CN110713557B - Process for extracting chondroitin by using meat and bone meal waste pulp through biological method - Google Patents
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- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/006—Heteroglycans, i.e. polysaccharides having more than one sugar residue in the main chain in either alternating or less regular sequence; Gellans; Succinoglycans; Arabinogalactans; Tragacanth or gum tragacanth or traganth from Astragalus; Gum Karaya from Sterculia urens; Gum Ghatti from Anogeissus latifolia; Derivatives thereof
- C08B37/0063—Glycosaminoglycans or mucopolysaccharides, e.g. keratan sulfate; Derivatives thereof, e.g. fucoidan
- C08B37/0069—Chondroitin-4-sulfate, i.e. chondroitin sulfate A; Dermatan sulfate, i.e. chondroitin sulfate B or beta-heparin; Chondroitin-6-sulfate, i.e. chondroitin sulfate C; Derivatives thereof
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Abstract
The invention discloses a process for extracting chondroitin by utilizing meat and bone meal waste pulp through a biological method, belonging to the technical field of biology. The method takes animal bone slurry as a raw material, integrates a modern biological enzymolysis technology and an efficient separation and extraction technology, and extracts and prepares the food-grade high-purity chondroitin, wherein a complex enzyme system adopted by the biological enzymolysis technology is one or a mixture of more of pancreatin powder, protease, trypsin and neutral protease. The chondroitin prepared by the process has high yield, has various biological activities such as anti-inflammation, anti-cancer, blood fat reduction, anticoagulant activity, arthritis prevention, food flavor improvement, color and moisture retention and the like, can be widely applied to various fields, and has remarkable market prospect.
Description
Technical Field
The invention relates to a process for extracting chondroitin by using meat and bone meal waste pulp through a biological method, in particular to a novel process for preparing a high-extraction-rate and high-purity chondroitin product by using a modern biological enzymolysis technology and an efficient separation and extraction technology, belonging to the technical field of biology.
Background
In recent years, fish meal yield has rapidly decreased due to the growing scarcity of fishery resources and the continuous implementation of capture-limiting policies in various countries. In China, the import quantity of fish meal is reduced year by year, so that the requirement of culture is difficult to meet. Under the background, meat meal rapidly rises in China as a substitute for fish meal, becomes an important component of feed industry, and the yield is rapidly improved. Meanwhile, meat meal processing produces a large amount of waste animal bone paste, and high-end products can be extracted from the animal bone paste: meat meal, high-purity protein, chondroitin; the chondroitin is mucopolysaccharide extracted from animal cartilage, has the functional activities of resisting inflammation, resisting cancer, reducing blood fat, resisting blood coagulation, preventing arthritis, improving food flavor, color and luster, preserving moisture and the like, and has important effects on preventing and treating cardiovascular diseases, arthropathy and the like. However, the recycling degree of the high-concentration waste pulp of the meat and bone meal processing by-product is low, the hidden danger of environmental pollution is large, and the harmless treatment cost is high, so that the high-concentration waste pulp becomes the most key bottleneck factor for restricting the development of the meat and bone meal industry and improving the quality and efficiency. The extraction and utilization of chondroitin from the meat and bone meal processing waste liquid has obvious driving and improving effects on improving the economic benefit of the meat and bone meal processing industry.
Chondroitin is glycosaminoglycan widely existing in animal connective tissues, has various physiological functions of relieving arthritis, reducing blood fat, resisting oxidation, resisting angiogenesis and the like, is widely applied to the fields of food, medicine, cosmetics and the like, and has wide development prospect. At present, the chondroitin raw material is mainly prepared from animal soft organs such as organs of cattle, chickens and ducks, the raw material cost is high, the yield is 12-15%, the cost of the chondroitin is high, and the supply on the market is not sufficient. The chondroitin extracted from the meat and bone meal waste pulp not only can realize high-value reutilization of the waste pulp, but also can promote the application of the chondroitin in the fields of food and medical health care.
