CN110066844A - A kind of preparation method with the U.S. rattan fruit dregs of rice biologically active peptide of anti-trioxypurine - Google Patents
A kind of preparation method with the U.S. rattan fruit dregs of rice biologically active peptide of anti-trioxypurine Download PDFInfo
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/34—Extraction; Separation; Purification by filtration, ultrafiltration or reverse osmosis
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/06—Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/54—Improvements relating to the production of bulk chemicals using solvents, e.g. supercritical solvents or ionic liquids
Abstract
The invention discloses a kind of preparation methods with the U.S. rattan fruit dregs of rice biologically active peptide of anti-trioxypurine, comprising: (1) U.S. rattan fruit dregs of rice pre-treatment: crushes, ungrease treatment, remove organic reagent, soak in water to obtain U.S. rattan fruit dregs of rice solution；(2) protease hydrolyzed: being heated to 40-60 DEG C, and adjustment pH is 2-11, and protease hydrolytic, then high temperature enzyme deactivation is added, and centrifugation takes supernatant to obtain thick enzymolysis liquid；(3) lactic acid bacillus DU-106 ferments: fermentation seed liquid being seeded to the U.S. thick enzymolysis liquid of rattan fruit albumen, stirs and evenly mixs, ferments 2-7 days at 25 DEG C -35 DEG C；(4) it neutralizes, filter: the pH of hydrolyzate after cooling being adjusted to neutrality, filters to obtain enzymolysis liquid；(5) UF membrane: crossing ultrafiltration membrane, is concentrated by evaporation, freeze-drying.The present invention utilizes protease hydrolyzed, lactic acid bacillus DU-106 fermentation and Ultrafiltration Membrane, prepare U.S. rattan fruit dregs of rice biologically active peptide, with significant anti-trioxypurine effect, the U.S. rattan fruit dregs of rice are the leftover bits and pieces generated after U.S. rattan fruit is extracted oil, and simple process, production cost are low, pollution-free.
Description
Technical field
The present invention relates to U.S. rattan fruit technical fields, specifically, it is raw to be related to a kind of U.S. rattan fruit dregs of rice with anti-trioxypurine
The preparation method of object active peptide.
Background technique
In recent years, with the change of the factors such as life style, eating habit and ambient enviroment, gout disease incidence is presented year by year
The trend of rising.Long-term high lithemia level can generate a series of adverse consequences to blood vessel, heart, kidney, increase and suffer from the heart
The risk of cranial vascular disease, hypertension, diabetes and kidney trouble etc..Common anti-trioxypurine drug such as allopurinol clinical at present,
Probenecid, Benzbromarone etc. all have the toxic side effects such as allergy, bone marrow suppression, hepatic disorder.Therefore, developing food products source
, the anti-trioxypurine active material of novel, less toxic side effect has great importance.
Existing numerous studies prove that flavonoids, phenolic acid class, alkaloids isoreactivity substance all have anti-trioxypurine effect, and
For biologically active peptide anti-trioxypurine effect research still in its infancy.
Small active peptides have remarkable result in terms of anti-trioxypurine treats gout.However, the peptide of anti-trioxypurine is main at present
Source is mostly marine fish, as the invention of patent CN201510094600.3, CN201510094901.6 uses saury through protease
Hydrolysis prepares anti-trioxypurine peptide；A kind of bonito stick protolysate with anti-trioxypurine of CN201410520344.5 Invention Announce
Preparation method；CN201810728741.X obtains a kind of anti-trioxypurine peptide from giant salamander cartilage enzymatic hydrolyzed extract；
CN201810215529.3, which is disclosed, prepares fish polypeptide dry powder, anti-trioxypurine peptide combinations, beverage containing anserine using tuna
Method.
As marine pollution is serious and marine resources are more in short supply, vegetable protein prepares biologically active peptide and also increasingly obtains
Concern.But compared with animal protein, the anti-trioxypurine peptide from vegetable protein, is reported still fewer: main patent at present
CN201310485124.9 discloses a kind of method for being prepared with walnut dregs and having effects that the biologically active peptide of anti-trioxypurine；
CN201711281385.3 discloses a kind of vegetable protein --- and sunflower disk small-molecular peptides combine Poria cocos, corn stigma, pueraria lobata, coix lacryma-jobi
Benevolence, antierythrite prepare the antipodagric drug of anti-trioxypurine.In addition to this there are also the anti-trioxypurines that a small amount of animal/vegetable protein is composed
Product, vegetable protein main source are soybean protein.
