CN107821922A - A kind of de- bitter enzymolysis of small oppossum shrimp of marine products prepares method and the application of full shrimp slurry - Google Patents
A kind of de- bitter enzymolysis of small oppossum shrimp of marine products prepares method and the application of full shrimp slurry Download PDFInfo
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Abstract
The method that the de- bitter enzymolysis of the small oppossum shrimp of marine products prepares the full shrimp slurry rich in small peptide, comprises the following steps：1）Granulating, will be using the full fish of fresh small fish of marine products as raw material fish, protein body of the classification grinding to full-size in below 3mm；2）Micronized, protein particle slurry more than ultra high shear to 150 mesh；3）10 20min are boiled in microminiaturization, 90 100 DEG C of boilings, are cooled to 55 DEG C~75 DEG C of microminiaturization temperature, and ultrasonic probe stretches into containing tank effect microminiaturization ultrasonic wave, and small corpuscular protein activation slurry is made；4）Enzymolysis, microwave surround be heated to 40 DEG C~65 DEG C of enzyme digestion reaction temperature, by raw material shrimp weigh 0.02~2% protease be added to particulate protein slurry in, microwave, ultrasonic wave, enzyme digestion reaction cycle alternation apply 10 20 minutes/time, total 20~60min of enzyme digestion reaction time, obtain enzymolysis protein slurry；Enzyme deactivation, it is 40 60wt% to be concentrated in vacuo to moisture, produces the small oppossum shrimp of marine products rich in small peptide and digests full shrimp slurry；The preparation method, forage protein source of the shrimp med as the economic feed of aquaculture can be substituted.
Description
Technical field
The present invention relates to a kind of protein digestion technical field, prepared by specially a kind of small oppossum shrimp enzymolysis of marine products is rich in small peptide
Full shrimp slurry preparation method and application.
Background technology
One in fishes and shrimps feed is important and raw material that cost is higher is protein.The abundant marine fishes of species are albumen
The main object of enzymolysis, after marine protein digests, one of its hydrolysate is small peptide.Small peptide should be the general name of small-molecular peptides, both
Include the molecules small peptide of 2~3 amino acid residues, the oligopeptides of 10 is also less than including amino acid residue, i.e., actually should
In, amino acid residue can be classified as small peptide product in the small peptide product of small molecule of 10 or so.Small peptide has good function
It characteristic, can not only quickly be absorbed in the form of complete by organism, directly participate in the synthesis of histone, and there is antibacterial
Physiological function with immunity is improved, realizes the high-valued of protein.Research has shown that small peptide substitutes a certain proportion of fish meal and made
Cultivation fish is fed for feed, in addition to it can promote to cultivate fish growth, can also strengthen the autoimmune function of feeding cultivation fish, drop
They low disease incident.
Oppossum shrimp, it is several to be commonly called as shrimp, is a kind of miniature shrimp of marine products, and the bodily form is slightly flat-sided, body surface gray, and body grows 2~15mm, at me
State Liaodong Peninsula, Zhejiang and the band of Coast of Guangdong Province one are distributed.The nutritive value of oppossum shrimp is high, protein, lipid and micro member
Cellulose content is extremely abundant.Because the bodily form is small, crust is meagre, fine and tender taste, and the phenomenon of self-dissolving occurs after oppossum shrimp disengaging seawater,
Freshness is difficult to maintain, can edible part it is less.Thus the oppossum shrimp overwhelming majority of fisherman's capture is purchased by shrimp med processing enterprise,
Shrimp med is processed into by processes such as boiling, squeezing, drying and crushing, is sold as fodder protein additive.
Therefore, how well using low value shrimp come to produce the shrimp products of high level be the striving direction that has very much social value.
It is exactly a wherein practical way that enzymolysis oppossum shrimp, which obtains the shrimp slurry rich in small peptide,.
However, the mechanism of proteolysis is extremely complex, except improving enzymolysis efficiency, remove bitter taste had turned into everybody pursuit already
Target.Problem proposes as follows：
1）Enzymolysis efficiency is low.Conventional water enzyme digestion needs 55-65 DEG C, more than 7 hours, wastes time and energy, is also easy to go bad；By
In the resolvase itself the defects of, enzyme digestion reaction is unfavorable for the production of polypeptide and prepare, such as:Resolvase acid, alkali, heat and
Mutability in the media such as organic solvent, activity are reduced or lost；Enzyme is remained in solution after the reaction, causes enzyme reaction to be difficult to connect
Continuousization and automation.
