CN105567773A - Preparation method of crocodile blood polypeptide powder and production and application of crocodile blood polypeptide powder - Google Patents
Preparation method of crocodile blood polypeptide powder and production and application of crocodile blood polypeptide powder Download PDFInfo
- Publication number
- CN105567773A CN105567773A CN201610032564.2A CN201610032564A CN105567773A CN 105567773 A CN105567773 A CN 105567773A CN 201610032564 A CN201610032564 A CN 201610032564A CN 105567773 A CN105567773 A CN 105567773A
- Authority
- CN
- China
- Prior art keywords
- crocodile blood
- polypeptide powder
- blood polypeptide
- crocodile
- enzymolysis
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 210000004369 blood Anatomy 0.000 title claims abstract description 63
- 239000008280 blood Substances 0.000 title claims abstract description 63
- 241000270722 Crocodylidae Species 0.000 title claims abstract description 58
- LNNWVNGFPYWNQE-GMIGKAJZSA-N desomorphine Chemical compound C1C2=CC=C(O)C3=C2[C@]24CCN(C)[C@H]1[C@@H]2CCC[C@@H]4O3 LNNWVNGFPYWNQE-GMIGKAJZSA-N 0.000 title claims abstract description 57
- 239000000843 powder Substances 0.000 title claims abstract description 44
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 39
- 229920001184 polypeptide Polymers 0.000 title claims abstract description 38
- 102000004196 processed proteins & peptides Human genes 0.000 title claims abstract description 38
- 238000002360 preparation method Methods 0.000 title claims abstract description 9
- 238000004519 manufacturing process Methods 0.000 title description 5
- 238000000034 method Methods 0.000 claims abstract description 22
- 238000002156 mixing Methods 0.000 claims abstract description 19
- 238000006047 enzymatic hydrolysis reaction Methods 0.000 claims abstract description 6
- 230000036039 immunity Effects 0.000 claims abstract description 6
- 235000013305 food Nutrition 0.000 claims abstract description 5
- 239000003814 drug Substances 0.000 claims abstract description 4
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 21
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 14
- 210000002784 stomach Anatomy 0.000 claims description 14
- 230000000694 effects Effects 0.000 claims description 13
- 239000000047 product Substances 0.000 claims description 10
- 238000006243 chemical reaction Methods 0.000 claims description 9
- 238000001976 enzyme digestion Methods 0.000 claims description 9
- 230000033001 locomotion Effects 0.000 claims description 8
- 102000004190 Enzymes Human genes 0.000 claims description 7
- 108090000790 Enzymes Proteins 0.000 claims description 7
- 239000006228 supernatant Substances 0.000 claims description 7
- 210000000936 intestine Anatomy 0.000 claims description 5
- 238000004088 simulation Methods 0.000 claims description 5
- 230000007071 enzymatic hydrolysis Effects 0.000 claims description 4
- 238000005507 spraying Methods 0.000 claims description 3
- 239000002253 acid Substances 0.000 claims description 2
- 239000003513 alkali Substances 0.000 claims description 2
- 239000002585 base Substances 0.000 claims description 2
- 150000007516 brønsted-lowry acids Chemical class 0.000 claims description 2
- 150000007528 brønsted-lowry bases Chemical class 0.000 claims description 2
- 238000001035 drying Methods 0.000 claims description 2
- 239000000758 substrate Substances 0.000 claims description 2
- 102000057297 Pepsin A Human genes 0.000 abstract description 6
- 108090000284 Pepsin A Proteins 0.000 abstract description 6
- 102000004142 Trypsin Human genes 0.000 abstract description 6
- 108090000631 Trypsin Proteins 0.000 abstract description 6
- 239000012588 trypsin Substances 0.000 abstract description 6
- 230000000386 athletic effect Effects 0.000 abstract 2
- 230000029087 digestion Effects 0.000 abstract 2
- 230000002496 gastric effect Effects 0.000 abstract 2
- 238000004925 denaturation Methods 0.000 abstract 1
- 230000036425 denaturation Effects 0.000 abstract 1
- 230000002708 enhancing effect Effects 0.000 abstract 1
- 230000007062 hydrolysis Effects 0.000 abstract 1
- 238000006460 hydrolysis reaction Methods 0.000 abstract 1
- 239000007788 liquid Substances 0.000 abstract 1
- 229940111202 pepsin Drugs 0.000 abstract 1
- 238000003304 gavage Methods 0.000 description 20
- DPXJVFZANSGRMM-UHFFFAOYSA-N acetic acid;2,3,4,5,6-pentahydroxyhexanal;sodium Chemical compound [Na].CC(O)=O.OCC(O)C(O)C(O)C(O)C=O DPXJVFZANSGRMM-UHFFFAOYSA-N 0.000 description 16
- 229920003123 carboxymethyl cellulose sodium Polymers 0.000 description 16
- 229940063834 carboxymethylcellulose sodium Drugs 0.000 description 16
- 241000699666 Mus <mouse, genus> Species 0.000 description 15
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 15
- 239000013642 negative control Substances 0.000 description 12
- 230000008569 process Effects 0.000 description 12
- 230000003301 hydrolyzing effect Effects 0.000 description 10
- 238000003756 stirring Methods 0.000 description 10
- 230000009182 swimming Effects 0.000 description 9
- 210000002966 serum Anatomy 0.000 description 7
- 229940088598 enzyme Drugs 0.000 description 6
- 239000004310 lactic acid Substances 0.000 description 6
- 235000014655 lactic acid Nutrition 0.000 description 6
- 235000002789 Panax ginseng Nutrition 0.000 description 5
- 101710151387 Serine protease 1 Proteins 0.000 description 5
- 102100032491 Serine protease 1 Human genes 0.000 description 5
- 101710119665 Trypsin-1 Proteins 0.000 description 5
- 238000013016 damping Methods 0.000 description 5
- 239000012530 fluid Substances 0.000 description 5
- 239000008363 phosphate buffer Substances 0.000 description 5
- 150000003016 phosphoric acids Chemical class 0.000 description 5
- 239000013641 positive control Substances 0.000 description 5
