CN105581281B - A method of material of spice is prepared using shrimp crab leftover bits and pieces - Google Patents
A method of material of spice is prepared using shrimp crab leftover bits and pieces Download PDFInfo
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Abstract
The invention discloses a kind of methods for preparing material of spice using shrimp crab leftover bits and pieces, comprising the following steps: Step 1: the thick enzyme of preparation shrimp head and shrimp head enzyme；Step 2: the thick enzyme of shrimp head digests shrimp shell；Step 3: shrimp head enzyme enzymatic hydrolysis crab shell and crab foot.The present invention utilizes the thick enzyme hydrolysis shrimp crab leftover bits and pieces of shrimp head of the extraction from shrimp head to prepare seafood taste seasoning.Containing a variety of amino acid necessary to human body in shrimp crab enzymolysis liquid of the present invention, nutritive value is high, the product being prepared into using it, and the nutriments such as amino acid and minerals in product are conducive to the absorption of human body.Enzymolysis liquid after freeze-drying can make an addition to varieties of food items, such as seafood condiment, edible pigment, children and the nutrient rice milk of pregnant woman.
Description
Technical field
The present invention relates to the technical fields of cultivation processing more particularly to a kind of utilization shrimp crab leftover bits and pieces to prepare material of spice
Method.
Background technique
China possesses the Gold Coast line for containing the abundant fishery resources of tremendous potential, and total length is more than 3.2 ten thousand kilometers
Positioned at the whole world the 4th.It is national more and more more to pay attention to quality of life and health, therefore, fish shrimp crab etc. with epoch progress
Health-oriented products demand is growing day by day, and the fishery in China has welcome rare development spring.Wherein, the by-product in fish production
Deep processing be China's Fisheries Development new direction and new opportunities.For example, being had accumulated big in the production products such as peeled shrimp and crab meat
Measure shrimp crab leftover bits and pieces.It is studied according to relevant speciality, shrimp head and the chitin in shrimp shell containing 15% or so, protein content 30%
Left and right, remaining is mineral matter element, fat-soluble A and vitamin E etc., and wherein the content of calcium is the abundantest up to 35% left side
Secondly it is 4%-5% that the right side is phosphorus.Protein in shrimp shell and shrimp head contains required eight kinds must amino acid.Except lysine
Content is slightly lower, other seven kinds must amino acid content reach human body must amino acid demand minimum requirements.Crab leftover bits and pieces
In the battalion such as protein, carbohydrate, unsaturated fatty acid, lecithin, carotenoid, cephalin, chitin, cellulose
A point rich content is formed, also there is the trace minerals such as calcium, magnesium, phosphorus and various amino acid.Therefore, shrimp crab leftover bits and pieces is fabulous
Industrial processes raw material can apply to various aspects, commercial exploitation with higher.If these processing can be got a foothold
Material comprehensively utilized rather than it is simple abandon, on the one hand can bring economic benefit to enterprise, on the other hand also can
It reduces environmental pollution.
With flourishing for shrimp crab industry, the recycling and reusing of shrimp crab by-product also becomes the hot spot of the research of all circles.
Current research is concentrated mainly on the albumen in shrimp crab leftover bits and pieces using various methods, pigment and grease etc., recycling and reusing,
Deep processing appropriate is reprocessed, high nutrition, inexpensive new product is made.
Chen Limei et al. citric acid and naoh treatment shrimp shell, preparation are free of the chitin of protein and calcium.Duan Shan
Et al. probe into the autolysis of lactic acid bacteria for the method for the Protein Recovery in shrimp head and shrimp shell.Islem Younes utilize from
The protein in alkali protease removal shrimp shell extracted in Rockfish internal organ is to prepare chitin.
The a large amount of crab leftover bits and pieces that can be generated when traditional industry processes the crabs products such as crab meat, crab cream, wherein quantity is most
Be crab foot, the dorsal shield of crab.The current scientific research hot spot when exploitation of crab leftover bits and pieces, many researchers are under crab
The product of heel exploitation purposes multiplicity.For example, Fan Jianfeng et al. using swimming crab leftover bits and pieces as raw material, adds in the differential responses stage
Add different commercial enzymes, to extract anti-oxidation peptide and antibacterial peptide in crab leftover bits and pieces.Wang Xiaoqian et al. probes into day in swimming crab
The extraction of right pigment, purification process, to substitute synthetic food color.And Tao Xueming is using dehydrated alcohol as solvent extraction
Raw material of the crab oil as health care product in swimming crab blanking.