2）There is bitter taste, the bitter taste of protein hydrolysate is mainly protein hydrolysis caused by the bitter peptides in hydrolysate
Matter destroys the structure of original albumen, makes to be hidden in intramolecular hydrophobic amino acid such as leucine, isoleucine, benzene originally
Alanine etc. is exposed to molecular surface, thus the hydrolysate of enzyme has different degrees of bitter taste to produce.These contain hydrophobicity
The bitter peptides of amino acid residue, its chain length is from 2,3 to more than ten amino acid.Bitter peptides turn into enzymolysis oppossum shrimp product and are applied to
One major obstacle of aquaculture.Shrimp slurry is good again, and rich in small peptide, but for feeding cultured prawn, prawn is not liked to eat, and this is also
Digest the main obstacle of oppossum shrimp product promotion.Bitter peptides can be also detected even if micromole, and the sensation of bitter taste also compares
Sweet taste, saline taste and tart flavour duration length, therefore simple bitter taste is the basic original of culturing fish and shrimp class food refusal bitter substance
Cause.
Hardship how is taken off, but also needs to consider that de- bitter operation does not interfere with range of hydrolysed peptides loss or destroyed.Conventional method is：
1）Filter off and remove, charcoal absorption, or ultrafiltration membrance filter remove.But it is not first during enzymatic isolation method production fish/shrimp slurry
With discoloration, activated carbon can only remove color, and the too fine cost of milipore filter is very high, therefore milipore filter and activated carbon and not apply to.
2）Cover, such as the bitter taste of caffeine, when adding sucrose, bitter taste is blanked is suppressed in other words, i.e. gustatory
Feel without so bitter.
In a word, how high efficiency digests, the de- hardship of low cost obtains the full shrimp slurry rich in small peptide, is that the technique of urgent need solution is difficult
Topic.
The content of the invention
In view of the above-mentioned problems of the prior art, the purpose of the present invention is high efficiency enzymolysis, clean de- hardship and does not damage peptide
Ground obtains the full shrimp slurry rich in small peptide, there is provided a kind of small oppossum shrimp enzymolysis of marine products prepare the full shrimp slurry rich in small peptide preparation method and
Using.
The present invention is achieved in that the method that a kind of de- bitter enzymolysis of the small oppossum shrimp of marine products prepares the full shrimp slurry rich in small peptide,
Comprise the following steps：
1）Granulating, it will be freeze-dried to water content in 50wt%-70wt%, be crushed to using the small oppossum shrimp of fresh marine products as raw material shrimp
Protein body of the full-size in below 3mm；
2）Micronized, according to protein body and water quality than 1:0.2 adds 0-3 DEG C of pure water, ultra high shear in protein body
Protein particle slurry more than to 150 mesh, it is 1 to add pure water to raw material shrimp and water quality ratio:0.5~5；
3）Microminiaturization, protein particle slurry carry out microwave and surround heating, and 90-100 DEG C of boiling boils 10-20min, be cooled to microminiaturization temperature
55 DEG C~75 DEG C, ultrasonic probe stretches into protein particle slurry and acts on microminiaturization ultrasonic wave so that the protein particle of protein particle slurry
Diameter is reduced to below micron order or micron order, and small corpuscular protein activation slurry is made；
4）Enzymolysis, slurry progress microwave encirclement is activated to small corpuscular protein and is heated to 40 DEG C~65 DEG C of enzyme digestion reaction temperature, by raw material
The protease that fish weighs 0.01~2wt% is fixed on dispersible carrier surface, then dispersible carrier is uniformly fixed on to multiple preset body surfaces
Face, microwave, ultrasound, small corpuscular protein activation slurry are immersed in preset body surface face enzyme digestion reaction cycle alternation and applied 10-20min/ times,
Total 20~60min of enzyme digestion reaction time, obtain enzymolysis protein slurry；
5）Selective Separation, enzymolysis protein is starched and carries out multi-stage booster filtering, Selective Separation goes out molecule from enzymolysis protein slurry
The oligopeptide in 1000-4000Da is measured, residue enzymolysis protein milk is de- bitter enzymolysis protein slurry；The de- bitter enzymolysis protein slurry is true
It is 40-60wt% that sky, which is concentrated into moisture, produces the small oppossum shrimp of marine products rich in small peptide and digests full shrimp slurry；
Step 1）-5）In remaining oxygen<Carried out in 5% nitrogen environmental protection.
