CN107267585A - A kind of selenium-rich corn small peptide with hepatoprotective effect and preparation method thereof - Google Patents
A kind of selenium-rich corn small peptide with hepatoprotective effect and preparation method thereof Download PDFInfo
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- CN107267585A CN107267585A CN201710571456.7A CN201710571456A CN107267585A CN 107267585 A CN107267585 A CN 107267585A CN 201710571456 A CN201710571456 A CN 201710571456A CN 107267585 A CN107267585 A CN 107267585A
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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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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
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- A61K38/011—Hydrolysed proteins; Derivatives thereof from plants
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- C07—ORGANIC CHEMISTRY
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- 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
- C07K1/16—Extraction; Separation; Purification by chromatography
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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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Abstract
The invention discloses a kind of selenium-rich corn small peptide with hepatoprotective effect and preparation method thereof, belong to biological technical field.It is characterized in that using zein as starting material, purify, be concentrated under reduced pressure by chelating enzymolysis, ultrafiltration, resin column, being spray-dried and obtain selenium-rich corn small peptide.Can be as primary raw material, adding other raw materials and auxiliary material, routinely the preparation that accordingly some are arranged is made in formulation method.The present invention using enzymolysis and rich micro- synchronous method carried out, with rich micro- efficiency high, the organic rate of element is high the characteristics of, selenium-rich corn small peptide hepatoprotective effect is notable.
Description
Technical field
The present invention relates to biological technical field, and in particular to a kind of with the selenium-rich corn small peptide of hepatoprotective effect and its preparation
Method.
Background technology
Liver is the maximum metabolic organ of human body, and the food that we eat is required for liver to be synthesized, decomposed.But liver
Dirty is also the organ for being easy to " sick ", and routine servicing is particularly significant.Liver is human body " chemical plant ".The major physiological work(of liver
Can probably there is kind more than 1500, it is human body " chemical plant ", the material needed for meeting synthetic protein, enzyme and various bodies.Liver
It is also the maximum digestive organs of human body, the digestive function of human body can be helped by hepatocytes secrete bile.Liver also has deposit work(
Can, energy and material necessary to the human body such as storage vitamin, fat, sugar.Liver " error " probability is high.Dry work is more, " goes out
The probability of mistake " is bigger, and liver function is more, task weight, is the organ of one easy " sick ".Best known is exactly liver
Inflammation, in addition to the virus hepatitis such as hepatitis A, hepatitis B, hepatitis, viral hepatitis type E, also alcoholic hepatitis, fat hepatitis, drug hepatitis,
Autoimmune liver disease etc..
Corn is the health-care good product in coarse food grain, and food corn is rather favourable to the health of human body.The nutritional ingredient of corn compares
Comprehensively, typically also contain containing protein 8.5%, fat 4.3%, carbohydrate 73.2%, calcium 0.022%, phosphorus .21%, iron 0.0016%
There are carrotene, vitamin B1, B2 and niacin and sitosterol, lecithin, vitamin E, lysine etc..The amino of corn peptide
In acid composition, branched-chain amino acid (leucine, isoleucine, valine) content is very high.Have been reported that, hepatic coma, hepatic encephalopathy are suffered from
Branched-amino acid content declines in person's blood, and input high content branched-chain amino acid can make the state of an illness be eased, and high side chain transfusion is extensive
Treatment applied to hepatic coma, hepatic sclerosis, serious hepatitis and chronic hepatitis.High branched-chain amino acid content is in this regard in corn peptide
There may be very high application prospect.
Peptide, is derived from albumen, the multi-functional compounds better than albumen, is current international most popular research topic and great
The function factor of development prospect.Peptide is that the polymer formed, egg are connected by covalent peptide bonds by two or more amino acid
The endless fully hydrolyzed product of white matter is also peptide, and usually by the peptide of 2 peptides~10, (, by 2 Amino acid profiles, n peptides are by n amino for 2 peptides
Acid constitute) or relative molecular mass be less than 2000D peptide be referred to as oligopeptides or small peptide (such as glutathione), by 10 with upper amino acid
The referred to as polypeptide of composition.Small peptide can be obtained by protein through modes such as enzymolysis, have advantages below compared with protein：1. small peptide
It is the courier for embodying information, with a variety of body metabolisms and physiological regulation function：Have promotion immune, it is hormone control, antibacterial, disease-resistant
Poison, lowering blood pressure and blood fat etc. are acted on；2. small peptide activity is higher, in the case of micro and low concentration, can play higher physiology
Effect；3. small peptide molecular weight is small, easy to digest to absorb, and edible safety is high.