Zhang Lianfu et al. is using shrimp shell protein enzymatic hydrolyzate as raw material, then adds the xylose and glucose of proper proportion and be made and have
The seasoning of dense shrimp flavor.Chen Haihua et al. prepares seafood by the optimum enzymolysis condition that Response Surface Method optimizes Mytilus galloprovincialis enzyme
Taste seasoning.The water of Ren Xiane et al. first extracts the albumen in shrimp shell, and remaining shrimp shell extracts albumen with papain again, extracts
Liquid is concentrated again to wait preparation seafood taste seasoning.
Although in recent years, achieving biggish progress for the comprehensive utilization of shrimp crab both at home and abroad, still there is processing to walk
It is rapid relatively complicated, it is converted into the disadvantages such as industrial production feasibility is low.Operational height is sought by experiment, step is simple, at low cost
Shrimp crab leftover bits and pieces prepare seafood taste seasoning production method, upgrading for fishery industry line and improve and have great meaning
Justice.In view of this, the present inventor's research and devise a kind of method for preparing material of spice using shrimp crab leftover bits and pieces, this case by
This is generated.
Summary of the invention
It is existing to solve the purpose of the present invention is to provide a kind of method for preparing material of spice using shrimp crab leftover bits and pieces
The problem of technology Prawn crab leftover bits and pieces is largely wasted.
To achieve the goals above, the technical scheme adopted by the invention to solve the technical problem is that:
A method of material of spice is prepared using shrimp crab leftover bits and pieces, comprising the following steps:
Step 1: the thick enzyme of preparation shrimp head and shrimp head enzyme:
Shrimp head is smashed to pieces, buffer is added, is centrifuged 20min at 12000r/min, 4 DEG C with refrigerated centrifuge, has been centrifuged
By 8 layers of filter-cloth filtering to remove fat, supernatant is taken；It is centrifuged at 12000r/min, 4 DEG C again after supernatant is stood 2 hours
The thick enzyme of shrimp is made in 20min；
By the obtained thick enzyme of shrimp, it is slowly added to the ammonium sulfate that saturation mass fraction is 80%, stands 2h at 4 DEG C
Afterwards, then at 12000r/min, 4 DEG C it is centrifuged 20min, abandons supernatant；A small amount of buffer solution is added in precipitating, uses 3.5Kda
The bag filter dialysis of pore size for 24 hours, is made shrimp head enzyme, saves backup at -20 DEG C；
Step 2: the thick enzyme of shrimp head digests shrimp shell:
Shrimp shell is thawed, after dried, crushed, the thick enzyme of shrimp head is added and is digested, enzymatic hydrolysis condition are as follows: time 4-8h,
PH3.5-6.5,30-50 DEG C of temperature, enzyme concentration 500-2300U/g, enzymolysis liquid carries out defishying after enzyme deactivation is lived, then divides admittedly through liquid
From sea food seasoning raw material is made after concentrate drying；
Step 3: shrimp head enzyme enzymatic hydrolysis crab shell and crab foot:
Crab shell and crab foot are thawed, after dried, crushed, shrimp head enzyme is added and is digested, enzymatic hydrolysis condition are as follows: time 5-
9h, pH3.5-6.5,30-50 DEG C of temperature, enzyme concentration 1475-1878U/g, enzymolysis liquid carries out defishying after enzyme deactivation is lived, then through liquid
Gu separation, after concentrate drying, is made sea food seasoning raw material.
As the preferred embodiment of embodiment, in the step 2, enzymatic hydrolysis condition are as follows: time 7h, pH6.0, add by 40 DEG C of temperature
Enzyme amount 1678.93U/g.
As the preferred embodiment of embodiment, in the step 3, enzymatic hydrolysis condition are as follows: time 7h, pH6.0, add by 40 DEG C of temperature
Enzyme amount 1678U/g.
The present invention utilizes the thick enzyme hydrolysis shrimp crab leftover bits and pieces of shrimp head of the extraction from shrimp head to prepare seafood taste seasoning.First
Using simply smashing to pieces and refrigerated centrifuge, the thick enzyme of endogenous shrimp extracted from shrimp head；Then it is digested respectively using shrimp head enzyme
Shrimp shell, crab shell and crab foot prepare various material of spice.Contain a variety of amino acid necessary to human body, nutrition in shrimp crab enzymolysis liquid
Value is high, the product being prepared into using it, and the nutriments such as amino acid and minerals in product are conducive to the absorption of human body.Freezing
Enzymolysis liquid after drying can make an addition to varieties of food items, such as: seafood condiment, edible pigment, children and the nutrient rice milk of pregnant woman.
