CN1195067C - Method for extracting protein and chitin from fly maggot by using enzyme hydrolysis as well as preparing chitosan from chitin - Google Patents
Method for extracting protein and chitin from fly maggot by using enzyme hydrolysis as well as preparing chitosan from chitin Download PDFInfo
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Abstract
The present invention relates to a method for extracting protein and chitin from fly maggots by using enzyme hydrolysis, and a method for preparing chitosan from chitin, which belongs to the technical field of enzymolyzing fly maggot protein by a biological technique. The present invention provides a technical skill that enzyme hydrolysis method is used for extracting protein and recovering chitin from fly maggots, the fly maggot hydrolysed protein is deodorized and added with fragrance, and chitin is used for preparing chitosan. The present invention has the technical scheme that the hydrolyzed protein is extracted by an enzyme hydrolysis method, and the hydrolyzed protein is deodorized by a Maillard reaction, and is modified with fragrance. Yellow objects and dark spots in fly maggot skins are removed through enzyme hydrolysis and sieving, and optimized process conditions are all provided for enzymolysis reaction, the Maillard reaction, deproteinization, decalcification and deacetylation. The quality of both the product protein and the chitosan reaches the food grade standard, and the present invention can be applied to industries, such as food product industry, health product industry, medicine industry, cosmetics industry, etc. which have high technical content requirements.
Description
Technical field
The present invention relates to a kind of method of from fly maggot, extracting protein and chitin, particularly a kind ofly from fly maggot, extract protein and chitin and prepare the method for chitosan, belong to biotechnology enzymolysis fly maggot protein matter technical field with chitin with enzyme hydrolysis method.
Background technology
Fly maggot is the insect that a kind of reproduction speed is fast, be of high nutritive value, and contains fine protein and chitin resource.Can form at present the level of large-scale farming fly maggot.But at present fly maggot is mainly as the substituted type feed, product with low content of technology, and deficiency in economic performance has seriously hindered the development of fly maggot breeding industry.In recent years, along with the deep understanding of people, the utilization of fly maggot has been turned to the deep level development of high added value purposes such as food, healthcare products, medicine, makeup by the substituted type feed to fly maggot nutritive value and functionally active.
Mechanical process or chemical method are mainly adopted in the production of Flyblow albumen powder at present, as with fly maggot wet pulverization, convection drying; Or will pulverize again after the fly maggot drying.The poorly water-soluble of this Flyblow albumen powder product, shade deviation also have heavier fishy smell, only are used for animal feed formulation or as animal feedstuff additive, Application Areas is subjected to bigger restriction.The extraction of relevant fly maggot chitin mainly utilizes Mechanical Crushing, cleans, and behind the acquisition fly maggot skin, handles with the diluted acid diluted alkaline again, removes technical process such as albumen, decolouring, decalcification.But because protein content is higher in the maggot skin that mechanical process makes, the following process complexity, long processing time, the protein resource waste is bigger, and production process is seriously polluted.The more important thing is does not still have the method for removing black oral cavity and yellow internal organ effectively in the course of processing, the polymerization degree, color and luster and impurity degree that these have all influenced chitin and chitosan product descend quality product.
Find out from pertinent literature, though existing fly maggot is used for the animal albumen powder prescription and as the open source literature report of animal protein feed additive, and the existing open source literature report that from fly maggot, extracts the fly maggot chitin with chemical method, as Chinese patent 971164908,981034462,991193911,001002651 etc., but do not see the report that from fly maggot, extracts protein and recovery chitin with enzyme hydrolysis method both at home and abroad.
Summary of the invention
(1) technical problem that will solve
The technical problem to be solved in the present invention is to utilize enzyme hydrolysis method to extract protein and recovery chitin from fly maggot; Fly maggot protolysate is taken off flavor and enhancing flavor; Utilize enzymatic hydrolysis and binding substances logos to sieve and wash, yellow in the fly maggot chitin and stain (fly maggot oral cavity) are removed; Prepare chitosan from the fly maggot chitin, make deacetylating degree of chitosan 〉=85%, reach food hygienic standard.
