CN111334551B - Production process of cow leather collagen peptide - Google Patents

Production process of cow leather collagen peptide Download PDF

Info

Publication number
CN111334551B
CN111334551B CN202010250674.2A CN202010250674A CN111334551B CN 111334551 B CN111334551 B CN 111334551B CN 202010250674 A CN202010250674 A CN 202010250674A CN 111334551 B CN111334551 B CN 111334551B
Authority
CN
China
Prior art keywords
enzymolysis
cow leather
temperature
product
collagen peptide
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.)
Active
Application number
CN202010250674.2A
Other languages
Chinese (zh)
Other versions
CN111334551A (en
Inventor
尹晓清
吴海
靳祯亮
王希搏
吴梦婷
王微
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Hubei Reborn Biotech Co ltd
Original Assignee
Hubei Reborn Biotech Co ltd
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Hubei Reborn Biotech Co ltd filed Critical Hubei Reborn Biotech Co ltd
Priority to CN202010250674.2A priority Critical patent/CN111334551B/en
Publication of CN111334551A publication Critical patent/CN111334551A/en
Application granted granted Critical
Publication of CN111334551B publication Critical patent/CN111334551B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Images

Classifications

    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12PFERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P21/00Preparation of peptides or proteins
    • C12P21/06Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07KPEPTIDES
    • C07K14/00Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/78Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin, cold insoluble globulin [CIG]

Abstract

The invention belongs to the technical field of enzymolysis of bovine collagen peptide, and particularly relates to a production process of bovine collagen peptide, which comprises S1 cleaning, S2 denaturation, S3 enzymolysis, S4 centrifugation, S5 enzyme inactivation, S6 impurity removal and S7 finished product. The invention has the following advantages and effects: the method adopts the combined action of the ultrasonic process, the denaturation process and the enzymolysis process, and can be conveniently suitable for the cowhide in various production places; the collagen peptide product produced by the process provided by the application has good solubility and dispersibility, good sensory quality and high yield, and a large amount of active polypeptide in the product enables the product to have strong antioxidant capacity; the method has mild reaction conditions, low production energy consumption and process conditions and process effects far superior to those of the prior art.