Detailed description of the invention
The SDS- polyacrylamid gel electrophoresis of Fig. 1 shrimp head enzyme solution；
Influence of Fig. 2 time for shrimp head enzyme solution degree of hydrolysis；
Influence of Fig. 3 enzyme concentration for shrimp enzymolysis liquid degree of hydrolysis；
The electrophoretic analysis figure of Fig. 4 difference enzyme concentration enzymolysis liquid；Wherein, M:Marker, 1: shrimp shell (before reaction), 2: shrimp head enzyme
Liquid (before reaction), 3: enzymolysis liquid (enzyme concentration 1229.475U/g), 4: enzymolysis liquid (enzyme concentration 1721.265U/g), 5: enzymatic hydrolysis
Liquid (enzyme concentration 2213.055U/g), 6: (shrimp shell is not added, after reaction) in blank enzymolysis liquid；
The influence of Fig. 5 pH prawn head enzyme solution degree of hydrolysis；
The influence of Fig. 6 temperature prawn head enzyme solution degree of hydrolysis；
The electrophoretogram of the thick enzyme hydrolysis shrimp shell of shrimp head under Fig. 7 optimum enzymolysis condition；Wherein, M:Marker, 1: shrimp head crude enzyme liquid,
2: shrimp shell enzymolysis liquid (under optimal conditions)；
Under the optimal enzymatic hydrolysis condition of Fig. 8, the electrophoretogram of the thick enzyme hydrolysis crab foot of shrimp head；Wherein, M:Marker, 1: shrimp head crude enzyme liquid
(before reaction), 2: crab foot (before reaction), 3: shrimp shell enzymolysis liquid (under optimal conditions).
Specific embodiment
Below in conjunction with the drawings and specific embodiments with the invention will be further described.Raw material used in embodiment: shrimp
Head: the fresh South America prawn bought by Xiamen City, the market of farm produce, Jimei District is stripped shrimp head collection and obtains；Shrimp shell: from good fortune
The Zhangzhou City the Jian Sheng development zone Zhangpu County Huang Cang Ying Feng Food Co., Ltd, peeled shrimp processing in generate shrimp shell leftover bits and pieces；Crab foot: it comes from
Crab leftover bits and pieces is generated in Zhangzhou City, the Fujian Province development zone Zhangpu County Huang Cang Ying Feng Food Co., Ltd, crab meat processing.
Embodiment 1 prepares the thick enzyme of shrimp head and shrimp head enzyme
Shrimp head is smashed to pieces, buffer is added, is centrifuged 20min at 12000r/min, 4 DEG C with refrigerated centrifuge, has been centrifuged
By 8 layers of filter-cloth filtering to remove fat, supernatant is taken；It is centrifuged at 12000r/min, 4 DEG C again after supernatant is stood 2 hours
The thick enzyme of shrimp is made in 20min；
By the obtained thick enzyme of shrimp, it is slowly added to the ammonium sulfate that saturation mass fraction is 80%, stands 2h at 4 DEG C
Afterwards, then at 12000r/min, 4 DEG C it is centrifuged 20min, abandons supernatant；A small amount of buffer solution is added in precipitating, uses 3.5Kda
The bag filter dialysis of pore size for 24 hours, is made shrimp head enzyme, saves backup at -20 DEG C.
Using the molecular mass of the method measurement shrimp head enzyme of SDS- polyacrylamide gel electrophoresis (SDS-PAGE), with standard
Albumen Marker (14.3-97.2kDa) is used as standard control.Wherein, the resolving gel concentration of SDS-PAGE is 12%, and concentration glue is dense
Degree is 5%.
From it is recognised that in molecular weight 14.3-97.5kDa, there are a plurality of in shrimp head endogenous protein crude enzyme liquid in Fig. 1
Band.Therefore, shrimp head enzyme crude liquid may be to be made of multiple protein enzyme, be worth continuing to study it for the water of shrimp shell and crab foot
Solve effect.
The thick enzyme of embodiment 2 shrimp head digests shrimp shell
Shrimp shell is thawed, after dried, crushed, the thick enzyme of shrimp head is added and is digested, enzymatic hydrolysis condition are as follows: time 4-8h,
PH3.5-6.5,30-50 DEG C of temperature, enzyme concentration 500-2300U/g, enzymolysis liquid carries out defishying after enzyme deactivation is lived, then divides admittedly through liquid
From sea food seasoning raw material is made after concentrate drying；
The present invention has investigated the influence of the thick enzyme hydrolysis degree of time prawn head, is 658.71U/g, pH 5 keeping enzyme concentration
Under the enzymatic hydrolysis condition for being 40 DEG C with temperature, chooses 6 different times (4,5,5.5,6,6.5,8h) and investigate it respectively for hydrolysis
The influence of degree.As shown in Figure 2, with the growth in reaction time, the substrate in system is reduced, and reaction product increases, substrate and enzyme
Between effective collision reduce, degree of hydrolysis changes after 6.5 hours slow.Therefore, choosing 6.5 hours is the reaction time.
