CN1297691A - Separation and extraction process of effective component in resource insect - Google Patents

Abstract

The present invention relates to especially the separation and extraction process of effective component in housefly maggot. Through shell/liquid separation, enzymolysis, oil/water separation, chemical extraction and other processes, protein hydrolysate, maggot oil and chitin are produced separately from housefly maggot. The present invention utilizes the housefly maggot resource fully without "three wastes" discharge by means of a reasonable and simple production process.

Description

Separation and extraction process of effective components of resource insects

The invention relates to a comprehensive processing technology for resource insects, in particular to a technology for separating and extracting effective components from housefly larvae (maggots).

The housefly is the most common and most abundant insect in most parts of my country. As a breeding development object, it has great production advantages: its growth cycle is short, its reproductive capacity is strong, and one generation can be completed every 14 to 18 days under suitable temperature conditions; each pair of houseflies produces nearly a thousand eggs. At present, the housefly larvae (fly maggots) can be mass-produced on a large scale and mechanized in China. At the same time, the nutritional advantages of houseflies are also relatively large. The dry matter of the larvae contains 59-65% of crude protein and 2.6-12% of fat. The protein contains various amino acids needed by animals. It is 43.83%, exceeding the reference value proposed by the World Food and Agriculture/Health Organization (FAO/WHO) by 40%. The reference value is 0.6.

At present, most of the development and utilization of fly maggots at home and abroad focus on the development of fly maggots as feed raw materials to replace fish meal. Although the product market is large, the added value of utilization is low. The invention patent with the patent publication number of CN1141130 uses fly maggots to develop and produce nutritionally active dry powder for human consumption, which provides a high-level application approach for the development and utilization of fly maggot resources. However, this invention only simply processes the fly maggots to make dry powder. This dry powder has no qualitative difference from fly maggots, and is not a real protein powder. It is difficult to popularize and apply it as food or food additives. . Furthermore, this invention does not redevelop and utilize the centrifuged residue or grease, which greatly wastes resources.

Animal hydrolyzed protein is a new type of food additive, mainly used for the production of high-grade condiments and nutritional fortification of food, and as the base material of functional food. Animal hydrolyzed protein is an unlimited food additive, which can be determined according to the desired flavor, amino acid content and ratio. In recent years, in developed countries, a large amount of animal hydrolyzed protein has been used in high-end condiments and soups. At present, the raw materials for producing animal hydrolyzed protein are mainly minced meat of livestock, poultry and aquatic animals, and some leftovers (such as blood, etc.) of livestock, poultry and aquatic animals are also used. The report of using insects as raw materials to produce hydrolyzed protein is only found in the use of silkworm chrysalis or Tenebrio molitor, and there is no report on using housefly or other insects as raw materials to produce hydrolyzed protein.

Among the fatty acids contained in fly maggot oil, the content of unsaturated fatty acids and essential fatty acids is relatively high, and its composition is close to that of fish oil. Drug efficacy experiments have confirmed that fly maggot oil has a better promoting effect on the healing of skin wounds. Fly maggot oil also has a good health care effect on the human body. How to extract fly maggot oil effectively has not seen any report yet.

Chitin is a natural high molecular polymer, which is natural, non-toxic, and has good biocompatibility, degradability, and unique molecular structure and physical, chemical, and biological properties. It has a wide range of uses in the fields of medicine, material science, chemical industry, food, cosmetics, printing and dyeing, textile, papermaking, agriculture and environmental protection. Using fly maggots to extract chitin has the advantages of cheap and easy-to-obtain raw materials, simple production process, and high-quality products.

The purpose of the present invention is to provide a new process for fully exploiting and utilizing fly maggots, that is to produce fly maggots hydrolyzed animal protein, chitin and fly maggot oil through two-step enzymolysis, separation and extraction of artificial large-scale production of fly maggots, This process can make full use of fly maggot resources, and no three wastes are discharged.

