CN105581281A - Method for preparing seasoning raw material from leftovers of shrimps and crabs - Google Patents
Method for preparing seasoning raw material from leftovers of shrimps and crabs Download PDFInfo
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- CN105581281A CN105581281A CN201510988027.0A CN201510988027A CN105581281A CN 105581281 A CN105581281 A CN 105581281A CN 201510988027 A CN201510988027 A CN 201510988027A CN 105581281 A CN105581281 A CN 105581281A
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Abstract
The invention discloses a method for preparing a seasoning raw material from leftovers of shrimps and crabs. The method comprises steps as follows: step one, preparation of shrimp head crude enzymes and shrimp head enzymes; step two, enzymolysis of shrimp shells by the shrimp head crude enzymes; step three, enzymolysis of crab shells and crab feet by the shrimp head enzymes. Seafood-flavored seasoning is prepared from leftovers of shrimps and crabs through hydrolysis by the aid of the shrimp head crude enzymes extracted from the shrimp heads. Shrimp and crab enzymatic hydrolysate contains multiple amino acids essential to human bodies and has high nutrient value, and amino acids, mineral substances and other nutrient substances contained in a product prepared from the shrimp and crab enzymatic hydrolysate are easily absorbed by the human bodies; the freeze-dried enzymatic hydrolysate can be added to various kinds of food such as seafood seasoning, edible coloring, rice cereal for children and pregnant women and the like.
Description
Technical field
The technical field that the present invention relates to cultivation processing, relates in particular to one and utilizes shrimp crab leftover bits and pieces to prepare material of spiceMethod.
Background technology
China has the Gold Coast line that enriches fishery resources that is containing tremendous potential, and total length exceedes 3.2 ten thousand kilometersBe positioned at the whole world the 4th. Along with the epoch are progressive, its people more and more more pay attention to quality of life and healthy, therefore, and fish shrimp crab etc.Health-oriented products demand grows with each passing day, and the fishery of China has welcome rare development spring. Wherein, the accessory substance in fish productionDeep processing be new direction and the new opportunity of China's Fisheries Development. For example, producing in the products such as peeled shrimp and crab meat, accumulated largeAmount shrimp crab leftover bits and pieces. According to relevant speciality research, the chitin that contains 15% left and right in shrimp head and shrimp shell, protein content is 30%Left and right, remaining is mineral matter element, fat-soluble A and vitamin E etc., wherein the abundantest 35% left side that reaches of the content of calciumThe right side, secondly for phosphorus is 4%-5%. Protein in shrimp shell and shrimp head contains eight kinds of essential necessary amino acid. Except lysineContent is lower slightly, other the seven kinds minimum requirements that must amino acid content all reach human body must amino acid whose demand. Crab leftover bits and piecesIn the battalion such as protein, carbohydrate, unrighted acid, lecithin, carotenoid, cephalin, chitin, celluloseForm a point rich content, have in addition trace mineral and each seed amino acids such as calcium, magnesium, phosphorus. Therefore, shrimp crab leftover bits and pieces is fabulousIndustrial processes raw material can apply to various aspects, there is higher commercial exploitation and be worth. If these processing can be got a footholdMaterial fully utilizes instead of simple abandoning, and can bring economic benefit to enterprise on the one hand, also can on the other handReduce environmental pollution.
Flourish along with shrimp crab industry, the recycling of shrimp crab accessory substance also becomes the focus of the research of all circles.Current research mainly concentrates on and adopts the whole bag of tricks by the albumen in shrimp crab leftover bits and pieces, pigment and grease etc., and recycling,Reprocess suitable deep processing, make high nutrition, cheaply new product.
The people such as Chen Limei do not contain the chitin of protein and calcium with citric acid and naoh treatment shrimp shell, preparation. Duan ShanProbe into the autolysis of lactic acid bacteria Deng people for the method for the Protein Recovery in shrimp head and shrimp shell. IslemYounes utilize fromProtein in the alkali protease removal shrimp shell extracting in Rockfish internal organ is to prepare chitin.