U.S. rattan fruit is the perennial bejuco of Euphorbiaceae, and protein content may be up to 27% in kernel, and after extracting oil
The protein content of the U.S. rattan fruit fruit dregs of rice may be up to 60%.Research finds that U.S. rattan fruit nutrient protein value is higher, is a kind of good plant
Albumen source, rich in 9 kinds of amino acid needed by human, content is up to 10.6%, accounts for total amino acid content 32.6%, threonine, figured silk fabrics
Propylhomoserin, glycine are significantly higher than soybean protein, are higher by 645,184,2110mg/100g respectively.It is easy to digest and absorb.U.S. rattan fruit
Argine Monohydrochloride composition is suitable, 18 kinds of amino acid Glutamic Acid content highests, followed by glycine, aspartic acid, arginine, bright
Propylhomoserin and tyrosine account for 4.02%, 3.71%, 3.57%, 2.66%, 2.28%, the 2.27% of seed respectively, it is necessary to smart ammonia in amino acid
Acid content is highest, and nonessential amino acid Glutamic Acid content highest, the minimum amino acid of content is methionine 0.3%.
U.S. rattan fruit is as a kind of new plant for introducing China, and the research about its albumen and polypeptide is at home and abroad all very
It is few, current study show that U.S. rattan fruit albumen has immunological regulation, promotes fermentation, anti-oxidation efficacy；And related U.S. rattan fruit anti-trioxypurine
Peptide preparation process and its bioactive functions research have not been reported.Therefore, develop U.S. rattan fruit anti-trioxypurine peptide, to the preparation of its polypeptide and
Functional activity is studied, and has great importance to U.S. rattan fruit polypeptide functional product is developed.
Summary of the invention
The primary purpose of the present invention is that providing a kind of preparation method of U.S. rattan fruit anti-trioxypurine peptide, which uses biological enzyme
Solution technology, which discharges, has the active ingredient of anti-trioxypurine in U.S. rattan fruit albumen, and has effects that the bacterial strain lactic acid of anti-trioxypurine using one plant
Bacillus DU-106 fermentation promotes the anti-trioxypurine activity of biologically active peptide in enzymolysis liquid, and it is living to be then enriched with target by ultrafiltration membrane
Property peptide.Anti-trioxypurine effect using U.S. rattan fruit protein peptides made from the method for the present invention is obvious, can significantly reduce serum uric acid level,
Economic value is high, is suitble to large-scale industrial production.
To achieve the goals above, the present invention adopts the following technical scheme:
A kind of preparation method with the U.S. rattan fruit dregs of rice biologically active peptide of anti-trioxypurine, includes the following steps:
(1) U.S. rattan fruit dregs of rice pre-treatment:
U.S. rattan fruit dregs of rice powder is subjected to ungrease treatment, volatilization eliminates organic reagent overnight, adds by weight the solid-liquid ratio of 1:10 ~ 1:25
Water impregnates to obtain U.S. rattan fruit dregs of rice solution；
(2) protease hydrolyzed:
U.S. rattan fruit dregs of rice solution is heated to 40-60 DEG C of required temperature of enzymatic hydrolysis, adjusts pH value of solution=2-11, protease hydrolytic 2- is added
6h, high temperature enzyme deactivation, centrifugation take supernatant, obtain the U.S. thick enzymolysis liquid of rattan fruit albumen；
(3) lactobacillus-fermented:
Lactic acid bacteria freeze drying strain transfer kind to MRS culture medium tube is activated into three generations, and activated spawn is expanded into culture medium in MRS
Thallus is collected in middle culture, centrifuge separation, is finally suspended with 0.85% physiological saline, and the ultimate density of fermentation seed liquid is 108−1010
CFU/ mL；According to 106-108Fermentation seed liquid is seeded to the U.S. thick enzymolysis liquid of rattan fruit albumen by the inoculum concentration of CFU/mL, and stirring is mixed
It is even, it ferments 2-7 days at 25 DEG C -35 DEG C；
(4) it neutralizes, filter:
Hydrolyzate after fermentation is adjusted to pH as neutrality；Then hydrolyzate is filtered using 0.22 μm of miillpore filter, is obtained
Clarification, sterile U.S. rattan fruit protein hydrolyzate；
(5) UF membrane:
By U.S. rattan fruit protein hydrolyzate by ultrafiltration membrane, permeate is taken to be concentrated by evaporation, is freeze-dried, obtains having effects that anti-trioxypurine
U.S. rattan fruit dregs of rice bioactivity peptide freeze-dried powder.
