CN104830937A - Method for preparing peptone by using chicken slaughtering by-products - Google Patents

Abstract

The invention discloses a method for preparing peptone by using chicken slaughter by-products. The preparation method comprises the following steps: first step, raw material pretreatment; second step, protease hydrolysis; third step, enzyme elimination, decolorization, and deodorization; second step Four steps, neutralization, filtration; fifth step, concentration, drying, pulverization, the beneficial effect of the present invention: the bone oil produced in the processing can be used to make oleic acid, stearic acid, soap, glycerin, high-grade lubricating oil and daily Chemical products, bone pellets and bone meal are mainly used as main raw materials such as feed and compound fertilizer. Due to the large number of applications in the fields of fermentation and edible fungus cultivation in biopharmaceuticals, the development of the feed industry and the demand for deep processing and finishing of chemical products, the demand for peptone produced by animal by-products will increase year by year, and the market application prospect is broad.

Description

A method for preparing peptone by utilizing chicken slaughter by-products

technical field

The invention relates to a method for preparing peptone, in particular to a method for preparing peptone by using chicken slaughter by-products.

Background technique

Peptone is a water-soluble mixture obtained by hydrolyzing protein with enzymes, acids, and alkalis. It is in a light yellow powder state and is the main component of microbial culture media. Because peptone contains high total nitrogen content and amino acid content, it provides a rich nitrogen source for the growth of microorganisms. It is a biological reagent widely used in fermentation industry, antibiotic industry, pharmaceutical industry, biochemical industry, etc. Peptone is often a complex mixture of polypeptides, soluble in water, does not precipitate in saturated ammonium sulfate but can be precipitated by protein precipitants. There are many existing literatures on the preparation methods of peptone, but these methods generally have the characteristics of poor product quality, long production cycle, and low yield. In particular, a single raw material is usually used for production and processing, and its nutritional components are limited. The range is narrow, and the technical performance indicators such as product clarity and acid-base stability are unstable, which has a negative impact on the application of the product.

With the implementation of "Animal Epidemic Prevention Law", "Regulations on the Management of Pig Slaughter" and other regulations, the treatment of a large amount of manure, sewage, harmful gas, stench, stomach contents and diseased meat produced by factory slaughtering of livestock and poultry has been significantly improved. The effective utilization of the by-products of livestock and poultry processing still needs to be developed urgently, and the fresh bones and organs of livestock and poultry are rich in protein and amino acids, which become good raw materials for the preparation of peptone, which can not only improve the utilization of by-products after animal slaughter, Saving resources and reducing production costs are of great significance to environmental protection and new product development.

Soybean meal is a by-product of soybean oil extraction. As a high-protein protein, soybean meal is the main raw material for making livestock and poultry feed, and can also be used as a raw material for cosmetics and antibiotics. A variety of amino acids contained in soybean meal are suitable for the nutritional requirements of microbial culture medium.

Contents of the invention

The purpose of the present invention is to provide a method for preparing peptone by utilizing the by-products of chicken slaughter and the comprehensive utilization of soybean meal by-products after chicken slaughter.

The method for preparing peptone provided by the invention by utilizing chicken slaughter by-products, its preparation method is as follows:

The first step, pretreatment of raw materials:

Chicken bone is preferred by weight: chicken liver: chicken heart: chicken gizzard = 20:2:2:2, the raw materials are crushed, wherein the chicken bone should be broken to 5-10cm, cleaned with running water, mixed according to the above ratio, added and mixed Put water 2-2.5 times the total weight of raw materials into a high-pressure cooking pot for cooking. The cooking temperature is 150-200°C and the cooking pressure is 0.1-0.2MPa. Cook for 4-6 hours. After cooking, reduce the pressure in the pot Side by side soup to remove foam and slick oil, separate oil and water from the liquid part, keep the bone broth, and use the remaining residue for other purposes;

The second step, protease hydrolysis:

Mix the bone broth and soybean milk prepared in the first step at a weight ratio of 2:1, control the temperature of the soup at 40-50°C, adjust the pH of the soup to 6.0-8.0 with a pH regulator, add the total soup Hydrolyze 2-4% trypsin, adjust the pH to about 7.5 after reacting for 3-5 hours, raise the temperature of the soup to 60-70°C, add compound protease with a total weight of 0.6-1.2% of the soup for the second step of hydrolysis, React for 2-3h, and use a 60-80rpm low-speed stirrer to stir during the two hydrolysis processes;