Further, the preset body is polyethylene board, and dispersible carrier is fixed on multiple preset body surface faces, and referring to will be described
Dispersible carrier even adhesion to thin polyethylene board upper and lower surface, it is finally that multiple thin polyethylene board compartments are multiple every assembling
The flase floor of concentric circles annular composition.
Further, the preset body is film, and the film upper surface has been spaced a plurality of isolation fin, it is described every
From the sine curve that fin is in same period, the film upper and lower surface even adhesion has dispersible carrier, and then film is outside axle
Side face is wound into the preset body coiling body of certain diameter.
Further, the microwave surrounds heating, refers to that tubular container perisporium interval is equipped with multiple microwave generators, to egg
White particulate slurry carries out microwave and surrounds heating.
Further, the effect microminiaturization ultrasonic wave, diameter 10-40mm ultrasonic probes stretch into tubular container in pipe
The depths of the heart 1/3, multiple ultrasonic probes arranged for interval on tubular container length direction, with frequency 20-40kHz, field strength 70-
100w/cm2, ultrasonication 3-10 minutes.
Further, it is to prepare Fe that the protease, which is fixed on dispersible carrier surface,3O4Magnetic nanoparticle, it is hydride modified
To Fe3O4Magnetic nanoparticle surface obtains micro- magnetic core, and protease covalent coupling is obtained into scattered silicon ribozyme to micro- magnetic core surface
Particle.
A kind of de- bitter enzymolysis of small oppossum shrimp of the marine products prepares the alternating action used in method of the full shrimp slurry rich in small peptide
Device, it is to pass through friendship that the microwave, ultrasound, small corpuscular protein, which activate slurry and be immersed in enzyme digestion reaction cycle alternation application in preset body,
Realized for acting device, the alternating action device includes enzyme immobilization reaction tube, microwave tube and the ultrasonic wave of head and the tail connection
Pipe, the flase floor or the preset body coiling body are fixed in enzyme immobilization reaction tube, and slurries are with certain pressure full of described
Alternating action device, and slurries in alternating action device per after certain intermittent time in alternating action device circulation primary；Between
Have a rest during the time, slurries receive microwave action in microwave tube, and slurries receive ul-trasonic irradiation, enzyme immobilization reaction in ultrasonic wave duct
Enzyme digestion reaction occurs in pipe.
A kind of de- bitter enzymolysis of small oppossum shrimp of the marine products prepares the multi-stage booster used in the method for the full shrimp slurry rich in small peptide
Filter, including back taper annular canister, back taper annular canister entrance, centre and the outlet be respectively fixed with first order filter membrane,
Second level filter membrane and third level filter membrane, internal cone wall are provided with recoil mouth, are slided on back taper annular canister internal cone wall axis and be provided with cone
Plug, tapered plug reach block the recoil mouth, are moved after tapered plug and decontrol the recoil mouth.
What a kind of method that small oppossum shrimp of the marine products takes off bitter full shrimp slurry of the enzymolysis preparation rich in small peptide obtained is rich in small peptide
The small oppossum shrimp of marine products digests full shrimp slurry, and the small oppossum shrimp of marine products digests full shrimp slurry and is in a liquid state pulpous state, and wherein amino acid residue is at 10
The small peptide content of following small molecule is 15~20 wt%.
A kind of small oppossum shrimp of the marine products rich in small peptide digests application of the full shrimp slurry in aquaculture, with accounting for fish meal gross weight
The full shrimp slurry replacement shrimp med of the small oppossum shrimp enzymolysis of the marine products for measuring at least 10wt% continuously feeds shrimps in culture.
By mainly improving, following what time to reach, " how high efficiency is digested the present invention, the de- hardship of low cost is obtained rich in small peptide
The purpose of full shrimp slurry ",
Improvement in terms of synergy：
1st, granulate->Micronized->Microminiaturization, the condition of small molecule is created for enzymolysis protein source
（1）Coordinate step by step and accomplish microminiaturization as far as possible
Proteolysis is at least divided into two stages, the first step, and enzyme molecule is connected with small protein body；Second step, hydrolysis occur,
Discharge great amount of soluble peptide and amino acid.