Description

Production process of cow leather collagen peptide
Technical Field
The invention belongs to the technical field of enzymolysis of bovine collagen peptide, and particularly relates to a production process of bovine collagen peptide.
Background
Collagen peptide is obtained by enzymolysis of collagen, and has small molecular weight, increased polar group and thus high solubility. After the collagen is degraded into the collagen peptide, because a large number of hydrophilic groups leak out, the water absorption and water retention of the collagen peptide are obviously higher than those of the collagen peptide, and the collagen peptide can keep skin moisture when being used in cosmetics, thereby improving the skin moisturizing effect. The emulsifying property and the foaming property of the collagen peptide also can be used as a food additive to improve the quality of food, so that the product has good taste and rich nutrition. Collagen peptide has not only the general functional properties of proteins but also many biological activities, such as antioxidant activity, angiotensin converting enzyme inhibitory activity, platelet aggregation inhibitory activity, antitumor activity, and the like.
Most of the collagen on the market today is mainly prepared from fish skin, because the collagen produced from cow skin generally has the following problems:
(1) the domestic cowhide raw materials are generally in various and miscellaneous situations, the production places of the cowhides are various, so that the quality of the cowhides is also uneven, the quality of products obtained in subsequent production is directly influenced, most manufacturers adopt a rough one-knife cutting treatment mode at present, a large amount of raw materials are wasted, and the quality of the products cannot be effectively guaranteed;
(2) the sterilization generally adopts an ultrahigh temperature mode, so that the color of the product is deepened, the activity of active polypeptide in the product is reduced and even lost, and the functionality of the product is influenced;
(3) the decolorization process generally adopts activated carbon powder adsorption, and irreversible loss of products (polypeptides) can be caused while pigment substances are removed;
(4) the produced collagen peptide has unpleasant peculiar smell and poor flavor.
Disclosure of Invention
The invention aims to provide a production process of bovine skin collagen peptide, which is a production method of bovine skin collagen peptide with high yield, good sensory effect and strong antioxidant activity.
The technical purpose of the invention is realized by the following technical scheme:
A production process of cow hide collagen peptide comprises the following steps:
s1 cleaning: cleaning 100 parts of cow leather by using clear water, soaking the cleaned cow leather in 2000 parts of water by weight, adding an alkaline regulator to regulate the pH value to be strong alkaline, adding 0.5-2 parts of a bactericide, and soaking for 48-240 hours;
s2 denaturation: cleaning the clean cow leather obtained in the step S1 to neutral pH, cutting and grinding the cleaned cow leather into cow leather slurry, adding water with the weight part of 4-6 times that of the cow leather slurry into the cow leather slurry, and preserving heat for a certain time to obtain modified cow leather protein slurry;
s3 enzymolysis: adding 2-5 parts by mass of alkaline protease into the denatured cow leather protein slurry obtained in the step S2, keeping a certain temperature and pH, reacting for a period of time, and performing enzymolysis with ultrasonic waves in an auxiliary manner in the enzymolysis process to finally obtain an enzymolysis solution;
s4 centrifugation: centrifuging the enzymolysis liquid obtained in the step S3, and taking supernate;
s5 enzyme deactivation: heating the supernatant obtained in the centrifugation in the step S4 to 95 ℃, and inactivating the enzyme for 10min to obtain an enzyme inactivation solution;
s6 impurity removal: filtering the enzyme deactivation liquid obtained in the step S5 at constant temperature by adopting a ceramic membrane to obtain a product dilute liquid; concentrating the dilute liquid product to solid content over 20% by using membrane equipment, and removing microorganisms from the concentrated liquid by using a filter element to obtain concentrated liquid;
S7 finished product: and (5) spray-drying the concentrated solution obtained in the step S6 to obtain a solid, and inspecting and packaging the solid product to obtain a finished product.
By adopting the technical scheme, the production conditions of bovine hide collagen decomposition are optimized, particularly bactericide and denaturation treatment are carried out, strong basicity and bactericide are used for one-time denaturation in the S1 cleaning process, so that the removal of unnecessary impurity components is facilitated, the toughness of the bovine hide is adjusted, the bovine hide is conveniently milled into slurry in the follow-up process, and the process can be quickly adjusted by carrying out small-scale pre-experiments according to different batches of bovine hides in different production places; in the denaturation process of S2, secondary denaturation is carried out, and the cow leather slurry is treated at a proper temperature for a period of time in advance for denaturation, so that the subsequent enzymolysis efficiency is greatly improved, and the enzymolysis requirement is reduced; thus milder conditions may be used in the S3 enzymatic step.
The invention is further provided with: the alkaline regulator in the step S1 comprises NaOH and Na2CO3、NaHCO3、K2CO3、KHCO3、CaCO3And one or more of CaO, and the pH value is adjusted to 10-14.
By adopting the technical scheme, the alkaline regulators have low cost on one hand, and even a small amount of ion residues do no harm to human bodies on the other hand.
The invention is further configured as follows: the bactericide in the step S1 is H2O2One or more of lysozyme, stable chlorine dioxide and sodium hypochlorite.
Through adopting above-mentioned technical scheme, the majority is strong oxidant, uses under the not good condition of cow hide quality, supplementary regulation cow hide toughness when disinfecting, makes things convenient for the homogenate, when the cow hide quality is better, can adopt comparatively mild lysozyme, furthest's protection collagen.
The invention is further configured as follows: and the heat preservation temperature in the S2 denaturation is set within 60-80 ℃, and the heat preservation time is set within 4-6 h.
By adopting the technical scheme, the denaturation treatment is performed in advance, and the subsequent enzymolysis is facilitated.
The invention is further provided with: in the step S3, the reaction temperature is set to be 50-65 ℃, the reaction pH is set to be 7.5-9.5, the reaction time is set to be 5-6h, and the power of the ultrasonic wave is 200-900 kW.
By adopting the technical scheme, the enzymolysis process is mild, ultrasonic enzymolysis is adopted in an auxiliary way, the enzymolysis efficiency is improved, and meanwhile, the content of ultra-small molecules is reduced by inhibiting the enzyme activity through the environment, so that the content of amino terminal residues is reduced, and the uncomfortable taste of small molecular peptides generated by decomposing bovine collagen is reduced.
The invention is further configured as follows: in the step S4 of centrifugation, a tubular centrifuge is adopted, the centrifugation speed is more than 10000r/min, and the temperature is kept between 40 and 60 ℃ during centrifugation.
The invention is further configured as follows: in the step S6, in the impurity removal, the aperture of the used ceramic membrane is set at 8000-14000Dal, and the feed liquid and the ambient temperature are controlled to be 60-80 ℃ during the filtration; the aperture of the membrane equipment is set at 500-600 Dal; the aperture of the filter element is set to be 0.22 μm.