The third step, inactivating enzymes, decolorizing, and deodorizing:

After the hydrolysis is completed, keep the temperature of the soup, add animal hair water with 0.5-0.8% of the total weight of the soup, use steam to increase the temperature of the soup to 90-95°C for 10-20min to inactivate the enzyme, and then add it after the temperature drops to 55-60°C The total weight of the soup is 0.2% activated carbon or activated kaolin for decolorization and deodorization;

The fourth step, neutralization and filtration:

Use the pH adjusting solution to adjust the pH of the hydrolyzed soup to 6.0-7.5, filter it with a plate and frame filter, separate the soup and impurity precipitates, and then use a microporous membrane with a pore size of 0.6-0.8 μm for fine filtration treatment until The soup is transparent and free of sediment;

The fifth step, concentration, drying, crushing:

Put the finely filtered filtrate obtained in the fourth step at 60-70°C for 3 hours, then put it into a stainless steel concentration pot for low-temperature continuous concentration, the temperature is ≤70°C, and concentrate it to a water content of 40-50%. Vacuum freeze-drying, the drying temperature is 70-90°C, the vacuum degree is 0.08-0.12MPa, and after drying to a water content of ≤2%, it is pulverized by a universal pulverizer and passed through a 100-140 mesh sieve to obtain a finished peptone product.

Preparation of trypsin: mince fresh, oil-free pure bovine pancreas with a meat grinder, add trichloroacetic acid with a mass fraction of 3%, extract with a stirrer for 24 hours, and prepare the extract by salting out and ultrafiltration Enzyme activity reaches 100,000-130,000 single trypsin.

The composite protease includes papain, flavor protease and neutral protease, and the ratio by weight is papain:flavor protease:neutral protease=1:1:1.

Preparation of the animal hair water: add 5 parts of pig hair and 3 parts of chicken feather to 50 parts of industrial hydrochloric acid with a concentration of 33-35% by weight, and store for 20 days.

Preparation of soybean milk: After cleaning the soybean meal, add water 3-5 times the weight of the soybean meal and soak for 5-7 hours, beat with a beater, and boil to obtain soybean milk.

The pH adjusting solution is hydrochloric acid or sodium hydroxide solution.

The filter cloth of the plate and frame filter is made of 4×4-8×8 pure cotton canvas, and the filter cloth is degreased with 0.2%-1.0% dilute alkali solution and rinsed before use.

Beneficial effects of the present invention:

The present invention uses protein-containing animal tissues as raw materials to produce peptone, enzymatically hydrolyzes with fresh trypsin to produce a variety of amino acids, and then supplements tryptophan, cystine and leucine that are lacking in proteins such as meat and bone with self-made animal hair water Amino acids such as acid and other amino acids are combined with the inherent amino acids in animal parts and tissues to form peptone, a highly nutritious medium product with more than 20 amino acids and multiple vitamins. Using trypsin and compound protease (papain, flavor protease, neutral protease) in the process of enzymolysis can improve the efficiency of enzymolysis and enhance the effect of enzymolysis. Timely adjustment of the pH ensures an acid-base environment, and the enzymolysis is carried out at an appropriate temperature and stirring speed. The enzymolysis time and degree of enzymolysis are moderate, which is conducive to improving the quality of peptone. The process route is reasonable, the cost of raw materials is low, the product quality is good, soluble in water, and has high nutritional value. It can be widely used in modern biomedical research, various bacterial detection, genetic engineering, bioengineering fermentation, edible fungus cultivation and other fields.

The invention uses chicken slaughter by-products and soybean by-products as raw materials, turns waste into treasure, improves the utilization rate of raw materials, reduces environmental pollution, and effectively solves the contradiction between resource utilization and environmental pollution. It provides new technical methods, improves the added value of the animal slaughter industry, and drives the development of animal husbandry and other related industries. The peptone product of the invention is mainly used in the fields of modern biomedical research, detection of various bacteria, genetic engineering, fermentation of bioengineering, cultivation of edible fungi and the like. The bone oil produced during the processing can be used to make oleic acid, stearic acid, soap, glycerin, high-grade lubricating oil and daily chemical products. The bone grains and bone meal are mainly used as main raw materials such as feed and compound fertilizer. Due to the large number of applications in the fields of fermentation and edible fungus cultivation in biopharmaceuticals, the development of the feed industry and the demand for deep processing and finishing of chemical products, the demand for peptone produced by animal by-products will increase year by year, and the market application prospect is broad.