At present, the extraction method of chondroitin mainly comprises dilute alkali, dilute alkali salt, dilute alkali enzyme and the like, and the method has the advantages of low yield of products, high content of protein and nitrogen impurities and the like. The chondroitin is extracted by an enzymolysis method, the protease can specifically hydrolyze proteins, the proteins combined with the chondroitin are decomposed into small molecular amino acids, and then the chondroitin is separated from other impurities according to the different solubility of the amino acids and the chondroitin in certain solvents, so that the yield of the chondroitin can be obviously improved. By adopting the complex enzymolysis method for extraction, the pollution of acid and alkali to the environment in the production process of a dilute alkali-enzymolysis extraction method and a concentrated alkali-enzymolysis extraction method can be avoided, and the influence of the acid and alkali on the functional activity of the chondroitin product is also avoided.
Disclosure of Invention
In order to overcome the defects of the prior art and realize the efficient extraction of functional active substances of the meat meal processing waste pulp, the application provides a process for extracting chondroitin from the meat and bone meal waste pulp by a biological method, integrates a modern biological enzymolysis technology and an efficient separation and extraction technology, extracts a high-purity and high-activity chondroitin product from the meat and bone meal waste pulp, meets the application requirements of the fields of health care, medical treatment, food and the like on the chondroitin, has high yield of the chondroitin product prepared by the process, has higher anti-inflammatory, anti-cancer, blood fat reduction, anticoagulant activity, arthritis prevention, food flavor improvement, color and moisture retention type effects, can be widely applied to various fields, particularly in the medical health field, and has obvious health care effect and application value; meanwhile, meat powder and high-purity protein powder products are separated, and the resource recycling rate and the economic benefit of the process are further improved.
The invention realizes the technical effects through the following technical scheme:
a process for extracting chondroitin by utilizing meat and bone meal waste pulp by a biological method comprises the following steps:
(1) preparing raw materials: injecting the animal bone slurry after oil slurry separation into an enzymolysis tank for later use;
(2) compound enzymolysis: adding pancreatin powder with the mass ratio of 1-2%, protease with the mass ratio of 2-3% and pig pancreas powder with the mass ratio of 1-2% into animal bone slurry, and performing enzymolysis for 6-12 hours at the temperature of 40-60 ℃;
(3) and (3) filtering: filtering the enzymolysis liquid after enzymolysis is finished to remove meat meal and grease;
(4) resin adsorption: adsorbing the filtrate by an ion adsorption resin column, washing with 0.5mol/L NaCl solution after the adsorption, and eluting with 1.8mol/L sodium chloride solution at the flow rate of 2L/h to obtain chondroitin stock solution;
(5) adding water for ultrafiltration: adding purified water with the same volume to the chondroitin stock solution, and then performing ultrafiltration to remove insoluble impurities;
(6) and (3) precipitation: adjusting the pH value of the filtrate to 6.8-7.2, adding more than 95% ethanol with the volume of 1.5 times of that of the filtrate, fully stirring, standing for more than 12 hours, and siphoning upper-layer ethanol to obtain chondroitin sulfate;
(7) and (3) purification: dissolving chondroitin in a sodium chloride solution with the concentration of 2-5%, adding a small amount of ethanol, standing at low temperature, centrifuging, adding a small amount of ethanol into supernatant, and repeating the steps to obtain high-purity chondroitin sulfate in sequence;
(8) dewatering and drying: dehydrating the product by using more than 95 percent of acetone, and then drying the product in vacuum to obtain the chondroitin product.
Preferably, the adding amount of the pancreatin powder, the protease and the pig pancreas powder in the composite enzymolysis process is respectively 2 percent, 2.5 percent and 1.3 percent of the mass ratio of the animal bone slurry.
Preferably, the protease can be one or a mixture of more of trypsin, pepsin, papain and flavourzyme.
Most preferably, the protease is a mixture of papain and pepsin in a mass ratio of 3: 2.
