CN107287270A - A kind of tower draws oligopeptide and its industrialized process for preparing and purposes - Google Patents

A kind of tower draws oligopeptide and its industrialized process for preparing and purposes Download PDF

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Publication number
CN107287270A
CN107287270A CN201710588063.7A CN201710588063A CN107287270A CN 107287270 A CN107287270 A CN 107287270A CN 201710588063 A CN201710588063 A CN 201710588063A CN 107287270 A CN107287270 A CN 107287270A
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China
Prior art keywords
tower
food
oligopeptide
enzyme activity
molecular weight
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CN201710588063.7A
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王昭日
杨胜杰
刘明川
刘敏
洪达
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Sinphar Tian Li Pharmaceutical Co Ltd
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Sinphar Tian Li Pharmaceutical Co Ltd
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Priority to CN201710588063.7A priority Critical patent/CN107287270A/en
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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
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/17—Amino acids, peptides or proteins
    • A23L33/18—Peptides; Protein hydrolysates
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/01—Hydrolysed proteins; Derivatives thereof
    • A61K38/011—Hydrolysed proteins; Derivatives thereof from plants
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs

Abstract

Oligopeptide and its industrialized process for preparing and purposes are drawn the present invention relates to a kind of tower.The peptide content of the oligomeric Gly-His-Lys accounts for more than 98% in more than 80wt%, molecular weight distribution in below 1500Dalton, and ash content is not higher than 5%.The method of the present invention is easy to operate, inexpensive in high yield, and product quality is stable, technique environmental protection, is adapted to large-scale production.The tower, which drags down poly- Gly-His-Lys, has preferable radicals scavenging effect, therefore can prepare the medicine, food or health products for treating relevant disease.

Description

A kind of tower draws oligopeptide and its industrialized process for preparing and purposes
Technical field
The present invention provides a kind of tower and draws oligopeptide and its industrialized producing technology and application thereof, belongs to food deep processing neck Domain.
Background technology
Tower draws (Caesalpinia spinose Kuntze) also known as Caesalpinia spinosaKuntze (Tara), is Caesalpiniaceae Caesalpinia, The main main country such as the Peru of the northwestward, Ecuador, Colombia in South America of distribution, abounds with Peru.Its dry heat resistance, Fast-growing, evergreen, perpetual bloom, are one of fine tree species of planting trees on barren hills.Its long lifespan abounds with the phase about 20 years up to more than 50 years, 3-4 per mu yields beanpod is up to more than 300 kilograms after cultivation.It is that worker employed in a plant nursery is few, invests small, income is big, the excellent economic forest tree of many benefits of a labor Kind.The beginning of the nineties in last century, Resources Insect Inst., Chinese Academy of Forestry Sciences has introduced the production of replacement Chinese gall from South America Supplement material plant-the Ta La of nutgall tannin and its series of products, and plant into Yunnan Province is xeothermic, dry-warm valley is regional Work(.To 2006, Yunnan was developed tower and furthered 1.5 ten thousand mu, and the provinces and regions such as Guizhou, Sichuan, Guangxi and Hainan also begin to introduce a fine variety.Tower is drawn Beanpod 55-61% containing talas of tannic acid, can produce hundreds of medicine and chemical products；Its benevolence glucomannoglycan containing gala is about 34%, the tara gum for having important use in terms of food, medicine, weaving, papermaking and oil drilling can be produced, while in benevolence Contain 13% aliphatic acid (linoleic acid accounts for the 52-54% of total fatty acids).Researcher is extracted stigmasterol and paddy steroid from Tara seeds (two alcohol account for the 90% of sterol total amount to alcohol, and the former is the primary raw material for producing cloud big 120, and the latter can produce hypolipidemic, are worth Up to more than 1000 dollars/kilogram).The beans purposes is wide, and nontoxic edible, is a kind of precious woody beans.Show Tara seeds according to the study In containing more than 13% protein, be a kind of important vegetable protein sources.Compared with albumen, its hydrolysate tower drags down poly- In addition to the characteristics of peptide has high-quality protein as amino acid constitutes and draws albumen with tower, also drawn with tower not available for albumen
The physicochemical property such as good dissolubility, stability, and with easily absorb and low-allergen, reduction blood fat and Cholesterol, reduce blood pressure, promote the different physiological roles such as mineral absorption and fat metabolism, enhancing athlete's physique, therefore opening Fa Tala oligopeptides have become the study hotspot in food and medicine field.Our A of early stage patent CN 105925648 disclose tower The preparation technology of albumen and its peptide is drawn, peptide content is not less than 70wt%, and more than 95% peptide molecular weight is less than 1500Dalton, ash Divide 10% or so.Oligopeptide is drawn in order to obtain the tower that peptide content is higher and ash content is lower, and applied to industrial production, this hair Bright to draw oligopeptide preparation technology there is provided a kind of more excellent tower, the tower obtained by preparation draws the peptide content of oligopeptide more than 80%, Ash content is controlled below 5%.Meanwhile, biologos experiment shows, the tower of this technique productions, which drags down poly- Gly-His-Lys, equally to be had more preferably Antioxidation activity, the medicine, food or health products for excessively causing relevant disease for treating free radical can be prepared.