By adopting the technical scheme, the peptide with ultra-small molecular weight can be removed, the average molecular weight is further improved, and the unpleasant odor is reduced; meanwhile, various impurities such as pigment and the like are removed, and the product quality is improved.
The invention is further provided with: the air inlet temperature of the spray drying is set to be 150-180 ℃, and the air outlet temperature is set to be 70-80 ℃.
The invention has the beneficial effects that:
1. the application provides a denaturation technology for handling cow hide in advance, reduces its difference in quality, has reduced follow-up cow hide processing degree of difficulty for even the cow hide raw materials of many sources also can be the processing of high efficiency low cost become the stable bovine collagen peptide product of quality homogeneity.
2. The method adopts the combined action of an ultrasonic process, a denaturation process and an enzymolysis process, and the denaturation process actively destroys the connection structure between proteins, so that the bovine skin collagen is more dispersed; ultrasonic waves destroy the space structure of collagen, expose more enzymolysis sites and increase the enzymolysis efficiency; under the combined action of the two, the reaction condition of enzymolysis can be milder, the energy consumption is reduced, the environment deviates from the optimal environment of protease, the enzyme digestion efficiency of the protease is inhibited, the product is more inclined to the peptide with larger molecular weight, the generation amount of the ultra-small molecular peptide is greatly reduced, and the ultra-small molecular peptide is the root cause of generating unpleasant temperature.
3. According to the method, the processes of membrane filtration and filter element filtration are adopted for impurity removal and sterilization, so that on one hand, the influence of high temperature on the activity of the small molecular peptide is avoided, and the small molecular peptide in the product has higher biological oxidation resistance activity; on the other hand, the Maillard reaction of the product is avoided without adopting high-temperature sterilization liquid, the generation of pigment is reduced, and the requirement on color removal is reduced, so that the requirement on color removal of the product can be met by membrane filtration, and the loss of protein peptide caused by adopting substances such as activated carbon is also avoided.
Drawings
In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the drawings needed to be used in the description of the embodiments will be briefly introduced below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art to obtain other drawings based on these drawings without creative efforts.
FIG. 1 is a graph showing the results of the oxidation resistance test according to the present invention
Detailed Description
The technical solutions in the examples will be clearly and completely described below. It is to be understood that the described embodiments are merely a few embodiments of the invention, and not all embodiments.
Example 1
A production process of bovine skin collagen peptide comprises the following steps:
s1 cleaning: cleaning 100 parts of cow leather with clear water, soaking the cleaned cow leather in 1400 parts of water, adding NaOH to adjust the pH value to 13, and adding 1.35 parts of H2O2Soaking for 78 h;
s2 denaturation: cleaning the clean cow leather obtained in the step S1 until the pH value is neutral, then cutting and grinding the clean cow leather to prepare cow leather slurry, adding 5 parts by weight of water into the cow leather slurry, setting the heat preservation temperature in denaturation to 72 ℃ and the heat preservation time to 5.5 hours to obtain denatured cow leather protein slurry;
s3 enzymolysis: adding 3.75 parts of alkaline protease into the denatured cow leather protein slurry obtained in the step S2, keeping the temperature at 57 ℃, keeping the pH at 8.2, reacting for 5.5 hours, and performing enzymolysis with ultrasonic waves in an auxiliary manner in the enzymolysis process, wherein the power of the ultrasonic waves is 700kW, so as to finally obtain an enzymolysis liquid;
s4 centrifugation: centrifuging the enzymolysis liquid obtained in the step S3, centrifuging at 20000r/min in a tubular centrifuge at 55 deg.C, and collecting supernatant;
s5 enzyme deactivation: heating the supernatant obtained in the centrifugation in the step S4 to 95 ℃, and inactivating the enzyme for 10min to obtain an enzyme inactivation solution;
s6 impurity removal: filtering the enzyme deactivation liquid obtained in the step S5 at constant temperature by adopting a ceramic membrane with the aperture of 10000Dal, and controlling the temperature of the liquid and the environment to be 72 ℃ during filtering to obtain a dilute liquid product; concentrating the dilute feed liquid product by membrane equipment with the aperture of 550Dal until the solid content is more than 20%, and removing microorganisms from the concentrated liquid by a filter element with the aperture of 0.22 mu m to obtain concentrated liquid;
S7 finished product: and (5) carrying out spray drying on the concentrated solution obtained in the step (S6) to obtain a solid, setting the air inlet temperature of a spray dryer to be 170 ℃ and the air outlet temperature to be 75 ℃, and inspecting and packaging the solid product to obtain a finished product.
Example 2
A production process of bovine skin collagen peptide comprises the following steps:
s1 cleaning: cleaning 100 parts of cow leather with clear water, immersing the cleaned cow leather in 1000 parts of water, adding NaOH and Na2CO3Adjusting the pH value to 10, adding 2 parts of stable chlorine dioxide, and soaking for 50 h;
s2 denaturation: cleaning the clean cow leather obtained in the step S1 until the pH value is neutral, then cutting and grinding the clean cow leather to prepare cow leather slurry, adding 4 parts by weight of water into the cow leather slurry, setting the heat preservation temperature in denaturation to be 60-80 ℃, and setting the heat preservation time to be 6h to obtain denatured cow leather protein slurry;
s3 enzymolysis: adding 2 parts of alkaline protease into the denatured cow leather protein slurry obtained in the step S2, keeping the temperature at 65 ℃, keeping the pH at 7.5, reacting for 6 hours, and performing enzymolysis with ultrasonic waves assisted in the enzymolysis process, wherein the power of the ultrasonic waves is 300kW, so as to finally obtain an enzymolysis liquid;
s4 centrifugation: centrifuging the enzymolysis liquid obtained in the step S3, centrifuging by a tubular centrifuge at the rotating speed of 22000r/min, keeping the temperature at 40 ℃ during centrifugation, and taking supernatant;
S5 enzyme deactivation: heating the supernatant obtained in the centrifugation in the step S4 to 95 ℃, and inactivating the enzyme for 10min to obtain an enzyme inactivation solution;
s6 impurity removal: filtering the enzyme deactivation liquid obtained in the step S5 at constant temperature by adopting a ceramic membrane with the aperture of 14000Dal, and controlling the temperature of the liquid and the environment to be 60 ℃ during filtering to obtain a dilute liquid product; concentrating the dilute feed liquid product by membrane equipment with the aperture of 600Dal until the solid content is more than 20%, and removing microorganisms from the concentrated liquid by a filter element with the aperture of 0.22 mu m to obtain concentrated liquid;
s7 finished product: and (5) carrying out spray drying on the concentrated solution obtained in the step (S6) to obtain a solid, setting the air inlet temperature of a spray dryer to be 150 ℃ and the air outlet temperature to be 80 ℃, and inspecting and packaging the solid product to obtain a finished product.