The step 1 of the present invention）-3）Purpose be exactly to obtain protein body as small as possible, step 1）First fraction
The broken full-size that arrives is in below 3mm, step 2）High speed shear is into particulate slurry, step 3）Ultrasonic vibration microminiaturization after boiling, ultrasound
Ripple is a kind of compressional wave that vibration frequency is more than 20KHz, and ultrasonication produces cavitation in liquid medium, and ultrasound can make liquid be situated between
Microvesicle, subsequent microbubble ruptures, beneficial to protein particle diameter microminiaturization are formed in matter.
（2）Inexpensive microminiaturization is to 150 mesh above is being not easily achieved.
Micronized realizes one's own small innovation, is indigenous method.Prior art accomplishes powder more than 150 mesh
Particle degree, it is necessary to buy Ultramicrofine crasher or colloid mill or fluidized bed air flow crusher, these equipment are very expensive.
And it is little Wei enterprises to do fish meal substitute, the machine on investment buying is so tall and big is unrealistic, but wants to reach this purpose,
What if.
High speed shear, small rotating vane, driven with import super high speed motor of the rotating speed more than 30000 revs/min, blade is at a high speed
The protein body of liquid dispersion is sheared, to increase shear effect, installs tapered sleeve additional around blade, tapered sleeve convection body, which has, gathers work
With the tapered sleeve inwall has fin, increases impact effects.After installing tapered sleeve additional, blade high speed agitation shearing, cone under the gathering of tapered sleeve
Liquid height is higher by one section than surrounding in set, forms the swabbing action to tapered sleeve peripheral liquid, and tapered sleeve periphery protein body is continuous
It is inhaled into tapered sleeve and is sheared crushing.Through laser detection, shearing grinding particle size is more than 150 mesh.That is, present invention superelevation
The scalp of speed motor solves high speed shear to the technical problem of 150 mesh granularities, low cost.
The key point of above-mentioned realization is that high rotating speed driving vanelets are sheared in tapered sleeve, can go out 5kg particulates per 5-10 minutes
Slurry, therefore output efficiency per minute is not high, has much room for improvement.
, first freeze after heat, the activation of small molecule of ultrasonic wave and microwave, produce from protein particle slurry->The activation of protein activation slurry
Effect
Step 1）Freezing, step 2）Middle microwave surrounds heating boiling and boiled, step 4）Microwave, ultrasound before enzyme digestion reaction are alternately applied
Add, heat after on the one hand first freezing, thermal shock, loose protein particulate institutional framework；On the other hand, microwave is utilized so that liquid
Quick orientation rotates and frictional heat the polar molecule of body medium repeatedly under microwave high-frequency electric field action, and this heating is from thing
Start inside matter, quickly reach required temperature；Ultrasonic vibration causes the repeatedly quick orientation of polar molecule to rotate, and also causes
Protein structure is looser, and ultrasonic wave not only causes protein particle diameter microminiaturization using cavitation, while high field intensity is super
More hydrophobic groups can be exposed to protein molecule surface by sound wave.The application point that can be more combined with enzyme is exposed,
Accelerate hydrolysis, degree of hydrolysis greatly improves
Therefore, the present invention is by hot after first freezing, the collective effect of ultrasonic wave and microwave, generate from protein particle slurry->Albumen is lived
The activation of slurrying.
, protease molecule contacted with liquid medium it is decentralized contact, enzymolysis time greatly reduces
Effect of both above has caused protein digestion to be provided with small and activation protein body, and this is significantly
The speed of protease hydrolyzed is accelerated, has been able to realize rapid enzymolysis.Periodicity stirring in enzymolysis process is exactly to be
The protease is allowed to spread more evenly across around the small molecule particle for more needing to hydrolyze.
Proteolysis is at least divided into two stages, the first step, and enzyme molecule is connected with small protein body；Second step, hydrolysis
Occur, discharge great amount of soluble peptide and amino acid.In order to quickly cause more enzyme molecules and small egg in the first step
White particle contact, it is then that carrier is dispersed and be fixed on preset body surface face present invention employs protease dispersible carrier
Method, such as flase floor or polyethylene rod both accomplish dispersing contact, can reclaim protease at any time again, accelerate enzyme digestion reaction
Speed, enzymolysis time were reduced to 20-60min by more than 7 hours of prior art.