The present invention is 6.5h, pH 5.5 in the retention time, under the enzymatic hydrolysis condition that temperature is 35 DEG C, has investigated enzyme concentration pair
In the influence of degree of hydrolysis.Experimental result is shown in Fig. 3.In enzymatic reaction, when the close saturation of the concentration of enzyme in substrate, reaction
Velocity variations will slow down.Related document report when using commercial enzyme, when the excessive concentration of enzyme, will lead to mutual between enzyme
Inhibit and hydrolyze, reduces enzyme activity.And in this experiment, since shrimp head enzyme only passes through coarse extraction, also contain in enzyme solution a large amount of
The albumen of shrimp head itself, causes degree of hydrolysis that can increase always with the increase of enzyme concentration.From figure 3, it can be seen that when enzyme
When amount is from 0-1000U/g, degree of hydrolysis rises rapid.When enzyme concentration is from 1229.475-2213.055U/g, degree of hydrolysis, which rises, to be become
It is slow.Therefore, maximum enzyme concentration should be between 1229.475-2213.055U/g, in order to further determine using SDS-PAGE
Enzymolysis liquid Degree of Enzymatic Hydrolysis is analyzed, as a result such as Fig. 4.With the increase of enzyme concentration, the albumen between 66.4-97.2kDa is digested,
Proteolysis wherein in the 4th swimming lane is the most thorough, and albumen occurs again in 29.0kDa or less in the 5th swimming lane, it may be possible to because of shrimp
Caused by head enzyme solution oneself protein enzymatic hydrolysis.Therefore, enzyme concentration is selected to be advisable for 1721.265U/g.
The present invention is 658.71U/g in enzyme concentration, and time 6.5h, temperature is to change system under 40 DEG C of enzymatic hydrolysis condition
PH, investigate its influence for degree of hydrolysis, experimental result is shown in Fig. 5.As seen from Figure 5, when pH is less than 5.5, degree of hydrolysis with
The increase of pH and increase, when pH be 6.5 when degree of hydrolysis reduce.Therefore, Optimal pH 5.5.
The present invention is under the enzymatic hydrolysis condition that enzyme concentration is 658.71U/g, enzymolysis time 6.5h, pH are 5.5, in not equality of temperature
Its influence for shrimp head enzyme hydrolysis situation is investigated under degree, as a result as shown in Figure 6.When from 30 DEG C to 35 DEG C, degree of hydrolysis increases
Rapidly, reach the first peak value at 35 DEG C.Since 40 DEG C, degree of hydrolysis is slowly increased as temperature increases.In experimentation
Middle discovery, the enzymolysis liquid under 35 DEG C of reaction conditions have denseer shrimp fragrance, and in 45 DEG C of items of degree of hydrolysis similar to 35 DEG C
Under part, there is stink.It and is optimal reaction temperature for the considerations of low-carbon emission reduction, choosing 35 DEG C in actual production.
The above single factor test condition optimizing result, it is determined that the reaction condition of a convenient shrimp head enzyme enzymatic hydrolysis shrimp shell: when
Between be 6.5h, pH 5.5, temperature is 35 DEG C and enzyme concentration is 1721.265U/g.But experiment of single factor can not investigate difference
Interaction between factor, for this purpose, the present inventor further uses response surface method to obtain optimal enzymatic hydrolysis condition.
On the basis of experiment of single factor result, using Box-Behnken design principle (BBD), with shrimp head crude enzyme liquid water
The degree of hydrolysis for solving the enzymolysis liquid of shrimp shell is response, and access time, pH, temperature, enzyme concentration four respond factors independently of each other, from
And carry out four factors, three horizontal analysis.Using response surface method, the result after optimization is the enzymatic hydrolysis optimal conditions are as follows: when
Between 7h, pH6.0,40 DEG C of temperature, enzyme concentration 1678.93U/g.
The present invention has carried out electrophoretic analysis in the case where digesting optimal conditions, to enzymolysis liquid, as shown in fig. 7, passing through swimming lane 1 and swimming
The comparison of band in road 2, it is known that the molecular weight in enzymolysis liquid is 97.5-44.3 or so by the hydrolysis of shrimp head enzyme solution
The hydrolyzed ratio of albumen more thoroughly.The protein band color of 2 middle-molecular-weihydroxyethyl from 44.3 to 14.3 or so of swimming lane is shallower.Therefore,
Under optimal enzymatic hydrolysis condition, the thick enzyme of shrimp head can efficiently hydrolyze shrimp shell, make the proteolysis in shrimp head and shrimp shell at amino acid
And small peptide.
