CN110846367A - Method for extracting small molecular peptide from fresh ox stick bone and application thereof - Google Patents
Method for extracting small molecular peptide from fresh ox stick bone and application thereof Download PDFInfo
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- CN110846367A CN110846367A CN201911228071.6A CN201911228071A CN110846367A CN 110846367 A CN110846367 A CN 110846367A CN 201911228071 A CN201911228071 A CN 201911228071A CN 110846367 A CN110846367 A CN 110846367A
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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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- 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
- C07K1/145—Extraction; Separation; Purification by extraction or solubilisation
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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
Abstract
The invention belongs to the technical field of biology, and particularly relates to a method for extracting small molecular peptides from fresh ox stick bones and application of the small molecular peptides. The method comprises a method for extracting small molecule peptide from fresh cattle clavus, and comprises the following steps: cleaning and crushing fresh ox stick bones, then cooking to obtain soup, removing oil and soup, and performing enzymolysis and purification step by step to obtain a small molecular peptide solution. The method has the advantages of simple operation process steps, easy control of each link and low processing cost, can be used for industrial production, and the obtained small molecular peptide can be widely applied to the fields of medicines, foods or cosmetics.
Description
Technical Field
The invention belongs to the technical field of biology, and particularly relates to a method for extracting small molecular peptides from fresh ox stick bones and application of the small molecular peptides.
Background
In recent years, the science holds that the human body absorption protein is mainly absorbed in the form of peptide, the peptide is a molecule consisting of amino acid, the molecular weight is between 5000-200Da, the peptide has extremely strong activity and diversity, and the physiological function and the biological activity of the peptide can be utilized to regulate the physiological function of each system and cell in the human body. Therefore, various peptides are important substances affecting human health.
Numerous studies have shown that: collagen polypeptide obtained by enzymolysis of animal bone protein has biological activity, such as hypotensive activity, anti-tumor activity, antioxidant activity, activity for preventing and treating osteoarthritis and osteoporosis, and most of active peptides exist in the form of small molecular peptides with relative molecular mass less than 5000, and the polypeptide has good food processing characteristics, and has better nutrition and absorption characteristics than amino acids and proteins.
China is the first major meat producing country in the world, byproducts are also produced in large quantities with the increasing slaughter quantity, and nearly 2000 million tons of livestock and poultry bones can be produced every year at present. The beef bone accounts for about 12-20% of the weight of the beef body, namely 12-20kg of beef bone is produced when 100kg of beef is produced. In the bone processing industry of China, most of livestock bones are used for producing animal feed, such as bone glue, bone oil, bone powder and the like, the added value is very low, and particularly, the deep processing and development research of the cattle bones is less. The bone has rich nutritive value, the content of crude protein in the bone is very close to that of meat, most of the crude protein is collagen, and the collagen accounts for more than 80 percent of the total amount of protein in the bone. The collagen is a protein with the largest content in animal bones, and the collagen is further subjected to enzymolysis to obtain collagen peptide with better performance, so that the collagen peptide is applied to the fields of beauty cosmetics, medicine, food industry and the like, and the added value of the collagen peptide can be fully improved.
The ossein polypeptide becomes a hot spot of the current research due to the rich nutritional value and physiological function, however, the bone enzymolysis is reported more at home and abroad, the bone enzymolysis is concentrated on pig bones, cow bones, fishbones and the like, and the research of obtaining the small molecular peptide by enzymolysis of fresh cow stick bones is not reported yet.
Disclosure of Invention
The invention aims to provide a method for extracting small molecular peptides from fresh cattle bone, which takes the fresh cattle bone as a raw material, extracts the small molecular peptides from the fresh cattle bone through cleaning, crushing, high-temperature cooking and fractional enzymolysis by special protease, turns the cattle bone into water-soluble active ingredients through biological enzymolysis, removes fishy smell and taste through active carbon, improves the taste of the small molecular peptides from the fresh cattle bone and is more beneficial to the absorption and utilization of the active ingredients.
Specifically, the invention discloses a method for extracting small molecule peptide from fresh cattle bone, which comprises the following steps:
cleaning and crushing fresh ox stick bones, then cooking to obtain soup, removing oil and soup, and performing enzymolysis and purification step by step to obtain a small molecular peptide solution.
It is to be understood that the present invention is not limited to the above-described steps and may include other additional steps without departing from the scope of the present invention.
Preferably, the washing and crushing step comprises: the fresh ox bone is washed several times and crushed.
In some preferred embodiments of the present invention, the fresh ox bone is washed for a plurality of times according to a set washing time period, and the washed fresh ox bone is crushed according to a set crushing size. The time range of the fixed cleaning is 20-30 minutes, the cleaning frequency is 2-3 times, and the crushing size range is 3-5 cm.