(2) technical scheme
A kind of from fly maggot method for extracting proteins.It is to use enzymatic hydrolysis process to extract fly maggot protein matter, utilizes Mei Lade (Maillard) reaction to take off flavor and enhancing flavor again.Its process flow diagram as shown in drawings.The present invention utilizes the biotechnology enzyme hydrolysis method to extract protolysate from fly maggot, uses enzymic hydrolysis high molecular weight protein is changed into water-soluble polypeptide and amino acid, simultaneously by selecting specific enzyme and condition optimizing to prevent the bitter taste that enzymolysis produces.To utilize Maillard reaction to carry out local flavor through the fly maggot protein matter that enzymolysis obtains and modify, eliminate fishy smell, can improve proteinic solvability and local flavor greatly like this.With fresh fly maggot is raw material, add water carry out glue gall broken, the heat tracing pre-treatment, the enzyme-added enzymatic hydrolysis reaction that carries out, the enzyme that goes out filters, filtrate is carried out Maillard reaction and is taken off flavor and enhancing flavor, concentrates or does not concentrate, spraying drying makes product fly maggot protolysate powder.
Enzyme hydrolysis method technology:
It is broken that glue galls: with 1 part of fly maggot add 1～3 part of water (w/w, weight ratio, down with) carry out glue gall broken, 2～4 parts of fly maggot slurries, can add water according to processing request and allocate the fly maggot slurry concentration.
Heat tracing pre-treatment: the fly maggot slurry in 60～90 ℃ of heat treated 10～60 minutes, is cooled to below 60 ℃.
Enzymatic hydrolysis reaction: how many size and bitter tastes with protein degree are index, select the enzyme of proper fly maggot protein hydrolysis from the multiple protein enzyme.Enzyme adopts single prozyme of planting enzyme or two kinds of enzymes of Sumizyme MP, AS.1398 proteolytic enzyme, papoid, trypsinase, food flavor enzyme, neutral protease.Processing condition are through orthogonal test analysis, and it is ratio＞pH at the bottom of temperature＞time＞enzyme that each the factor primary and secondary that influences degree of hydrolysis is closed.Enzymatic hydrolysis condition is 30～55 ℃ of temperature, 1～4 hour time, at the bottom of the enzyme than (u/g) 50～700, pH6.5～8.5.Reaction finishes to go out enzyme.
Filter: filtrate is fly maggot protein liquid, and filter residue is the fly maggot skin.Filtrate is carried out Maillard reaction and is taken off flavor and enhancing flavor.
Enzymolysis protein liquid takes off flavor and enhancing flavor: adopt Maillard reaction to carry out back and modify, reach flavouring and except that the purpose of raw meat.Amino acid and adding small amounts of sugars that reaction utilizes enzymic hydrolysis to produce are carried out according to following proportioning.
Raw material and proportioning thereof
| Raw material | Consumption (w) |
| Fly maggot hydrolyzed solution amino acid glucose | 100 0.05～1.5 0.2～0.5 |
Reaction conditions: 100～125 ℃ of temperature, 30～100 minutes time.
The preparation protein powder: reaction solution concentrates, and spraying drying makes product fly maggot protolysate powder.Protein content in the product 〉=60%.
Also can enzymolysis solution go out behind the enzyme without filtration, directly enzymolysis solution is carried out Maillard reaction and take off flavor and enhancing flavor under the same process condition, concentrate or do not concentrate, spraying drying makes fly maggot protolysate powder, protein content 〉=54%.