Embodiments of the present invention are as follows:

The separation and extraction process of the active ingredients of resource insects provided by the invention is characterized in that the active ingredients include fly maggot hydrolyzed protein, fly maggot oil and fly maggot chitin, and the process steps are as follows:

1. Fly maggot shell/liquid separation

1.1 Place the cleaned fly maggots in a 0.08-0.1mol/L sodium hydroxide solution with 4-5 times the volume of the fly maggots, grind and pulverize them in a colloid mill;

1.2 Collect the ground maggot pulp, put it into a nylon sieve device, carry out centrifugation and separation, and collect the filter residue and filtrate respectively;

2. Preparation of fly maggot hydrolyzed protein and fly maggot oil

2.1 pump the above-mentioned filtrate into the stirred reactor, heat up to 40-45°C, stir, and then press the alkaline protease to the fresh maggot amount in a weight ratio of 1: 180-250 (enzyme activity: 100,000 units/ml) Add alkaline protease in proportion, and carry out enzymolysis at a constant temperature of 40-45°C. After 2-3 hours of enzymolysis, when the pH drops to 8.2-8.6, the weight ratio of neutral protease to fresh maggots is 1:40- Add neutral protease at 50 (enzyme activity 50,000 units/g), continue enzymatic hydrolysis at 40-45°C for 4-5 hours, when the pH drops to 7.0-7.5, heat up and boil to inactivate the enzyme;

2.2 Cool the enzymolyzed protein solution after the enzyme-inactivating treatment to room temperature, then centrifuge to collect the upper layer of fly maggot oil, and concentrate the remaining filtrate under reduced pressure, spray dry and sterilize to obtain the fly maggot hydrolyzed protein powder ;

3. Extraction of chitin

3.1 Pour the fly maggot filter residue obtained in the above step 1.2 into a stirring reaction pot, add 3% hydrochloric acid 2-3 times the total weight of the filter residue, stir continuously, soak for 24 hours at room temperature, and then carry out suction filtration , wash the filter residue with water until the pH value is neutral.

3.2 Put the filter residue washed to neutral into a heating mixing tank, add 10% sodium hydroxide solution 2-3 times the total weight of the filter residue, heat to 70-80°C for 3 hours, keep stirring, and carry out alkali boiling , remove the protein in the filter residue.

3.3 Collect the above alkali boiled matter and wash until neutral, put it into a stirring and bleaching pot, add 2-3% potassium permanganate solution 3-5 times the weight of the alkali boiled matter, soak for 1 hour, filter and wash, and continue vacuum drying. That is, the white solid product chitin is obtained.

Described alkaline protease is 2709 alkaline protease, also can be the mixture of 2709 alkaline protease and H ₂ NCONH ₂ , (NH ₄ )SO ₄ for accelerating hydrolysis; Described neutral protease is AS1398 neutral protease or is Mixture of AS1398 neutral protease and 166 neutral protease.

Compared with prior art, feature of the present invention mainly lies in:

1. Improve the comprehensive utilization of fly maggots. The method provided by the invention can simultaneously prepare fly maggot hydrolyzed protein, fly maggot oil and chitin deep-processing industrial products, and there is no discharge of three wastes, which provides a new method for the deep-level processing and utilization of fly maggots. way.

2. The fly maggot hydrolyzed protein, fly maggot oil and chitin products prepared by the method provided by the invention are of good quality and low cost.

3. The process of the present invention is reasonable, the process is connected and compact, and the production operation is safe.

According to the separation and extraction process of effective components of resource insects provided by the invention, 152-210 g of fly maggot hydrolyzed protein powder, 15-22 g of chitin, and 30-50 ml of fly maggot oil can be prepared per kilogram of fresh fly maggots. The quality detection indexes of prepared fly maggot hydrolyzed protein powder, chitin, and fly maggot oil are shown in Table 1, Table 2, and Table 3 respectively:

Table 1. Amino acid content in hydrolyzed protein powder (%) amino acid content amino acid content aspartic acid 7.10～8.10 Isoleucine 3.50～4.27 threonine 2.71～3.67 Leucine 5.50～6.20 serine 2.21～3.10 Tyrosine 4.90～5.77 glutamic acid 10.20～11.10 Phenylalanine 4.40～5.20 Glycine 3.54～4.32 Lysine 4.40～5.57 Alanine 2.71～3.80 arginine 3.38～4.28 cystine 0.30～0.67 Histidine 1.50～2.30 Valine 3.57～4.21 proline 3.12～3.83 Methionine 1.95～2.70