In food in addition to containing large amount of organic matter, also containing more rich inorganic constituents.These inorganic constituents are maintained The normal physiological function of human body, has highly important effect in terms of constituting tissue.Food is remained after high temperature sintering Inorganic substances be referred to as ash content.Ash content is mainly the mineral salt or inorganic salts in food, and it is to evaluate food to determine ash content in food One of index of quality.For food service industry, ash content is the important quality index of a body.For example, adding in flour In work, flour grade is often evaluated with total ash content, because 20 times higher than endosperm or so of the content of ashes of wheat bran, because This, the machining accuracy of flour is higher, and content of ashes is lower.When producing the gelled products such as pectin, gelatin, this can be explained in total ash The jelly performance of a little products；Water-soluble ass then largely shows the fruit content in the fruit products such as jam, jelly； And the increase of acid-insoluble ash then imply that pollution and adulterate.This is to ensureing that food quality is highly important.Food and medicine The control of ash content has following significances in product：
(1) total ash content of food is the important evidence for controlling food products or semi-manufactured goods quality.Such as：In milk Content of the total ash in milk is constant.Typically in 0.68%--0.74%, average value closely 0.70%, therefore can It is whether adulterated to determine milk with the method for determining total ash in milk, if water mixing, ash content reduction.It can in addition contain judge concentration Than if measuring milk ash content 1.4% or so, illustrating that milk concentrates one times.Wheat bran content of ashes in and for example superior wheat flour, wheat Height, and protein content is high in endosperm, the content of the ash rate endosperm of wheat bran is high 20 times, that is the precision in flour is high, then Ash content is just low.
(2) whether evaluation food is hygienic, either with or without pollution.Different food, because of raw materials used, processing method and measure bar The difference of part, the composition and content of various ash contents are also differed, after these conditions are determined, the ash content of certain food is often certain In the range of.If content of ashes has exceeded normal range (NR), illustrate to have used the original departing from sanitary standard requirement in food production Material or food additives, or food are contaminated in processing, transporting procedures.Therefore, determine ash content may determine that food by The degree of pollution
(3) judge whether food is adulterated.
(4) reference index (can be by surveying various elements) of nutrition is evaluated.
(5) machining accuracy and food quality of food can be judged.Inorganic salts are one of six big nutritional factors, are mankind's lifes The indispensable material of life activity, will correctly evaluate certain nutritive value of food, and its inorganic salt content is an evaluation index.Example As usual to evaluate flour grade with total ash content, superior wheat flour is 0.3-0.5%；Standard flour is 0.6-0.9%.Produce pectin, bright During the colloid product of glue etc, ash content is the mark of the jelly performance of these products.
The content of the invention
Oligopeptide and its industrial preparation process are drawn the invention discloses a kind of tower, it is intended to ensure the premise of product yield Under there is provided the industrial preparation process that a kind of peptide content is higher and ash content is lower tower draws oligopeptide, technique is more easy, green Environmental protection, is adapted to large-scale production.The tower of this technique productions, which drags down poly- Gly-His-Lys, has more preferable antioxidation activity, can be used for treatment Or medicine, food or the health food of the excessive caused symptom of prevention free radical.
