CN111235201A - Deer bone peptide and extraction method thereof - Google Patents
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- C—CHEMISTRY; METALLURGY
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- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
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- C07K1/36—Extraction; Separation; Purification by a combination of two or more processes of different types
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- 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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Abstract
The invention discloses a method for extracting deer bone peptide, which comprises the following steps: 1) taking deer bones and taking out bone marrow for later use; pulverizing Os Cervi for use; 2) dispersing bone marrow into purified warm water, adding zymophyte agent, fermenting; after the fermentation is finished, taking supernatant for later use; 3) blowing the crushed deer bones by high-pressure steam to remove grease, cleaning, then softening the deer bones by using the high-pressure steam again, boiling the deer bones in water after the softening treatment, keeping stirring, obtaining deer bone soup after the boiling, adding an enzyme preparation, heating to inactivate the enzyme preparation after the hydrolysis is finished, obtaining deer bone hydrolysate, standing for precipitation, and taking a hydrolysate liquid part for later use; circularly filtering the filtrate obtained after each filtration, and filtering for multiple times to obtain a clear solution; 4) mixing the supernatant obtained in 2) and the clarified solution obtained in 3), stirring, and concentrating to obtain deer bone peptide concentrated solution. The deer bone peptide prepared by the invention is easy to absorb, and the utilization rate is improved.
Description
Technical Field
The invention belongs to the technical field of peptide preparation, and particularly relates to a deer bone peptide extraction method and deer bone peptide prepared by the method.
Technical Field
Deer bone is a traditional Chinese medicine with the efficacy of tonifying deficiency and strengthening bones. The deer bone has effects of tonifying deficiency, strengthening tendons and bones, and is suitable for treating asthenia due to long-term illness, insufficiency of marrow and essence, anemia, rheumatism, pain of limbs, etc.
Medical works are well documented:
① Bian Lu (miscellaneous records): 'Antai, Xie Qi'
② Qianjin dietotherapy, governing internal deficiency, continuing to stop injury, and tonifying bone, it can be used as wine "
③ Tang Ben Cao (Tang materia Medica) for treating consumptive disease, including wine, wind-evil deficiency, and bone marrow supplementation "
④ compendium for infants who should be taken with ash-burning water and have diarrhea "
⑤ recorded in Sichuan Chinese materia medica, for treating rheumatism, limbs pain, cold arthralgia of muscles and bones "
The deer bone recorded in compendium of materia medica has slight heat, sweet and nontoxic taste, has the efficacies of tonifying deficiency, strengthening the bone and the like, is a rare body tonifying treasure, contains rich mineral substances such as calcium, phosphorus and the like, has the effect of reuniting bones and tendons, and particularly has obvious effect on senile osteoporosis. The compendium of materia medica considers that the deer bone is durable after being taken for a long time, can provide better nutrition supply and health maintenance aiming at the old and weak body, and is a high-grade tonic for the Yinfu family.
The deer bone has extremely similar components with human skeleton, wherein calcium accounts for 30%, collagen accounts for 30%, and other minerals and vitamins account for about 30%, so that the deer bone five-collagen peptide can supplement calcium and bone. The deer bone five-collagen peptide comprehensively relieves the symptoms of waist soreness, backache, leg cramp, deadlimb, hunchback, hyperosteogeny, cervical vertebra pain, lumbar disc herniation, scapulohumeral periarthritis, arthritis, growth retardation of children, chondropathy and the like caused by bone deficiency through three-in-one of bone formation, bone protection and bone nourishing, and constructs a healthier bone. Deer bone is rich in calcium, phosphorus, zinc and iron which are needed by human body, a large amount of protein, phospholipid, chondroitin, mucopolysaccharide, deer bone formation protein, a plurality of trace elements and amino acids which can not be synthesized by human body and the like, and is rich in natural active vitamin.
Deer blood in deer bone has an important position in clinical traditional Chinese medicine and is widely applied in folk. Recently, animal experiments and clinical researches prove that the deer blood really has multiple treatment and health-care effects of beautifying, treating anemia, regulating immunity, delaying senility, improving memory, resisting fatigue, improving sexual function and the like. The deer blood is originally recorded in Tang Dynasty Sun Simiao (Qianjin Aifang) dietotherapy, to Ming Dynasty Li Shizhen (compendium of materia Medica) to detail the medical action of deer blood, which is mainly used for treating impotence, deficiency, lumbago, epistaxis, traumatic injury, rabies injury, and the immediate cure of people who want to drink the deer blood for treating consumptive lung disease, hematemesis, metrorrhagia, leukorrhagia and other qi pains. Tonify deficiency, nourish essence and blood, and remove pox and drug toxicity.