Table 2. Quality standard of fly maggot chitin project standard indicators project standard indicators Exterior light yellow powder Insolubles ≤2% granularity 100 mesh arsenic <0.5ppm viscosity 100～1500mpa·s lead <1ppm pH 6.5～7.5 HG <0.3ppm Degree of deacetylation >75% Total number of bacteria 1×10 ^3pcs /g moisture ≤10% coliform bacteria <40 Ash ≤2%

table 3. Fatty acid content of maggot oil (%) fatty acid content fatty acid content myristic acid 1.9～2.7 Linoleic acid 32.0～33.1 palmitic acid 19.2～20.6 Linolenic acid 2.9～3.8 stearic acid 1.9～2.8 saturated fatty acid 26.8～27.9 Palmitic acid 12.1～13.3 unsaturated fatty acid 67.9～69.1 Oleic acid 17.2～18.7 essential fatty acids 35.5～36.5

Embodiment 1: use the separation and extraction process of the active ingredients of resource insects provided by the present invention to prepare maggot hydrolyzed protein, maggot oil and maggot chitin as raw materials with maggots, and the process steps are as follows:

1. Fly maggot shell/liquid separation:

1.1 Place 3 kg of cleaned fly maggots in 0.08mol/L sodium hydroxide solution of 4 times the volume of fly maggots, and grind and pulverize them in a colloid mill;

1.2 Collect the ground maggot pulp, put it into a nylon sieve device with a sieve hole of 200 mesh, carry out centrifugation through an electric centrifuge, and collect the filter residue and filtrate respectively;

2. Preparation of fly maggot hydrolyzed protein and fly maggot oil:

2.1 Pump the above-mentioned filtrate into the enzymolysis tank, heat up to 45°C, stir with a mixer, and then add alkaline protease in a ratio of 1:180 (enzyme activity: 100,000 units/ml) to fresh maggots Alkaline protease, carry out enzymolysis at a constant temperature of 40°C, observe the pH changes regularly, when the pH drops to 8.2 in 2 hours after enzymolysis, the weight ratio of neutral protease and fresh maggots is 1: 40 (enzyme activity 5 10,000 units/gram) by adding neutral protease, and continue enzymatic hydrolysis at 40°C for 4 hours, when the pH drops to 7.5, heat up and boil for 15 minutes to inactivate the enzyme;

The alkaline protease is 2709 alkaline protease, and it can also be a mixture of 2709 alkaline protease and H ₂ NCONH ₂ , (NH ₄ )SO ₄ to accelerate hydrolysis; the neutral protease is AS1398 neutral protease or AS1398 Mixture of dispase and 166 dispase.

2.2 Cool the enzymolyzed protein solution after the enzyme-inactivating treatment to room temperature, then centrifuge at 3500rpm for 15 minutes, collect the upper layer of fly maggot oil, and concentrate the remaining filtrate under reduced pressure, spray dry and sterilize to obtain the fly Maggot hydrolyzed protein powder; present embodiment makes fly maggot oil 90ml, fly maggot hydrolyzed protein powder 527g;

3. Extraction of chitin:

3.1 Pour the fly maggot filter residue obtained in the above step 1.2 into the stirring reaction pot, add 3% hydrochloric acid twice the total weight of the filter residue, stir constantly, soak for 24 hours at room temperature, then carry out suction filtration, and use Wash the filter residue with water until the pH value is neutral.

3.2 Put the filter residue washed to neutral into a heating mixing tank, add 10% sodium hydroxide solution twice the total weight of the filter residue, heat to 70°C for 3 hours, and keep stirring to remove the protein in the filter residue.

3.3 Collect the above alkali boiled matter and wash until neutral, put it into a stirring bleaching pot, add 2% potassium permanganate solution 3 times the weight of the alkali boiled matter, soak for 1 hour, filter and wash, and continuously vacuum dry to obtain white The solid product chitin is 62g.