Example 3
A production process of cow hide collagen peptide comprises the following steps:
s1 cleaning: cleaning 100 parts of cowhide by using clear water, soaking the cleaned cowhide in 2000 parts of water, adding NaOH and CaO, adjusting the pH value to 14, adding 0.5 part of sodium hypochlorite, and soaking for 200 hours;
s2 denaturation: cleaning the clean cow leather obtained in the step S1 until the pH value is neutral, then cutting and grinding the clean cow leather to prepare cow leather slurry, adding 6 parts by weight of water into the cow leather slurry, setting the heat preservation temperature in denaturation to be 80 ℃, and setting the heat preservation time to be 4 hours to obtain denatured cow leather protein slurry;
S3 enzymolysis: adding 5 parts of alkaline protease into the denatured cow leather protein slurry obtained in the denaturation step S2, keeping the temperature at 50 ℃, keeping the pH at 9.5, reacting for 5 hours, and carrying out enzymolysis with ultrasonic waves in an auxiliary manner in the enzymolysis process, wherein the power of the ultrasonic waves is 850kW, so as to finally obtain an enzymolysis solution;
s4 centrifugation: centrifuging the enzymolysis liquid obtained in the step S3, performing tubular centrifuge at a rotation speed of 12000r/min, keeping the temperature at 60 ℃ during centrifugation, and taking supernate;
s5 enzyme deactivation: heating the supernatant obtained in the centrifugation in the step S4 to 95 ℃, and inactivating the enzyme for 10min to obtain an enzyme inactivation solution;
s6 impurity removal: filtering the enzyme deactivation liquid obtained in the step S5 at constant temperature by adopting a ceramic membrane with the aperture of 8000Dal, and controlling the temperature of the liquid and the environment to be 80 ℃ during filtering to obtain a dilute liquid product; concentrating the dilute feed liquid product by membrane equipment with the aperture of 500Dal until the solid content is more than 20%, and then removing microorganisms from the concentrated liquid by a filter element with the aperture of 0.22 mu m to obtain concentrated liquid;
s7 finished product: and (5) carrying out spray drying on the concentrated solution obtained in the step (S6) to obtain a solid, setting the air inlet temperature of a spray dryer to be 180 ℃ and the air outlet temperature to be 70 ℃, and inspecting and packaging the solid product to obtain a finished product.
Comparative example 1
Comparative example 1 is different from example 1 in that only the denaturation of S2 is not performed, and the specific scheme is as follows:
s1 cleaning: cleaning 100 parts of cow leather with clear water, soaking the cleaned cow leather in 2000 parts of water, adding NaOH to adjust the pH value to 13, and adding 1.35 parts of H2O2Soaking for 78h, and cleaning the cowhide to be neutral;
s2 enzymolysis: adding 3.75 parts of alkaline protease into the clean cowhide obtained in the step S1, keeping the temperature at 57 ℃, keeping the pH at 8.2, reacting for 5.5h, and performing enzymolysis by using ultrasonic waves with the ultrasonic wave power of 700kW in the enzymolysis process to finally obtain an enzymolysis liquid;
s3 centrifugation: centrifuging the enzymolysis liquid obtained in the step S3, centrifuging at 20000r/min in a tubular centrifuge at 55 deg.C, and collecting supernatant;
s4 enzyme deactivation: heating the supernatant obtained in the centrifugation in the step S4 to 95 ℃, and inactivating the enzyme for 10min to obtain an enzyme inactivation solution;
s5 impurity removal: filtering the enzyme deactivation liquid obtained in the step S5 at constant temperature by adopting a ceramic membrane with the aperture of 10000Dal, and controlling the temperature of the liquid and the environment to be 72 ℃ during filtering to obtain a dilute liquid product; concentrating the dilute feed liquid product by membrane equipment with the aperture of 550Dal until the solid content is more than 20%, and removing microorganisms from the concentrated liquid by a filter element with the aperture of 0.22 mu m to obtain concentrated liquid;
S6 finished product: and (5) carrying out spray drying on the concentrated solution obtained in the step (S6) to obtain a solid, setting the air inlet temperature of a spray dryer to be 170 ℃ and the air outlet temperature to be 75 ℃, and inspecting and packaging the solid product to obtain a finished product.
Comparative example 2
Compared with example 1, the difference between comparative example 1 and example 1 is that no ultrasonic wave is used only in the enzymolysis of S3, and the specific scheme is as follows:
s1 cleaning: cleaning 100 parts of cow leather with clear water, soaking the cleaned cow leather in 1400 parts of water, adding NaOH to adjust the pH value to 13, and adding 1.35 parts of H2O2Soaking for 78 h;
s2 denaturation: cleaning the clean cow leather obtained in the step S1 until the pH value is neutral, then cutting and grinding the clean cow leather to prepare cow leather slurry, adding 5 parts by weight of water into the cow leather slurry, setting the heat preservation temperature in denaturation to 72 ℃ and the heat preservation time to 5.5 hours to obtain denatured cow leather protein slurry;
s3 enzymolysis: to the denatured bovine hide protein slurry obtained in the denaturation in step S2, 3.75 parts of alkaline protease was added, the temperature was maintained at 57 ℃, the pH was maintained at 8.2, and the reaction was carried out for 5.5 hours.
S4 centrifugation: centrifuging the enzymolysis liquid obtained in the step S3, centrifuging at 20000r/min in a tubular centrifuge at 55 deg.C, and collecting supernatant;
S5 enzyme deactivation: heating the supernatant obtained in the centrifugation in the step S4 to 95 ℃, and inactivating the enzyme for 10min to obtain an enzyme inactivation solution;
s6 impurity removal: filtering the enzyme deactivation liquid obtained in the step S5 by adopting a ceramic membrane with the aperture of 10000Dal at constant temperature, and controlling the temperature of the liquid and the environment to be 72 ℃ during filtering to obtain a dilute liquid product; concentrating the dilute feed liquid product by membrane equipment with the aperture of 550Dal until the solid content is more than 20%, and then removing microorganisms from the concentrated liquid by a filter element with the aperture of 0.22 μm to obtain concentrated liquid;
s7 finished product: and (5) carrying out spray drying on the concentrated solution obtained in the step (S6) to obtain a solid, setting the air inlet temperature of a spray dryer to be 170 ℃ and the air outlet temperature to be 75 ℃, and inspecting and packaging the solid product to obtain a finished product.
Comparative example 3
Compared with the example 1, the difference of the comparative example 1 is that the filter element sterilization is not used when the S6 is subjected to impurity removal, and the ultrahigh temperature sterilization with the temperature of 180 ℃ lasting for 30S is adopted, and the specific scheme is as follows:
s1 cleaning: cleaning 100 parts of cow leather with clear water, soaking the cleaned cow leather in 1400 parts of water, adding NaOH to adjust the pH value to 13, and adding 1.35 parts of H2O2Soaking for 78 h;
s2 denaturation: cleaning the clean cow leather obtained in the step S1 until the pH value is neutral, then cutting and grinding the clean cow leather to prepare cow leather slurry, adding 5 parts by weight of water into the cow leather slurry, setting the heat preservation temperature in denaturation to 72 ℃ and the heat preservation time to 5.5 hours to obtain denatured cow leather protein slurry;