- 230000001681 protective effect Effects 0.000 description 5
- 230000001105 regulatory effect Effects 0.000 description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 5
- 239000000463 material Substances 0.000 description 4
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 3
- 102000019197 Superoxide Dismutase Human genes 0.000 description 3
- 108010012715 Superoxide dismutase Proteins 0.000 description 3
- PNNCWTXUWKENPE-UHFFFAOYSA-N [N].NC(N)=O Chemical compound [N].NC(N)=O PNNCWTXUWKENPE-UHFFFAOYSA-N 0.000 description 3
- 230000008901 benefit Effects 0.000 description 3
- 210000004027 cell Anatomy 0.000 description 3
- 239000008103 glucose Substances 0.000 description 3
- 210000003205 muscle Anatomy 0.000 description 3
- 210000000663 muscle cell Anatomy 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- 150000003254 radicals Chemical class 0.000 description 3
- 239000002994 raw material Substances 0.000 description 3
- 229940032362 superoxide dismutase Drugs 0.000 description 3
- UFTFJSFQGQCHQW-UHFFFAOYSA-N triformin Chemical compound O=COCC(OC=O)COC=O UFTFJSFQGQCHQW-UHFFFAOYSA-N 0.000 description 3
- 102000004420 Creatine Kinase Human genes 0.000 description 2
- 108010042126 Creatine kinase Proteins 0.000 description 2
- 241000938605 Crocodylia Species 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 241000699670 Mus sp. Species 0.000 description 2
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 2
- 230000005779 cell damage Effects 0.000 description 2
- 208000037887 cell injury Diseases 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 230000006378 damage Effects 0.000 description 2
- 230000003834 intracellular effect Effects 0.000 description 2
- 230000002503 metabolic effect Effects 0.000 description 2
- 230000004060 metabolic process Effects 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 230000036542 oxidative stress Effects 0.000 description 2
- 239000001301 oxygen Substances 0.000 description 2
- 229910052760 oxygen Inorganic materials 0.000 description 2
- 241000208340 Araliaceae Species 0.000 description 1
- 102000001554 Hemoglobins Human genes 0.000 description 1
- 108010054147 Hemoglobins Proteins 0.000 description 1
- 102000004316 Oxidoreductases Human genes 0.000 description 1
- 108090000854 Oxidoreductases Proteins 0.000 description 1
- 235000005035 Panax pseudoginseng ssp. pseudoginseng Nutrition 0.000 description 1
- 235000003140 Panax quinquefolius Nutrition 0.000 description 1
- 102000035195 Peptidases Human genes 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- OUUQCZGPVNCOIJ-UHFFFAOYSA-M Superoxide Chemical compound [O-][O] OUUQCZGPVNCOIJ-UHFFFAOYSA-M 0.000 description 1
- 208000027418 Wounds and injury Diseases 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 125000003275 alpha amino acid group Chemical group 0.000 description 1
- 230000006538 anaerobic glycolysis Effects 0.000 description 1
- 230000002929 anti-fatigue Effects 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 210000004556 brain Anatomy 0.000 description 1
- 210000005252 bulbus oculi Anatomy 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 239000002360 explosive Substances 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 238000011207 functional examination Methods 0.000 description 1
- 235000008434 ginseng Nutrition 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- 238000012423 maintenance Methods 0.000 description 1
- 210000001640 nerve ending Anatomy 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 238000002331 protein detection Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 210000002027 skeletal muscle Anatomy 0.000 description 1
- 230000000638 stimulation Effects 0.000 description 1
Classifications
-
- 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
- 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
- A61K38/012—Hydrolysed proteins; Derivatives thereof from animals
- A61K38/017—Hydrolysed proteins; Derivatives thereof from animals from blood
-
- 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
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Zoology (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Immunology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- Pharmacology & Pharmacy (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Molecular Biology (AREA)
- Biotechnology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- General Chemical & Material Sciences (AREA)
- Microbiology (AREA)
- Medicinal Chemistry (AREA)
- Hematology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Genetics & Genomics (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The invention relates to the preparation method of crocodile blood polypeptide powder. The method is characterized by including the following steps that A, crocodile blood is taken and homogenized so that cells can be broken; B, obtained homogenized liquid is subjected to enzymolysis in an enzymatic hydrolysis reaction system simulating a human body gastrointestinal digestion system, hydrolysis is conducted in the enzymatic hydrolysis reaction system simulating the human body gastrointestinal digestion system through pepsin and trypsin in sequence under appropriate enzymolysis conditions at blending rotating speed; C, thermal denaturation is conducted after enzymolysis, centrifuging is conducted, supernate is taken and dried, and obtained crocodile blood polypeptide powder is collected. The invention further relates to crocodile blood polypeptide powder prepared through the method. Crocodile blood polypeptide powder has certain functions of resisting fatigue, improving body immunity and athletic ability, and the like, and can be used for preparing food or heath care products or medicine for resisting fatigue and improving body athletic ability or enhancing immunity.