, microwave, ultrasound, enzymolysis alternating action
For step 4）Enzymolysis, devise alternating action device, activating slurry to small corpuscular protein carries out microwave, ultrasound and enzymolysis
Alternating apply, the enzyme immobilization that the alternating action device is included head and the tail connection by the alternating action device that connects from beginning to end is anti-
Ying Guan, microwave tube and ultrasonic wave duct, the alternating action unit efficiency is high, substantially increases the enzymolysis degree of slurries so that small peptide contains
Amount reaches 15-20wt%.
The improvement of de- bitter aspect：
1st, back taper annular canister entrance, centre, outlet dress three-level filter membrane plate
Back taper annular canister entrance, centre, outlet dress three-level filter membrane plate, liquid are acted on ingress filtering film with certain pressure, led to
The fluid pressure for crossing filter membrane necessarily reduces, and in order to make up this pressure decline, devises back taper annular canister, back taper annular canister makes
Three-stage filtration cross section reduce step by step, before the first order, the first order to the second level, the osmotic pressure of the second level to the third level step by step
Increase.
, the recoil mouth of internal cone wall, tapered plug coordinate
By setting the recoil mouth of internal cone wall, tapered plug to coordinate so that filter can wet sterilization, also can will retain Liquid backwash
Wash, be from the third level, the second level, the first order filter membrane plate during backwash, the cross section passed through increases step by step, pressure needed for recoil by
Level reduce, greatly facilitate backwash so that beneficial to recycle, greatly reduce separation costs, compared with purchase ultrafiltration apparatus,
The expense of hard work sequence is only taken off, it is per ton to reduce 500-700 members using ultra filtration equipment than purchase.
The small oppossum shrimp enzymolysis of marine products prepares the preparation method of the full shrimp slurry rich in small peptide, and enzymolysis efficiency is high, production cost
It is low, shrimp med can be substituted and be used for aquaculture, can be as the forage protein source of the economic feed of aquaculture.
The present invention takes full advantage of the living marine resources of China's abundant using the small oppossum shrimp of marine products of low value as raw material,
The small peptide product of pulpous state of high value is prepared using zymolysis technique, energy consumption is low and technological process is short, remains the nutritional ingredient of full fish；
Transition for China's fish meal processing industry product opens new approach.
Brief description of the drawings
Fig. 1 is the alternating action device signal for the method that the small oppossum shrimp enzymolysis of invention marine products prepares the full shrimp slurry rich in small peptide
Figure.
Fig. 2 is the transversal of the preset body coiling body for the method that the small oppossum shrimp enzymolysis of invention marine products prepares the full shrimp slurry rich in small peptide
Face figure.
Fig. 3 is the expansion of the preset body coiling body for the method that the small oppossum shrimp enzymolysis of invention marine products prepares the full shrimp slurry rich in small peptide
Figure.
Fig. 4 is the multi-stage booster filter for the method that the small oppossum shrimp enzymolysis of marine products of the present invention prepares the full shrimp slurry rich in small peptide
The main sectional view in pressurizing filter state.
Fig. 5 is the multi-stage booster filter for the method that the small oppossum shrimp enzymolysis of marine products of the present invention prepares the full shrimp slurry rich in small peptide
In decompression backwash state main sectional view.
Fig. 6 is the multi-stage booster filter for the method that the small oppossum shrimp enzymolysis of marine products of the present invention prepares the full shrimp slurry rich in small peptide
Schematic diagram when rinsing well.
1 enzyme immobilization reaction tube, 2 microwave tubes, 3 ultraphonic pipes, 4 microwave generators, 5 preset body coiling bodies, 6 ultrasounds
Probe, 7 inlet pumps, 8 circulating pumps, 9 drain pumps, 10 intermediate valves, 11 inlet valves, 12 outlet valves, 13 intake valves, 14 rows
Air valve,
5.1 axles, 5.2 films, 5.3 isolation fins, 5.4 dispersible carriers,
20 multi-stage booster filters, 21 back taper annular canisters, 22 first order filter membranes, 23 second level filter membranes, 24 the 3rd
Level filter membrane, 25 tapered plugs, 26 guides, 27 liquid-in pipes, 28 drain pipes, 29 recoil tubes, 30 first backwash mouths, 31 the
Two backwash mouths,
32 close water plug.
Embodiment
Embodiments of the invention are elaborated below in conjunction with accompanying drawing, but are not limited to the scope of the present invention.