In the preparation process that tower draws oligopeptide, due in protein extraction and enzymolysis process, to reach biological enzyme reaction Condition, constantly adds hydrochloric acid and sodium hydroxide, causes the content of salt high, so as to cause content of ashes high.Higher ash content, shadow Peptide content in product is rung.Because the molecular weight of salt is smaller, the purification process of early stage peptide is all the milipore filter of larger molecular weight, most The permeate for milipore filter collected afterwards, therefore peptide, amino acid and salt are in permeate, it is impossible to salt is removed.
The ash content of numerous food is controlled in 8wt%, national standard (the GBT 22492- of such as soy peptide powder in the prior art 2008, its content is fully incorporated and referred to herein).But using the application preparation method, ash content can be controlled in 5wt% with Under.
To reach above-mentioned purpose, the technical solution adopted by the present invention is：
A kind of tower draws oligopeptide, it is characterised in that：Using GB/T 22492-2008 appendix As and the detection method of Appendix B, Peptide content is measured in more than 80wt%, molecular weight accounts for more than 98% less than below 1500Dalton, and its molecular weight distribution is as follows：
Wherein described ash content is less than 5wt%；
Preferably, peptide content is more than 82.3wt%, and molecular weight accounts for more than 98.5% less than below 1500Dalton.
Preferably, peptide content is more than 85wt%, and molecular weight accounts for more than 98% less than below 1500Dalton.
Preferably, peptide content is more than 90wt%, and molecular weight accounts for more than 98% less than below 1500Dalton.
Preferably, peptide content is more than 95wt%, and molecular weight accounts for more than 98% less than below 1500Dalton.
Present invention also offers the preparation method that the tower drags down poly- Gly-His-Lys, comprise the steps：
(1) Tara seeds are crushed, crosses 40 mesh with upper screen cloth to obtain Tara powder；
(2) Tara powder and purified water mixs for 1: 10~1: 20 in mass ratio, addition bean powder quality 0.1%~ 1% Polyose degradation enzyme, adjusts pH to 5~8,40 DEG C of 1~2h of stirring of constant temperature, after the completion of, centrifugal filtration, filtrate is stand-by；
(3) filter residue in step (2) is mixed with its mass ratio for 1: 10~1: 20 purified water, pH is to 8~10 for regulation, 40 DEG C of 1~2h of extraction of constant temperature, centrifugal filtration after the completion of extraction discards filter residue, and filtrate is stand-by；
(4) filtrate in combining step (2) and (3), regulation pH is 3~5, stands 1~2h, and abandoning supernatant obtains egg White sediment；
(5) purified water that volume ratio is 1: 5~1: 15 will be added in the albumen precipitation thing in step (4), stir shape Liquid is redissolved into albumen；
(6) albumen in step (5) is redissolved into liquid and is heated to 40~55 DEG C, add 0.5~2% egg of Tara powder quality After white enzyme, 4~6h of stirring enzymolysis, adjustment pH is 2~6, boils 10~30min of inactivation, centrifugal filtration, filtrate is standby；
(7) filtrate in step (6) is filtered using aperture for less than 0.5 μm of microfiltration membranes, permeate is again through 100 After~400Dalton nanofiltration membrane treatments, after trapped fluid is concentrated and dried, obtains tower and drag down poly- Gly-His-Lys.
Described preparation method, it is characterised in that：Described method for concentration can for film concentration, be concentrated under reduced pressure or normal pressure is dense Contracting；Described drying means can be spray drying, vacuum drying, heat drying or freeze-drying.
Described preparation method, it is characterised in that：Described Polyose degradation enzyme is selected from the 1,4 beta-glucanase (enzyme activity of food-grade Power >=2000U/g) and mannase (enzyme activity >=2000U/g) in a kind of or their mixture, it is preferred to use 1,4 beta-glucanase.
The tower draws the industrialized process for preparing of oligopeptide, it is characterised in that：The biology enzyme is selected from the neutrality of food-grade Protease (enzyme activity >=300,000 u/g), papain (enzyme activity >=400,000 u/g), the bromelain (u/ of enzyme activity >=300,000 G), alkali protease (enzyme activity >=200,000 u/g), pepsin (enzyme activity >=500,000 u/g), pancreatin (enzyme activity >=3000u/ G) a kind of or their mixture in, it is preferred to use neutral proteinase.