S3 enzymolysis: adding 3.75 parts of alkaline protease into the denatured cow leather protein slurry obtained in the denaturation step S2, keeping the temperature at 57 ℃, keeping the pH at 8.2, reacting for 5.5 hours, and carrying out enzymolysis with ultrasonic waves in an auxiliary manner in the enzymolysis process, wherein the power of the ultrasonic waves is 700kW, so as to finally obtain an enzymolysis solution;
s4 centrifugation: centrifuging the enzymolysis liquid obtained in the step S3, centrifuging at 20000r/min in a tubular centrifuge at 55 ℃, and taking supernatant;
s5 enzyme deactivation: heating the supernatant obtained in the centrifugation in the step S4 to 95 ℃, and inactivating the enzyme for 10min to obtain an enzyme inactivation solution;
s6 impurity removal: filtering the enzyme deactivation liquid obtained in the step S5 by adopting a ceramic membrane with the aperture of 10000Dal at constant temperature, and controlling the temperature of the liquid and the environment to be 72 ℃ during filtering to obtain a dilute liquid product; concentrating the dilute feed liquid product by membrane equipment with the aperture of 550Dal until the solid content is more than 20%, and then sterilizing at the ultra-high temperature of 180 ℃ for 30s to obtain concentrated solution;
s7 finished product: and (5) carrying out spray drying on the concentrated solution obtained in the step (S6) to obtain a solid, setting the air inlet temperature of a spray dryer to be 170 ℃ and the air outlet temperature to be 75 ℃, and inspecting and packaging the solid product to obtain a finished product.
After all production is finished, the final finished product is taken, the oxidation resistance, the product molecular weight distribution, the product yield and the product sensory contrast are detected, and the detection method and the result are as follows:
1. oxidation resistance assay
The currently generally used in-vitro antioxidant activity index generally adopts a spectrophotometry method, and a spectrophotometer is used for measuring the change of the content of various color components. The spectrophotometry method is simple and convenient to operate, and does not need special detection equipment.
DPPH shows more stable radical scavenging ability in purple color in methanol or ethanol solution, and has stronger ultraviolet absorption peak at 517 nm. When an antioxidant is present in DPPH.solution, lone pair electrons in DPPH.are paired, the color of the solution changes from purple to yellow, the absorption value at 517nm decreases,
first, 1.5mL of sample was added to 1.5mL of 0.1mmol/L DPPH (95% ethanol), mixed well and incubated at 25 ℃ for 30 min. The absorbance was measured at 517 nm. The Vc solution was used as a control. DPPH-scavenging capacity W (%) was calculated as follows:
Figure GDA0002486179120000081
wherein A0 is the absorbance value for 1.5mL of distilled water and 1.5mL of 95% ethanol containing 0.1mmol/L DPPH, A1 is the absorbance value for 1.5mL of hydrolysate containing 0.1mmol/L DPPH, and A2 is the absorbance value for 1.5mL of hydrolysate and 1.5mL of 95% ethanol.
The detection results are shown in fig. 1:
as shown in the figure, the DPPH removing capacity of the collagen increases greatly along with the increase of the concentration of the bovine collagen, and finally the DPPH removing capacity approaches 80% at 126.4mg/mL, which indicates that the bovine collagen has stronger antioxidant capacity, and examples 1, 2 and 3 all approach to the upper removing limit at about 63 mg/mL.
In contrast, comparative example 1 has no denaturation treatment and has fewer enzyme cutting sites, so that the active components are fewer and the antioxidant capacity is lower; in the comparative example 2, the enzymolysis is not assisted by ultrasonic waves, and the tertiary quaternary structure of the protein still exists, so that enzyme cutting sites are fewer, and the method is similar to the comparative example 1; the use of ultra-high temperature flash sterilization in comparative example 3 resulted in a significant reduction in product activity.
Compared with similar products, the bovine collagen peptide DPPH produced by the process has stronger removing capability, higher activity and better oxidation resistance.
2. Determination of product molecular weight distribution and product yield
The obtained product was sampled and analyzed by chromatography, the analysis method refers to appendix A in GB/T22729-2008, the ratio of the protein hydrolysate with the relative molecular mass less than 1000u (high performance gel filtration chromatography) was obtained, the molecular weight range and the number average molecular weight were obtained, the yield was calculated from the yield and the input, and the final results are shown in Table 1
TABLE 1 Table of the results of the molecular weight and yield measurements
Figure GDA0002486179120000082
Figure GDA0002486179120000091
The method can easily see that the product yield is improved to a certain extent by simultaneously using ultrasonic-assisted enzymolysis and denaturation processes, and the products with ultra-small molecular weight in the blended production process are few, so that the yield is greatly improved; although comparative example 3 only changes the subsequent treatment into ultrahigh-temperature instantaneous sterilization, and the extraction process is not changed too much, the yield is about the same as that of examples 1, 2 and 3, but the yield is always slightly lower than that of other methods according to the application, and comparative example 3 also has a serious browning phenomenon in the actual production, and the results of data in a table and the actual production phenomenon show that the extracting solution contains reducing sugar, comparative example 3 has serious browning after being subjected to ultrahigh-temperature sterilization, and excessive peptides and reducing sugar undergo Maillard reaction.
3. Product sensory comparison
Taking the off-flavor of clear water as 0 and the off-flavor of a 10% solution of commercially available bovine collagen peptide as 10, 10 professional evaluators were found for off-flavor evaluation, and the results are shown in table 2, after removing one highest score and one lowest score, the average value was taken:
TABLE 2 sense organ examination table for products
Example 1 Example 2 Example 3 Comparative example 1 Comparative example 2 Comparative example 3
Evaluator 1 7 7 6 9 9 9
Evaluator 2 6 7 8 10 9 7
Evaluator 3 7 7 7 9 9 8
Evaluator 4 6 7 7 9 9 8
Evaluator 5 6 7 8 9 8 7
Evaluator 6 8 7 7 9 10 8
Evaluator 7 7 7 7 9 10 8
Evaluator 8 6 7 8 9 9 8
Evaluator 9 7 7 7 10 8 8
Evaluator 10 7 7 7 8 9 8
Mean value 6.6 7.0 7.3 9.1 9.0 7.9
It can be seen from the table that the odor is more obvious in comparative example 1 and comparative example 2 without adjusting the production conditions, because the production conditions are not adjusted, the enzyme cutting sites are less, and the small molecular peptides extracted by the prior art scheme are more biased, while the scorched odor is more obvious in comparative example 3, and the product taste is lower. The new process adopts a denaturation process and ultrasonic waves to expose more enzyme cutting sites and simultaneously improve the enzymolysis speed, and the reaction environment inhibits the enzyme activity and reduces excessive cutting, so that the obtained product has proper length distribution, the ultra-small molecular peptides generating odor and peculiar smell are greatly reduced, the uncomfortable taste of the product is reduced, and the product quality is improved.
In conclusion, after the novel method provided by the application is adopted for production, the extracted bovine collagen peptide has high yield, good sense, high oxidation resistance activity, extremely low browning rate and high product quality.
All other embodiments, which can be derived by a person skilled in the art from the embodiments of the present invention without any inventive step, are within the scope of the present invention.

