CN102718832A

CN102718832A - Marine organism collagen peptide

Info

Publication number: CN102718832A
Application number: CN2012100628897A
Authority: CN
Inventors: 蒋鹏
Original assignee: Individual
Current assignee: Individual
Priority date: 2012-03-05
Filing date: 2012-03-05
Publication date: 2012-10-10

Abstract

According to a method provided by the invention, the body wall of a marine organism Asterias amurensis is used as a material to extract collagen, then the collagen protein is prepared into marine organism collagen peptide. First, body wall of Asterias amurensis is treated with enzymolysis by protease to extract collagen; and then the extracted collagen is degraded by complex enzyme to obtain a product with a molecular weight of approximately 100-10000 Da, namely the collagen polypeptide. Collagen peptide is a biological material with a variety of biological activities and excellent antioxidant performance, and has wide medical purposes of resisting oxidation, aging and ulcer, lowering blood pressure, preventing osteoporosis, promoting repair and growth of epithelial cell and improving the human immunity, etc.

Description

The sea life collagen polypeptide

One, technical field

The present invention relates to that a kind of to improve that human body immunity of organisms, anti-oxidant, anti-ageing, antiulcer agent, hypotensive, preventing osteoporosis, promotion epithelium cytothesis and growth etc. possess the collagen peptide goods of multiple excellent anti oxidation physiologically active, particularly relate to a kind of be that base mateiral extracts collagen polypeptide and method of manufacture thereof with sea life.

Two, background technology

Many sour jujubes starfish is a kind of common ocean invertebral living creature, the Echinodermata Stelleroidea, and as snackfoods, but the overwhelming majority goes out of use with it in the minority coastland.Its pharmaceutical use receives the medical circle concern gradually in recent years; Be rich in many sour jujubes starfish body and multiplely have the active organic components of good biological and comprise saponin(e, sterol, vegeto-alkali, polysaccharide etc.; Also contain abundant collagen protein in its body wall in addition; Collagen protein is one of most important protein in the animal body, has special mechanical property and biological nature, and collagen protein has been widely used in fields such as medical device, medicine, healthcare products as a kind of emerging material in recent years; And collagen polypeptide has than collagen protein superior bioactive and oxidation-resistance more, also more is prone to be absorbed by the body.Along with going deep into to the research of collagen series products; The collagen protein of marine source is very obvious as sanitary prods basic raw material advantage; Possess characteristics such as no animal immune originality, no cytotoxicity, so it will become the research field of the most active, tool bright prospects.

Three, summary of the invention

Collagen protein obtains having high absorbance after the compound protease degradation treatment, the product of the about 100-10000Da of molecular weight, be collagen peptide.Collagen peptide is in the reflection proteinaceous nutrient; Possess antiulcer agent, anti-oxidant, antianaphylaxis, anti-ageing, hypotensive, promote wound healing, promote characteristics such as skin collagen metabolism, immunomodulatory; Therefore collagen peptide possesses excellent more biological activity, and its a lot of activity are all relevant with its oxidation-resistance.

Four, practical implementation method

The purpose of this invention is to provide a kind of method for preparing collagen polypeptide of from many sour jujubes starfish body wall, extracting, this method comprises with proteolytic enzyme extracts collagen protein from many sour jujubes starfish body wall, the collagen protein degraded is prepared the process of collagen polypeptide again.

1, the sea life collagen protein extracts method of manufacture

Earlier many sour jujubes starfish body wall is cleaned with 2% salt soak, made it take off bone, use 3% citric acid solution acid-swellable again, the starfish of the many sour jujubes after swelling body wall homogenate; 4 ℃ of papain enzymolysis 24 hours, whizzer 10000r/min is centrifugal, gets supernatant; Again throw out is repeated enzymolysis 2 times to improve the collagen extraction yield, merge 3 times supernatant, regulate pH value=7 with acetate; Get the collagen deposition with 65% ammonium sulfate precipitation, distill water dialysis 16h gets pure collagen liquid.

2, the sea life collagen polypeptide extracts method of manufacture

Collagen liquid is dissolved in the 0.5mol/L acetate; With prozyme (flavor protease, neutral protease, trypsinase) enzymolysis 4h (enzyme material ratio is 2: 1000); Place 95 ℃ of water-baths enzyme 10min that goes out; The centrifugal 15min of 4000r/min gets collagen peptide solution, and vacuum freeze drier-80 a ℃ vacuum lyophilization is processed collagen polypeptide lyophilized powder goods.