Preferably, step (1) described degreasing is to adopt in the preparation method of above-mentioned U.S. rattan fruit dregs of rice biologically active peptide
With petroleum ether or supercritical extract.
Preferably, step (2) protease is stomach in the preparation method of above-mentioned U.S. rattan fruit dregs of rice biologically active peptide
One or more of protease, alkali protease, papain, neutral proteinase, trypsase it is compound.
Preferably, step (5) ultrafiltration membrane is film in the preparation method of above-mentioned U.S. rattan fruit dregs of rice biologically active peptide
Flux is the ultrafiltration membrane of 3000Da.
Preferably, step (3) lactic acid bacteria is lactic acid bud in the preparation method of above-mentioned U.S. rattan fruit dregs of rice biologically active peptide
Spore bacillus DU-106 is preserved in Guangdong Province's Culture Collection on March 22nd, 2019, address: GuangZhou, China city is first
5 building, the building of compound the 59th of strong Road 100, deposit number are GDMCC 60621.Systematic name isBacillus lactics。
Compared with prior art, the invention has the following beneficial effects:
(1) present invention prepares U.S. rattan fruit anti-trioxypurine peptide using protease hydrolyzed, and safe and healthy, product property is stablized, dynamic through rat
Object experiment shows that the anti-trioxypurine peptide using the preparation of this patent method can significantly reduce the serum uric acid level of rat, has significant drop
Uric acid effect.
(2) utilize reported the lactic acid bacillus DU-106 with anti-trioxypurine to the U.S. rattan fruit egg after enzymatic hydrolysis before this
White hydrolysate carries out glycolysis, and product anti-trioxypurine activity further increases, and molecular weight is small, and absorption of human body utilization efficiency is more preferable.
(3) the U.S. rattan fruit dregs of rice are the leftover bits and pieces generated after U.S. rattan fruit is extracted oil, prepare biologically active peptide technique using the U.S. rattan fruit dregs of rice
It is easy to operate, production cost is low, without any pollution, realize the effective use of resource, meet Green Sustainable idea.
Detailed description of the invention
Fig. 1 is U.S. rattan fruit dregs of rice degree of hydrolysis after different Protease Treatments, sour hydrolysis process；
Fig. 2 is 1 graph of molecular weight distribution of comparative example；
Fig. 3 is 1 graph of molecular weight distribution of embodiment；
Fig. 4 is 2 graph of molecular weight distribution of embodiment；
Fig. 5 is 3 graph of molecular weight distribution of embodiment；
Fig. 6 is mice serum uric acid content figure after administration 4 weeks.
Specific embodiment
Below with reference to embodiment, the invention will be further described, and embodiment is served only for explaining the present invention, will not be to this hair
It is bright to constitute any restriction.
Comparative example 1: preparation has effects that the U.S. rattan fruit dregs of rice biologically active peptide of anti-trioxypurine
(1) U.S. rattan fruit dregs of rice pre-treatment
The U.S. rattan fruitcake dregs of rice are crushed and then cross 40 meshes, organic solvent is dried in oven overnight after supercritical extract degreasing and removes
To the greatest extent, by weight the U.S. rattan fruit dregs of rice: distilled water=1:25 solid-liquid ratio soaks in water 3 hours to obtain U.S. rattan fruit dregs of rice solution；
(2) protease hydrolyzed
It selects trypsase that the U.S. rattan fruit dregs of rice are hydrolyzed and prepares polypeptide.U.S. rattan fruit dregs of rice solution is heated to trypsin digestion most
55 DEG C of thermophilic degree, 5% tryptose is added in hydrochloric acid or sodium hydrate regulator solution pH=9 using 0.1mol/L, after hydrolyzing 6h,
90 DEG C of high temperature enzyme deactivation 15min, 4000 r/min centrifugation 10min, take the U.S. rattan fruit dregs of rice enzymolysis liquid of supernatant
(3) UF membrane
The ultrafiltration membrane for being 3000Da by membrane flux by U.S. rattan fruit dregs of rice enzymolysis liquid, takes permeate to be concentrated by evaporation, and is freeze-dried, obtains
Has effects that the U.S. rattan fruit protolysate 1 of anti-trioxypurine.