Claims (5)

1. The production process of the cow skin collagen peptide is characterized by comprising the following steps:
s1 cleaning: cleaning 100 parts of cow leather by using clear water, soaking the cleaned cow leather in 2000 parts of water of 1000 times, adding an alkaline regulator to regulate the pH value to be alkaline, adding 0.5-2 parts of a bactericide, and soaking for 48-240 hours;
s2 denaturation: cleaning the clean cow leather obtained in the step S1 to neutral pH, cutting and grinding the cleaned cow leather into cow leather slurry, adding water with the weight part of 4-6 times that of the cow leather slurry into the cow leather slurry, and preserving heat for a certain time to obtain modified cow leather protein slurry;
s3 enzymolysis: adding 2-5 parts by mass of alkaline protease into the denatured cow leather protein slurry obtained in the step S2, keeping a certain temperature and pH, reacting for a period of time, and performing enzymolysis with ultrasonic waves in an auxiliary manner in the enzymolysis process to finally obtain an enzymolysis solution;
s4 centrifugation: centrifuging the enzymolysis liquid obtained in the step S3, and taking supernate;
s5 enzyme deactivation: heating the supernatant obtained in the centrifugation in the step S4 to 95 ℃, and inactivating the enzyme for 10min to obtain an enzyme inactivation solution;
s6 impurity removal: filtering the enzyme deactivation liquid obtained in the step S5 at constant temperature by adopting a ceramic membrane to obtain a product dilute liquid; concentrating the dilute liquid of the product by membrane equipment, and then removing microorganisms from the concentrated liquid by a filter element to obtain concentrated liquid;
S7 finished product: spray drying the concentrated solution obtained in the step S6 to obtain a solid, and inspecting and packaging the solid product to obtain a finished product;
the temperature for heat preservation in denaturation of S2 is set within 60-80 ℃, the time for heat preservation is set within 4-6h, the aperture of the ceramic membrane used in impurity removal in the step S6 is set at 8000-14000Dal, and the temperature of the feed liquid and the environment is controlled to be 60-80 ℃ during filtration; the aperture of the membrane equipment is set at 500-600Dal, and the dilute liquid is concentrated to the solid content of more than 20 percent; the aperture of the filter element is set to be 0.22 mu m;
in the step S3, the reaction temperature is set to be 50-65 ℃, the reaction pH is set to be 7.5-9.5, the reaction time is set to be 5-6h, and the power of the ultrasonic wave is 200-900 kW.
2. The process for producing bovine skin collagen peptide according to claim 1, wherein: the alkaline regulator in the step S1 is NaOH, and the pH value is regulated to 10-14.
3. The process for producing bovine skin collagen peptide according to claim 1, wherein: the bactericide in the step S1 is H2O2One or more of lysozyme, stable chlorine dioxide and sodium hypochlorite.
4. The process for producing bovine skin collagen peptide according to claim 1, wherein: in the step S4, a tubular centrifuge is adopted, the centrifugal speed is more than 10000r/min, and the temperature is kept between 40 and 60 ℃ during centrifugation.
5. The process for producing bovine skin collagen peptide according to claim 1, wherein: the air inlet temperature of the spray drying is set to be 150-180 ℃, and the air outlet temperature is set to be 70-80 ℃.
CN202010250674.2A 2020-04-01 2020-04-01 Production process of cow leather collagen peptide Active CN111334551B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202010250674.2A CN111334551B (en) 2020-04-01 2020-04-01 Production process of cow leather collagen peptide

