection is deducted background OD value, and (complete medium adds tetrazolium bromide (MTT), no
Cell) or blank medicine hole OD value (complete medium, plus the different dilution factors of test medicine, plus tetrazolium bromide (MTT), no thin
Born of the same parents), the OD value of each repeating hole takes mean ± SD；
The survival rate of cell is represented with T/C%, and T is the OD value of dosing cell, and C is the OD value of compared with control cells；
Cell survival rate %=(dosing cell OD/ compared with control cells OD) × 100
Obtain drug level (IC during T/C=50%50).
Show from result：Chickpea alexin YZDCM has very strong inhibitory action to human colon adenocarcinoma cell Caco-2, its
IC50Value be 1.74 μ g/mL.
SEQUENCE LISTING
<110>Xinjiang physics and chemistry institute of the Chinese Academy of Sciences
<120>A kind of alexinic preparation method and application of chickpea
<160> 1
<210> 1
<211> 15
<212> PRT
<213>Chickpea（Cicer arietinum L.）
<220>
<221>Chickpea alexin ZYDCM
<400> 1
Ala Cys Cys Glu Asn Lue Ala Asp Thr Tyr Arg Gly Pro Cys Phe
1 5 10 15
Claims (3)
1. a kind of alexinic preparation method of chickpea it is characterised in that the raw material in the method is chickpea, press by concrete operations
The following step is carried out：
A, chickpea is pulverized, sieve, and with normal hexane or petroleum ether soak degreasing；
B, by the olecranon Semen Glycines powder of step a gained defat with phosphate buffer dissolve, 4 DEG C of temperature, ultrasonic 30min, centrifugation, discard
Precipitation, obtains chickpea polypeptide extracting solution；
C, will in the chickpea polypeptide extracting solution of step b gained add ammonium sulfate so as to saturation be 30%, stirring and dissolving is simultaneously quiet
After putting 24 h, precipitation, 4 DEG C of temperature, rotating speed 12000 rpm is centrifuged 15 min, takes out precipitation desalination and is dried, obtains thick polypeptide；
D, by diethylaminoethyl cellulose soak 8-10 h after, successively with NaOH the and HCl process of 0.5 N, Ran Houbao
Exist in water, the diethylaminoethyl cellulose handled well is loaded in 25 × 150 mm pillars, first rinsed with water, then use acetic acid
The thick polypeptide of ammonium buffer solution 0.05 mol/L, pH=9.0 dissolving step c gained, 4 DEG C of temperature, rotating speed 10000 rpm is centrifuged 5min,
Remove precipitation, the diethylaminoethyl cellulose absorption after supernatant is processed, take diethylaminoethyl cellulose unadsorbed
Component；
After e, 8-10 h that cation exchange resin CM-TSK-650M is soaked in water, carried out with NaOH and HCl of 0.5 N successively
Process, be then saved in water, the cation exchange resin handled well CM-TSK-650M is loaded in 15 × 100 mm pillars,
First rinsed with water, use 0.05 mol/L, the ammonium acetate buffer solution balance of pH=5.0, then by step d gained diethyllaminoethyl
PH adjusted by cellulose unadsorbed component acetic acid is 5.0, is carried out point with the cation exchange resin CM-TSK-650M pillar after processing
From, take CM binding site dialysis desalting, dry；
F, the sample tri-distilled water dissolving that step e is obtained so as to concentration is 1 mg/mL, in 4 DEG C of temperature, rotating speed 10000 rpm
It is centrifuged 5 min, remove precipitation, supernatant is added in the ultra-filtration centrifuge tube of molecular cut off 10 kDa, 4 DEG C of temperature, rotating speed
3000 rpm are centrifuged 3 min, collect and are concentrated to give infiltration component；
G, by the infiltration component of step f gained using HW-55F gel resin carry out separate, purification, obtain with SEQ ID NO：
Aminoacid sequence shown in 1, Ala Cys Cys Glu Asn Lue Ala Asp Thr Tyr Arg Gly Pro Cys Phe
Chickpea alexin.
2. the chickpea alexin that a kind of method as claimed in claim 1 obtains is preparing anti-Staphylococcus aureus or large intestine bar
Purposes in the medicine of bacterium.
3. the chickpea alexin that a kind of method as claimed in claim 1 obtains is in preparation treatment human colon adenocarcinoma cell Caco-2
Medicine in purposes.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104761617A (en) * | 2015-04-21 | 2015-07-08 | 中国科学院新疆理化技术研究所 | Preparation method and application of chickpea polypeptide part |
| CN104761617B (en) * | 2015-04-21 | 2020-04-10 | 中国科学院新疆理化技术研究所 | Preparation method and application of chickpea polypeptide part |
| CN110577587A (en) * | 2018-06-07 | 2019-12-17 | 瑞鑫百奥生物科技（深圳）有限公司 | Isolated plant defensin polypeptide and preparation method and application thereof |
| CN110577587B (en) * | 2018-06-07 | 2022-07-12 | 瑞鑫百奥生物科技（深圳）有限公司 | Isolated plant defensin polypeptide and preparation method and application thereof |
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