Description
Technical field
The application relates to the fields such as food, healthcare products or medicine, is specifically related to preparation method of a kind of crocodile blood polypeptide powder and products thereof and application.
Background technology
Crocodile is a class animal of reptilia, Crocodilia, Crocodylidae, has very high economy and medical value.Crocodile can hide in water when preying on prey for a long time, can carry a large amount of oxygen supply bodies and use, make crocodile have powerful explosive power and snap-in force in its blood.There are some researches show, crocodile blood hemoglobin amino acid chain has very peculiar structure, makes the oxyphorase oxygen carrying content of crocodile exceed other animals more than 100 times.To the exploitation of crocodile blood, contribute to studying the product useful to human body, by supplementary corresponding product, the immunity of organisms of the crowd of having a delicate constitution can be improved, or improve the effect of motion function and resisting fatigue, to raising people live and work level, there is good facilitation effect.
Therefore focus and focus will be become to the development and application of crocodile blood, and also huge economic benefit and social benefit can be produced simultaneously.
Summary of the invention
The invention provides a kind of preparation method of crocodile blood polypeptide powder, it is characterized in that comprising following steps: A: get crocodile blood, homogenate makes cytoclasis; B: by the homogenate that obtains enzymolysis in the enzyme digestion reaction system of simulation human intestines and stomach Digestive tract, the enzyme digestion reaction system of described simulation human intestines and stomach Digestive tract is included in priority stomach en-under suitable enzymatic hydrolysis condition, trypsinase and is hydrolyzed mixing under rotating speed; C: after enzymolysis, thermally denature, centrifuging and taking supernatant liquor, dry, collect the crocodile blood polypeptide powder of gained.In some embodiments, described crocodile blood can be after fresh collection, be kept at-80 degrees Celsius stand-by.In further embodiment, the system total mass in described enzyme digestion reaction system: substrate quality: enzyme quality=1000:(20 ~ 100): (5 ~ 0.1).In other embodiments, the time of wherein said pepsic enzymolysis can be 1 ~ 5 hour, and described tryptic enzymolysis time is 1 ~ 5 hour.In preferred embodiments, described suitable enzymatic hydrolysis condition can comprise: for stomach en-, pH2.0-3.0, temperature 37 degrees Celsius; For trypsinase, pH7.0-7.4, temperature 37 degrees Celsius; Mixing rotating speed is 50-300 revolutions per second, preferably 100 revolutions per seconds.In further embodiment, the pH regulator in described enzyme digestion reaction system can adopt bronsted lowry acids and bases bronsted lowry to regulate, and wherein, described acid is preferably hydrochloric acid, and described alkali is preferably NaOH or KOH.In further embodiment, the mode of described drying can be lyophilize or spraying dry.
Present invention also offers a kind of crocodile blood polypeptide powder, described crocodile blood polypeptide powder is prepared by according to method of the present invention.
Present invention also offers, according to crocodile blood polypeptide powder of the present invention, there is in preparation the immunity of organisms improving the crowd that has a delicate constitution, or the purposes in the food of raising motion function and resisting fatigue effect, healthcare products and medicine.
Enzyme digestion reaction system of the present invention builds for simulation human intestines and stomach Digestive tract, has advantages such as crocodile blood high molecular weight protein thorough enzymolysis, favorable repeatability.The crocodile blood polypeptide powder of gained is standby by mixed enzymolysis legal system, and molecular weight is little, and absorption easy to digest, biological activity is higher.
Crocodile blood polypeptide powder of the present invention has the immunity of organisms that can improve the crowd of having a delicate constitution, or improves the effect of motion function and resisting fatigue.