Description of drawings

Fig. 1 is the flow chart of the preparation process of the method of the present invention.

Detailed ways

Embodiment one:

The method for preparing peptone provided by the invention by utilizing chicken slaughter by-products, its preparation method is as follows:

The first step, pretreatment of raw materials:

Weigh 20g of chicken bone, 2g of chicken liver, 2g of chicken heart, and 2g of chicken gizzard, and crush the raw materials. The fresh bone should be crushed to 5-10cm. Put it into a high-pressure cooking pot for cooking, and cook for 5 hours under the conditions of 0.1MPa and 150°C. After cooking, the pressure in the pot is reduced and the soup is drained to remove foam and slick oil, and the liquid part is separated from oil and water to keep the bone broth. The remaining residue can be used as bone meal for additional processing and utilization.

The second step, protease hydrolysis:

Mix the bone broth and soybean milk prepared in the first step at a mass ratio of 2:1, control the temperature of the soup at 50°C, adjust the pH of the soup to 7.0 with a pH regulator, and add 3.0% of the total weight of the soup Trypsin hydrolysis, adjust the pH to 7.5 after reacting for 3.5 hours, raise the temperature of the soup to 70°C, add a compound protease with a total weight of 1.2% of the soup (papain: flavor protease: neutral protease = 1:1:1) The second step of hydrolysis, reaction 2h. A low-speed stirrer at 60-80 rpm was used to stir during the two hydrolysis processes.

The third step, inactivating enzymes, decolorizing, and deodorizing:

After the hydrolysis is complete, keep the temperature of the soup, add animal hair water with 0.6% of the total weight of the soup, use steam to raise the temperature of the soup to 90°C for 20 minutes to kill the enzyme, and add 0.2% of the total weight of the soup with activated carbon after the temperature drops to 55°C Decolorization and deodorization.

The fourth step, neutralization and filtration:

Adjust the hydrolyzed soup to 7.0 with a pH adjusting solution, filter it with a plate and frame filter, separate the soup and impurities, and then use a microporous membrane with a pore size of 0.6-0.8 μm for fine filtration until the soup is Transparent state, and no sediment.

The fifth step, concentration, drying, crushing:

After fine filtration, the filtrate was left to stand at 70°C for 3 hours, then placed in a stainless steel concentrator for low-temperature continuous concentration at a temperature of ≤70°C, concentrated to a water content of 40-50%, and vacuum-dried at a drying temperature of 80°C. The degree of vacuum is 0.10 MPa, and after drying to a water content of ≤2%, it is pulverized by a universal pulverizer and sieved with 100 meshes to obtain the finished peptone.

According to the above process steps, the product peptone of the present invention can be obtained with a yield of 91%.

Embodiment two:

The method for preparing peptone provided by the invention by utilizing chicken slaughter by-products, its preparation method is as follows:

The first step, pretreatment of raw materials:

Weigh 18g of chicken bone, 2g of chicken liver, 2g of chicken heart, and 2g of chicken gizzard, and crush the raw materials. The fresh bone should be crushed to 5-10cm. Put it into a high-pressure cooking pot for cooking, and cook for 6 hours under the conditions of 0.1MPa and 180°C. After cooking, the pressure in the pot is reduced and the soup is drained to remove foam and slick oil, and the liquid part is separated from oil and water to keep the bone broth. The remaining residue can be used as bone meal for additional processing and utilization.

The second step, protease hydrolysis:

Mix the bone broth and soybean milk prepared in the first step at a mass ratio of 2:1, control the temperature of the soup at 50°C, adjust the pH of the soup to 7.5 with a pH regulator, and add 4.0% of the total weight of the soup Trypsin hydrolysis, adjust the pH to 7.5 after reacting for 3 hours, raise the temperature of the soup to 60°C, add 1.0% of the total weight of the soup (papain: flavor protease: neutral protease = 1:1:1) for the second Two-step hydrolysis, reaction 3.5h. A low-speed stirrer at 60-80 rpm was used to stir during the two hydrolysis processes.