Preferably, the cooking step comprises: heating water in a cooking device, adding fresh ox bone while heating, stirring, keeping the temperature after heating, stopping stirring after keeping the temperature, and cooking to form primary soup; pumping the primary soup into a storage tank, adding water into the residual bone residues, heating while stirring, keeping the temperature after heating, stopping stirring after keeping the temperature, and steaming to obtain secondary soup.
In some embodiments of the present invention, the temperature range set in the cooking step is 100-.
Preferably, the step of removing oil and draining soup comprises the following steps: standing for degreasing, and standing for 2-3 h.
In some embodiments of the present invention, the primary soup obtained in the cooking step is extracted by an extraction device and pumped into a primary soup storage device for standing and degreasing, and the secondary soup after the secondary extraction is pumped into another storage device for standing and degreasing. The extraction device is specifically a vacuum extraction tank.
Preferably, the step-by-step enzymolysis step comprises: carrying out enzymolysis reaction on the soup after oil removal; the enzyme comprises trypsin and flavourzyme, and the enzyme activity is inactivated after the enzymolysis is finished, so that hydrolysate after the enzyme inactivation is obtained.
In some embodiments of the present invention, the first soup and the second soup after separating the oil layer are both pumped into a hydrolysis device, cooled to a set enzymolysis temperature, added with trypsin and flavourzyme for hydrolysis, kept for a set enzymolysis time, heated to a set enzyme deactivation temperature after the enzymolysis is finished, and kept for a set enzyme deactivation time, so as to form an enzyme-deactivated hydrolysate. The set enzymolysis program is that flavor white enzyme with the solid content of the feed liquid of 0.1-2% is firstly added for enzymolysis for 1-2h, and then trypsin with the solid content of the feed liquid of 0.2-3% is added for hydrolysis for 2-3 h; wherein the set enzymolysis temperature range is 52-54 ℃; the set enzymolysis duration range is 3-4 hours; setting the enzyme deactivation temperature range to be 85-90 ℃; the set enzyme deactivation time period ranges from 15 minutes to 20 minutes.
Preferably, the method further comprises a step of decoloring and deodorizing before the step of purifying, and the method comprises the following specific steps: adding active carbon into the obtained hydrolysate after enzyme deactivation, standing, decoloring and removing fishy smell, wherein the adding amount of the active carbon is 0.15-0.2% of the weight of the raw materials, and standing for 3-4 h.
Preferably, the purification step comprises a filtration step, and the small molecule peptide solution is obtained after purification.
In some embodiments of the invention, filtration is performed using a multifunctional inorganic ceramic membrane. The method comprises the following specific steps: and filtering the hydrolysate obtained after decolorization and deodorization by using a filtering device to a clear liquid storage device, filtering by using a multifunctional inorganic ceramic membrane in the process, and not adding any auxiliary agent in the filtering process to obtain the small molecular peptide solution.
Preferably, the method further comprises the step of concentrating and drying the obtained molecular peptide solution to obtain small molecular peptide powder. Further, the concentration and drying step sequentially comprises the steps of filtrate concentration, high-speed tubular separation and high-speed centrifugal spray drying.
In some embodiments of the invention, the filtrate concentrating step comprises: and (3) carrying out double-effect vacuum energy-saving concentration on the small molecular peptide solution to a set Baume degree to form a clear concentrated solution. Wherein the first effect vacuum degree and the temperature are respectively controlled to be-0.06 MPa and 70-80 ℃, the second effect vacuum degree and the temperature are respectively controlled to be-0.08 MPa and 50-60 ℃, the Baume degree is taken as an index, and the Baume degree value is set to be 10-15.
The high-speed tubular separation step comprises: separating the obtained concentrated solution by a tubular separator. The rotating speed value of the tubular separator is set to 16000 r/min.
The high-speed centrifugal spray drying step comprises the following steps: heating to 80 deg.C to obtain concentrated solution with sterilization effect, cooling to 60-70 deg.C, and spray drying according to set air inlet temperature and set air outlet temperature to obtain Os bovis Seu Bubali small molecule peptide powder; the air inlet temperature range is set to be 200-220 ℃, and the air outlet temperature range is set to be 90-100 ℃.
The second aspect of the invention discloses the small molecule peptide prepared by the method.
The third aspect of the invention discloses the application of the method or the small molecule peptide in the fields of medicine, chemistry and food. The small molecular peptide obtained by the method has great development and utilization values. Preferably, the chemical field comprises the field of cosmetic cosmetics.