Trace element is also known as trace element, although content in human body is simultaneously few, but the existence and health with people cease
Manner of breathing is closed, and the life of people is served vital.Their excess intake, deficiency, imbalance or shortage all can different journeys
Degree ground causes the exception of Human Physiology or occurs disease.Trace element most prominent effect be it is closely related with vitality, only
Size or less amount just can play huge physiological action only as match end.It is worth noting that these trace elements are logical
Directly or indirectly it must be supplied in the case of often by soil, but most people tend not to obtain enough micro members by diet
Element.This every kind of trace element has its special physiological function.Although they are minimum in people's in-vivo content, they are to maintaining people
Some conclusive metabolism in body are very necessary.Once having lacked these required trace elements, human body is just
Disease, or even threat to life occurs.Compare it is clear that about 30% disease is directly microelement deficiencies or uneven institute
Cause.
1973, WHO and international nutritive issue confirmed that selenium is behaved and necessary trace element in animal body.The nutrition master of selenium
If particularly being combined performance antioxidation with zymoprotein by protein.Selenium-supply can make liver cancer incidence decline 35%, make
There is the liver cancer family history person incidence of disease to decline 50%, selenium-supply can inhibition of hepatitis b virus carrier occur liver cancer and hepatitis B in crowd
In propagation and improve immune function of human body, prevent liver virus disease, hepatic sclerosis, liver canceration, fatty liver, alcoholic liver etc..
Selenium is classified as 15 kinds of daily diet nutritionals by " the daily diet nutrient quantity delivered " of Chinese Soclety of Nutrition's in October, 1998 revision
One of element, proposes that suitable intake of the adult daily to selenium is 50-250 micrograms.
Selenium-rich corn small peptide can not only further improve the absorptivity of selenium element, and can also make one body two class things of intake
Matter (small peptide and trace element), effect that is respective and cooperateing with performance liver protection.
The content of the invention
The first object of the present invention is to provide a kind of selenium-rich corn small peptide with hepatoprotective effect, the second mesh of the invention
Be to provide the selenium-rich corn small peptide prepare for liver protection food, health food and medicine in application.
The purpose of the object of the invention first is realized in：
A kind of selenium-rich corn small peptide with hepatoprotective effect, it is characterised in that prepared using following processing step：
The preparation of selenium-rich corn small peptide：Sodium selenite (or L- selenium-methyl selenium substituted aminothiopropionics) is dissolved in into deionized water to match somebody with somebody
The solution that concentration is 4~6mg/ml is made, in solution and zein ratio 8~12:1 ratio adds zein, stirring
Uniformly, heating boil 10~15 minutes, be cooled to about 30~50 DEG C, with watery hydrochloric acid adjust pH value 1.0~3.0, add 0.5~
2% pepsin, is incubated 1~3 hour, adjusts pH value 7.0~9.0, adds 0.5~2% trypsase, is incubated 3~5 hours,
Boil 10~15 minutes, centrifuge, precipitation is discarded, supernatant is added by the milipore filter ultrafiltration that relative molecular mass is 3KD, permeate
In on DA201-C large pore resin absorption columns, with water elution, water lotion is discarded, then with 50~90% (V/V) ethanol elutions, collect
Ethanol eluate, is recovered under reduced pressure ethanol and continues to be concentrated into relative density for 1.10~1.15 (60 DEG C), spray drying (enters wind-warm syndrome
Spend for 180~200 DEG C, leaving air temp is 90~100 DEG C), spray powder is collected, selenium-rich corn small peptide is produced.After testing, selenium-rich is beautiful
Rice small peptide peptide content is 60%~90%, the accumulation rate ＞ 95% of selenium.