At present, deer bones are mostly used as traditional Chinese medicines or wine, but the treatment mode is not favorable for the effective components to be exerted and utilized.
Disclosure of Invention
The invention aims to provide a method for extracting deer bone peptide, which enriches the utilization of deer bones, and the deer bone peptide prepared by the method has the characteristic of easy absorption.
The specific scheme is as follows:
a method for extracting deer bone peptide comprises the following steps:
1) raw material treatment
Taking fresh or frozen deer bone, removing residual meat, and taking out bone marrow for later use; pulverizing Os Cervi, and washing with clear water;
2) bone marrow processing
Dispersing bone marrow into purified warm water, and performing ultrasonic treatment to break bone marrow cells to obtain bone marrow solution; after cooling to room temperature, adding a zymophyte agent, and performing fermentation treatment; after fermentation is finished, inactivating the zymophyte agent, cooling, centrifuging and taking supernatant for later use;
3) deer bone treatment
Blowing the crushed deer bones with high-pressure steam to remove grease, cleaning, then softening the deer bones with high-pressure steam again, boiling in water after softening, keeping stirring, obtaining deer bone soup after boiling, and then standing;
after standing, sterilizing the deer bone soup at high temperature, cooling to hydrolysis temperature and keeping at a constant temperature after sterilization, adding enzyme preparation, heating to inactivate enzyme preparation after hydrolysis is finished, obtaining deer bone hydrolysate, standing for precipitation, and taking hydrolysate liquid part for later use; precipitating the liquid part of the hydrolysate with anhydrous ethanol, filtering, and concentrating; diluting with water, adding diatomite and/or active carbon, standing, filtering to obtain filtrate, circularly filtering the filtrate obtained after each filtration, and filtering for multiple times to obtain clear solution;
4) late stage preparation
Mixing the supernatant obtained in 2) and the clarified solution obtained in 3), stirring for 5-7h, and concentrating to obtain deer bone peptide concentrated solution.
In a specific embodiment of the invention, 1) in the raw material treatment, the crushed size of the deer bone is controlled to be 10-25 mm.
In a specific embodiment of the present invention, 2) the temperature of the purified warm water is controlled to 40 to 45 ℃ in the bone marrow treatment.
In the bone marrow treatment, the ultrasonic frequency is 400-500KHz, and the time duration is controlled to be 1-1.5 min.
In a specific embodiment of the invention, 2) in the bone marrow treatment, the fermentation bacteria agent is selected from one or more of lactobacillus, streptococcus, micrococcus, lactobacillus and pediococcus acidilactici; the fermentation time is not less than 10 h.
The invention specifically relates to a method for processing deer bones, which comprises the following steps of 3) boiling in water for 10-13 hours, and controlling the temperature at 80-90 ℃.
In the embodiment of the invention, 3) the standing treatment time is 3-7h in the deer bone treatment.
In a specific embodiment of the present invention, 3) the deer bone treatment, the enzyme preparation is selected from animal proteases.
In the embodiment of the invention, 3) the hydrolysis temperature is 50-65 ℃ and the time is 5-7h in the deer bone treatment.
The invention also provides deer bone peptide, which is prepared by any one of the extraction methods of the deer bone peptide.
Research shows that deer bone peptide not only has good digestion and absorption characteristics, but also has more effective physiological function than collagen, and since Japanese scholars confirm that collagen polypeptide has the effect of lowering blood pressure in 1979, people continuously discover that the polypeptide also has various physiological functions such as bone formation promotion, skin collagen metabolism promotion and the like.
The polypeptide has good nutritional characteristics and excellent processing characteristics, can be applied to the food industry to develop health-care food with beauty effect and osteoporosis prevention, and can be directly added into the food, or the concentrated solution prepared by the invention is added into auxiliary materials acceptable in the food or health-care product industry to prepare corresponding additive products, and the auxiliary materials can adopt any products disclosed in the prior art. For example, deer bone peptide is added into functional wine to enhance the effects of nourishing, resisting fatigue, etc. After the polypeptide is added, the physical and chemical property indexes of the product can be improved, and the wine is beautiful and clear.