On the basis of the common general knowledge in the field, the above-mentioned preferred conditions can be combined arbitrarily without departing from the concept and the protection scope of the invention.
Compared with the prior art, the invention has the following remarkable advantages and effects:
according to the invention, small molecular peptides are extracted from fresh ox stick bones, so that the additional value of the ox stick bones is greatly increased, and the obtained small molecular peptides have great development and utilization values and can be widely applied to the fields of beauty cosmetics, medicine, food industry and the like.
The method disclosed by the invention adopts high-quality fresh ox stick bones as raw materials, achieves oil-liquid separation in a physical mode, does not add any chemical reagent, has a simple process flow, and can be used for mass production in workshops at present.
Drawings
FIG. 1 is a schematic process flow diagram of the extraction of small molecule peptides from fresh bovine clavus in the example of the present invention.
Detailed Description
The technical solutions of the present invention are described in detail below with reference to the drawings and the embodiments, but the present invention is not limited to the scope of the embodiments.
The experimental methods without specifying specific conditions in the following examples were selected according to the conventional methods and conditions, or according to the commercial instructions. The reagents and starting materials used in the present invention are commercially available.
Example 1
The embodiment discloses a method for extracting small molecule peptide from fresh ox stick bone, a process flow chart of which is shown in figure 1, and the method specifically comprises the following steps:
(1) cleaning and crushing: cleaning fresh ox stick bones for multiple times according to the set cleaning time, and crushing the cleaned fresh ox stick bones according to the set crushing size. The time range of the fixed cleaning is 20-30 minutes, the cleaning frequency is 2-3 times, and the crushing size range is 3-5 cm.
(2) Putting water into a cooking device, adding a certain amount of fresh beef stick bones while heating, synchronously stirring, stopping heating after heating to a set heat preservation range, preserving heat according to set heat preservation duration, stopping stirring after heat preservation is finished, cooking according to set standing duration to form primary soup, pumping the primary soup into a storage tank, adding a certain amount of water into residual bone residues, stopping heating after heating to the set heat preservation range, preserving heat according to set heat preservation duration, stopping stirring after heat preservation is finished, and cooking according to set standing duration to form secondary soup. The set heat preservation range is 100-120 ℃, the set heat preservation time range of the primary extraction is 4-6 hours, and the set heat preservation time range of the secondary extraction is 2-3 hours.
(3) Removing oil and soup: and (3) extracting the primary soup obtained in the step (2) by using an extraction device, pumping the primary soup into a primary soup storage device, standing for degreasing, and pumping the secondary soup obtained after secondary extraction into another storage device for standing for degreasing. Standing for 2-3h, and the extraction device is a vacuum extraction tank.
(4) Step-by-step enzymolysis: pumping the primary soup and the secondary soup obtained after the oil layer separation in the step (3) into a hydrolysis device, cooling to a set enzymolysis temperature, adding trypsin and flavourzyme for hydrolysis, keeping for a set enzymolysis time, heating to a set enzyme deactivation temperature after the enzymolysis is finished, and keeping for a set enzyme deactivation time to form a hydrolysate after enzyme deactivation. The set enzymolysis mode is that flavor white enzyme with the solid content of the feed liquid of 0.1-2% is added for enzymolysis for 1-2h, and then trypsin with the solid content of the feed liquid of 0.2-3% is added for hydrolysis for 2-3 h; the set enzymolysis temperature range is 52-54 ℃; the set enzymolysis duration range is 3-4 hours; setting the enzyme deactivation temperature range to be 85-90 ℃; the set enzyme deactivation time period ranges from 15 minutes to 20 minutes.
(5) Decoloring and deodorizing: and (4) adding active carbon into the hydrolysate seed after enzyme deactivation obtained in the step (4), standing, decoloring and removing fishy smell, wherein the adding amount of the active carbon is 0.15-0.2% of the weight of the raw materials, and the standing time is 3-4 h.
(6) And (3) filtering by using a multifunctional inorganic ceramic membrane: and (3) filtering the hydrolysate obtained after standing in the step (5) to a clear liquid storage device by using a filtering device, wherein a multifunctional inorganic ceramic membrane is used for filtering in the process, and no auxiliary agent is added in the filtering process.
(7) Concentrating the filtrate: and (4) performing double-effect vacuum energy-saving concentration on the clear liquid filtered in the step (6) to set Baume degree to form clear concentrated liquid. Wherein the first effect vacuum degree and the temperature are respectively controlled to be-0.06 MPa and 70-80 ℃, the second effect vacuum degree and the temperature are respectively controlled to be-0.08 MPa and 50-60 ℃, the Baume degree is taken as an index, and the Baume degree value is set to be 10-15.