The third step, inactivating enzymes, decolorizing, and deodorizing:

After the hydrolysis is complete, keep the temperature of the soup, add animal hair water with 0.7% of the total weight of the soup, use steam to increase the temperature of the soup to 92°C for 15 minutes to kill the enzyme, and add 0.2% of the total weight of the soup with activated carbon after the temperature drops to 65°C Decolorization and deodorization.

The fourth step, neutralization and filtration:

Adjust the hydrolyzed soup to 7.5 with a pH regulator, filter it with a plate-and-frame filter to separate the soup and impurities, and then use a microporous membrane with a pore size of 0.6-0.8 μm for fine filtration until the soup is Transparent state, and no sediment.

The fifth step, concentration, drying, crushing:

After fine filtration, the filtrate was left to stand at 60°C for 3 hours, then placed in a stainless steel concentrator for low-temperature continuous concentration at a temperature of ≤70°C, concentrated to a water content of 40-50%, and vacuum-dried at a drying temperature of 75°C. The degree of vacuum is 0.08MPa, after drying to a water content of ≤2%, it is pulverized by a universal pulverizer, and sieved with 140 meshes to obtain the finished peptone.

According to the above process steps, the product peptone of the present invention can be obtained with a yield of 93%.

Embodiment three:

The method for preparing peptone provided by the invention by utilizing chicken slaughter by-products, its preparation method is as follows:

The first step, pretreatment of raw materials:

Weigh 18g of chicken bone, 4g of chicken liver, 4g of chicken heart, and 4g of chicken gizzard, and crush the raw materials. The fresh bone should be crushed to 5-10cm. Put it into a high-pressure cooking pot for cooking, and cook for 5 hours under the conditions of 0.2MPa and 180°C. After cooking, the pressure in the pot is reduced and the soup is drained to remove foam and slick oil, and the liquid part is separated from oil and water to keep the bone broth. The remaining residue can be used as bone meal for additional processing and utilization.

The second step, protease hydrolysis:

Mix the bone broth and soybean milk prepared in the first step at a mass ratio of 2:1, control the temperature of the soup at 60°C, adjust the pH of the soup to 7.0 with a pH regulator, and add 3.0% of the total weight of the soup Trypsin hydrolysis, adjust the pH to 7.5 after reacting for 4 hours, raise the temperature of the soup to 70°C, add 0.8% of the total weight of the soup (papain: flavor protease: neutral protease = 1:1:1) for the second Two-step hydrolysis, reaction 5h. A low-speed stirrer at 60-80 rpm was used to stir during the two hydrolysis processes.

The third step, inactivating enzymes, decolorizing, and deodorizing:

After the hydrolysis is completed, keep the temperature of the soup, add 0.8% of the total weight of the soup to hydrolyze the animal hair water, use steam to raise the temperature of the soup to 92°C for 18 minutes to inactivate the enzyme, and add 0.2% of the total weight of the soup after the temperature drops to 60°C Kaolin for decolorization and deodorization.

The fourth step, neutralization and filtration:

Adjust the hydrolyzed soup to 7.5 with a pH regulator, filter it with a plate-and-frame filter to separate the soup and impurities, and then use a microporous membrane with a pore size of 0.6-0.8 μm for fine filtration until the soup is Transparent state, and no sediment.

The fifth step, concentration, drying, crushing:

After fine filtration, the filtrate was left to stand at 70°C for 3 hours, then placed in a stainless steel concentrator for low-temperature continuous concentration at a temperature of ≤70°C, concentrated to a water content of 40-50%, and vacuum-dried at a drying temperature of 85°C. The degree of vacuum is 0.08MPa, and after drying to a water content of ≤2%, it is pulverized by a universal pulverizer and sieved with 100 meshes to obtain the finished peptone.

According to the above process steps, the product peptone of the present invention can be obtained with a yield of 90%.

The physical and chemical indicators of the product prepared by the above method are as follows:

See the table below for the percentage content of amino acids in this product:

The prepared peptone was prepared into nutrient agar plate for microbial culture test, and the selected strains were porcine pathogenic Escherichia coli, Bacillus subtilis, Staphylococcus aureus and Salmonella typhi. The test shows that the growth conditions of the four kinds of bacteria and the shape of the bacteria are good, indicating that the product of the present invention can be used as a culture substrate for microorganisms such as bacteria.