The deer bone peptide has great market development potential and can be further used as a cosmetic product. The addition method can be added according to the related requirements of cosmetics.
Compared with the prior art, the invention has the beneficial effects that:
the deer bone peptide prepared by the invention is easy to absorb, improves the utilization rate, can effectively prevent the damage of free radicals to human cell membranes, slows down the aging process of human cells, resists oxidation and effectively enhances the activity of SOD. Can effectively increase the content of PGI2 in human body, prevent platelet aggregation and protect heart and cerebral vessels. The deer bone peptide has functions of increasing hemoglobin and preventing anemia. The product has effects in tonifying kidney, strengthening kidney, promoting marrow production, tonifying marrow, relieving bone marrow aging, recovering young state of bone marrow, preventing and treating various bone and joint diseases, lubricating joints, providing nutrients for human skeleton, strengthening tendons and bones, enhancing bone density, preventing hyperosteogeny and osteoporosis; delaying aging and improving immunity; enhancing muscle elasticity and collagen formation.
Detailed Description
The technical solutions of the present invention will be described clearly and completely below, and it should be understood that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. As used herein, the term "and/or" includes any and all combinations of one or more of the associated listed items.
Example 1
A method for extracting deer bone peptide comprises the following steps:
1) raw material treatment
Taking fresh or frozen deer bone, removing residual meat, and taking out bone marrow for later use; pulverizing Os Cervi into small pieces of 10-25mm, and washing with clear water;
2) bone marrow processing
Dispersing bone marrow into purified warm water with temperature controlled at 40 deg.C, with frequency of 400KHz, and ultrasonic treating for 1min to break bone marrow cells to obtain bone marrow solution; after cooling to room temperature, adding a zymophyte agent, and fermenting for 10 h; after fermentation is finished, inactivating the zymophyte agent, cooling, centrifuging and taking supernatant for later use; the zymophyte agent is selected from one or more of lactobacillus, streptococcus, micrococcus, lactobacillus and pediococcus acidilactici;
3) deer bone treatment
Blowing the crushed deer bones with high-pressure steam to remove grease, cleaning, then softening the deer bones with high-pressure steam again, placing the deer bones in water with the temperature controlled at 80 ℃ after softening, boiling for 10-13h, keeping stirring, obtaining deer bone soup after boiling, and standing for 3 h;
after standing, performing high-temperature sterilization treatment on the deer bone soup by a conventional method, cooling to hydrolysis temperature after sterilization, keeping the temperature constant, adding an enzyme preparation, wherein the hydrolysis temperature is 50 ℃, the time is 5 hours, heating to more than 85 ℃ after hydrolysis, inactivating the enzyme preparation to obtain deer bone hydrolysate, standing for precipitation for 3 hours, and taking a hydrolysate liquid part for later use; precipitating the liquid part of the hydrolysate with anhydrous ethanol, filtering, and concentrating; diluting with water, adding diatomite and/or active carbon, standing, filtering to obtain filtrate, circularly filtering the filtrate obtained after each filtration, and filtering for multiple times to obtain clear solution; the enzyme preparation is selected from animal protease;
4) late stage preparation
Mixing the supernatant obtained in 2) and the clarified solution obtained in 3), stirring for 5h, and concentrating to obtain deer bone peptide concentrated solution.