Embodiment 1: preparation has effects that the U.S. rattan fruit dregs of rice biologically active peptide of anti-trioxypurine
(1) U.S. rattan fruit dregs of rice pre-treatment
The U.S. rattan fruitcake dregs of rice are crushed and then cross 40 meshes, organic solvent is dried in oven overnight after supercritical extract degreasing and removes
To the greatest extent, by the U.S. rattan fruit dregs of rice: distilled water=1:25 feed liquid weight ratio soaks in water 3 hours to obtain U.S. rattan fruit dregs of rice solution；
(2) protease hydrolyzed
It selects neutral proteinase that the U.S. rattan fruit dregs of rice are hydrolyzed and prepares polypeptide.U.S. rattan fruit dregs of rice solution is heated to neutral proteinase enzyme
45 DEG C of optimum temperature of solution, 5% neutral proteinase, water is added in hydrochloric acid and sodium hydrate regulator solution pH=7 using 0.1mol/L
After solving 6h, 90 DEG C of high temperature enzyme deactivation 15min, 4000 r/min centrifugation 10min take the U.S. rattan fruit dregs of rice enzymolysis liquid of supernatant；
(3) lactic acid bacillus DU-106 ferments
The lactic acid bacillus DU-106 freeze-drying lactobacillus that laboratory saves is turned to be seeded to MRS culture medium tube activation three generations, and
Activated spawn is expanded in culture medium in MRS and is cultivated, thallus is collected in centrifuge separation, is finally suspended, will be sent out with 0.85% physiological saline
The ultimate density of ferment seed liquor is 108−1010 CFU/ mL.According to 108The inoculum concentration of CFU/mL, fermentation seed liquid is seeded to
The U.S. thick enzymolysis liquid of rattan fruit albumen, stirs and evenly mixs, and ferments 7 days at 25 DEG C.
(4) it neutralizes, filter
It is neutral that the hydrolyzate after fermentation, which is adjusted to pH, with appropriate sodium hydroxide solution or sodium carbonate liquor;By the hydrolysis after neutralization
Liquid is filtered using 0.22 μm of miillpore filter, is clarified, sterile U.S. rattan fruit protein hydrolyzate；
(5) UF membrane
The ultrafiltration membrane for being 3000Da by membrane flux by U.S. rattan fruit protein hydrolyzate, takes permeate to be concentrated by evaporation, and is freeze-dried, obtains
To having effects that the U.S. rattan fruit protolysate 2 of anti-trioxypurine.
Embodiment 2: preparation has effects that the U.S. rattan fruit dregs of rice biologically active peptide of anti-trioxypurine
(1) U.S. rattan fruit dregs of rice pre-treatment
The U.S. rattan fruitcake dregs of rice are crushed and then cross 40 meshes, organic solvent is dried in oven overnight after supercritical extract degreasing and removes
To the greatest extent, by the U.S. rattan fruit dregs of rice: distilled water=1:25 feed liquid weight ratio soaks in water 3 hours to obtain U.S. rattan fruit dregs of rice solution；
(2) protease hydrolyzed
It selects alkali protease that the U.S. rattan fruit dregs of rice are hydrolyzed and prepares polypeptide.U.S. rattan fruit dregs of rice solution is heated to alkali protease enzyme
50 DEG C of optimum temperature of solution, 5% alkali protease is added in hydrochloric acid or sodium hydrate regulator solution pH=10 using 0.1mol/L,
After hydrolyzing 6h, 90 DEG C of high temperature enzyme deactivation 15min, 4000 r/min centrifugation 10min take the U.S. rattan fruit dregs of rice enzymolysis liquid of supernatant.