A kind of from fly maggot the method for chitin extraction (Chitin).It is as mentioned above, and fly maggot is raw material with the filter residue behind the filtering separation fly maggot protein liquid through the fly maggot skin of enzymic hydrolysis after enzymic hydrolysis, through deproteinated and decalcification, and preparation fly maggot chitin finished product.Its process flow diagram as shown in drawings.After enzymic hydrolysis extraction fly maggot protein matter, main component is a chitin in the fly maggot skin, also have a small amount of protein, fat and ash content (carbonate salt) etc., compare with shrimp and crab shells, chitin content height in the fly maggot skin, the content of protein and calcium salt is low, and it is bigger different that this just determines that fly maggot chitin extraction process and shrimp and crab shells chitin extraction process have.Therefore, adopting rational deproteinated and decalcification processing condition is extremely important to the quality that improves chitin.The Applied Biotechnology enzyme hydrolysis method is except that protein in the fly maggot skin is removed in hydrolysis, and the also mouthpart and the xanchromatic internal organ of black in the fly maggot of the hydrolysis simultaneously skin can effectively be removed impurity in the product chitin through the physics method again.The technology of chitin extraction is: the fly maggot skin is sieved and washed, and the soda boiling deproteinated cleans to neutral, and the acidleach decalcification is cleaned to neutral, and drying makes fly maggot chitin finished product.
Deproteinated: carry out deproteinated with NaOH solution, concentration of lye 2～6% (w/w), 60～100 ℃ of temperature, liquid-solid ratio (ml/g) 3～10,1～5 hour reaction times.
Decalcification: the fly maggot skin is through deproteinated, after sieving and washing decon, the residue that stays cleans to neutral, after centrifugal the anhydrating, carries out decalcification with dilute hydrochloric acid solution, concentration of hydrochloric acid 0.5～1.5mol/L, liquid-solid ratio 2.0～9.0ml/g, in 20～60 minutes reaction times, temperature is a room temperature, clean to neutral with pure water, drying obtains the fly maggot chitin at low temperatures again.
A kind of method for preparing chitosan (Chitosan) from the fly maggot chitin.It is that the fly maggot chitin that extracts with enzymatic hydrolysis process as mentioned above is a raw material, carries out deacetylation and the chitosan product for preparing in high alkali liquid.
Chitosan is a kind of cationic straight chain glycosaminoglycan, is that chitin is sloughed the ethanoyl in the molecule and the product that forms.The preparation process of chitosan, reality be the saccharoidal alkaline hydrolysis process of this acid amides of chitin just, i.e. deacetylation.The reaction formula of deacetylation is as follows:
At present the preparation raw material of chitosan is mainly shrimp crab chitin, normally adopts highly basic at high temperature to handle chitin and makes it remove most of ethanoyl to make.Compare with shrimp crab chitin, fly maggot chitin ash content is few, and heavy metal content is low, stay in grade.Deacetylation is one of main performance index of chitosan.The deacetylation that the food grade chitosan is had relatively high expectations, i.e. deacetylation 〉=85%.
The present invention prepares chitosan, is that the fly maggot chitin that adopts method as mentioned above to extract is a raw material, through basic hydrolysis, i.e. and deacetylation and the chitosan product for preparing.Raw material adopts the fly maggot chitin of not pulverizing, and alkali lye adopts NaOH solution, alkali concn 30～55% (w/w), and 30～140 ℃ of temperature of reaction, in 2～30 hours reaction times, liquid-solid ratio is 5～20ml/g.Optimizing under the processing condition, the deacetylation of product can reach more than 88%.
(3) beneficial effect
The present invention by studying fly maggot protein matter in great detail the enzyme hydrolysis process condition and the preparation process condition of chitin and chitosan, for the deep processing and the comprehensive utilization of fly maggot resource provides a new way.
By the relatively discovery of proteinic extraction yield and solubility rate, proteinic extraction yield and solubility rate reach more than 92% and 80% after the enzymatic treatment, and the mechanical process separation only is 64% and 24%.Enzyme process has significantly improved proteinic solvability, has improved proteinic solubility rate, has increased proteinic extraction yield.
Adopt Maillard reaction, the purpose that protein liquid reaches flavouring and removes raw meat is being optimized product aromatic flavour under the processing condition, and fishy smell disappears.
Protein content is lower than the fly maggot skin that Mechanical Crushing obtains in the fly maggot skin after the enzymic hydrolysis.The consumption and the reaction times of alkali in the deproteinated operation have been saved greatly.Black mouthpart and yellow internal organ are also by enzymolysis in the fly maggot skin after the enzymic hydrolysis, again through sieving and washing, wherein greatly part can be removed easily, and the impurity of small amount of residual in the fly maggot skin is sieved and washed after alkaline purification also further to be removed, and the microscopy stain rate of removing reaches more than 98%.