The raw material of use involved in the present invention and process for processing environment meet the hygienic requirements of concerned countries regulation to food, healthcare products or pharmaceutical raw material and production environment.
Accompanying drawing explanation
Fig. 1 is mouse swimming time.Negative control group is with 0.3mL0.5% carboxymethylcellulose sodium solution gavage.Low, in and high dose group be respectively the application's crocodile blood polypeptide powder of 0.15g, 0.3g and 0.45g be dissolved in 0.3mL0.5% carboxymethylcellulose sodium solution, gavage after mixing.Positive controls is gavage after 0.3g red ginseng powder mixes with 0.3mL0.5% carboxymethylcellulose sodium solution.
Fig. 2 is SOD activity level in mouse blood, SOD level condition in serum after the swimming of reflection mouse.Negative control group is with 0.3mL0.5% carboxymethylcellulose sodium solution gavage.Low, in and high dose group be respectively the application's crocodile blood polypeptide powder of 0.15g, 0.3g and 0.45g be dissolved in 0.3mL0.5% carboxymethylcellulose sodium solution, gavage after mixing.Positive controls is gavage after 0.3g red ginseng powder mixes with 0.3mL0.5% carboxymethylcellulose sodium solution.
Fig. 3 is TG, GLU, BUN level in mouse blood, and reflection mouse power exhausts TG, GLU and BUN level condition in the rear serum of swimming.Negative control group is with 0.3mL0.5% carboxymethylcellulose sodium solution gavage.Low, in and high dose group be respectively the application's crocodile blood polypeptide powder of 0.15g, 0.3g and 0.45g be dissolved in 0.3mL0.5% carboxymethylcellulose sodium solution, gavage after mixing, positive controls is gavage after 0.3g red ginseng powder mixes with 0.3mL0.5% carboxymethylcellulose sodium solution.
Fig. 4 is LDH and CK level in mouse blood, reflection mouse power exhaust LDH and CK after swimming changing conditions (* *, compared with negative control group, P<0.01; *, compared with negative control group, P<0.05).Negative control group is with 0.3mL0.5% carboxymethylcellulose sodium solution gavage.Low, in and high dose group be respectively the application's crocodile blood polypeptide powder of 0.15g, 0.3g and 0.45g be dissolved in 0.3mL0.5% carboxymethylcellulose sodium solution, gavage after mixing.Positive controls is gavage after 0.3g red ginseng powder mixes with 0.3mL0.5% carboxymethylcellulose sodium solution.
Embodiment
To be described in further detail the application by way of example below, put into practice the application to enable those skilled in the art.Should be appreciated that and can adopt other embodiments, and suitable change can be made and do not depart from the spirit or scope of the application.In order to avoid for enabling those skilled in the art put into practice details unnecessary the application, specification sheets may eliminate some known to those skilled in the art information.Therefore, below describe in detail and should not understand with restrictive meaning, and scope of the present invention is only defined by claims.
Following embodiment is convenient to understand the application better, but is not used for limiting the scope of the application.
The each raw material used in following embodiment, except particularly pointing out, all can by commercially available acquisition.
The crocodile blood used in following embodiment 1-4 is blood stand-by at-80 degrees Celsius of freezen protective after fresh collection.
Embodiment 1
1) get the crocodile blood of freezen protective, thaw under room temperature.Homogenate makes cytoclasis, then with homogenate and phosphate buffer 1: the volume ratio of 25 adds phosphoric acid salt (Chemical Reagent Co., Ltd., Sinopharm Group, the analytical pure) damping fluid of pH7;
2) with concentrated hydrochloric acid (Chemical Reagent Co., Ltd., Sinopharm Group, analytical pure) by the 1st) pH regulator to 2.0 of the solution of step gained, add the stomach en-(Pepsin1:3000fromPorcineStomach of massfraction 1 ‰, raw work is biological), 37 degrees Celsius are hydrolyzed 5 hours, set to mix rotating speed 100 revolutions per seconds in hydrolytic process and carry out mixing stirring, pH to 7.0 is regulated afterwards with NaOH, add the trypsin Trypsin1:3000fromPorcineStomach of massfraction 1 ‰, raw work is biological), 37 degrees Celsius are hydrolyzed 5 hours, set to mix rotating speed 100 revolutions per seconds in hydrolytic process and carry out mixing stirring,
3) after enzymolysis, 90 degrees Celsius of thermally denatures 10 minutes, 7000 revolutions per seconds centrifugal 10 minutes to remove the high molecular weight protein of proteolytic enzyme and non-enzymolysis.Get supernatant liquor, after lyophilize, obtain crocodile blood polypeptide powder.Gained powder is formulated as 1mg/mL, and recording crocodile blood peptide concentration with BCA protein detection kit (Applygen, Beijing) is 925-973ug/mL.