(8) High-speed tubular separation: and (4) separating the concentrated solution obtained in the step (7) by a tubular separator. The rotating speed value of the high-speed tube type separator is set to 16000 r/min.
(9) High-speed centrifugal spray drying: heating to 80 deg.C to obtain concentrated solution with sterilization effect, cooling to 60-70 deg.C, and spray drying according to set air inlet temperature and set air outlet temperature to obtain fresh Os bovis Seu Bubali small molecule peptide powder; the air inlet temperature range is set to be 200-220 ℃, and the air outlet temperature range is set to be 90-100 ℃.
(10) And (3) product packaging: and (5) adopting sealed packaging.
The method disclosed by the embodiment adopts high-quality fresh ox stick bones as raw materials, achieves oil-liquid separation in a physical mode, does not add any chemical reagent, is simple in process flow, and can be used for mass production in workshops.
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.
Claims (10)
1. A method for extracting small molecule peptide from fresh ox stick bone is characterized by comprising the following steps: cleaning and crushing fresh ox stick bones, then cooking to obtain soup, removing oil and soup, and performing enzymolysis and purification step by step to obtain a small molecular peptide solution.
2. The method of claim 1, wherein the step of washing and crushing comprises: the fresh ox bone is washed several times and crushed.
3. The method of claim 1, wherein the cooking step comprises: heating water in a cooking device, adding fresh ox bone while heating, stirring, keeping the temperature after heating, stopping stirring after keeping the temperature, and cooking to form primary soup; pumping the primary soup into a storage tank, adding water into the residual bone residues, heating while stirring, keeping the temperature after heating, stopping stirring after keeping the temperature, and steaming to obtain secondary soup.
4. The method of claim 1, wherein the step of removing oil and draining comprises: standing for degreasing.
5. The method of claim 1, wherein the step-wise enzymatic step comprises: carrying out enzymolysis reaction on the soup after oil removal; the enzyme comprises trypsin and flavourzyme, and the enzyme activity is inactivated after the enzymolysis is finished, so that hydrolysate after the enzyme inactivation is obtained.
6. The method according to claim 1, further comprising a step of decolorizing and deodorizing before the purification step, in particular as follows: adding active carbon into the obtained hydrolysate after enzyme deactivation, standing, decoloring and removing fishy smell, wherein the adding amount of the active carbon is 0.15-0.2% of the weight of the raw materials.
7. The method according to claim 1, wherein the purification step comprises a filtration step, and the small molecule peptide solution is obtained after purification.
8. The method according to claim 1, further comprising a step of concentrating and drying the obtained molecular peptide solution to obtain small molecular peptide powder.
9. A small molecule peptide produced by the method of any one of claims 1-8.
10. Use of the method according to any one of claims 1-8 or the small molecule peptide according to claim 9 in the fields of medicine, chemistry and food.
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| CN201911228071.6A CN110846367A (en) | 2019-12-04 | 2019-12-04 | Method for extracting small molecular peptide from fresh ox stick bone and application thereof |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN111685221A (en) * | 2020-06-22 | 2020-09-22 | 拓普河北生物科技有限公司 | Small molecule yak bone total nutrient peptide and preparation method thereof |
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| CN105795363A (en) * | 2016-03-25 | 2016-07-27 | 东北农业大学 | Amino acid-enriched bovine bone powder and preparation method thereof |
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| WO2020113974A1 (en) * | 2018-12-03 | 2020-06-11 | 无锡金农生物科技有限公司 | Preparation method and use for rice protein peptide |
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2019
- 2019-12-04 CN CN201911228071.6A patent/CN110846367A/en active Pending
Patent Citations (5)
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| CN101822308A (en) * | 2010-04-30 | 2010-09-08 | 中国肉类食品综合研究中心 | Enzymatic animal bone salty compound peptide and preparation method thereof |
| CN103783254A (en) * | 2013-07-02 | 2014-05-14 | 吴长海 | Preparation method of yak bone collagen peptide |
| CN106480142A (en) * | 2015-09-02 | 2017-03-08 | 于海峰 | A kind of preparation method of ox bone collagen protein peptide |
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| CN111685221A (en) * | 2020-06-22 | 2020-09-22 | 拓普河北生物科技有限公司 | Small molecule yak bone total nutrient peptide and preparation method thereof |
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Effective date of registration: 20200515 Address after: No. 402, gate 1, building 6, dongshengyuan apartment dormitory, Haidian District, Beijing 100083 Applicant after: Wu Wei Address before: 031800 Nihezhang Village, Jicheng Town, Yushe County, Jinzhong City, Shanxi Province Applicant before: Shanxi Native Peptide Technology Co.,Ltd. |
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