Five, the sea life collagen polypeptide is removed oxyradical ability mensuration

Remove the oxyradical ability and measure the pyrogallol autoxidation method that adopts.Get collagen polypeptide sample solution 0.2ml, add pH8.2,50mmol/L Tris-HCL damping fluid 4.5mL, deionized water 4mL; Be mixed, 25 ℃ of insulation 10min are as first liquid; Get 0.3mL with 10mmol/L HCl solution preparation 3mmol/L pyrogallol solution, 25 ℃ of insulation 10min are as second liquid; With the mixed solution of the HCl solution of first liquid and 0.3mL 10mmol/L as the blank zeroing after, first liquid is mixed rapidly with second liquid, every 30s measures absorbancy at the 320nm place, in 4min, asks absorbancy variation slope K ₁Replace the collagen polypeptide sample to repeat said process with zero(ppm) water, calculate the slope that absorbancy changes as control sample.

Six, the sea life collagen polypeptide is measured the mouse interior antioxidation action

Through setting up the acute liver model of oxidation of mouse alcohol research collagen polypeptide to the mouse interior antioxidation action.Get Kunming mouse, be divided into 6 groups, be respectively normal control group, VC positive controls, negative control (ethanol damage) group, the high, medium and low dose groups of collagen polypeptide by body weight, 10 every group, male and female half and half.Wherein, the collagen polypeptide group give respectively low (50mg/kgd), in (100mg/kgd), high (200mg/kgd) dosage, the VC positive controls gives VC (75mg/kgd), normal control group, negative control group give equal-volume zero(ppm) water.Continuous 7d, after the last administration 2 hours, except that the blank group, all give 50% alcohol 12mL/kg, set up acute ethanol liver model of oxidation.Take off neck behind the jejunitas 12h and put to death whole mouse, get liver and clean back preparation 10% liver tissue homogenate's liquid, press the test kit explanation and measure tissue SOD, GSH-PX activity and MDA content with ice-cold saline water.

Claims

One kind be used to remove oxyradical excessive in the human body, suppress lipid peroxidation, anti-ageing, antiulcer agent, hypotensive, the marine collagen polypeptide that improves human immunity; It is characterized in that this collagen polypeptide by sea life extract, no animal immune originality, through take off bone, acid-swellable, enzymolysis, centrifugal, dialysis, degree of depth enzymolysis step is processed.
2. according to the marine collagen polypeptide of claim 1, wherein said marine collagen polypeptide is from many sour jujubes of sea life starfish body wall, to extract.
3. the method for manufacture of a marine collagen polypeptide, this method comprise:

(1) many sour jujubes starfish body wall is cleaned, immersed and soak 12h in 2% hydrochloric acid soln and take off bone and handle;

(2) with 3% citric acid solution swelling;

(3) starfish of the many sour jujubes behind acid-swellable body wall is blended homogenate with high-speed tissue mashing machine, in 4 ℃ of following enzymolysis 24h;

(4) the centrifugal 20min of supercentrifuge (10000r/min) gets supernatant;

(5) with supernatant with 65% ammonium sulfate precipitation 24 hours, the collagen deposition;

(6) with collagen deposition with the dialysis tubing 48h that dialyses, enzymatic property collagen protein;

(7) with collagen solution with prozyme degree of depth degraded 4h, the centrifugal 15min of supercentrifuge (4000r/min) collagen peptide solution;

(8) vacuum freeze drier-80 ℃ is the collagen peptide lyophilized powder with the freeze-drying of collagen peptide solution.
4. according to the method for claim 3, the employed proteolytic enzyme of its preparation collagen is papoid (enzyme material ratio is 1: 100).
5. according to the method for claim 3, its step (3) comprises with NaoH transfers PH=10, leaves standstill the step that 24h makes enzyme deactivation.
6. according to the method for claim 3, its step (6) comprises dialysis tubing inserted in the 1mmol/L EDTA-2Na solution uses pretreatment technology behind the heating 10min.
7. according to the method for claim 3, the said prozyme of its step (7) is flavor protease, neutral protease, trypsinase, and three's ratio is 1: 1: 1, and wherein enzyme material ratio is 2: 1000.
8. according to the method for claim 3, its step (7) comprises inserts 10min in 95 ℃ of water-baths with collagen peptide solution, makes the step of enzyme deactivation.