Example 2
A method for extracting deer bone peptide comprises the following steps:
1) raw material treatment
Taking fresh or frozen deer bone, removing residual meat, and taking out bone marrow for later use; pulverizing Os Cervi into small pieces of 10-25mm, and washing with clear water;
2) bone marrow processing
Dispersing bone marrow into purified warm water with temperature controlled at 43 deg.C, with frequency of 450KHz, and ultrasonic treating for 1min to break bone marrow cells to obtain bone marrow solution; after cooling to room temperature, adding a zymophyte agent, and fermenting for 12 h; after fermentation is finished, inactivating the zymophyte agent, cooling, centrifuging and taking supernatant for later use; the zymophyte agent is selected from one or more of lactobacillus, streptococcus, micrococcus, lactobacillus and pediococcus acidilactici;
3) deer bone treatment
Blowing the crushed deer bones by high-pressure steam, removing grease, cleaning, then softening the deer bones by using the high-pressure steam again, placing the deer bones in water with the temperature controlled at 85 ℃ after the softening treatment, boiling for 11 hours, keeping stirring, obtaining deer bone soup after the boiling is finished, and standing for 5 hours;
after standing, performing high-temperature sterilization treatment on the deer bone soup by a conventional method, cooling to hydrolysis temperature after sterilization, keeping the temperature constant, adding an enzyme preparation, wherein the hydrolysis temperature is 60 ℃, the duration is 6 hours, heating to more than 85 ℃ after hydrolysis is finished to inactivate the enzyme preparation, obtaining deer bone hydrolysate, standing for precipitation for 5 hours, and taking a hydrolysate liquid part for later use; precipitating the liquid part of the hydrolysate with anhydrous ethanol, filtering, and concentrating; diluting with water, adding diatomite and/or active carbon, standing, filtering to obtain filtrate, circularly filtering the filtrate obtained after each filtration, and filtering for multiple times to obtain clear solution; the enzyme preparation is selected from animal protease;
4) late stage preparation
Mixing the supernatant obtained in 2) and the clarified solution obtained in 3), stirring for 6h, and concentrating to obtain deer bone peptide concentrated solution.
Example 3
A method for extracting deer bone peptide comprises the following steps:
1) raw material treatment
Taking fresh or frozen deer bone, removing residual meat, and taking out bone marrow for later use; pulverizing Os Cervi into small pieces of 10-25mm, and washing with clear water;
2) bone marrow processing
Dispersing bone marrow into purified warm water with temperature controlled at 45 deg.C, with frequency of 500KHz, and ultrasonic treating for 1.5min to break bone marrow cells to obtain bone marrow solution; after cooling to room temperature, adding a zymophyte agent, and fermenting for 13 h; after fermentation is finished, inactivating the zymophyte agent, cooling, centrifuging and taking supernatant for later use; the fermentation inoculum is selected from the composition of lactobacillus and streptococcus in any mass ratio;
3) deer bone treatment
Blowing the crushed deer bones by high-pressure steam, removing grease, cleaning, then softening the deer bones by using the high-pressure steam again, placing the deer bones in water with the temperature controlled at 90 ℃ after the softening treatment, boiling for 13 hours, keeping stirring, obtaining deer bone soup after the boiling is finished, and then standing for 7 hours;
after standing, performing high-temperature sterilization treatment on the deer bone soup by a conventional method, cooling to hydrolysis temperature after sterilization, keeping the temperature constant, adding an enzyme preparation, wherein the hydrolysis temperature is 65 ℃, the duration is 7 hours, heating to more than 85 ℃ after hydrolysis, inactivating the enzyme preparation to obtain deer bone hydrolysate, standing for precipitation for 6 hours, and taking a hydrolysate liquid part for later use; precipitating the liquid part of the hydrolysate with anhydrous ethanol, filtering, and concentrating; diluting with water, adding diatomite and/or active carbon, standing, filtering to obtain filtrate, circularly filtering the filtrate obtained after each filtration, and filtering for multiple times to obtain clear solution; the enzyme preparation is selected from animal protease;
4) late stage preparation
Mixing the supernatant obtained in 2) and the clarified solution obtained in 3), stirring for 7h, and concentrating to obtain deer bone peptide concentrated solution.
Comparative example 1
The difference from example 1 is that bone marrow and deer bone are combined and treated according to the deer bone treatment method, i.e., one operation without bone marrow treatment, and the rest is the same.
Comparative example 2
The difference from the example 1 is that 3) the two standing treatment processes are eliminated in the deer bone treatment, and the rest operations are the same.
The method for measuring the hydrolysis degree adopted by the invention comprises the following steps:
NH formed after hydrolysis2The amount of the base is measured by an amino nitrogen measuring method, namely a double indicator formaldehyde titration method to obtain N1, and the total nitrogen amount of the sample is measured by a micro Kjeldahl nitrogen method to obtain N2;
the degree of hydrolysis was N1/N2 x 100%.
The degree of hydrolysis of each example and comparative example is shown in table 1 below.