Embodiment 2: use the separation and extraction process of the active ingredients of resource insects provided by the present invention to prepare maggot hydrolyzed protein, maggot oil and maggot chitin as raw materials with maggots, and the process steps are as follows:

1. Fly maggot shell/liquid separation:

1.1 Place 2 kg of cleaned fly maggots in 0.1mol/L sodium hydroxide solution with 5 times the volume of fly maggots, and grind and pulverize them in a colloid mill;

1.2 Collect the ground maggot pulp, put it into a nylon sieve device with a sieve hole of 200 mesh, carry out centrifugation through an electric centrifuge, and collect the filter residue and filtrate respectively;

2. Preparation of fly maggot hydrolyzed protein and fly maggot oil:

2.1 Pump the above-mentioned filtrate into the enzymatic hydrolysis tank, heat up to 40°C, stir with a mixer, and then add alkaline protease in a ratio of 1:250 (enzyme activity: 100,000 units/ml) by weight to fresh maggots Alkaline protease, carry out enzymolysis at constant temperature 45 ℃, regularly observe pH change, when 3 hours after enzymolysis, when pH drops to 8.6, be 1: 50 by the weight ratio of neutral protease and fresh maggot amount (enzyme activity 5 10,000 units/gram) by adding neutral protease, and continue enzymatic hydrolysis at 45°C for 5 hours, when the pH drops to 7.3, heat up and boil for 15 minutes to inactivate the enzyme;

The alkaline protease is 2709 alkaline protease, and it can also be a mixture of 2709 alkaline protease and H ₂ NCONH ₂ , (NH ₄ )SO ₄ to accelerate hydrolysis; the neutral protease is AS1398 neutral protease or AS1398 Mixture of dispase and 166 dispase.

2.2 Cool the enzymolyzed protein solution after the enzyme-inactivating treatment to room temperature, then centrifuge at 3500rpm for 15 minutes, collect the upper layer of fly maggot oil, and concentrate the remaining filtrate under reduced pressure, spray dry and sterilize to obtain the fly Maggot hydrolyzed protein powder; present embodiment makes fly maggot oil 75ml, fly maggot hydrolyzed protein powder 348g;

3. Extraction of chitin:

3.1 Pour the fly maggot filter residue obtained in the above step 1.2 into the stirring reaction pot, add 3% hydrochloric acid which is 3 times the total weight of the filter residue, stir constantly, soak for 24 hours at room temperature, then carry out suction filtration, and use Wash the filter residue with water until the pH value is neutral.

3.2 Put the filter residue washed to neutral into a heating mixing tank, add 10% sodium hydroxide solution 3 times the total weight of the filter residue, heat to 80°C for 3 hours, and keep stirring to remove the protein in the filter residue.

3.3 Collect the above alkali boiled matter and wash until neutral, put it into a stirring bleaching pot, add 3% potassium permanganate solution 5 times the weight of the alkali boiled matter, soak for 1 hour, filter and wash, and continuously vacuum dry to obtain white Solid product chitin 50g.

Embodiment 3: use the separation and extraction process of the active ingredients of resource insects provided by the present invention to prepare fly maggot hydrolyzed protein, fly maggot oil and fly maggot chitin as raw materials with fly maggots, and its process steps are as follows:

1. Fly maggot shell/liquid separation:

1.1 Place 3 kg of cleaned fly maggots in 0.09mol/L sodium hydroxide solution of 4.5 times the volume of fly maggots, and grind and pulverize them in a colloid mill;

1.2 Collect the ground maggot pulp, put it into a nylon sieve device with a sieve hole of 200 mesh, carry out centrifugation through an electric centrifuge, and collect the filter residue and filtrate respectively;

2. Preparation of fly maggot hydrolyzed protein and fly maggot oil:

2.1 Pump the above-mentioned filtrate into the enzymatic hydrolysis tank, heat up to 43°C, stir with a mixer, and then add alkaline protease in a ratio of 1:200 (enzyme activity: 100,000 units/ml) to fresh maggots Alkaline protease, carry out enzymolysis at constant temperature 43 ℃, regularly observe pH change, when 2.5 hours after enzymolysis, when pH drops to 8.4, be 1: 45 by the weight ratio of neutral protease and fresh maggot amount (enzyme activity 5 10,000 units/gram) by adding neutral protease, and continue enzymatic hydrolysis at 43°C for 4.5 hours, when the pH drops to 7.0, heat up and boil for 15 minutes to inactivate the enzyme;

The alkaline protease is 2709 alkaline protease, and it can also be a mixture of 2709 alkaline protease and H ₂ NCONH ₂ , (NH ₄ )SO ₄ to accelerate hydrolysis; the neutral protease is AS1398 neutral protease or AS1398 Mixture of dispase and 166 dispase.