It is highly preferred that the preparation method, which is included in purification procedures, uses 100~400Dalton nanofiltration membrane treatments The step of.
Present invention also offers a kind of composition, it contains above-mentioned tower and dragged down can connect on poly- Gly-His-Lys, and medicine or food The auxiliary agent received.
The formulation of the composition be selected from plain piece, thin membrane coated tablet, sugar coated tablet, casing piece, dispersible tablet, capsule, granule, Oral administration solution or oral administration mixed suspension.
Tower of the present invention drags down poly- Gly-His-Lys, composition, all excessively caused for preparing treatment or prevention free radical The application of the medicine of symptom, food or health products.Improve the Cardiovascular Toxicity as caused by PM2.5 particulate matters for preparing；Reduction The particulate matters of PM 2.5 are to macrophage inhibitory action；Strengthen phagocytic function of the macrophage to the particulate matters of PM 2.5；Promote enteron aisle Application to the secretory medicine, food or health products of the particulate matters of PM 2.5.The oligomeric Gly-His-Lys of described Tara seeds are in Property protease prepare.
Embodiment
Below by embodiment, the invention will be further described.It should be understood that methods described of the embodiment of the present invention It is only used for the explanation present invention, rather than limitation of the present invention, to preparation side of the invention under the concept thereof of the present invention The simple modifications of method belong to the scope of protection of present invention.Unless otherwise instructed, all raw materials for being used in embodiment and Solvent is commercially available prod.The amount of activity product and positive control medicine of the present invention used in implementing, according to experiment acceptor Maximum tolerated dose is added.
Comparative example：
The Gly-His-Lys that the A of patent CN 105925648 preparation method is obtained：By Tara seeds (Deposits, Yimen, Yunnan medicinal material market) powder It is broken, 40 mesh are crossed with upper screen cloth to obtain Tara powder.100kg Tara powders are mixed for 1: 10 in mass ratio with purified water, 0.1% 1,4 beta-glucanase (enzyme activity 3000U/g) of bean powder quality is added, pH to 6,40 DEG C of extraction 2h of constant temperature is adjusted；Extract After the completion of, centrifugal filtration, filtrate is stand-by；Filter residue is mixed with its mass ratio for 1: 15 purified water again, pH to 9, constant temperature is adjusted 40 DEG C of extraction 2h；Centrifugal filtration after the completion of extraction, discards filter residue, and filtrate is stand-by；Stand-by filtrate more than finally merging, adjusting pH is 3.5,1h is stood, abandoning supernatant adds the purified water that volume ratio is 1: 10 into sediment, is uniformly mixing to obtain albumen redissolution Liquid.Albumen redissolution liquid is heated to 45 DEG C, 1% neutral proteinase (enzyme activity is 300,000 u/g) of Tara powder quality is added, it is permanent It is 7 to determine pH, after stirring enzymolysis 4h, boils inactivation 30min, centrifuges, supernatant is protein enzymatic hydrolyzate.Protein enzymatic hydrolyzate is used Aperture is filtered for 0.5 μm of microfiltration membranes, and permeate is again after 5000Dalton ultrafiltration membrane treatments, and permeate is dense at 50 DEG C When being reduced to solid contain for 6.5wt%, it is spray-dried, obtains faint yellow tower and draw oligopeptide (lot number is TL-1), yield is 5.9wt%, measures peptide content for 71.32wt%, at least 95.77% peptide molecular weight is distributed in below 1500Dalton.