TABLE 1
| Degree of hydrolysis | |
| Example 1 | 21.6％ |
| Example 2 | 23.2％ |
| Example 3 | 22.1％ |
| Comparative example 1 | 17.3％ |
| Comparative example 2 | 16.8％ |
According to the scheme, the bone marrow is subjected to fermentation treatment and then combined with the deer bone in an enzymolysis method, so that the hydrolysis degree of the prepared deer bone peptide can be improved to a certain extent.
The application the extraction process is the defect, can effectively improve the quality of the prepared deer bone peptide through the synergistic effect of a plurality of steps, thereby leading the deer bone peptide to be absorbed more easily, further effectively preventing the damage of free radicals to human cell membranes relatively, slowing down the aging process of human cells, resisting oxidation, effectively enhancing the activity of SOD and preventing or improving the occurrence of osteoporosis. Of course, the improvement of the above effects is not limited, and the enhancement in the relative sense of other effects is also included.
The foregoing is illustrative of the preferred embodiments of this invention, and it is to be understood that the invention is not limited to the precise form disclosed herein and that various other combinations, modifications, and environments may be resorted to, falling within the scope of the concept as disclosed herein, either as described above or as apparent to those skilled in the relevant art. And that modifications and variations may be effected by those skilled in the art without departing from the spirit and scope of the invention as defined by the appended claims.
Claims (10)
1. The method for extracting the deer bone peptide is characterized by comprising the following steps of:
1) raw material treatment
Taking fresh or frozen deer bone, removing residual meat, and taking out bone marrow for later use; pulverizing Os Cervi, and washing with clear water;
2) bone marrow processing
Dispersing bone marrow into purified warm water, and performing ultrasonic treatment to break bone marrow cells to obtain bone marrow solution; after cooling to room temperature, adding a zymophyte agent, and performing fermentation treatment; after fermentation is finished, inactivating the zymophyte agent, cooling, centrifuging and taking supernatant for later use;
3) deer bone treatment
Blowing the crushed deer bones with high-pressure steam to remove grease, cleaning, then softening the deer bones with high-pressure steam again, boiling in water after softening, keeping stirring, obtaining deer bone soup after boiling, and then standing;
after standing, sterilizing the deer bone soup at high temperature, cooling to hydrolysis temperature and keeping at a constant temperature after sterilization, adding enzyme preparation, heating to inactivate enzyme preparation after hydrolysis is finished, obtaining deer bone hydrolysate, standing for precipitation, and taking hydrolysate liquid part for later use; precipitating the liquid part of the hydrolysate with anhydrous ethanol, filtering, and concentrating; diluting with water, adding diatomite and/or active carbon, standing, filtering to obtain filtrate, circularly filtering the filtrate obtained after each filtration, and filtering for multiple times to obtain clear solution;
4) late stage preparation
Mixing the supernatant obtained in 2) and the clarified solution obtained in 3), stirring for 5-7h, and concentrating to obtain deer bone peptide concentrated solution.
2. The method for extracting deer bone peptide according to claim 1, wherein: 1) in the raw material treatment, the crushed size of the deer bone is controlled to be 10-25 mm.
3. The method for extracting deer bone peptide according to claim 1, wherein: 2) in the bone marrow treatment, the temperature of the purified warm water is controlled at 40-45 ℃.
4. The method for extracting deer bone peptide according to claim 1, wherein: 2) in the bone marrow treatment, the ultrasonic frequency is 400-500KHz, and the time duration is controlled to be 1-1.5 min.
5. The method for extracting deer bone peptide according to claim 1, wherein: 2) in the bone marrow treatment, the zymophyte agent is selected from one or more of lactobacillus, streptococcus, micrococcus, lactobacillus and pediococcus acidilactici; the fermentation time is not less than 10 h.
6. The method for extracting deer bone peptide according to claim 1, wherein: 3) in the deer bone treatment, the boiling time in water is 10-13h, and the temperature is controlled at 80-90 ℃.
7. The method for extracting deer bone peptide according to claim 1, wherein: 3) in the deer bone treatment, the standing treatment time is 3-7 h.
8. The method for extracting deer bone peptide according to claim 1, wherein: 3) in the deer bone treatment, the enzyme preparation is selected from animal protease.
9. The method for extracting deer bone peptide according to claim 1, wherein: 3) in the deer bone treatment, the hydrolysis temperature is 50-65 deg.C, and the time is 5-7 h.
10. A deer bone peptide, which is characterized in that: is prepared by the method for extracting deer bone peptide as described in any one of claims 1-9.
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