Preferably, the meat meal and the grease filtered in the step (3) can be reused for producing meat and bone meal, so that the yield of the meat and bone meal is improved.
Preferably, the solution after the resin adsorption in the step (4) can be subjected to nanofiltration, double-effect evaporation and spray drying to prepare the protein powder.
Preferably, the ultrafiltration membrane used in the step (5) is a hollow fiber type ultrafiltration membrane, a roll type ultrafiltration membrane or a flat plate type ultrafiltration membrane.
The invention provides a process for extracting chondroitin by using meat and bone meal waste pulp by a biological method, the obtained chondroitin is white powder, the ChS content is more than 95%, the nitrogen content is less than 2%, and the chondroitin has high anti-inflammatory, anti-cancer, blood fat reducing, anticoagulant activity, arthritis preventing, food flavor, color and moisture-keeping effects, can be widely applied to various fields, particularly in the medical and health care field, and has obvious health care effect and application value.
The invention provides a process for extracting chondroitin by utilizing meat and bone meal waste pulp by a biological method, which has the following remarkable advantages compared with the prior art:
(1) the invention uses animal bone slurry as raw material, realizes high-value utilization and harmless treatment of animal bone slurry which is waste in meat and bone powder processing by a biological enzymolysis technology, can extract high-purity chondroitin product, and can co-produce meat powder and protein powder;
(2) the method extracts the chondroitin by an enzymolysis method, the protease can hydrolyze the protein specifically, the protein combined with the chondroitin is decomposed into small-molecular amino acid, and then the chondroitin is separated from other impurities according to the difference of the solubility of the amino acid and the solubility of the chondroitin in certain solvents, and the yield and the nutritive value of the chondroitin can be obviously improved by the enzymolysis extraction method; the test result of the invention shows that the composition of the protease system in the compound enzyme preparation has obvious influence on the nutritional characteristics of the chondroitin sulfate;
(3) the method adopts the complex enzymolysis method for extraction, avoids the pollution of acid and alkali environment in the production process, simultaneously avoids the influence of acid and alkali on the functional activity of the chondroitin product, and the chondroitin product refined by the resin is obviously superior to similar products in the market in quality, performance and function and has wide market application value;
(4) the method realizes the industrialization of high-added-value products such as chondroitin and the like, realizes the full utilization of harmless waste resources, provides an effective way for developing extraction resources of chondroitin, realizing high-valued utilization of byproducts and improving the processing degree of meat and bone meal, effectively solves the problem of environmental pollution in meat meal processing, changes waste into valuable, and generates good economic, social and ecological environmental benefits.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments.
Example 1
A process for extracting chondroitin by utilizing meat and bone meal waste pulp by a biological method comprises the following steps:
(1) preparing raw materials: injecting the animal bone slurry after oil slurry separation into an enzymolysis tank for later use;
(2) compound enzymolysis: adding pancreatin powder accounting for 2 percent of the mass ratio of the raw materials, protease accounting for 2.5 percent of the mass ratio of the raw materials and pig pancreas powder accounting for 1.3 percent of the mass ratio of the raw materials into animal bone slurry, and performing enzymolysis for 9 hours at the temperature of 40-60 ℃; the protease is a mixture of papain and pepsin, and the mass ratio of the papain to the pepsin is 3: 2;
(3) and (3) filtering: filtering the enzymolysis liquid after enzymolysis is finished to remove meat meal and grease;
(4) resin adsorption: adsorbing the filtrate by an ion adsorption resin column, washing with 0.5mol/L NaCl solution after the adsorption, and eluting with 1.8mol/L sodium chloride solution at the flow rate of 2L/h to obtain chondroitin stock solution;
(5) adding water for ultrafiltration: adding purified water with the same volume to the chondroitin stock solution, and then performing ultrafiltration to remove insoluble impurities;
(6) and (3) precipitation: adjusting the pH value of the filtrate to 6.8-7.2, adding more than 95% ethanol with the volume of 1.5 times of that of the filtrate, fully stirring, standing for more than 12 hours, and siphoning upper-layer ethanol to obtain chondroitin sulfate;
(7) and (3) purification: dissolving chondroitin in a sodium chloride solution with the concentration of 2-5%, adding a small amount of ethanol, standing at low temperature, centrifuging, adding a small amount of ethanol into supernatant, and repeating the steps to obtain high-purity chondroitin sulfate in sequence;
(8) dewatering and drying: dehydrating the product by using more than 95 percent of acetone, and then drying the product in vacuum to obtain the chondroitin product.