A large amount of crab leftover bits and pieces that can produce in the time of the crab products such as traditional industry processing crab meat, crab oil, wherein quantity is maximumBe crab pin, the shell on the back of crab. The current scientific research focus when exploitation of crab leftover bits and pieces, many researchers are under crabThe various product of pin material exploitation purposes. Such as, the people such as Fan Jianfeng, taking swimming crab leftover bits and pieces as raw material, add in the differential responses stageAdd different commercial enzymes, in order to extract anti-oxidation peptide and the antibacterial peptide in crab leftover bits and pieces. The people such as Wang Xiaoqian probe into sky in swimming crabSo extraction, purification process of pigment, in order to substitute synthetic food color. And Tao Xueming utilizes absolute ethyl alcohol as solvent extractionCrab oil in swimming crab blanking is as the raw material of health products.
The people such as Zhang Lianfu are taking shrimp glutelin enzymolysis liquid as raw material, then the xylose and glucose that adds proper proportion is made and hadThe flavoring of dense shrimp local flavor. The optimum enzymolysis condition that the people such as Chen Haihua optimize Mytilus galloprovincialis enzyme by Response Surface Method is prepared seafoodTaste flavoring. The people such as the Ren Xiane first water extraction of use get the albumen in shrimp shell, and remaining shrimp shell extracts albumen with papain again, extractLiquid is again through the concentrated extra large delicate flavour sauce of preparation that waits.
Although in recent years, obtain larger progress for the comprehensive utilization of shrimp crab both at home and abroad, still had the work step of addingSuddenly comparatively loaded down with trivial details, be converted into the shortcomings such as industrial production feasibility is low. Seek by experiment operability high, step is simple, and cost is lowShrimp crab leftover bits and pieces prepare seafood taste production of condiments method, for the upgrading of fishery industry line with improve and have great meaningJustice. In view of this, inventor research and designed a kind of method of utilizing shrimp crab leftover bits and pieces to prepare material of spice, this case byThis generation.
Summary of the invention
The object of the present invention is to provide a kind of method of utilizing shrimp crab leftover bits and pieces to prepare material of spice, existing to solveThe problem that technology Prawn crab leftover bits and pieces is wasted in a large number.
To achieve these goals, the present invention solves the technical scheme that its technical problem takes and is:
Utilize shrimp crab leftover bits and pieces to prepare a method for material of spice, comprise the following steps:
Step 1, the preparation thick enzyme of shrimp head and a shrimp enzyme:
Shrimp head is smashed to pieces, added buffer solution, with refrigerated centrifuge centrifugal 20min at 12000r/min, 4 DEG C, centrifugal completeWith except degrease, get supernatant by 8 layers of filter-cloth filtering; Supernatant is left standstill after 2 hours at 12000r/min, 4 DEG C centrifugal again20min, makes the thick enzyme of shrimp head;
By the thick enzyme of prepared shrimp head, slowly adding saturated mass fraction is 80% ammonium sulfate, leaves standstill 2h at 4 DEG CAfter, then at 12000r/min, 4 DEG C centrifugal 20min, abandon supernatant; In precipitation, add a small amount of buffer solution to dissolve, use 3.5KdaThe bag filter dialysis 24h of pore size, makes a shrimp enzyme, at-20 DEG C, saves backup;
Step 2, the thick enzyme enzymolysis of shrimp head shrimp shell:
Shrimp shell is thawed, after dried, crushed, add the thick enzyme of shrimp head to carry out enzymolysis, enzymatic hydrolysis condition is: time 4-8h,PH3.5-6.5, temperature 30-50 DEG C, enzyme concentration 500-2300U/g, enzymolysis liquid go out enzyme live after defishying, then through liquid-solid pointFrom, after concentrate drying, make sea food seasoning raw material;
Step 3, shrimp enzyme enzymolysis crab shell and crab pin:
Crab shell and crab pin are thawed, after dried, crushed, add a shrimp enzyme to carry out enzymolysis, enzymatic hydrolysis condition is: time 5-9h, pH3.5-6.5, temperature 30-50 DEG C, enzyme concentration 1475-1878U/g, enzymolysis liquid go out enzyme live after defishying, then through liquidGu separate, after concentrate drying, make sea food seasoning raw material.