The Gly-His-Lys obtained by the inventive method：Tara seeds (Deposits, Yimen, Yunnan medicinal material market) are crushed, crosses more than 40 mesh and sieves Net is so as to obtain Tara powder.100kg Tara powders are mixed for 1: 10 in mass ratio with purified water, bean powder quality is added 0.1% 1,4 beta-glucanase (enzyme activity 3000U/g), adjusts pH to 6,40 DEG C of extraction 2h of constant temperature；After the completion of extraction, centrifuged Filter, filtrate is stand-by；Filter residue is mixed with its mass ratio for 1: 15 purified water again, pH to 9,40 DEG C of extraction 2h of constant temperature is adjusted；Carry Centrifugal filtration after the completion of taking, discards filter residue, and filtrate is stand-by；Stand-by filtrate more than finally merging, regulation pH is 3.5, stands 1h, abandons Supernatant is removed, the purified water that volume ratio is 1: 10 is added into sediment, albumen is uniformly mixing to obtain and redissolves liquid.Albumen is redissolved Liquid is heated to 45 DEG C, adds 1% neutral proteinase (enzyme activity is 300,000 u/g) of Tara powder quality, constant pH is 7, stirring Digest after 4h, pH is to 5 for regulation, boil inactivation 30min, centrifuge, supernatant is protein enzymatic hydrolyzate.Protein enzymatic hydrolyzate is used into hole Footpath is filtered for 0.5 μm of microfiltration membranes, and permeate is again after 100Dalton ultrafiltration membrane treatments, and trapped fluid liquid is dense at 50 DEG C When being reduced to solid contain for 6.2wt%, it is spray-dried, obtains faint yellow tower and draw oligopeptide (lot number is TL-2), yield is 7.1wt%, measures peptide content for 82.3wt%, at least 98.5% peptide molecular weight is distributed in below 1500Dalton.
BIOLOGICAL ACTIVITY EXAMPLES 1：
1.DPPH radicals scavengings are tested：
The preparation of 1.1 DPPH ethanol solutions：Precision weighs DPPH 4mg, is placed in 100mL brown volumetric flasks, adds 50mL ethanol, ultrasonic 30s, scale is settled to ethanol, is shaken up, stand-by.This product must be now with the current.
The preparation of 1.2 need testing solutions：Precision weighs appropriate tower and dragged down after poly- Gly-His-Lys, plus ethanol dissolving, constant volume, dilution Into required concentration, you can.
1.3 operating procedure：Accurate absorption 2mL need testing solutions and 2mL DPPH solution are well mixed；It is accurate to draw 2mL Need testing solution and 2mL ethanol are well mixed；Accurate absorption 2mLDPPH solution and 2mL ethanol are well mixed, and room temperature is placed 30min, determines absorbance, and calculate free radical scavenging activity according to following calculation formula at 515nm wavelength：
IR%=[1- (Ai-Aj)/A0] * 100%；
Wherein, Ai represents the absorbance of solution after solution to be measured and DPPH mixing；
Aj represents the absorbance of solution after solution to be measured and solvent mixing；
A0 represents the absorbance of solution after DPPH and solvent mixing.
2. ABTS+Radicals scavenging is tested：
The preparation of 2.1 PBSs：Weigh sodium chloride 8g, potassium chloride 0.2g, potassium dihydrogen phosphate 0.24g, 12 hydrations Disodium hydrogen phosphate 3.62g, is placed in 1000mL beakers, and adding 800mL distilled water , Jiao Ban dissolves it, with hydrochloric acid or hydroxide Sodium adjusts pH to 7.4, is transferred in 1000mL volumetric flasks, plus distilled water diluting is to scale, shakes up, stand-by.
2.2 ABTS+The preparation of stock solution：Precision weighs ABTS+78mg or so, is placed in 20mL brown volumetric flasks, plus Enter 15mL distilled water, ultrasonic 5min is settled to scale with distilled water, shaken up.Precision weighs potassium peroxydisulfate 76mg or so, is placed in In 2mL brown volumetric flasks, 1mL distilled water is added, ultrasound dissolves it, scale is settled to distilled water, shakes up.It is accurate to draw 352 μ L potassium persulfate solutions are added into ABTS solution, are shaken up, are stood overnight.
2.3 ABTS+The preparation of working solution：It is accurate to draw stock solution 1mL, 65mL or so PBS is added, is shaken It is even.
The preparation of 2.4 need testing solutions：Precision weighs appropriate tower and drawn after oligopeptide, plus PBS dissolving, and constant volume is dilute It is interpreted into required concentration, you can.
2.5 operating procedure：Accurate absorption 0.5mL need testing solutions and 5mL ABTS working solutions are well mixed；It is accurate to inhale Take 0.5mL need testing solutions and 5mL PBSs well mixed；It is accurate to draw 5mL ABTS working solutions and 0.5mL PBS Buffer solution is well mixed, and determines absorbance at 734nm immediately, and calculate free radical scavenging activity according to below equation：
IR%=[1- (Ai-Aj)/A0] * 100%；
Wherein, Ai represents the absorbance of solution after solution to be measured and ABTS mixing；
Aj represents the absorbance of solution after solution to be measured and solvent mixing；
A0 represents the absorbance of solution after ABTS and solvent mixing.