Fly maggot chitosan is prepared by the fly maggot chitin.Optimizing under the processing condition, the deacetylation of chitosan and yield are all very high.Through infrared spectra and X-ray diffraction analysis, the conclusive evidence products obtained therefrom is a chitosan.
Fly maggot protein matter has very high nutritive value.To the analysis of fly maggot nutritive ingredient, fly maggot (butt) contains crude protein 54～65% according to both at home and abroad, fat 10～14%, and chitin 8～10% and the VITAMIN of enriching, trace element etc., nutritive ingredient is more comprehensive.Amino acid is more complete in the fly maggot protein matter, and the amino acid that is provided all can satisfy the amino acid requirement of children and adult's suggestion.A large amount of experimentation on animalies show: the Flyblow albumen powder that is made by fly maggot has tangible immunoregulation effect, antifatigue effect, radiation resistance, liver protection effect and delaying senility function.Use the fly maggot protolysate powder that enzymic hydrolysis makes high molecular weight protein is changed into water-soluble polypeptide and amino acid, easilier after edible absorbed by human body.
The fly maggot protein matter of enzyme hydrolysis method preparation also owing to have good solubility, water-retentivity, emulsifying property and oxidation-resistance etc., can be widely used in industries such as food, medicine, makeup except that having very high nutritive value.
In foodstuffs industry, protolysate is a kind of flavouring base material that can strengthen the food natural flavor well, it can be applicable to the nutrition-fortifying agent of various flavor type food flavourings and food, can also improve simultaneously Nutritive value of food and storage security, can be widely used among the various food as a kind of natural endless foodstuff additive, as meat product processing, protein drinks, baked goods etc.In cosmetic industry, protolysate has good emulsifying, oilness and surfactivity effect, is one of high-quality base-material of skin care product, treatment.In medicine industry, oral hydrolyzed protein can be applicable to need the special population of edible high protein, and for lacking proteinic patient, and because of functions of intestines and stomach decline, serious burn, severe loss of blood can not oral persons, injection protolysate is effective means very.
Chitosan is one of most important derivative of chitin, because of containing free-NH2 active group in its molecule, when being dissolved in acidic solution, can be protonated and the lotus that becomes positively charged, it is unique alkaline polysaccharide in the natural polysaccharide, thereby have many special physicochemical character and physiology, pharmacological function, be described as necessary the 6th vital principle of human body by many scholars.
Studies confirm that in a large number; chitin and chitosan have multiple function, as reducing cholesterol, protection liver, enhance immunity power, reduce the heat, antibacterial anticorrosion etc. of uric acid in the body, gastric acid inhibitory secretion, adjustment body trace element level, reduction food.With chitin and chitosan be the protective foods of main component by extensive exploitation, its nourishing function has the immunity of adjusting, regulating blood fat, lowering blood glucose etc.
In sum, the present invention has important economic benefit and social benefit.
Description of drawings
Accompanying drawing Enzymatic Extraction fly maggot protein matter and chitin and prepare the process flow diagram of chitosan with chitin.
Embodiment:
Embodiment 1
Enzyme hydrolysis method extracts protolysate and chitin from fly maggot.Fresh fly maggot galls broken through glue, after the slurry heat treated, adopt AS.1398 proteolytic enzyme, at the bottom of the enzyme than 500u/g, 45 ℃ and pH7.0 under, hydrolysis 3 hours.This moment, degree of hydrolysis was 18%, and the protein solubility rate is 81%, and hydrolyzed solution does not have bitter taste.
The enzyme hydrolyzate enzyme that goes out filters, and filtrate is through Maillard reaction, and 120 ℃, reaction are in the time of 60 minutes, and the aromatic flavour of product, fishy smell disappear.Spraying drying obtains Flyblow albumen powder.
Filter residue fly maggot skin is used for chitin extraction, and method is with following embodiment 4.