Embodiment 2
1) get the crocodile blood of freezen protective, thaw under room temperature.Homogenate makes cytoclasis, then with homogenate and phosphate buffer 1: the volume ratio of 50 adds phosphoric acid salt (Chemical Reagent Co., Ltd., Sinopharm Group, the analytical pure) damping fluid of pH7;
2) with concentrated hydrochloric acid (Chemical Reagent Co., Ltd., Sinopharm Group, analytical pure) by the 1st) pH regulator to 3.0 of the solution of step gained, add the stomach en-(Pepsin1:3000fromPorcineStomach of massfraction 5 ‰, raw work is biological), 37 degrees Celsius are hydrolyzed 1 hour, set to mix rotating speed 300 revolutions per seconds in hydrolytic process and carry out mixing stirring, pH to 7.0 is regulated afterwards with NaOH, add the trypsin Trypsin1:3000fromPorcineStomach of massfraction 5 ‰, raw work is biological), 37 degrees Celsius are hydrolyzed 1 hour, set to mix rotating speed 300 revolutions per seconds in hydrolytic process and carry out mixing stirring,
3), after enzymolysis, 90 degrees Celsius of thermally denatures 10 minutes, 7000 revolutions per seconds centrifugal 10 minutes, and get supernatant liquor, lyophilize obtains crocodile blood polypeptide powder.
Embodiment 3
1) get the crocodile blood of freezen protective, thaw under room temperature.Homogenate makes cytoclasis, then with homogenate and phosphate buffer 1: the volume ratio of 20 adds phosphoric acid salt (Chemical Reagent Co., Ltd., Sinopharm Group, the analytical pure) damping fluid of pH7;
2) with concentrated hydrochloric acid (Chemical Reagent Co., Ltd., Sinopharm Group, analytical pure) by the 1st) pH value of solution of step gained is adjusted to 2.5, add the stomach en-(Pepsin1:3000fromPorcineStomach of massfraction 3 ‰, raw work is biological), 37 degrees Celsius are hydrolyzed 3 hours, set to mix rotating speed 200 revolutions per seconds in hydrolytic process and carry out mixing stirring, pH to 7.0 is regulated afterwards with NaOH, add the trypsin Trypsin1:3000fromPorcineStomach of massfraction 3 ‰, raw work is biological), 37 degrees Celsius are hydrolyzed 3 hours, set to mix rotating speed 200 revolutions per seconds in hydrolytic process and carry out mixing stirring,
3), after enzymolysis, 90 degrees Celsius of thermally denatures 10 minutes, 7000 revolutions per seconds centrifugal 10 minutes, and get supernatant liquor, spraying dry obtains crocodile blood polypeptide powder.
Embodiment 4
1) get the crocodile blood of freezen protective, thaw under room temperature.Homogenate makes cytoclasis, then with homogenate and phosphate buffer 1: the volume ratio of 10 adds phosphoric acid salt (Chemical Reagent Co., Ltd., Sinopharm Group, the analytical pure) damping fluid of pH7;
2) with concentrated hydrochloric acid (Chemical Reagent Co., Ltd., Sinopharm Group, analytical pure) by the 1st) pH regulator to 2.0 of the solution of step gained, add the stomach en-(Pepsin1:3000fromPorcineStomach of massfraction 2 ‰, raw work is biological), 37 degrees Celsius are hydrolyzed 4 hours, set to mix rotating speed 150 revolutions per seconds in hydrolytic process and carry out mixing stirring, pH to 7.0 is regulated afterwards with NaOH, add the trypsin Trypsin1:3000fromPorcineStomach of massfraction 2 ‰, raw work is biological), 37 degrees Celsius are hydrolyzed 4 hours, set to mix rotating speed 150 revolutions per seconds in hydrolytic process and carry out mixing stirring,
3), after enzymolysis, 90 degrees Celsius of thermally denatures 10 minutes, 7000 revolutions per seconds centrifugal 10 minutes, and get supernatant liquor, lyophilize obtains crocodile blood polypeptide powder.
Embodiment 5
1) get the crocodile blood that 50kg is fresh, homogenate makes cytoclasis, then with homogenate and phosphate buffer 1: the volume ratio of 25 adds phosphoric acid salt (Chemical Reagent Co., Ltd., Sinopharm Group, the analytical pure) damping fluid of pH7;
2) with concentrated hydrochloric acid (Chemical Reagent Co., Ltd., Sinopharm Group, analytical pure) by the 1st) pH regulator to 2.0 of the solution of step gained, add the stomach en-(Pepsin1:3000fromPorcineStomach of massfraction 3 ‰, raw work is biological), 37 degrees Celsius are hydrolyzed 5 hours, set to mix rotating speed 300 revolutions per seconds in hydrolytic process and carry out mixing stirring, pH to 7.0 is regulated afterwards with NaOH, add the trypsin Trypsin1:3000fromPorcineStomach of massfraction 3 ‰, raw work is biological), 37 degrees Celsius are hydrolyzed 5 hours, set to mix rotating speed 300 revolutions per seconds in hydrolytic process and carry out mixing stirring.