(3) lactic acid bacillus DU-106 ferments
The lactic acid bacillus DU-106 freeze-drying lactobacillus that laboratory saves is turned to be seeded to MRS culture medium tube activation three generations, and
Activated spawn is expanded in culture medium in MRS and is cultivated, thallus is collected in centrifuge separation, is finally suspended, will be sent out with 0.85% physiological saline
The ultimate density of ferment seed liquor is 108−1010 CFU/ mL.According to 108The inoculum concentration of CFU/mL, fermentation seed liquid is seeded to
The U.S. thick enzymolysis liquid of rattan fruit albumen, stirs and evenly mixs, and ferments 7 days at 25 DEG C.
(4) it neutralizes, filter
It is neutral that the hydrolyzate after fermentation, which is adjusted to pH, with appropriate sodium hydroxide solution or sodium carbonate liquor;By the hydrolysis after neutralization
Liquid is filtered using 0.22 μm of miillpore filter, is clarified, sterile U.S. rattan fruit protein hydrolyzate；
(5) UF membrane
The ultrafiltration membrane for being 3000Da by membrane flux by U.S. rattan fruit protein hydrolyzate, takes permeate to be concentrated by evaporation, and is freeze-dried, obtains
To having effects that the U.S. rattan fruit protolysate 3 of anti-trioxypurine.
Embodiment 3: preparation has effects that the U.S. rattan fruit dregs of rice biologically active peptide of anti-trioxypurine
(1) U.S. rattan fruit dregs of rice pre-treatment
The U.S. rattan fruitcake dregs of rice are crushed and then cross 40 meshes, organic solvent is dried in oven overnight after supercritical extract degreasing and removes
To the greatest extent, by the U.S. rattan fruit dregs of rice: distilled water=1:25 feed liquid weight ratio soaks in water 3 hours to obtain U.S. rattan fruit dregs of rice solution；
(2) protease hydrolyzed
It selects trypsase that the U.S. rattan fruit dregs of rice are hydrolyzed and prepares polypeptide.U.S. rattan fruit dregs of rice solution is heated to trypsin digestion most
55 DEG C of thermophilic degree, 5% trypsase is added in hydrochloric acid or sodium hydrate regulator solution pH=9 using 0.1mol/L, hydrolyzes 6h
Afterwards, 90 DEG C of high temperature enzyme deactivation 15min, 4000 r/min centrifugation 10min, take the U.S. rattan fruit dregs of rice enzymolysis liquid of supernatant；
(3) lactic acid bacillus DU-106 ferments
The lactic acid bacillus DU-106 freeze-drying lactobacillus that laboratory saves is turned to be seeded to MRS culture medium tube activation three generations, and
Activated spawn is expanded in culture medium in MRS and is cultivated, thallus is collected in centrifuge separation, is finally suspended, will be sent out with 0.85% physiological saline
The ultimate density of ferment seed liquor is 108−1010 CFU/ mL.According to 108The inoculum concentration of CFU/mL, fermentation seed liquid is seeded to
The U.S. thick enzymolysis liquid of rattan fruit albumen, stirs and evenly mixs, and ferments 7 days at 25 DEG C.
(4) it neutralizes, filter
It is neutral that the hydrolyzate after fermentation, which is adjusted to pH, with appropriate sodium hydroxide solution or sodium carbonate liquor;By the water after neutralization
Solution liquid is filtered using 0.22 μm of miillpore filter, is clarified, sterile U.S. rattan fruit protein hydrolyzate；
(5) UF membrane
The ultrafiltration membrane for being 3000Da by membrane flux by U.S. rattan fruit protein hydrolyzate, takes permeate to be concentrated by evaporation, and is freeze-dried, obtains
To having effects that the U.S. rattan fruit protolysate 4 of anti-trioxypurine.