The selenium-rich corn small peptide prepared by the above method, for being prepared into following preparations：
(1) selenium-rich corn small peptide is pressed:Corn oil:Beeswax:Lecithin is 10:35:4:1 mixed material, ground, system
It is standby into soft capsule；
(2) selenium-rich corn small peptide is pressed:Polyethylene glycol 400:Propane diols ratio is 10:38:2 mixed materials, it is ground, prepare
Into soft capsule；
(3) selenium-rich corn small peptide, adds customary adjuvant, is prepared into oral liquid, capsule, tablet, granule.
But the invention is not restricted to above-mentioned formulation.Auxiliary material used in preparation process can be replaced with other conventional auxiliary materials, such as
Starch, dextrin, Icing Sugar, honey, glycerine, sodium carboxymethylcellulose etc.；Selenium-rich corn small peptide can also be shared with other medical substances,
The effect of liver protection is reached jointly.
The second object of the present invention is to provide the purposes of above-mentioned selenium-rich corn small peptide, and it has the effect of liver protection, and it can use
In the food, health food and the medicine that prepare liver protection.
The beneficial effects of the invention are as follows：Selenium-rich corn small peptide of the present invention, prepares simplicity, with low cost；Corn small peptide with it is micro-
Secondary element collaboration plays liver-protective effect, while having the advantages that toxic side effect is small, having no drug resistance, being difficult residual, reaches
The pure natural requirement of pollution-free food, can obtain good Social benefit and economic benefit.
The test of pesticide effectiveness
For the effect of the further liver protection of checking selenium-rich corn small peptide of the present invention, inventor has carried out pharmacodynamics test and tested
Card, test method and result are as follows：
To mouse polarity alcoholic liver injury protective effect
(1) test medicine and animal
Selenium-rich corn small peptide, self-control, small peptide containing corn (with total phthaleinometer) is 83.4%, containing selenium 1.10%.Adopt before use
Finite concentration decoction, gastric infusion are made into aseptic deionized water.
Corn small peptide, self-control, small peptide containing corn (with total phthaleinometer) 83.1%.It is made into before use using aseptic deionized water
Finite concentration decoction, gastric infusion.
Sodium selenite.Finite concentration decoction, gastric infusion are made into using aseptic deionized water before use.
SPF grades of Kunming mouses, male, 18~22g of body weight, purchased from Shanghai Slac Experimental Animal Co., Ltd.,
Animal quality certification number：2015000515110.
(2) test method and result
Mouse is taken, 5 groups is randomly divided into, every group 10, is respectively：Selenium-rich corn small peptide group (30mg/kg), corn small peptide group
(30mg/kg), sodium selenite group (1.5mg/kg), Normal group (physiological saline), model control group (physiological saline), gavage
It is administered, 1 time/d, continuous 21d, after last dose 1h, in addition to Normal group, every group of ethanol of difference gavage 50%, 0.012ml/
G, causes acute hepatic injury model.Mouse fasting be can't help after water 24h, and every eyeball of mouse takes blood, and blood was stood after a period of time
Serum is prepared with refrigerated centrifuge 2500rpm centrifugations 10min, by ALT, AST, TBIL kit explanations are divided with biochemical instruments
Analysis.It the results are shown in Table 1.
Table 1 is to mice serum ALT, AST, TBIL influence (X ± s)
Note：Compared with model control group, * P<0.O5, * * P<0.O1.
Above result of the test shows：Selenium-rich corn small peptide can significantly improve the liver function of acute alcohol-induced hepatic injury mouse.