Embodiment 1
A kind of small oppossum shrimp enzymolysis of marine products prepares the preparation method of the full shrimp slurry rich in small peptide, comprises the following steps：
1）Granulating, will be freeze-dried to water content in 50wt%-70wt%, fraction using the small oppossum shrimp of fresh marine products as raw material shrimp
It is broken to below 3mm protein body；
2）Micronized, according to protein body and water quality than 1:0.2 adds 0-3 DEG C of pure water, ultra high shear in protein body
Protein particle slurry more than to 150 mesh, it is 1 to add pure water to raw material shrimp and water quality ratio:0.5~5, the protein particle is starched
It is put into containing tank；More preferably, it is 1 to expect water quality ratio:0.5~2.
Specifically, for shear-blade rotating diameter in 20-40mm, motor speed is provided with cone more than 30000 turns around blade
Shape gathers cover, and cone-shaped hood inwall is interval with collision fin, and fin is higher by inner conical surface 3-5mm.Laser particle size detects, in particulate slurry
Maximum length is in below 0.1mm, i.e., more than 150 mesh.From the point of view of distribution of particles, fish-bone, fish scale are due to harder, and grinding particle size is most
It is small, concentrate on more than 200 mesh, and fish protein, because it softens in water and granularity is slightly larger, and it is in the majority to concentrate on 150-170 mesh.
3）Microminiaturization, protein particle slurry carry out microwave and surround heating, and 90-100 DEG C of boiling boils 10-20min, be cooled to microminiaturization
55 DEG C~75 DEG C of temperature, ultrasonic probe stretch into containing tank effect microminiaturization ultrasonic wave so that the protein particle of protein particle slurry is straight
Footpath is reduced to below micron order or micron order, and small corpuscular protein activation slurry is made；
4）Enzymolysis, slurry progress microwave encirclement is activated to small corpuscular protein and is heated to 40 DEG C~65 DEG C of enzyme digestion reaction temperature, by raw material
The protease that shrimp weighs 0.01~2% is fixed on dispersible carrier surface, and dispersible carrier then is fixed on into multiple preset body surface faces, micro-
Granule protein activation slurry immerses preset body enzyme digestion reaction, microwave, ultrasonic wave cycle alternation and applies 10-20min, during total enzyme digestion reaction
Between 20~60min, obtain enzymolysis protein slurry；
5) Selective Separation, enzymolysis protein is starched and carries out multi-stage booster filtering, Selective Separation goes out molecule from enzymolysis protein slurry
The oligopeptide in 1000-4000Da is measured, residue enzymolysis protein milk is de- bitter enzymolysis protein slurry；Being concentrated in vacuo to moisture is
40-60wt%, produce the small oppossum shrimp of marine products rich in small peptide and digest full shrimp slurry；
Step 1）-5）In remaining oxygen<Carried out in 5% nitrogen environmental protection.
For step 4）, the protease dispersible carrier, prepare the Fe of 30-500nm particle diameters3O4Magnetic nanoparticle, it is right
Fe3O4Magnetic nano-particle is surface-treated, and Fe is arrived using hydride modified3O4Magnetic nanoparticle surface obtains micro- magnetic core, tool
Body is such as：3- aminopropyls-triethoxysilane.By the use of double sulphur cyanobenzenes or glutaraldehyde as coupling agent, by protease covalent coupling
Scattered silicon ribozyme particle is obtained to micro- magnetic core surface.
Preset body is flase floor, and dispersible carrier is fixed on multiple grid plate surfaces, referred to, the silicon ribozyme particle is uniform
The upper and lower surface of thin polyethylene board is bonded to, finally by multiple thin polyethylene board compartments every assembling flase floor.Between flase floor
Gauge is from 5-8mm.Thermosetting resin is applied in polyethylene thin plate upper and lower surface, silicon ribozyme particle is carried same electrostatic charge, such as
Positive charge, powered silicon ribozyme particle tile onto thermosetting resin, and due to powered silicon ribozyme particle, identical charges repel each other, contributes to silicon core
Enzyme granulate is uniformly distributed on polyethylene thin plate, increases its specific surface area.Thermosetting resin is heated, by silicon ribozyme particle after solidification
It is fixed.With the concentration of bicinchoninic acid method measure immobilized enzyme, reach 2-3 μ g/cm2, in practice it has proved that the immobilized enzyme has good
Activity.