Embodiment 2
Enzyme hydrolysis method extracts protolysate and chitin from fly maggot.Adopt AS.1398 proteolytic enzyme and food flavor enzyme, both carry out enzymolysis with the prozyme that 1: 1 (w/w) forms, and all the other conditions are with embodiment 1.Degree of hydrolysis after the hydrolysis and protein solubility rate also can reach satisfied result.
Embodiment 3
Enzyme hydrolysis method extracts protolysate from fly maggot.Through behind the enzymolysis, directly through the Maillard reaction flavouring, spraying drying obtains Flyblow albumen powder to enzymolysis solution, protein content 〉=60% at fly maggot.
Embodiment 4.
Enzyme hydrolysis method is chitin extraction from fly maggot.After the maggot skin that obtains after the enzymic hydrolysis sieved and washed impurity elimination, deproteinated 5%NaOH solution, 100 ℃ were boiled 3 hours, and liquid-solid ratio 9ml/g cleans to neutral; Decalcification 1mol/L hydrochloric acid soln, liquid-solid ratio 6ml/g reacted 50 minutes, cleaned to neutral.
Chitin behind the deproteinated proves that through amino acid analysis the protein removal rate is 97% (in total amino acid content).The ash oontent of chitin is 0.2% after the decalcification.
Embodiment 5
The fly maggot de-acetyl chitin prepares chitosan.The fly maggot chitin is carried out deacetylated reaction in concentrated base.Adopt 120 ℃ of temperature, 3 hours time, alkali concn 45%, liquid-solid ratio 10ml/g.This moment, the deacetylation of product was 88%, and yield is 75%.
Claims (1)
1. method for extracting proteins from fly maggot, it is characterized in that using enzymatic hydrolysis process and extract fly maggot protein matter, utilize Maillard reaction to take off flavor and enhancing flavor again, its technology is to be raw material with fresh fly maggot, adding water carries out glue and galls broken, the heat tracing pre-treatment, the enzyme-added enzymatic hydrolysis reaction that carries out, enzyme goes out, filter, filtrate is carried out Maillard reaction and is taken off flavor and enhancing flavor, and spraying drying makes product fly maggot protolysate powder; Filter residue cleans to neutral through the soda boiling deproteinated, and the acidleach decalcification is cleaned to neutral, and drying makes fly maggot chitin finished product; The concrete technology of extracting fly maggot protein matter is:
(1) enzyme hydrolysis method technology:
It is broken that glue galls: with 1 part of fly maggot add 1～3 part of water carry out glue gall broken, 2～4 parts of fly maggot slurries, add water and allocate the fly maggot slurry concentration;
Heat tracing pre-treatment: the fly maggot slurry 60～90 ℃ of heat treated 10～60 minutes, is cooled to below 60 ℃;
Enzymatic hydrolysis reaction: the prozyme that enzyme adopts AS.1398 proteolytic enzyme or AS.1398 proteolytic enzyme and food flavor enzyme to form at 1: 1 with mass ratio, enzymatic hydrolysis condition is 30～55 ℃ of temperature, 1～4 hour time, ratio 50～700u/g at the bottom of the enzyme, pH6.5～8.5, reaction finishes to go out enzyme;
Filter, filtrate is fly maggot protein liquid, and filter residue is the fly maggot skin;
(2) enzymolysis protein liquid takes off flavor and enhancing flavor:
Filtrate is carried out Maillard reaction and is taken off flavor and enhancing flavor, and raw material and proportioning thereof are as follows:
Raw material consumption (w)
Fly maggot hydrolyzed solution 100
Amino acid 0.05～1.5
Glucose 0.2～0.5
Reaction conditions: 100～125 ℃ of temperature, 30～100 minutes time;
(3) preparation protein powder:
Reaction solution concentrates or does not concentrate, and spraying drying makes product fly maggot protolysate powder.
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| CN 02138647 CN1195067C (en) | 2002-11-20 | 2002-11-20 | Method for extracting protein and chitin from fly maggot by using enzyme hydrolysis as well as preparing chitosan from chitin |
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