Claims (7)

1. a method utilizing chicken slaughter by-products to prepare peptone is characterized in that: its preparation method is as follows: The first step, pretreatment of raw materials: Chicken bone is preferred by weight: chicken liver: chicken heart: chicken gizzard = 20:2:2:2, the raw materials are crushed, wherein the chicken bone should be broken to 5-10cm, cleaned with running water, mixed according to the above ratio, added and mixed Put water 2-2.5 times the total weight of raw materials into a high-pressure cooking pot for cooking. The cooking temperature is 150-200°C and the cooking pressure is 0.1-0.2MPa. Cook for 4-6 hours. After cooking, reduce the pressure in the pot Side by side soup to remove foam and slick oil, separate oil and water from the liquid part, keep the bone broth, and use the remaining residue for other purposes; The second step, protease hydrolysis: Mix the bone broth and soybean milk prepared in the first step at a weight ratio of 2:1, control the temperature of the soup at 40-50°C, adjust the pH of the soup to 6.0-8.0 with a pH regulator, add the total soup Hydrolyze 2-4% trypsin, adjust the pH to about 7.5 after reacting for 3-5 hours, raise the temperature of the soup to 60-70°C, add compound protease with a total weight of 0.6-1.2% of the soup for the second step of hydrolysis, React for 2-3h, and use a 60-80rpm low-speed stirrer to stir during the two hydrolysis processes; The third step, inactivating enzymes, decolorizing, and deodorizing: After the hydrolysis is completed, keep the temperature of the soup, add animal hair water with 0.5-0.8% of the total weight of the soup, use steam to increase the temperature of the soup to 90-95°C for 10-20min to inactivate the enzyme, and then add it after the temperature drops to 55-60°C The total weight of the soup is 0.2% activated carbon or activated kaolin for decolorization and deodorization; The fourth step, neutralization and filtration: Use the pH adjusting solution to adjust the pH of the hydrolyzed soup to 6.0-7.5, filter it with a plate and frame filter, separate the soup and impurity precipitates, and then use a microporous membrane with a pore size of 0.6-0.8 μm for fine filtration treatment until The soup is transparent and free of sediment; The fifth step, concentration, drying, crushing: Put the finely filtered filtrate obtained in the fourth step at 60-70°C for 3 hours, then put it into a stainless steel concentration pot for low-temperature continuous concentration, the temperature is ≤70°C, and concentrate it to a water content of 40-50%. Vacuum freeze-drying, the drying temperature is 70-90°C, the vacuum degree is 0.08-0.12MPa, and after drying to a water content of ≤2%, it is pulverized by a universal pulverizer and passed through a 100-140 mesh sieve to obtain a finished peptone product. 2. a kind of method utilizing chicken slaughter by-products to prepare peptone according to claim 1 is characterized in that: the preparation of described trypsin: fresh, degreased pure cow pancreas is minced with a meat grinder, Adding trichloroacetic acid with a mass fraction of 3%, stirring and extracting with a stirrer for 24 hours, salting out and ultrafiltration of the extract to prepare trypsin with an enzyme activity of 100,000-130,000 units. 3. a kind of method utilizing chicken slaughter by-products to prepare peptone according to claim 1 is characterized in that: described composite protease comprises papain, flavor protease and neutral protease, is papain by weight ratio: Flavor protease: neutral protease = 1:1:1. 4. a kind of method utilizing chicken slaughter by-products to prepare peptone according to claim 1 is characterized in that: the preparation of described animal hair water: add in 50 parts by weight concentration to be 33-35% industrial hydrochloric acid 5 parts of pig feathers and 3 parts of chicken feathers can be stored for 20 days. 5. A kind of method utilizing chicken slaughter by-products to prepare peptone according to claim 1, is characterized in that: the preparation of described soya-bean milk: after soya-bean meal is cleaned up, add the clear water of 3-5 times of soya-bean meal weight to soak 5- 7h, beat with a beater, and boil to obtain soybean milk. 6. A method for preparing peptone by utilizing chicken slaughter by-products according to claim 1, characterized in that: said pH adjusting solution is hydrochloric acid or sodium hydroxide solution. 7. a kind of method utilizing chicken slaughter by-products to prepare peptone according to claim 1 is characterized in that: the filter cloth of described plate-and-frame filter selects the pure cotton canvas of 4 * 4-8 * 8 for use, filter cloth Use after degreasing with 0.2%-1.0% dilute alkaline solution and rinsing.