3), after enzymolysis, 90 degrees Celsius of thermally denatures 10 minutes, 7000 revolutions per seconds centrifugal 10 minutes, and get supernatant liquor, lyophilize obtains crocodile blood polypeptide powder.
The functional examination of embodiment 6-crocodile blood polypeptide powder
Raising motion function and antifatigue are tested--and power exhausts swimming model:
Mouse eight components are five groups: negative control group, gavage 0.3mL0.5% carboxymethylcellulose sodium solution; Low dose group, gavage after the crocodile blood polypeptide powder obtained in 0.15g embodiment 3 and 0.3mL0.5% Xylo-Mucine (Aladdin, Shanghai) solution mix; Middle dosage group, gavage after the crocodile blood polypeptide powder obtained in 0.3g embodiment 3 mixes with 0.3mL0.5% carboxymethylcellulose sodium solution; High dose group, gavage after the crocodile blood polypeptide powder obtained in 0.45g embodiment 3 mixes with 0.3mL0.5% carboxymethylcellulose sodium solution; Positive controls, gavage after 0.3g red ginseng powder (Korea Ginseng Corporation) mixes with 0.3mL0.5% carboxymethylcellulose sodium solution.
As described above after gavage, each group of mouse is placed in the tank that water temperature is room temperature 25 degrees Celsius, depth of water 40cm, water is constantly stirred that mouse must not be stopped transport is dynamic simultaneously, treat that mouse power records the time after exhausting.
Result as shown in Figure 1, takes the mouse of the polypeptide powder of crocodile of the application, and the swimming time in warm water is significantly higher than negative control group, and presents certain dosage effect.
Get five groups of mouse equally, gavage arranges the same, terminates after gavage, fasting one night, allows its swimming half an hour.Eyeball gets blood subsequently, after obtaining serum, measures the content of triglyceride level TG, blood urea nitrogen BUN, glucose GLU, creatine phosphokinase CK, lactic acid LDH and superoxide-dismutase SOD wherein.Result is shown in Fig. 2-Fig. 4.
Glycolysis-is ubiquitous a kind of metabolic way in organism.When muscle acutely shrinks, because starting anaerobic glycolysis for hypoxgia, produce a large amount of lactic acid (LDH).Lactic acid is piled up more in muscle, and degree of fatigue is more serious, so can reduce the material of Serum lactic acid content, just can play the effect lessened fatigue.In addition, Glu is the topmost energy supply material of body, the too low tired maincenter that will stimulate brain of the concentration of Glu, is to cause tired major reason; TG is lipometabolic product, is important energy supply material, especially mainly participates in the material of energy supply during muscle cell aerobic repiration, and it is large that TG consumes, and proves that the sport efficiency of skeletal muscle in moving process promotes; BUN is the product that protein or nucleic acid participate in production capacity metabolic breakdown, and the abnormal production capacity metabolism just can carried out when this production capacity metabolism is the energy supply deficiency of body, the concentration of BUN directly can reflect the situation of organism fatigue.CK and LDH is intracellular enzyme, CK only has in muscle cell and just has, LDH exists in all cells, when body carries out strenuous exercise, cell injury may be caused, now intracellular enzyme wherein enters the recycle system, and when the vigor of both CK and LDH reflects motion, the situation of damage occurs for muscle cell or other cells, and the major reason of sore muscle when the stimulation of cell injury to nerve ending is motion.SOD superoxide-dismutase, be the antioxidase removing superoxide radical, body carries out aerobic repiration in a large number can produce a large amount of free radical, and the accumulation of free radical can change the oxidative stress status of body, affects the function of histoorgan; The increased activity of antioxidant reductase, represents body stronger to the tolerance of oxidative stress, can when body carry out strenuous exercise produce a large amount of free radical, better maintain the redox state in body, be conducive to continuity and the maintenance of kinestate.Therefore, the changing conditions of the composition such as triglyceride level TG, glucose Glu, blood urea nitrogen BUN, creatine phosphokinase CK, lactic acid LDH and superoxide-dismutase SOD in blood after study movement, can protelytic decomposition, the degree of injury of cells of organs film and the relation of exercise tolerance and anoxic.
As can be seen from Figure 3, take the mouse of the application's crocodile blood polypeptide powder (high, in and low dosage) of various dose after power exhausts swimming, blood urea nitrogen BUN in its serum and triglyceride level TG level are significantly lower than negative control group, and the level of glucose GLU is higher than negative control group.
And illustrate at Fig. 2, the SOD enzyme activity taken in the mice serum of the application's crocodile blood polypeptide powder of high dosage significantly raises (P<0.01) relative to negative control group.