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202010250674.2A CN111334551B (en) 2020-04-01 2020-04-01 Production process of cow leather collagen peptide

Publications (2)

Publication Number Publication Date
CN111334551A CN111334551A (en) 2020-06-26
CN111334551B true CN111334551B (en) 2022-06-28

Family

ID=71178691

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202010250674.2A Active CN111334551B (en) 2020-04-01 2020-04-01 Production process of cow leather collagen peptide

Country Status (1)

Country Link
CN (1) CN111334551B (en)

Families Citing this family (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114395598A (en) * 2021-12-31 2022-04-26 兆鑫堂(山东)生物科技有限公司 Production method of collagen peptide
CN114271503A (en) * 2022-01-04 2022-04-05 兆鑫堂(山东)生物科技有限公司 A multi-active food containing collagen peptide and its preparation method
CN114304644A (en) * 2022-01-04 2022-04-12 兆鑫堂(山东)生物科技有限公司 Collagen peptide food capable of rapidly supplementing protein required by exercise and preparation method thereof
CN114736288B (en) * 2022-04-26 2023-08-01 河北昊鹿翔生物科技有限公司 Collagen oligopeptide and extraction process thereof

Citations (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1597742A (en) * 2004-08-19 2005-03-23 四川大学 Preparation method of water soluble undenatured natural collagen
CN1948412A (en) * 2006-11-06 2007-04-18 四川大学 Method of extracting undenatured natural collagen from tanning waste leather offcut
CN101186786A (en) * 2006-11-06 2008-05-28 四川大学 Method for extracting unmodified natural collagen from tanning waste leather scrap
WO2013054363A1 (en) * 2011-10-14 2013-04-18 Council Of Scientific And Industrial Research Peptides from fish gelatine
CN107236778A (en) * 2017-08-08 2017-10-10 北京华信佳音医疗科技发展有限责任公司 A kind of extracting method of water-soluble collagen
CN107446981A (en) * 2017-09-27 2017-12-08 湖北瑞邦生物科技有限公司 A kind of grading extraction technique of bovine collagen protein peptides
CN109136316A (en) * 2018-09-11 2019-01-04 余碧芝 Collagen peptidase hydrolyzed method and its application with targeting repair function
CN109295145A (en) * 2018-10-31 2019-02-01 西藏央金生态农牧科技有限公司 A kind of preparation method of Ultra-low molecular weight yak collagen peptide
CN110024901A (en) * 2019-04-30 2019-07-19 集美大学 A kind of collagen peptide and its production method and process units and application