Embodiment
Embodiment is set forth below, the present invention is further illustrated, each embodiment is merely to illustrate the present invention, is not intended to limit this hair
It is bright：
Embodiment 1
The preparation of selenium-rich corn small peptide：Sodium selenite is dissolved in deionized water and is configured to the solution that concentration is 5mg/ml, is pressed
Solution and zein ratio 10:1 ratio adds zein, stirs, and heating is boiled 15 minutes, is cooled to about 45
DEG C, pH value 2.0 is adjusted with watery hydrochloric acid, 1.5% pepsin is added, 2 hours are incubated, pH value 8.0 is adjusted, 1.0% pancreas egg is added
White enzyme, is incubated 4 hours, boils 15 minutes, centrifuges, and precipitation is discarded, and supernatant is surpassed by relative molecular mass for 3KD milipore filter
Filter, permeate is added on DA201-C large pore resin absorption columns, with water elution, discards water lotion, then wash with 75% (V/V) ethanol
It is de-, ethanol eluate is collected, ethanol is recovered under reduced pressure and continues to be concentrated into relative density for 1.12 (60 DEG C), (air intake is spray-dried
Temperature is 180~200 DEG C, and leaving air temp is 90~100 DEG C), spray powder is collected, selenium-rich corn small peptide is produced.
Embodiment 2
The preparation of selenium-rich corn small peptide：Sodium selenite is dissolved in deionized water and is configured to the solution that concentration is 4mg/ml, is pressed
Solution and zein ratio 8:1 ratio adds zein, stirs, and heating is boiled 10 minutes, is cooled to about 50
DEG C, pH value 3.0 is adjusted with watery hydrochloric acid, 2% pepsin is added, 3 hours are incubated, pH value 7.5 is adjusted, 2% trypsase is added,
Insulation 5 hours, boils 10 minutes, centrifuges, precipitation discards, and supernatant is by the milipore filter ultrafiltration that relative molecular mass is 3KD, thoroughly
Cross liquid to be added on DA201-C large pore resin absorption columns, with water elution, discard water lotion, then with 60% (V/V) ethanol elution, receive
Collect ethanol eluate, ethanol and continuing is recovered under reduced pressure and is concentrated into relative density for 1.15 (60 DEG C), (EAT is for spray drying
180~200 DEG C, leaving air temp is 90~100 DEG C), spray powder is collected, selenium-rich corn small peptide is produced.
Embodiment 3
Selenium-rich corn small peptide 100g obtained by the method for Example 1, by selenium-rich corn small peptide:Corn oil:Beeswax:Lecithin
Ratio is 10:35:4:1 mixed material, ground 30 minutes, 100 are pressed with gelatin, purified water, glycerine, iron oxide black:100:40:
1 ratio making soft capsule hide glue gelatin, 40~50 DEG C of control glue box temperature, 35~45 DEG C of heat insulation tank, 30~40 DEG C of sprinkler body temperature,
Soft capsule is pressed into, per the tolerant heavy 0.45g of intragranular, 17 DEG C~27 DEG C 3~5h of cold air drying sizings wash ball, drying, select ball, bag
Dress, is prepared into soft capsule finished product.
Embodiment 4
Selenium-rich corn small peptide 100g obtained by the method for Example 1, by selenium-rich corn small peptide:Polyethylene glycol 400:Propane diols
Ratio is 10:38:2 mixed materials, ground 30 minutes, 100 are pressed with gelatin, purified water, glycerine, iron oxide black:100:40:1
Ratio making soft capsule hide glue gelatin, 40~50 DEG C of control glue box temperature, 35~45 DEG C of heat insulation tank, 30~40 DEG C of sprinkler body temperature,
Soft capsule is pressed into, per the tolerant heavy 0.45g of intragranular, 17 DEG C~27 DEG C 3~5h of cold air drying sizings wash ball, drying, select ball, bag
Dress, is prepared into soft capsule finished product.
Embodiment 5
Selenium-rich corn small peptide 100g obtained by the method for Example 2, plus starch to total amount are 200g, are well mixed, fluidisation
Bed granulation, loads capsule, and packaging is prepared into capsule finished product.