Example 2
A process for extracting chondroitin by utilizing meat and bone meal waste pulp by a biological method comprises the following steps:
(1) preparing raw materials: injecting the animal bone slurry after oil slurry separation into an enzymolysis tank for later use;
(2) compound enzymolysis: adding pancreatin powder accounting for 2 percent of the mass ratio of the raw materials, protease accounting for 2.5 percent of the mass ratio of the raw materials and pig pancreas powder accounting for 1.3 percent of the mass ratio of the raw materials into animal bone slurry, and performing enzymolysis for 9 hours at the temperature of 40-60 ℃; the protease is papain;
(3) and (3) filtering: filtering the enzymolysis liquid after enzymolysis is finished to remove meat meal and grease;
(4) resin adsorption: adsorbing the filtrate by an ion adsorption resin column, washing with 0.5mol/L NaCl solution after the adsorption, and eluting with 1.8mol/L sodium chloride solution at the flow rate of 2L/h to obtain chondroitin stock solution;
(5) adding water for ultrafiltration: adding purified water with the same volume to the chondroitin stock solution, and then performing ultrafiltration to remove insoluble impurities;
(6) and (3) precipitation: adjusting the pH value of the filtrate to 6.8-7.2, adding more than 95% ethanol with the volume of 1.5 times of that of the filtrate, fully stirring, standing for more than 12 hours, and siphoning upper-layer ethanol to obtain chondroitin sulfate;
(7) and (3) purification: dissolving chondroitin in a sodium chloride solution with the concentration of 2-5%, adding a small amount of ethanol, standing at low temperature, centrifuging, adding a small amount of ethanol into supernatant, and repeating the steps to obtain high-purity chondroitin sulfate in sequence;
(8) dewatering and drying: dehydrating the product by using more than 95 percent of acetone, and then drying the product in vacuum to obtain the chondroitin product.
Example 3
A process for extracting chondroitin by utilizing meat and bone meal waste pulp by a biological method comprises the following steps:
(1) preparing raw materials: injecting the animal bone slurry after oil slurry separation into an enzymolysis tank for later use;
(2) compound enzymolysis: adding pancreatin powder accounting for 2 percent of the mass ratio of the raw materials, protease accounting for 2.5 percent of the mass ratio of the raw materials and pig pancreas powder accounting for 1.3 percent of the mass ratio of the raw materials into animal bone slurry, and performing enzymolysis for 9 hours at the temperature of 40-60 ℃; the protease is pepsin;
(3) and (3) filtering: filtering the enzymolysis liquid after enzymolysis is finished to remove meat meal and grease;
(4) resin adsorption: adsorbing the filtrate by an ion adsorption resin column, washing with 0.5mol/L NaCl solution after the adsorption, and eluting with 1.8mol/L sodium chloride solution at the flow rate of 2L/h to obtain chondroitin stock solution;
(5) adding water for ultrafiltration: adding purified water with the same volume to the chondroitin stock solution, and then performing ultrafiltration to remove insoluble impurities;
(6) and (3) precipitation: adjusting the pH value of the filtrate to 6.8-7.2, adding more than 95% ethanol with the volume of 1.5 times of that of the filtrate, fully stirring, standing for more than 12 hours, and siphoning upper-layer ethanol to obtain chondroitin sulfate;
(7) and (3) purification: dissolving chondroitin in a sodium chloride solution with the concentration of 2-5%, adding a small amount of ethanol, standing at low temperature, centrifuging, adding a small amount of ethanol into supernatant, and repeating the steps to obtain high-purity chondroitin sulfate in sequence;
(8) dewatering and drying: dehydrating the product by using more than 95 percent of acetone, and then drying the product in vacuum to obtain the chondroitin product.