Embodiment 4: Efficacy experiments
1, the measurement of degree of hydrolysis: formol titration
2, the measurement of hydrolysate molecular mass: the molecular vibrational temperature of protein peptides is measured using gel permeation chromatography.Chromatography
Column be TSKgel G2000 SWxL(300 mm × 7.8 mm, 5 μm), mobile phase be acetonitrile-water-trifluoroacetic acid (volume ratio 45:
55: 0.1), being that 214 nm are measured sample with Detection wavelength under conditions of 30 DEG C of column temperature, 0.5 mL/min of flow velocity.
3, animal experiment.Anti-trioxypurine effect animal experiment method of the embodiment of the present invention is as follows:
(1) experimental material
Animal: SPF grades SD rat 70, male, weight 200 ± 20 (g), the offer of Traditional Chinese Medicine University Of Guangzhou's Experimental Animal Center.
Reagent: allopurinol tablet；Oteracil Potassium；Sodium carboxymethylcellulose；Uric acid detection kit.
(2) experimental method
Animal packet and modeling: 70 rats are randomly divided into Normal group (10) and modeling group (60), modeling group rat
Stomach stomach-filling Oteracil Potassium one week (once a day) utilizes 3% yellow Jackets (30 mgkg-1) intraperitoneal injection of anesthesia, after eye socket
Veniplex is taken a blood sample (0.5 ml), with 4 DEG C, 3000rpm, centrifugation 15min, upper serum is taken to measure uric acid content；Normal group
Rat oral gavage gives the solvent of isometric(al).Uric acid content is higher than 110 μm of olL-1Rat be determined as modeling success.It will make
The successful rat of mould is randomly divided into model control group (isometric(al) solvent), U.S. rattan fruit protolysate 1 by uric acid content
(200mgkg-1), U.S. rattan fruit protolysate 2(200mgkg-1), U.S. rattan fruit protolysate 3(200mgkg-1), beauty
Rattan fruit protolysate 4(200mgkg-1) and Allopurinol group (25mgkg-1), every group rat 10, gastric infusion (administration
Volume 10ml/kg), rat model gives the distilled water of isometric(al).After above-mentioned sample treatment 28d, in last dose 50min, 3%
Yellow Jackets (30mgkg-1) after intraperitoneal injection of anesthesia, retroorbital venous clump takes blood 0.5ml, measures serum uric acid.
Present invention selection is compared with above-described embodiment 1,2,3 and comparative example 1, compares the degree of hydrolysis of hydrolysate, relative molecular weight point
Uric acid content in cloth and zoopery.
The present invention is raw material using the U.S. rattan fruit dregs of rice, hydrolyzes preparation beauty under each protease optimum condition using different protease
Rattan fruit dregs of rice polypeptide the experiment has found that neutral proteinase, trypsase and alkali protease tentatively digest the U.S. rattan fruit dregs of rice, obtain
The enzymolysis liquid degree of hydrolysis arrived is up to 16 ~ 25%, and wherein trypsin hydrolysis degree highest, release small-molecular peptides ability are stronger；Embodiment exists
On the basis of selecting different protease to carry out initial hydrolysis to the U.S. rattan fruit dregs of rice, continues to use lactic acid bacillus DU-106 and it is carried out
Fermentation, the zymolyte degree of hydrolysis after fermenting increase in various degree, reach as high as 30% or more, improve 20% than comparative example, release
More new peptide fragments out.
It is less than the zymolyte of 3000kDa to the further concentration and separation molecular weight of hydrolyzate using the ultrafiltration membrane of 3000Da, it is real
It applies shown in example 1,2,3 and 1 graph of molecular weight distribution of comparative example, 1 average molecular weight Mw=1746 of comparative example, 1 average molecular weight of embodiment
Mw=872,2 average molecular weight Mw=1056 of case, 3 molecular weight of product of embodiment < 1500kDa peptide fragment account for 90% or more, further
Illustrate that protease combination DU-106 fermentation is higher than single use protease to U.S. rattan fruit dregs of rice hydrolysis efficiency and is hydrolyzed.