3.SRSA ultra-oxygen anion free radicals remove experiment：
The preparation of 3.1 NBT solution：Precision weighs NBT (NBT) 24.5mg, is placed in 100mL browns appearance In measuring bottle, 50mL distilled water is added, ultrasonic 30s is settled to scale with distilled water, shaken up, stand-by.
The preparation of 3.2 NADH solution：Precision weighs NADH reduced Coenzyme I (NADH) 33.2mg, is placed in 100mL brown volumetric flasks, adds 50mL distilled water, and ultrasonic 30s is settled to scale with distilled water, shaken up, It is stand-by.
The preparation of 3.3 PMS solution：Precision weighs phenazine methosulfate (PMS) 4.59mg, is placed in 250mL brown volumetric flasks In, 150mL distilled water is added, ultrasonic 30s is settled to scale with distilled water, shaken up, stand-by.
The preparation of 3.4 Tris-HCl cushioning liquid：Precision weighs trishydroxymethylaminomethane (Tris) 3.025g, is placed in In 500mL beakers, 200mL distilled water is added, stirring dissolves it, adjust pH to 8.0 with 1M HCl, be transferred to 250mL capacity In bottle, scale is settled to distilled water, is shaken up, it is stand-by.In use, taking the above-mentioned solution of 16mL to be placed in 100mL volumetric flasks, plus steam Distilled water is diluted to scale.
The preparation of 3.3 need testing solutions：Precision weighs appropriate tower and drawn after oligopeptide, plus Tris-HCl buffer solutions, fixed Hold, be diluted to required concentration, you can.
The detection of 3.4 light absorption values and the calculating of free radical scavenging activity：It is accurate to draw 1mL NADH solution, 1mL NBT solution and 3mL need testing solutions, control plus 3mLTris-HCl cushioning liquid, are well mixed respectively, respectively add 1mL PMS solution to mix, room temperature 5min is placed, absorbance is determined at 558nm, free radical scavenging activity calculation formula is as follows：
IR% (SRSA)=(1-Ai/A0) * 100%
Wherein, Ai represents the absorbance of solution after solution to be measured and detection reagent mixing；
A0 represents the absorbance of solution after blank solution and detection reagent mixing.
Preparing the tower of not be the same as Example respectively, to drag down poly- peptide product (lot number is TL-1 and TL-2) concentration be 1mg/mL, to tie up Raw element C is positive control (concentration is 0.1mg/mL), and test result is as shown in table 1：
The tower of table 1 drags down poly- Gly-His-Lys antioxidation activity
As shown in Table 1, the tower prepared by two methods drags down poly- Gly-His-Lys has stronger scavenging action to ABTS free radicals, Clearance rate is shown as medium more than 90% to the clearance rate of DPPH free radicals and superoxide anion (SRSA) more than 50% Intensity scavenging action.And removing of the oligopeptide (TL-2) to three kinds of free radicals is drawn by the tower prepared by this patent disclosed method Effect is preferable, thus it is speculated that reason is higher peptide content.It can be seen that the tower prepared by the inventive method of use this patent drags down poly- Gly-His-Lys It is equally applicable to prepare medicine, food or the health food for treating or preventing the excessive caused symptom of free radical.

Claims (10)

1. a kind of tower draws oligopeptide, it is characterised in that：Using GB/T 22492-2008 appendix As and the detection method of Appendix B, survey Peptide content is obtained in more than 80wt%, molecular weight accounts for more than 98% less than 1500 below Dalton, and its molecular weight distribution is as follows：
Wherein described ash content is less than 5wt%；
Preferably, peptide content is more than 82.3wt%, and molecular weight accounts for more than 98.5% less than 1500 below Dalton.
2. tower described in claim 1 draws the industrialized process for preparing of oligopeptide, it is characterised in that comprise the steps：It is pure separating Change and 100~400 Dalton nanofiltration membrane treatments are used in step.