As the optimal way of embodiment, in described step 2, enzymatic hydrolysis condition is: time 7h, and pH6.0,40 DEG C of temperature, addEnzyme amount 1678.93U/g.
As the optimal way of embodiment, in described step 3, enzymatic hydrolysis condition is: time 7h, and pH6.0,40 DEG C of temperature, addEnzyme amount 1678U/g.
The thick enzyme hydrolysis shrimp of the shrimp head crab leftover bits and pieces of the extraction of utilization of the present invention from shrimp head is prepared extra large delicate flavour sauce. FirstUtilize and simply smash to pieces and refrigerated centrifuge, the thick enzyme of endogenous shrimp head extracting from shrimp head; Then adopt shrimp enzyme enzymolysis respectivelyShrimp shell, crab shell and crab pin are prepared various material of spice. In shrimp crab enzymolysis liquid, contain the necessary several amino acids of human body, nutritionBe worth highly, utilize its product being prepared into, the nutriment such as amino acid and mineral matter in product is beneficial to the absorption of human body. FreezingDried enzymolysis liquid can make an addition to varieties of food items, as: seafood condiment, food coloring, children and pregnant woman's Nutritive Rice paste etc.
Brief description of the drawings
The SDS-polypropylene amine gel electrophoresis of Fig. 1 shrimp enzyme liquid;
Fig. 2 time is for the impact of a shrimp enzyme liquid degree of hydrolysis;
Fig. 3 enzyme concentration is for the impact of shrimp enzymolysis liquid degree of hydrolysis;
The electrophoretic analysis figure of the different enzyme concentration enzymolysis liquids of Fig. 4; Wherein, M:Marker, 1: shrimp shell (before reaction), 2: a shrimp enzymeLiquid (before reaction), 3: enzymolysis liquid (enzyme concentration is 1229.475U/g), 4: enzymolysis liquid (enzyme concentration is 1721.265U/g), 5: enzymolysisLiquid (enzyme concentration is 2213.055U/g), 6: blank enzymolysis liquid (not adding shrimp shell, after reaction);
The impact of Fig. 5 pH prawn head enzyme liquid degree of hydrolysis;
The impact of Fig. 6 temperature prawn head enzyme liquid degree of hydrolysis;
The electrophoretogram of the thick enzyme hydrolysis shrimp of shrimp head shell under Fig. 7 optimum enzymolysis condition; Wherein, M:Marker, 1: a shrimp crude enzyme liquid,2: shrimp shell enzymolysis liquid (under optimal conditions);
Under the optimum enzymatic hydrolysis condition of Fig. 8, the electrophoretogram of the thick enzyme hydrolysis crab of shrimp head pin; Wherein, M:Marker, 1: a shrimp crude enzyme liquid(before reaction), 2: crab pin (before reaction), 3: shrimp shell enzymolysis liquid (under optimal conditions).
Detailed description of the invention
Below in conjunction with the drawings and specific embodiments so that the invention will be further described. The raw material that embodiment is used: shrimpHead: the fresh South America prawn of being bought by the market of farm produce, Jimei District, Xiamen City, obtains through peeling off the collection of shrimp head; Shrimp shell: come from good fortuneZhangpu County Huang Cang development zone, Jian Sheng Zhangzhou City Ying Feng Food Co., Ltd, produces shrimp shell leftover bits and pieces in peeled shrimp processing; Crab pin: fromIn Zhangpu County Huang Cang development zone, Zhangzhou City of Fujian Province Ying Feng Food Co., Ltd, in crab meat processing, produce crab leftover bits and pieces.