Claims (9)
1. a kind of selenium-rich corn small peptide with hepatoprotective effect, it is characterised in that prepared using following processing step：
The preparation of selenium-rich corn small peptide：Sodium selenite is dissolved in deionized water and is configured to the solution that concentration is 4~6mg/ml, by molten
Liquid and zein ratio 8~12:1 ratio adds zein, stirs, and heating is boiled 10~15 minutes, to be cooled
To about 30~50 DEG C, pH value 1.0~3.0 is adjusted with watery hydrochloric acid, 0.5~2% pepsin is added, 1~3 hour is incubated, pH is adjusted
Value 7.0~9.0, adds 0.5~2% trypsase, is incubated 3~5 hours, boils 10~15 minutes, centrifuges, and precipitation is discarded, supernatant
Liquid is added on DA201-C large pore resin absorption columns by the milipore filter ultrafiltration that relative molecular mass is 3KD, permeate, to wash
It is de-, water lotion is discarded, then with the ethanol elution of volume ratio 50~90%, ethanol eluate is collected, ethanol is recovered under reduced pressure and continues concentration
It is 1.10~1.15 to determine relative density to 60 DEG C, and spray drying EAT is 180~200 DEG C, and leaving air temp is 90~100
DEG C, spray powder is collected, selenium-rich corn small peptide is produced.
2. the selenium-rich corn small peptide according to claim 1 that there is protection liver to act on, it is characterised in that selenium-rich corn is small
Peptide peptide content is 60%~90%, and the accumulation rate of selenium is more than 95%.
3. the selenium-rich corn small peptide according to claim 1 that there is protection liver to act on, it is characterised in that sodium selenite can
To be replaced with yeast selenium, L- selenium-methyl selenium substituted aminothiopropionics etc..
4. the selenium-rich corn small peptide according to claim 1 that there is protection liver to act on, it is characterised in that the salting liquid
Concentration be 4~6mg/ml.
5. the selenium-rich corn small peptide according to claim 1 that there is protection liver to act on, it is characterised in that the salting liquid
It is 8~10 with wheat gluten ratio:1.
6. the selenium-rich corn small peptide according to claim 1 that there is protection liver to act on, it is characterised in that the milipore filter
Retention relative molecular mass be 3000Da.
7. the selenium-rich corn small peptide according to claim 1 that there is protection liver to act on, it is characterised in that the spraying is done
Dry EAT is 180~200 DEG C, and leaving air temp is 90~100 DEG C.
8. the selenium-rich corn small peptide according to claim 1 that there is protection liver to act on, it is characterised in that be prepared into following
Preparation：
（1）By selenium-rich corn small peptide:Corn oil:Beeswax:Lecithin is 10:35:4:1 mixed material, it is ground, it is prepared into
Soft capsule；
（2）By selenium-rich corn small peptide:Polyethylene glycol 400:Propane diols ratio is 10:38:2 mixed materials, it is ground, it is prepared into soft
Capsule；
（3）Selenium-rich corn small peptide, adds customary adjuvant, is prepared into oral liquid, capsule, tablet, granule.
9. the selenium-rich corn small peptide described in claim 1 is preparing the application in being used to protect liver drug.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109077325A (en) * | 2018-04-24 | 2018-12-25 | 恩施德源健康科技发展有限公司 | A kind of selenium-enriched plant peptides products of efficient strengthen immunity and its preparation method and application |
| CN109486886A (en) * | 2018-10-12 | 2019-03-19 | 中国食品发酵工业研究院有限公司 | A kind of maize oligopeptide selenium and its preparation method and application |
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| AU2007249806A1 (en) * | 2006-05-12 | 2007-11-22 | Hill's Pet Nutrition, Inc. | Compositions and methods for preventing or treating chronic renal failure in felines with hyperthyroidism |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109077325A (en) * | 2018-04-24 | 2018-12-25 | 恩施德源健康科技发展有限公司 | A kind of selenium-enriched plant peptides products of efficient strengthen immunity and its preparation method and application |
| CN109486886A (en) * | 2018-10-12 | 2019-03-19 | 中国食品发酵工业研究院有限公司 | A kind of maize oligopeptide selenium and its preparation method and application |
| CN109486886B (en) * | 2018-10-12 | 2021-04-13 | 中国食品发酵工业研究院有限公司 | Corn oligopeptide selenium and preparation method and application thereof |
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