Example 4
A process for extracting chondroitin by utilizing meat and bone meal waste pulp by a biological method comprises the following steps:
(1) preparing raw materials: injecting the animal bone slurry after oil slurry separation into an enzymolysis tank for later use;
(2) compound enzymolysis: adding pancreatin powder accounting for 2 percent of the mass ratio of the raw materials, protease accounting for 2.5 percent of the mass ratio of the raw materials and pig pancreas powder accounting for 1.3 percent of the mass ratio of the raw materials into animal bone slurry, and performing enzymolysis for 9 hours at the temperature of 40-60 ℃; the protease is a mixture of papain and pepsin, and the mass ratio of the papain to the pepsin is 2: 3;
(3) and (3) filtering: filtering the enzymolysis liquid after enzymolysis is finished to remove meat meal and grease;
(4) resin adsorption: adsorbing the filtrate by an ion adsorption resin column, washing with 0.5mol/L NaCl solution after the adsorption, and eluting with 1.8mol/L sodium chloride solution at the flow rate of 2L/h to obtain chondroitin stock solution;
(5) adding water for ultrafiltration: adding purified water with the same volume to the chondroitin stock solution, and then performing ultrafiltration to remove insoluble impurities;
(6) and (3) precipitation: adjusting the pH value of the filtrate to 6.8-7.2, adding more than 95% ethanol with the volume of 1.5 times of that of the filtrate, fully stirring, standing for more than 12 hours, and siphoning upper-layer ethanol to obtain chondroitin sulfate;
(7) and (3) purification: dissolving chondroitin in a sodium chloride solution with the concentration of 2-5%, adding a small amount of ethanol, standing at low temperature, centrifuging, adding a small amount of ethanol into supernatant, and repeating the steps to obtain high-purity chondroitin sulfate in sequence;
(8) dewatering and drying: dehydrating the product by using more than 95 percent of acetone, and then drying the product in vacuum to obtain the chondroitin product.
Example 5
A process for extracting chondroitin by utilizing meat and bone meal waste pulp by a biological method comprises the following steps:
(1) preparing raw materials: injecting the animal bone slurry after oil slurry separation into an enzymolysis tank for later use;
(2) compound enzymolysis: adding pancreatin powder accounting for 2 percent of the mass ratio of the raw materials, protease accounting for 2.5 percent of the mass ratio of the raw materials and pig pancreas powder accounting for 1.3 percent of the mass ratio of the raw materials into animal bone slurry, and performing enzymolysis for 9 hours at the temperature of 40-60 ℃; the protease is trypsin;
(3) and (3) filtering: filtering the enzymolysis liquid after enzymolysis is finished to remove meat meal and grease;
(4) resin adsorption: adsorbing the filtrate by an ion adsorption resin column, washing with 0.5mol/L NaCl solution after the adsorption, and eluting with 1.8mol/L sodium chloride solution at the flow rate of 2L/h to obtain chondroitin stock solution;
(5) adding water for ultrafiltration: adding purified water with the same volume to the chondroitin stock solution, and then performing ultrafiltration to remove insoluble impurities;
(6) and (3) precipitation: adjusting the pH value of the filtrate to 6.8-7.2, adding more than 95% ethanol with the volume of 1.5 times of that of the filtrate, fully stirring, standing for more than 12 hours, and siphoning upper-layer ethanol to obtain chondroitin sulfate;
(7) and (3) purification: dissolving chondroitin in a sodium chloride solution with the concentration of 2-5%, adding a small amount of ethanol, standing at low temperature, centrifuging, adding a small amount of ethanol into supernatant, and repeating the steps to obtain high-purity chondroitin sulfate in sequence;
(8) dewatering and drying: dehydrating the product by using more than 95 percent of acetone, and then drying the product in vacuum to obtain the chondroitin product.