Model group rats are after Oteracil Potassium is handled 4 weeks as shown in Figure 5, and serum uric acid obviously increases, with model group
Comparison, after embodiment group, comparative example group and medicine group are handled 4 weeks, play the role of reducing rat model serum uric acid (p <
0.05), embodiment group and comparative example group anti-trioxypurine effect are superior to medicine group, and 1,2,3 anti-trioxypurine effect of embodiment is superior to pair
Ratio 1, wherein 3 anti-trioxypurine effect of embodiment is best.The result shows that being had using the U.S. rattan fruit dregs of rice polypeptide of U.S. rattan fruit dregs of rice hydrolysis preparation
There is anti-trioxypurine effect, further increased using anti-trioxypurine effect after DU-106 fermentation process, for preparing anti-trioxypurine effect series
Peptide product with good application prospect.
Claims (5)
1. a kind of preparation method with the U.S. rattan fruit dregs of rice biologically active peptide of anti-trioxypurine, it is characterised in that including walking as follows
It is rapid:
(1) U.S. rattan fruit dregs of rice pre-treatment:
U.S. rattan fruit dregs of rice powder is subjected to ungrease treatment, volatilization eliminates organic reagent overnight, adds by weight the solid-liquid ratio of 1:10 ~ 1:25
Water impregnates to obtain U.S. rattan fruit dregs of rice solution；
(2) protease hydrolyzed:
U.S. rattan fruit dregs of rice solution is heated to 40-60 DEG C of required temperature of enzymatic hydrolysis, adjusts pH value of solution=2-11, protease hydrolytic 2- is added
6h, high temperature enzyme deactivation, centrifugation take supernatant, obtain the U.S. thick enzymolysis liquid of rattan fruit albumen；
(3) lactic acid bacillus DU-106 ferments:
By lactic acid bacillus DU-106 freeze-drying lactobacillus turn be seeded to MRS culture medium tube activation three generations, and by activated spawn in
MRS expand culture medium in cultivates, centrifuge separation collect thallus, finally with 0.85% physiological saline suspension, fermentation seed liquid it is final
Concentration is 108−1010CFU/ mL；According to 106-108Fermentation seed liquid is seeded to U.S. rattan fruit albumen by the inoculum concentration of CFU/mL
Thick enzymolysis liquid, stirs and evenly mixs, and ferments 2-7 days at 25 DEG C -35 DEG C；
(4) it neutralizes, filter:
Hydrolyzate after fermentation is adjusted to pH as neutrality；Then hydrolyzate is filtered using 0.22 μm of miillpore filter, is obtained
Clarification, sterile U.S. rattan fruit protein hydrolyzate；
(5) UF membrane:
By U.S. rattan fruit protein hydrolyzate by ultrafiltration membrane, permeate is taken to be concentrated by evaporation, is freeze-dried, obtains having effects that anti-trioxypurine
U.S. rattan fruit dregs of rice bioactivity peptide freeze-dried powder.
2. the preparation method of U.S. rattan fruit dregs of rice biologically active peptide as described in claim 1, which is characterized in that step (1) is described de-
Rouge is using petroleum ether or supercritical extract.
3. the preparation method of U.S. rattan fruit dregs of rice biologically active peptide as described in claim 1, which is characterized in that step (2) described egg
White enzyme is the compound of one or more of pepsin, alkali protease, papain, neutral proteinase, trypsase.
4. the preparation method of U.S. rattan fruit dregs of rice biologically active peptide as described in claim 1, which is characterized in that step (5) is described super
Filter membrane is the ultrafiltration membrane that membrane flux is 3000Da.
5. the preparation method of U.S. rattan fruit dregs of rice biologically active peptide as described in claim 1, which is characterized in that step (3) described cream
Sour bacterium is lactic acid bacillus DU-106.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910320558.0A CN110066844A (en) | 2019-04-20 | 2019-04-20 | A kind of preparation method with the U.S. rattan fruit dregs of rice biologically active peptide of anti-trioxypurine |
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| CN111961125A (en) * | 2020-08-27 | 2020-11-20 | 华南农业大学 | Plukenetia volubilis linneo immune active peptide and preparation method thereof |
| CN112080539A (en) * | 2020-08-25 | 2020-12-15 | 华中农业大学 | Kidney bean bioactive peptide and application thereof in preparation of uric acid reducing medicines |
| CN113287649A (en) * | 2021-03-10 | 2021-08-24 | 咀香园健康食品（中山）有限公司 | Composition with function of reducing uric acid and honey-suckle fruit polypeptide biscuit |
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Application publication date: 20190730 |