As Figure 1-3, in order to realize the alternating application of enzymolysis, microwave, ultrasonic wave, alternating action device is devised, is wrapped
Enzyme immobilization reaction tube 1, microwave tube 2 and the ultrasonic wave duct 3 of head and the tail connection are included, slurries are provided between microwave tube 2 and ultrasonic wave duct 3
Inlet tube, injection point pipe are provided with inlet valve 11 and inlet pump 7, gone out between enzyme immobilization reaction tube 1, microwave tube 2 provided with slurries
Mouth pipe, serum outlet pipe are provided with outlet valve 12 and drain pump 9, circulating pump are provided between ultrasonic wave duct 3 and enzyme immobilization reaction tube 1
8.The ultrasonic perisporium of wave duct 3 is interval with ultrasonic probe 6, and the perisporium of microwave tube 2 is interval with multiple microwave generators 5.Microwave tube 2 enters
Mouth is provided with intermediate valve 10, and the both sides of intermediate valve 10 are respectively equipped with nitrogen inlet duct and blast pipe, and nitrogen inlet duct is provided with intake valve 13
With air bleeding valve 14.Flase floor is assembled into cylindrical and is fitted into enzyme immobilization reaction tube 1.
During work, 1. nitrogen charging, first make it that remaining oxygen is less than 5% in alternating action device.Concrete operations：Close intermediate valve
10, intake valve 13 is opened and is filled with nitrogen, and air bleeding valve 14 is opened, and until detecting, remaining oxygen is less than 5% in blast pipe, closes air inlet
Valve 13 and air bleeding valve 14.
2. filling slurry, intermediate valve 10, inlet valve 11 are opened, and air bleeding valve 14 is opened, and start inlet pump 7, by step 4）It is small
Corpuscular protein activation slurry is filled with alternating action device, while circulating pump 8 starts, and exhaust is then closed until there is slurries to be filled with blast pipe
Valve 14, inlet pump 7, circulating pump 8 are closed when pressure reaches 1-5MPa in alternating action device.
3. alternating action, open microwave generator 4 and microwave encirclement heating is carried out to microwave tube 2, open ultrasonic probe 6
Generator in ultrasonic wave duct slurries carry out ultrasonic activation processing, while in immobilization reactor 1 digest slurries disperse connect
Touch protease and carry out enzyme digestion reaction；For total enzymolysis time 60min technique, microwave, ultrasonic wave, enzymolysis act on 20min respectively
Afterwards, one cycle pump 8 is started, the slurries that enzymolysis immobilization reactor 1 can be calculated according to pipe flow all flow to microwave tube 2
In, the slurries in microwave tube 2 are all flowed in ultrasonic wave duct 3, and the slurries in ultrasonic wave duct 3 all flow to enzymolysis immobilized reactant
Device 1, the cycle time T needed for such circulation primary, circulating pump 8 is closed after cycle time T.After starting circulating pump 8 three times, i.e., just
Beginning fills to starch to flow to again in enzymolysis immobilization reactor 1 in the slurries in digesting immobilization reactor 1 to be digested for the second time, then is opened
After dynamic circulating pump 8 three times, initially fill slurries third time of the slurry in immobilization reactor 1 is digested and flow to enzymolysis immobilization reactor
Third time is digested in 1, and total enzymolysis time reaches 60min, obtains enzymolysis protein slurry.For total enzymolysis time 20min technique,
After microwave, ultrasonic wave, enzymolysis act on 10min respectively after pump circulation, slurry is initially filled after pump circulation three times in enzymolysis immobilization
Slurries in reactor 1 are flowed in enzymolysis immobilization reactor 1 and digested for the second time again, and total enzymolysis time reaches 20min.For
Slurries secondary enzymolysis is realized, microwave, ultrasonic wave, enzymolysis cycle alternation apply at least 3 pump circulations.
4. emptying, intermediate valve 10 is closed, outlet valve 12 is opened, and starts drain pump 9, intake valve 13 is opened after half a minute, directly
Slurries can not be taken out to drain pump 9, then completes the task of the enzymolysis protein slurry emptying in alternating action device, closes outlet valve
12, stop drain pump 9.
So carry out subsequent cycle 1.->②->③->④.