Fig. 4 to illustrate in the mice serum of the application's polypeptide powder of crocodile (high, middle dosage) taking various dose LDH, CK activity comparatively negative control group significantly reduce (P<0.01 or P<0.05).
To sum up, we find, the crocodile blood polypeptide powder of the application can make mouse swimming time extend, and reduce BUN and lactic acid content in blood, and can reduce LDH, CK enzyme activity, improve SOD enzyme activity.
In sum; these are only the preferred embodiment of the application, be not intended to limit the protection domain of the application, therefore; the any amendment done within all spirit in the application and principle, equivalent replacement, improvement etc., within the protection domain that all should be included in the application.
Claims (9)
1. a preparation method for crocodile blood polypeptide powder, is characterized in that comprising following steps:
A: get crocodile blood, homogenate makes cytoclasis;
B: by the homogenate that obtains enzymolysis in the enzyme digestion reaction system of simulation human intestines and stomach Digestive tract, the enzyme digestion reaction system of described simulation human intestines and stomach Digestive tract is included in priority stomach en-under suitable enzymatic hydrolysis condition, trypsinase and is hydrolyzed mixing under rotating speed;
C: after enzymolysis, thermally denature, centrifuging and taking supernatant liquor, dry, collect the crocodile blood polypeptide powder of gained.
2. method according to claim 1, wherein said crocodile blood is after fresh collection, be kept at-80 degrees Celsius stand-by.
3. method according to claim 1, the system total mass in wherein said enzyme digestion reaction system: substrate quality: enzyme quality=1000:(20 ~ 100): (5 ~ 0.1).
4. method according to claim 1, the time of wherein said pepsic enzymolysis is 1 ~ 5 hour, and described tryptic enzymolysis time is 1 ~ 5 hour.
5. the method according to any one of claim 1-4, wherein said suitable enzymatic hydrolysis condition comprises: for stomach en-, pH2.0-3.0, temperature 37 degrees Celsius; For trypsinase, pH7.0-7.4, temperature 37 degrees Celsius; And mixing rotating speed is 50-300 revolutions per second.
6. method according to claim 5, the pH regulator in wherein said enzyme digestion reaction system adopts bronsted lowry acids and bases bronsted lowry to regulate, and described acid is preferably hydrochloric acid, and described alkali is preferably NaOH or KOH.
7. method according to claim 1, the mode of wherein said drying is lyophilize or spraying dry.
8. a crocodile blood polypeptide powder, the method preparation of described crocodile blood polypeptide powder according to any one of claim 1-7.
9. crocodile blood polypeptide powder as claimed in claim 8 has the immunity of organisms improving the crowd that has a delicate constitution in preparation, or the purposes in the food of raising motion function and resisting fatigue effect, healthcare products and medicine.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610032564.2A CN105567773A (en) | 2016-01-18 | 2016-01-18 | Preparation method of crocodile blood polypeptide powder and production and application of crocodile blood polypeptide powder |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610032564.2A CN105567773A (en) | 2016-01-18 | 2016-01-18 | Preparation method of crocodile blood polypeptide powder and production and application of crocodile blood polypeptide powder |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN105567773A true CN105567773A (en) | 2016-05-11 |
Family
ID=55878391
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610032564.2A Pending CN105567773A (en) | 2016-01-18 | 2016-01-18 | Preparation method of crocodile blood polypeptide powder and production and application of crocodile blood polypeptide powder |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105567773A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107970437A (en) * | 2017-12-08 | 2018-05-01 | 苟春虎 | Cordyceps sinensis gram oncogene peptide |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1990875A (en) * | 2005-12-30 | 2007-07-04 | 中国科学院大连化学物理研究所 | Globin zymolyte and its preparation and application |
| CN101550178A (en) * | 2008-03-31 | 2009-10-07 | 四川省中医药科学院 | Blood polypeptide and preparation method and application thereof |
| CN101878849A (en) * | 2010-06-13 | 2010-11-10 | 厦门大学 | Polypeptide powder of crocodile and preparation method and application thereof |
| CN102296100A (en) * | 2011-09-09 | 2011-12-28 | 江南大学 | Preparation method of casein antihypertensive peptides |
| CN102329845A (en) * | 2011-09-28 | 2012-01-25 | 常熟市金城食品添加剂有限公司 | Preparation method of hypotensive peptide of lactalbumin |
| CN102342407A (en) * | 2011-09-30 | 2012-02-08 | 华南理工大学 | Anti-fatigue drink and preparation method thereof |
| CN104263789A (en) * | 2014-09-29 | 2015-01-07 | 江苏大学 | Method for preparing navodon septentrionalis skin anti-oxidative peptide liquid through in vitro simulation of gastrointestinal digestion |