Embodiment 1 prepares the thick enzyme of shrimp head and a shrimp enzyme
Shrimp head is smashed to pieces, added buffer solution, with refrigerated centrifuge centrifugal 20min at 12000r/min, 4 DEG C, centrifugal completeWith except degrease, get supernatant by 8 layers of filter-cloth filtering; Supernatant is left standstill after 2 hours at 12000r/min, 4 DEG C centrifugal again20min, makes the thick enzyme of shrimp head;
By the thick enzyme of prepared shrimp head, slowly adding saturated mass fraction is 80% ammonium sulfate, leaves standstill 2h at 4 DEG CAfter, then at 12000r/min, 4 DEG C centrifugal 20min, abandon supernatant; In precipitation, add a small amount of buffer solution to dissolve, use 3.5KdaThe bag filter dialysis 24h of pore size, makes a shrimp enzyme, at-20 DEG C, saves backup.
Adopt the method for SDS-polyacrylamide gel electrophoresis (SDS-PAGE) to measure the molecular mass of a shrimp enzyme, with standardAlbumen Marker (14.3-97.2kDa) is as standard control. Wherein, the resolving gel concentration of SDS-PAGE is 12%, and concentrated glue is denseDegree is 5%.
From Fig. 1, can know, in molecular weight 14.3-97.5kDa, in a shrimp endogenous protein crude enzyme liquid, have manyBand. Therefore, the thick liquid of a shrimp enzyme may be made up of multiple protein enzyme, is worth continuing its water for shrimp shell and crab pin of researchSeparate effect.
The thick enzyme enzymolysis of embodiment 2 shrimp head shrimp shell
Shrimp shell is thawed, after dried, crushed, add the thick enzyme of shrimp head to carry out enzymolysis, enzymatic hydrolysis condition is: time 4-8h,PH3.5-6.5, temperature 30-50 DEG C, enzyme concentration 500-2300U/g, enzymolysis liquid go out enzyme live after defishying, then through liquid-solid pointFrom, after concentrate drying, make sea food seasoning raw material;
The present invention has investigated the impact of the thick enzyme hydrolysis degree of time prawn head, is that 658.71U/g, pH are 5 keeping enzyme concentrationWith temperature be under the enzymatic hydrolysis condition of 40 DEG C, choose 6 different times (4,5,5.5,6,6.5,8h) and investigate respectively it for hydrolysisThe impact of degree. As shown in Figure 2, along with the growth in reaction time, the substrate in system reduces, and product increases, substrate and enzymeBetween effective collision reduce, degree of hydrolysis changed slowly after 6.5 hours. Therefore, choosing 6.5 hours is the reaction time.
The present invention is that 6.5h, pH are 5.5 in the retention time, and temperature is, under the enzymatic hydrolysis condition of 35 DEG C, to have investigated enzyme concentration pairIn the impact of degree of hydrolysis. Experimental result is shown in Fig. 3. In the middle of enzymatic reaction, when the concentration of enzyme in substrate approaches when saturated, its reactionVelocity variations will slow down. Related documents report, while adopting commercial enzyme, in the time of the excessive concentration of enzyme, can cause between enzyme mutuallySuppress and hydrolysis, enzyme is lived and reduce. And in this experiment, because a shrimp enzyme only extracts through thick, in enzyme liquid, also contain a large amount ofThe albumen of shrimp head self causes degree of hydrolysis to increase along with the increase of enzyme concentration always. As can be seen from Figure 3, when enzyme-addedWhile measuring from 0-1000U/g, degree of hydrolysis rises rapidly. When enzyme concentration is during from 1229.475-2213.055U/g, degree of hydrolysis rises and becomesSlow. Therefore, maximum enzyme concentration should, between 1229.475-2213.055U/g, in order further to determine, adopt SDS-PAGEAnalyze enzymolysis liquid Degree of Enzymatic Hydrolysis, result is as Fig. 4. Along with the increase of enzyme concentration, be positioned at albumen between 66.4-97.2kDa by enzymolysis,Wherein the proteolysis in the 4th swimming lane is the most thorough, and in the 5th swimming lane, albumen has appearred again in 29.0kDa below, may be because shrimpDue to enzyme liquid oneself protein enzymolysis. Therefore, selecting enzyme concentration is that 1721.265U/g is advisable.