Example 6
A process for extracting chondroitin by utilizing meat and bone meal waste pulp by a biological method comprises the following steps:
(1) preparing raw materials: injecting the animal bone slurry after oil slurry separation into an enzymolysis tank for later use;
(2) compound enzymolysis: adding pancreatin powder accounting for 2 percent of the mass ratio of the raw materials, protease accounting for 2.5 percent of the mass ratio of the raw materials and pig pancreas powder accounting for 1.3 percent of the mass ratio of the raw materials into animal bone slurry, and performing enzymolysis for 9 hours at the temperature of 40-60 ℃; the protease is flavourzyme;
(3) and (3) filtering: filtering the enzymolysis liquid after enzymolysis is finished to remove meat meal and grease;
(4) resin adsorption: adsorbing the filtrate by an ion adsorption resin column, washing with 0.5mol/L NaCl solution after the adsorption, and eluting with 1.8mol/L sodium chloride solution at the flow rate of 2L/h to obtain chondroitin stock solution;
(5) adding water for ultrafiltration: adding purified water with the same volume to the chondroitin stock solution, and then performing ultrafiltration to remove insoluble impurities;
(6) and (3) precipitation: adjusting the pH value of the filtrate to 6.8-7.2, adding more than 95% ethanol with the volume of 1.5 times of that of the filtrate, fully stirring, standing for more than 12 hours, and siphoning upper-layer ethanol to obtain chondroitin sulfate;
(7) and (3) purification: dissolving chondroitin in a sodium chloride solution with the concentration of 2-5%, adding a small amount of ethanol, standing at low temperature, centrifuging, adding a small amount of ethanol into supernatant, and repeating the steps to obtain high-purity chondroitin sulfate in sequence;
(8) dewatering and drying: dehydrating the product by using more than 95 percent of acetone, and then drying the product in vacuum to obtain the chondroitin product.
Example 7
1. Index measuring method
The extraction rate of the chondroitin sulfate is measured according to the method of Xiefrui phoenix 'research on the separation and purification of shark chondroitin sulfate and the antioxidant function thereof'; the chondroitin pair S in the present application was determined according to the test method described in Zhangxiu Juan et al, influence of shark cartilage powder on immune cell modulating function of S180 tumor-bearing mice180Influence of splenic lymphocyte proliferation function and phagocytic activity of phagocytic cells of tumor-bearing mice.
2. Test method and grouping
The mice were randomly divided into 8 groups, and the chondroitin in the experimental group was administered 100 mg-kg by intragastric administration-1The negative control group was given the same volume of physiological saline, 10 of each group. Well-growing S from day 7 of inoculation was aseptically aspirated180Ascites (viable cell count) in tumor-bearing mice>97%) of normal saline, diluted 1: 3, and inoculated subcutaneously in the right anterior axilla of mice at a rate of 0.2 mL/mouse. Administering after 24 h of inoculation, 1 time/d, continuously administering for 10d, stopping administration, and collecting blood by eyeball on the next day;
the grouping method comprises the following steps: examples 1-6 groups are the same as example 1 except that chondroitin described in examples 1-6 is separately subjected to intragastric administration, and chondroitin without protease is added in the enzymolysis process of the positive control group by intragastric administration; perfusing distilled water with the same volume as the perfused blank control group; the test methods are the same;
3. test results
TABLE 1 Effect of different Process conditions chondroitin extraction yield and Immunity enhancing anti-tumor Capacity
| Test group | Extraction ratio (%) | A570Value of | A490Value of |
| Blank control group | — | 0.2257 | 0.5649 |
| Example 1 test group | 10.21 | 0.5846 | 1.1879 |
| Example 2 test group | 9.27 | 0.4893 | 0.9056 |
| Example 3 test group | 8.74 | 0.4587 | 0.9452 |
| Example 4 test group | 8.29 | 0.4623 | 0.9731 |
| Example 5 test group | 6.29 | 0.4893 | 0.9905 |
| Example 6 test group | 8.51 | 0.4029 | 0.9121 |
| Positive control group | 6.32 | 0.4127 | 0.7893 |
The experimental results show that the embodiments of the invention have good chondroitin extraction rate, and the obtained chondroitin has good anti-tumor immunity enhancing activity and high nutritive value, and can be widely applied to the fields of food, medical treatment, health care and the like. In addition, the experimental result also shows that the composition of the protease in the composite enzymolysis process has obvious influence on the extraction rate and the immune enhancement activity of the product, wherein the protease comprises the following components: the effect is best when the pepsin adding ratio is 3:2, and the embodiment 1 is the best embodiment of the invention.