Patent Citations (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1597742A (en) * 2004-08-19 2005-03-23 四川大学 Preparation method of water soluble undenatured natural collagen
CN1948412A (en) * 2006-11-06 2007-04-18 四川大学 Method of extracting undenatured natural collagen from tanning waste leather offcut
CN101186786A (en) * 2006-11-06 2008-05-28 四川大学 Method for extracting unmodified natural collagen from tanning waste leather scrap
WO2013054363A1 (en) * 2011-10-14 2013-04-18 Council Of Scientific And Industrial Research Peptides from fish gelatine
CN107236778A (en) * 2017-08-08 2017-10-10 北京华信佳音医疗科技发展有限责任公司 A kind of extracting method of water-soluble collagen
CN107446981A (en) * 2017-09-27 2017-12-08 湖北瑞邦生物科技有限公司 A kind of grading extraction technique of bovine collagen protein peptides
CN109136316A (en) * 2018-09-11 2019-01-04 余碧芝 Collagen peptidase hydrolyzed method and its application with targeting repair function
CN109295145A (en) * 2018-10-31 2019-02-01 西藏央金生态农牧科技有限公司 A kind of preparation method of Ultra-low molecular weight yak collagen peptide
CN110024901A (en) * 2019-04-30 2019-07-19 集美大学 A kind of collagen peptide and its production method and process units and application

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
超声波辅助法提取牛皮胶原蛋白;瞿怡等;《皮革科学与工程》;20180430;第28卷(第2期);5-9 *
酶法提取牛皮胶原蛋白的研究;李彦春等;《中国皮革》;20021231;第31卷(第23期);6-9 *

Also Published As

Publication number Publication date
CN111334551A (en) 2020-06-26

Similar Documents

Publication Publication Date Title
CN111334551B (en) Production process of cow leather collagen peptide
JP3466189B2 (en) Rice bran stabilization method and rice bran product
US9974321B2 (en) Fish protein oligopeptide with low allergenicity and slight fishiness and industrial preparation method and application thereof
Wallace et al. Studies on the processing and properties of soymilk: II.—Effect of processing conditions on the trypsin inhibitor activity and the digestibility in vitro of proteins in various soymilk preparations
CN101928742B (en) Whey protein active peptide with antioxidant activity and preparation method thereof
CN101948900B (en) Method for extracting hydrolyzed collagen from bovine cartilage
CN106455625A (en) Hypoallergenic, bitterness-reduced soybean oligopeptide, preparation method for same, and applications thereof
EP0153310A4 (en) Soymilk process.
Kwok et al. Heat inactivation of trypsin inhibitors in soymilk at ultra‐high temperatures
CN113826703A (en) Preparation method of oat milk beverage
CN107278208B (en) Chitin, hydrolysate and production of at least one target product from insects by means of enzymatic hydrolysis
CN110734948A (en) extraction device and process for extracting selenium polypeptide from soybeans
CN110973342A (en) Apostichopus japonicus oligopeptide and preparation method and application thereof
CN107723329B (en) Preparation method of immunocompetent peanut peptide
CN109234345A (en) A method of enzymatic hydrolysis pigskin prepares anti-oxidation peptide
CN109777849B (en) Preparation method for extracting proteolysis polypeptide from debitterized peach kernel
CN111087447B (en) Crocodile antioxidant peptide compound and preparation method and application thereof
US7297512B2 (en) Method for producing amino acid components by enzymatic hydrolysis of fish egg skin
JP4227984B2 (en) Composition for improving flavor and taste of food
CN107475331A (en) A kind of method that extrusion processing solid fermentation prepares purple perilla seed active peptide
Wang et al. Study on optimization of extraction process and resistance to oxidation of Polypeptide from sea cucumber waste liquid
CN109497531B (en) Fermented collagen peptide and preparation method thereof
CN110973341A (en) Method for preparing blue clam-rich small molecule peptide
CN111349676A (en) Preparation method of goat milk whey high-F-value oligopeptide
KR100492371B1 (en) Method for manufacturing mild water-soluble collagen

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant
PE01 Entry into force of the registration of the contract for pledge of patent right

Denomination of invention: Production process of a collagen peptide from cowhide

Effective date of registration: 20230323

Granted publication date: 20220628

Pledgee: China Construction Bank Corporation Jingzhou Yuqiao Branch

Pledgor: HUBEI REBORN BIOTECH CO.,LTD.

Registration number: Y2023980035961

PE01 Entry into force of the registration of the contract for pledge of patent right
PC01 Cancellation of the registration of the contract for pledge of patent right

Granted publication date: 20220628

Pledgee: China Construction Bank Corporation Jingzhou Yuqiao Branch

Pledgor: HUBEI REBORN BIOTECH CO.,LTD.

Registration number: Y2023980035961