-
2016
- 2016-01-18 CN CN201610032564.2A patent/CN105567773A/en active Pending
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1990875A (en) * | 2005-12-30 | 2007-07-04 | 中国科学院大连化学物理研究所 | Globin zymolyte and its preparation and application |
| CN101550178A (en) * | 2008-03-31 | 2009-10-07 | 四川省中医药科学院 | Blood polypeptide and preparation method and application thereof |
| CN101878849A (en) * | 2010-06-13 | 2010-11-10 | 厦门大学 | Polypeptide powder of crocodile and preparation method and application thereof |
| CN102296100A (en) * | 2011-09-09 | 2011-12-28 | 江南大学 | Preparation method of casein antihypertensive peptides |
| CN102329845A (en) * | 2011-09-28 | 2012-01-25 | 常熟市金城食品添加剂有限公司 | Preparation method of hypotensive peptide of lactalbumin |
| CN102342407A (en) * | 2011-09-30 | 2012-02-08 | 华南理工大学 | Anti-fatigue drink and preparation method thereof |
| CN104263789A (en) * | 2014-09-29 | 2015-01-07 | 江苏大学 | Method for preparing navodon septentrionalis skin anti-oxidative peptide liquid through in vitro simulation of gastrointestinal digestion |
Non-Patent Citations (1)
| Title |
|---|
| 张蓓: "鳄鱼血的新功效", 《中国体育教练员》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107970437A (en) * | 2017-12-08 | 2018-05-01 | 苟春虎 | Cordyceps sinensis gram oncogene peptide |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JP4704377B2 (en) | Fermentation and culture method, plant fermented extract, plant fermented extract powder and blended plant fermented extract | |
| CN103820517B (en) | The preparation method of the yak skin micromolecular collagen of regulation and control human body energy metabolism | |
| Mzengereza et al. | Growth performance, growth-related genes, digestibility, digestive enzyme activity, immune and stress responses of de novo Camelina meal in diets of red seabream (Pagrus major) | |
| CN107095312A (en) | A kind of krill polypeptide formulations with reducing blood lipid ability and preparation method thereof | |
| CN103005466B (en) | Application for drone pupae polypeptide extract | |
| CN103462864A (en) | Embryo water extract extracted from animal embryo internal organs, and extraction method and applications thereof | |
| CN101356954A (en) | Preparation method of anti-stress medicine earthworm small-peptide chelated chromium | |
| Zakaria et al. | Fermented soybean meal (FSBM) in African catfish (Clarias gariepinus) diets: effects on growth performance, fish gut microbiota analysis, blood haematology, and liver morphology | |
| CN104706543A (en) | Plant polypeptide composition and application thereof in cosmetics | |
| Chen et al. | Recovery of chitin from Antarctic krill (Euphausia superba) shell waste by microbial deproteinization and demineralization | |
| Caipang et al. | Host-derived probiotics for finfish aquaculture | |
| Huang et al. | Effect of dietary vitamin B6 supplementation on growth and intestinal microflora of juvenile golden pompano (Trachinotus ovatus) | |
| CN102652533A (en) | Toxin producing algae detoxification technology, product and application of product | |
| Rodrigues et al. | Enzymatic hydrolysis of the Eisenia andrei earthworm: Characterization and evaluation of its properties | |
| Riolo et al. | Cytoprotective and antioxidant effects of hydrolysates from black soldier fly (Hermetia illucens) | |
| Zhang et al. | The effects of dietary fermented soybean meal supplementation on the growth, antioxidation, immunity, and mTOR signaling pathway of juvenile coho salmon (Oncorhynchus kisutch) | |
| Zhang et al. | Growth performance, antioxidant and immunity capacity were significantly affected by feeding fermented soybean meal in juvenile coho salmon (Oncorhynchus kisutch) | |
| CN105567773A (en) | Preparation method of crocodile blood polypeptide powder and production and application of crocodile blood polypeptide powder | |
| Fan et al. | Protein sparing effect of carbohydrate on growth performance, digestion ability of common carp (Cyprinus carpio) at different feeding frequencies | |
| Huang et al. | Effects of fishmeal replacement by Clostridium Autoethanogenum protein meal on cholesterol bile acid metabolism, antioxidant capacity, hepatic and intestinal health of pearl gentian grouper (Epinephelus Fuscoguttatus♀× Epinephelus Lanceolatus♂) | |
| CN101690595A (en) | Ocean fish stuffing and preparation method thereof | |
| CN103462865A (en) | Embryo water extract extracted from animal embryo skin and muscle, and extraction method and applications thereof | |
| CN103045546A (en) | Method for producing feeding compound enzyme by taking lotus seed shells as raw material | |
| Yang et al. | Effects of replacing soybean meal protein with Chlorella vulgaris powder on the growth and intestinal health of grass carp (Ctenopharyngodon idella) | |
| Wang et al. | Dietary Lactobacillus casei K17 improves lipid metabolism, antioxidant response, and fillet quality of Micropterus salmoides |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20160511 |
|
| RJ01 | Rejection of invention patent application after publication |