The above embodiments are only used for illustrating the technical solution of the present invention, and not for limiting the same; although the invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that various changes may be made and equivalents may be substituted for elements thereof; such modifications and substitutions do not depart from the spirit and scope of the present invention as set forth in the appended claims.
Claims (5)
1. A process for extracting chondroitin by utilizing meat and bone meal waste pulp by a biological method is characterized by comprising the following steps:
(1) preparing raw materials: injecting the animal bone slurry after oil slurry separation into an enzymolysis tank for later use;
(2) compound enzymolysis: adding pancreatin powder with the mass ratio of 1-2%, protease with the mass ratio of 2-3% and pig pancreas powder with the mass ratio of 1-2% into animal bone slurry, and performing enzymolysis for 6-12 hours at the temperature of 40-60 ℃; the protease is a mixture of papain and pepsin, and the mass ratio of the papain to the pepsin is 3: 2;
(3) and (3) filtering: filtering the enzymolysis liquid after enzymolysis is finished to remove meat meal and grease;
(4) resin adsorption: adsorbing the filtrate by an ion adsorption resin column, washing with 0.5mol/L NaCl solution after the adsorption, and eluting with 1.8mol/L sodium chloride solution at the flow rate of 2L/h to obtain chondroitin stock solution;
(5) adding water for ultrafiltration: adding purified water with the same volume to the chondroitin stock solution, and then performing ultrafiltration to remove insoluble impurities;
(6) and (3) precipitation: adjusting the pH value of the filtrate to 6.8-7.2, adding more than 95% ethanol with the volume of 1.5 times of that of the filtrate, fully stirring, standing for more than 12 hours, and siphoning upper-layer ethanol to obtain chondroitin sulfate;
(7) and (3) purification: dissolving chondroitin in a sodium chloride solution with the concentration of 2-5%, adding a small amount of ethanol, standing at low temperature, centrifuging, adding a small amount of ethanol into supernatant, and repeating the steps to obtain high-purity chondroitin sulfate in sequence;
(8) dewatering and drying: dehydrating the product by using more than 95 percent of acetone, and then drying the product in vacuum to obtain the chondroitin product.
2. The process for extracting chondroitin from meat and bone meal waste pulp by using a biological method as claimed in claim 1, wherein the addition amounts of pancreatin powder, protease and porcine pancreas powder in the composite enzymolysis process are respectively 2%, 2.5% and 1.3% of the mass ratio of the animal bone pulp.
3. The process for extracting chondroitin from meat and bone meal waste pulp by using a biological method as claimed in claim 1, wherein the meat meal and the grease filtered in the step (3) can be reused for meat and bone meal production, so as to improve the yield of the meat and bone meal.
4. The process for extracting chondroitin from meat and bone meal waste pulp by using a biological method as claimed in claim 1, wherein the solution after resin adsorption in the step (4) can be subjected to nanofiltration, double effect evaporation and spray drying to prepare protein powder.
5. The process for extracting chondroitin from meat and bone meal waste pulp by using a biological method according to claim 1, wherein the ultrafiltration membrane used in the step (5) is a hollow fiber type ultrafiltration membrane, a roll type ultrafiltration membrane or a flat plate type ultrafiltration membrane.
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Citations (4)
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