The present invention is 658.71U/g at enzyme concentration, and the time is 6.5h, and temperature is under the enzymatic hydrolysis condition of 40 DEG C, change systemPH, investigate its impact for degree of hydrolysis, experimental result is shown in Fig. 5. As seen from Figure 5, in the time that pH is less than 5.5, degree of hydrolysis withThe increase of pH and increasing, in the time that pH is 6.5, degree of hydrolysis reduces. Therefore, best pH is 5.5.
The present invention is that 658.71U/g, enzymolysis time are that 6.5h, pH are under 5.5 enzymatic hydrolysis condition at enzyme concentration, in equality of temperature notUnder degree, investigated its impact for a shrimp enzyme hydrolysis situation, result as shown in Figure 6. During from 30 DEG C to 35 DEG C, degree of hydrolysis increasesIn the time of 35 DEG C, reach the first peak value rapidly. Since 40 DEG C, degree of hydrolysis is along with temperature increases and slowly increases. At experimentationMiddle discovery, the enzymolysis liquid under 35 DEG C of reaction conditions has denseer shrimp fragrance, and to 45 DEG C of bars of 35 DEG C of similar degree of hydrolysisUnder part, frowziness. And for the consideration of low-carbon emission reduction in actual production, choose 35 DEG C for optimal reaction temperature.
Above single factor condition optimizing result, determine the reaction condition of a more suitable shrimp enzyme enzymolysis shrimp shell: timeBetween for 6.5h, pH are 5.5, temperature is that 35 DEG C and enzyme concentration are 1721.265U/g. But experiment of single factor cannot be investigated differenceInteraction between factor, for this reason, the inventor further adopts response surface method to obtain best enzymatic hydrolysis condition.
On the basis of experiment of single factor result, adopt Box-Behnken design principle (BBD), with a shrimp crude enzyme liquid waterThe degree of hydrolysis of separating the enzymolysis liquid of shrimp shell is response, access time, pH, temperature, four separate response factors of enzyme concentration, fromAnd carry out four factor three horizontal analysis. Adopt response surface method, the result after optimization is that described enzymolysis optimal conditions is: timeBetween 7h, pH6.0,40 DEG C of temperature, enzyme concentration 1678.93U/g.
The present invention, under enzymolysis optimal conditions, has carried out electrophoretic analysis to enzymolysis liquid, as shown in Figure 7, and by swimming lane 1 and swimmingThe contrast of the band in road 2, can know the hydrolysis through a shrimp enzyme liquid, and the molecular weight in enzymolysis liquid is 97.5-44.3 left and rightAlbumen be hydrolyzed more thorough. In swimming lane 2, the protein band color of molecular weight from 44.3 to 14.3 left and right is more shallow. Therefore,Under optimum enzymatic hydrolysis condition, the thick enzyme of shrimp head can be hydrolyzed shrimp shell efficiently, makes shrimp head become amino acid with the proteolysis in shrimp shellWith little peptide.
Embodiment 3 shrimps enzyme enzymolysis crab shell and crab pin
Crab shell and crab pin are thawed, after dried, crushed, add a shrimp enzyme to carry out enzymolysis, enzymatic hydrolysis condition is: time 5-9h, pH3.5-6.5, temperature 30-50 DEG C, enzyme concentration 1475-1878U/g, enzymolysis liquid go out enzyme live after defishying, then through liquidGu separate, after concentrate drying, make sea food seasoning raw material;
In like manner, inventor's prawn head enzyme enzymolysis crab shell and crab pin have also carried out single factor experiment and response surface analysis,Determine that eventually described enzymolysis optimal conditions is: time 7h, pH6.0,40 DEG C of temperature, enzyme concentration 1678U/g. With this understanding, SDS-PAGE testing result, as shown in Figure 8.