2.2 Cool the enzymolyzed protein solution after the enzyme-inactivating treatment to room temperature, then centrifuge at 3500rpm for 15 minutes, collect the upper layer of fly maggot oil, and concentrate the remaining filtrate under reduced pressure, spray dry and sterilize to obtain the fly Maggot hydrolyzed protein powder; present embodiment makes fly maggot oil 118ml, fly maggot hydrolyzed protein powder 5.4kg;

3. Extraction of chitin:

3.1 Pour the fly maggot filter residue obtained in the above step 1.2 into the stirring reaction pot, add 3% hydrochloric acid 2.5 times the total weight of the filter residue, stir constantly, soak for 24 hours at room temperature, then carry out suction filtration, and use Wash the filter residue with water until the pH value is neutral.

3.2 Put the filter residue washed to neutral into a heating mixing tank, add 10% sodium hydroxide solution 2.5 times the total weight of the filter residue, heat to 75°C for 3 hours, and keep stirring to remove the protein in the filter residue.

3.3 Collect the above alkali boiled matter and wash until neutral, put it into a stirring bleaching pot, add 2.5% potassium permanganate solution 4 times the weight of the alkali boiled matter, soak for 1 hour, filter and wash, and continuously vacuum dry to obtain white The solid product chitin is 530g.

Claims (3)

1. the separation of a resource insect effective ingredient, extraction process is characterized in that, effective ingredient comprises fly maggot protein hydrolysate, fly maggot oil and fly maggot chitin, and its processing step is as follows:

1) maggot shell/liquid separates

(1) fly maggot that cleans up is placed the 0.08-0.1mol/L sodium hydroxide solution of fly maggot 4-5 times volume, in colloid mill, grind;

(2) maggot of collecting through grinding is starched, and puts into the nylon mesh device, carries out centrifugal, separation, collects its filter residue and filtrate respectively;

2) preparation of fly maggot protein hydrolysate and fly maggot oil

(1) above-mentioned filtrate is pumped into stirred autoclave, be warming up to 40-45 ℃, stir, weight ratio in alkali protease and bright maggot amount is 1 then: the ratio of 180-250 (enzyme 100,000 units per ml alive) adds alkali protease, under 40-45 ℃ of constant temperature, carry out enzymolysis, behind enzymolysis 2-3 hour, when pH drops to 8.2-8.6, weight ratio by neutral proteinase and bright maggot amount is 1: 40-50 (enzyme 50,000 units/gram alive) adds neutral proteinase, continued enzymolysis 4-5 hour down at 40-45 ℃, when pH reduced to 7.0-7.5, intensification was boiled, and the enzyme that goes out is handled;

(2) will be through the enzymolysis protein liquid cool to room temperature after the enzyme that goes out is handled, centrifugal then, collect upper strata fly maggot oil, and residual filtrate is concentrated through reducing pressure, spray-drying, sterilization, promptly make fly maggot protein hydrolysate powder;

3) chitinous extraction.

(1) with above-mentioned 1) the fly maggot filter residue of (2) step operation gained pour in the stirring reaction pot, add filter residue gross weight 2-3 3% hydrochloric acid doubly, constantly stir, at ambient temperature, soaked 24 hours, carry out suction filtration then, washing filter residue to pH value with water is neutrality,

(2) will be washed till neutral filter residue and put into a mixing bowl of heating, add filter residue gross weight 2-3 10% sodium hydroxide solution doubly, and be heated to 70-80 ℃ and kept 3 hours, constantly stir, carry out soda boiling, remove the albumen in the filter residue.

(3) collect above soda boiling thing and be washed till neutrality, put into and stir clay kettle, add soda boiling thing weight 3-5 2-3% liquor potassic permanganate doubly, soaked 1 hour, filter and clean, continuous vacuum drying promptly makes white solid product chitin.

2. by separation, the extraction process of the described resource insect effective ingredient of claim 1, it is characterized in that: described alkali protease is 2709 alkali proteases or is 2709 alkali proteases and H ₂NCONH ₂, (NH ₄) SO ₄Mixture.

3. by separation, the extraction process of the described resource insect effective ingredient of claim 1, it is characterized in that: described neutral proteinase is the AS1398 neutral proteinase or is the mixture of AS1398 neutral proteinase and 166 neutral proteinases.