3 shrimp of embodiment head enzyme enzymatic hydrolysis crab shell and crab foot
Crab shell and crab foot are thawed, after dried, crushed, shrimp head enzyme is added and is digested, enzymatic hydrolysis condition are as follows: time 5-
9h, pH3.5-6.5,30-50 DEG C of temperature, enzyme concentration 1475-1878U/g, enzymolysis liquid carries out defishying after enzyme deactivation is lived, then through liquid
Gu separation, after concentrate drying, is made sea food seasoning raw material；
Similarly, the present inventor's prawn head enzyme enzymatic hydrolysis crab shell and crab foot have also carried out single factor experiment and response surface analysis, most
The enzymatic hydrolysis optimal conditions is determined eventually are as follows: time 7h, pH6.0,40 DEG C of temperature, enzyme concentration 1678U/g.With this condition, SDS-
PAGE testing result, as shown in Figure 8.
Wherein, swimming lane 1 and 2 is shrimp head enzyme and crab foot before reacting, and swimming lane 3 is shrimp head enzyme enzymolysis liquid.Pass through this three swimming lanes
Comparison, it can be found that hydrolysis albumen be concentrated mainly between molecular weight 40-120kDa.
The production technology of multiple products is reasonably combined in this process flow, so that shrimp crab processing fent is filled
The utilization divided.On the one hand, which takes full advantage of the endogenous enzymes itself contained in shrimp processing fent and produced
Enzymolysis process in journey, thus greatly reduces production cost；On the other hand, it is former to be all made of enzymatic isolation method production flavouring by the present invention
Material, compared with solvent extraction method in the prior art and extraction, condition is more mild, and will not generate secondary dirt
Dye, more suitable for industrial production.
All deformations that those skilled in the art directly can export or associate from the disclosure of invention, should all
It is considered protection scope of the present invention.
Claims (1)
1. a kind of method for preparing material of spice using shrimp crab leftover bits and pieces, it is characterised in that: the following steps are included:
Step 1: the thick enzyme of preparation shrimp head and shrimp head enzyme:
Shrimp head is smashed to pieces, buffer is added, 20min is centrifuged at 12000r/min, 4 DEG C with refrigerated centrifuge, through 8 after being centrifuged
Layer filter-cloth filtering takes supernatant to remove fat；It is centrifuged 20min at 12000r/min, 4 DEG C again after supernatant is stood 2 hours, is made
Obtain the thick enzyme of shrimp；
By the obtained thick enzyme of shrimp, it is slowly added to the ammonium sulfate that saturation mass fraction is 80%, after standing 2h at 4 DEG C,
20min is centrifuged at 12000r/min, 4 DEG C again, abandons supernatant；A small amount of buffer solution is added in precipitating, with the hole 3.5Kda
The bag filter dialysis of diameter size for 24 hours, is made shrimp head enzyme, saves backup at -20 DEG C；
Step 2: the thick enzyme of shrimp head digests shrimp shell:
Shrimp shell is thawed, after dried, crushed, the thick enzyme of shrimp head is added and is digested, enzymatic hydrolysis condition are as follows: time 7h, pH6.0, temperature
40 DEG C, enzyme concentration 1678.93U/g of degree, enzymolysis liquid carry out defishying after enzyme deactivation is lived, then through solid-liquor separation, after concentrate drying, system
At sea food seasoning raw material；
Step 3: shrimp head enzyme enzymatic hydrolysis crab shell and crab foot:
Crab shell and crab foot are thawed, after dried, crushed, shrimp head enzyme is added and is digested, enzymatic hydrolysis condition are as follows: time 7h,
PH6.0,40 DEG C of temperature, enzyme concentration 1678U/g, enzymolysis liquid carries out defishying after enzyme deactivation is lived, then through solid-liquor separation, is concentrated and dried
Afterwards, sea food seasoning raw material is made.
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Cited By (1)
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| RU2795474C2 (en) * | 2020-02-11 | 2023-05-03 | Денис Владимирович Полещук | Method for processing waste obtained after cutting crabs |
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| CN106360623A (en) * | 2016-11-19 | 2017-02-01 | 朱莉 | Comprehensive aquatic product leftover utilization method |
| CN106942647A (en) * | 2017-01-23 | 2017-07-14 | 岱山县通衢水产食品有限公司 | One breeder tail crab pin preparation technology |
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