Wherein, swimming lane 1 and 2 is the front shrimp enzyme of reaction and crab pin, and swimming lane 3 is a shrimp enzyme enzymolysis liquid. By these three swimming lanesContrast, can find hydrolysis albumen mainly concentrate between molecular weight 40-120kDa.
This technological process is reasonably combined the production technology of multiple products, and shrimp crab processing fent is filledThe utilization dividing. On the one hand, this technological process takes full advantage of the endogenous enzymes self containing in shrimp processing fent and producedEnzymolysis process in journey, thus greatly reduce production cost; On the other hand, it is former that the present invention all adopts enzymatic isolation method production flavouringMaterial, it is compared with extraction with solvent extraction method of the prior art, and condition is more gentle, and can not produce secondary dirtDye, be more suitable in industrial production.
All distortion that those of ordinary skill in the art can directly derive or associate from the disclosure of invention, all shouldThink protection scope of the present invention.
Claims (3)
1. utilize shrimp crab leftover bits and pieces to prepare a method for material of spice, it is characterized in that: comprise the following steps:
Step 1, the preparation thick enzyme of shrimp head and a shrimp enzyme:
Shrimp head is smashed to pieces, is added buffer solution, with refrigerated centrifuge centrifugal 20min at 12000r/min, 4 DEG C, centrifugal complete after through 8Layer filter-cloth filtering, with except degrease, got supernatant; Supernatant is left standstill after 2 hours to centrifugal 20min at 12000r/min, 4 DEG C, system againObtain the thick enzyme of shrimp head;
By the thick enzyme of prepared shrimp head, slowly adding saturated mass fraction is 80% ammonium sulfate, leave standstill after 2h at 4 DEG C,Centrifugal 20min at 12000r/min, 4 DEG C, abandons supernatant again; In precipitation, add a small amount of buffer solution to dissolve, use 3.5Kda holeThe bag filter dialysis 24h of footpath size, makes a shrimp enzyme, at-20 DEG C, saves backup;
Step 2, the thick enzyme enzymolysis of shrimp head shrimp shell:
Shrimp shell is thawed, after dried, crushed, add the thick enzyme of shrimp head to carry out enzymolysis, enzymatic hydrolysis condition is: time 4-8h, pH3.5-6.5, temperature 30-50 DEG C, enzyme concentration 500-2300U/g, enzymolysis liquid go out enzyme live after defishying, then through liquid-solid separation, denseAfter contracting is dry, make sea food seasoning raw material;
Step 3, shrimp enzyme enzymolysis crab shell and crab pin:
Crab shell and crab pin are thawed, after dried, crushed, add a shrimp enzyme to carry out enzymolysis, enzymatic hydrolysis condition is: time 5-9h,PH3.5-6.5, temperature 30-50 DEG C, enzyme concentration 1475-1878U/g, enzymolysis liquid go out enzyme live after defishying, then through liquid-solidSeparate, after concentrate drying, make sea food seasoning raw material.
2. a kind of method of utilizing shrimp crab leftover bits and pieces to prepare material of spice as claimed in claim 1, is characterized in that: described inIn step 2, enzymatic hydrolysis condition is: time 7h, pH6.0,40 DEG C of temperature, enzyme concentration 1678.93U/g.
3. a kind of method of utilizing shrimp crab leftover bits and pieces to prepare material of spice as claimed in claim 1, is characterized in that: described inIn step 3, enzymatic hydrolysis condition is: time 7h, pH6.0,40 DEG C of temperature, enzyme concentration 1678U/g.
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