3. tower draws the industrialized process for preparing of oligopeptide according to claim 2, it is characterised in that comprise the steps：
(1) Tara seeds are crushed, crosses 40 mesh with upper screen cloth to obtain Tara powder；
(2) Tara powder and purified water mixs for 1: 10~1: 20 in mass ratio, the 0.1%~1% of addition bean powder quality Polyose degradation enzyme, adjusts pH to 5~8,40 DEG C of 1~2h of stirring of constant temperature, after the completion of, centrifugal filtration, filtrate is stand-by；
(3) filter residue in step (2) is mixed with its mass ratio for 1: 10~1: 20 purified water, adjusts pH to 8~10, constant temperature 40 DEG C of 1~2h of extraction, centrifugal filtration after the completion of extraction discards filter residue, and filtrate is stand-by；
(4) filtrate in combining step (2) and (3), regulation pH is 3~5, stands 1~2h, and abandoning supernatant obtains albumen and sunk Starch；
(5) purified water that volume ratio is 1: 5~1: 15 will be added in the albumen precipitation thing in step (4), stirs to form egg It is white to redissolve liquid；
(6) albumen in step (5) is redissolved into liquid and is heated to 40~55 DEG C, add 0.5~2% albumen of Tara powder quality After enzyme, 4~6h of stirring enzymolysis, adjustment pH is 2~6, boils 10~30min of inactivation, centrifugal filtration, filtrate is standby；
(7) filtrate in step (6) is filtered using aperture for less than 0.5 μm of microfiltration membranes, permeate again through 100~ After 400 Dalton nanofiltration membrane treatments, after trapped fluid is concentrated and dried, obtains tower and drag down poly- Gly-His-Lys.
4. preparation method according to claim 3, it is characterised in that：Described method for concentration can be film concentration, decompression Concentration or normal pressure concentration；Described drying means can be spray drying, vacuum drying, heat drying or freeze-drying.
5. preparation method according to claim 3, it is characterised in that：Described Polyose degradation enzyme is selected from β-Portugal of food-grade A kind of or their mixture in dextranase (enzyme activity >=2000U/g) and mannase (enzyme activity >=2000U/g), 1,4 beta-glucanase is preferably used.
6. tower draws the industrialized process for preparing of oligopeptide according to claim 3, it is characterised in that：The biology enzyme is selected from food The neutral proteinase (enzyme activity >=300,000 u/g) of grade, papain (enzyme activity >=400,000 u/g), bromelain (enzyme activity The u/g of power >=300,000), alkali protease (enzyme activity >=200,000 u/g), pepsin (enzyme activity >=500,000 u/g), pancreatin (enzyme activity Power >=3000u/g) in a kind of or their mixture, it is preferred to use neutral proteinase.
7. a kind of tower draws oligopeptide, it is characterised in that：Using GB/T 22492-2008 appendix As and the detection method of Appendix B, survey Peptide content is obtained in more than 80wt%, molecular weight accounts for more than 98% less than 1500 below Dalton, and its molecular weight distribution is as follows：
Wherein described ash content is less than 5wt%；
Preferably, peptide content is more than 82.3wt%, and molecular weight accounts for more than 98.5% less than 1500 below Dalton.
The tower draw oligopeptide to be prepared by any described methods of claim 2-6.
8. a kind of composition, it is characterised in that：Being dragged down containing the tower described in claim 1 can on poly- Gly-His-Lys, and medicine or food The auxiliary agent of receiving.
9. composition according to claim 8, it is characterised in that：Its formulation be selected from plain piece, thin membrane coated tablet, sugar coated tablet, Casing piece, dispersible tablet, capsule, granule, oral administration solution or oral administration mixed suspension.
10. the tower described in claim 1 drag down poly- Gly-His-Lys be used for prepare treat or prevent the medicine of the excessive caused symptom of free radical The application of thing, food or health products；Improve the Cardiovascular Toxicity as caused by the particulate matters of PM 2.5 for preparing；Reduce PM 2.5 Grain thing is to macrophage inhibitory action；Strengthen phagocytic function of the macrophage to the particulate matters of PM 2.5；Promote enteron aisle to PM 2.5 The application of the secretory medicine of particulate matter, food or health products.
CN201710588063.7A 2017-07-18 2017-07-18 A kind of tower draws oligopeptide and its industrialized process for preparing and purposes Pending CN107287270A (en)

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