For step 5）, as shown in figure 3, multi-stage booster filtering is：Setting back taper annular canister, energy molecular cut off are big
Back taper annular canister entrance is arranged in 4000Da first order filter membrane, can second level filtering of the molecular cut off more than 2000Da
Film is arranged among back taper annular canister, and third level filter membrane of the energy molecular cut off more than 1000Da is arranged on back taper annular canister and gone out
Mouthful, the step 4）Obtained enzymolysis protein is starched by the back taper annular canister, and molecular weight M is 2000Da≤M≤4000Da's
Oligopeptide is trapped between first order filter membrane and second level filter membrane, the third level of the molecular weight M in 1000Da≤M≤2000Da
The oligopeptide of filter membrane is trapped between second level filter membrane and third level filter membrane；From the enzymolysis of back taper annular canister outlet outflow
Protein milk is mixed into not by the enzymolysis protein slurry of back taper annular canister；Due to the entrance of back taper annular canister, centre and the horizontal stroke of outlet
Sectional area reduces step by step, and enzymolysis protein slurry inlet pressure is 0.2Mpa, then middle and exit pressure increases step by step, ensure that
The larger of filter membrane hour passes through pressure.
A kind of small oppossum shrimp enzymolysis of the marine products prepares the multi-stage booster mistake that the preparation method of the full shrimp slurry rich in small peptide uses
Device 20 is filtered, including back taper annular canister 21, the back taper annular canister 21 include the pitch cone of first segment cone cylinder 21.1 and second of split
21.2 assembled in sequence of cylinder form, and the entrance of first segment cone cylinder 21.1 is fixed with first order filter membrane 22, the second pitch cone cylinder 21.2
Outlet is fixed with third level filter membrane 24, and second level filter membrane is fixed between the pitch cone cylinder 21.2 of first segment cone cylinder 21.1 and second
23, tapered plug 25 is combined with back taper annular canister 21, and the head of tapered plug 25 and afterbody are slidably fitted in guide 26.First pitch cone
The entrance connection liquid-in pipe 27 of cylinder 21.1, the outlet connection drain pipe 28 of the second pitch cone cylinder 21.2, the connection recoil of the collateral branch of drain pipe 8
Pipe 29.
The inwall of first segment cone cylinder 21.1 is provided with the first backwash mouth 30, and the inwall of the second pitch cone cylinder 21.2 is provided with the second backwash mouth
31.First order filter membrane 22, second level filter membrane 23 and third level filter membrane 24 are arranged in screen pack framework.
The multi-stage booster filter has two kinds of working conditions, pressurizing filter state and decompression backwash state, is pressurized
Filter state, tapered plug reach, blocks the first backwash mouth 30 and the second backwash mouth 31, and recoil tube 29 closes water, and enzymolysis protein slurry is from entering liquid
Pipe 27 flows into back taper annular canister 21, and molecular weight M is trapped in the He of first order filter membrane 22 in 2000Da≤M≤4000Da oligopeptide
Between second level filter membrane 23, molecular weight M is trapped in second in the oligopeptide of 1000Da≤M≤2000Da third level filter membrane
Between level filter membrane 23 and third level filter membrane 24；It is mixed from the enzymolysis protein slurry of back taper annular canister outlet outflow after the completion of filtering
Enter not by the enzymolysis protein slurry of back taper annular canister, obtaining de- bitter enzymolysis protein slurry.
Backwash state to be depressurized, is blocked using closing water plug 32 before first order filter membrane 22, the use of drain pipe 28 is closed water plug 32 and blocked,
Moved after tapered plug, recoil tube 29 is opened, and is washed by water and is gone out retention protein milk from the first backwash mouth 30 and the second backwash mouth 31.From outlet
To entrance, because cross section where three-stage filtration film increases step by step, pressure reduces step by step, forms the decompression punching to back taper annular canister
Wash.Until state as shown in Figure 3 is that three-stage filtration film is rinsed totally.
Workflow is：Recoil->Draining->Wet sterilization->Cooling->Filtering->Backwash plasma discharge；
Other step details are explained as follows：
The classification grinding crushes for two level, and the first order crushes send the small fish of marine products into first disintegrator for conveyer belt,
First disintegrator delivery outlet is not recycled to first disintegrator charge door by material, second is output to by material through 4 mesh sieve net filtrations
Pulverizer, the second pulverizer delivery outlet are defeated by expecting not by expecting to be recycled to the second pulverizer charge door through 6 mesh sieve net filtrations
Go out for qualified protein body；
It is to ensure that Fish protein is fully denatured opening space structure that 10-20min is boiled in 90-100 DEG C of boiling before microminiaturization, is easy to digest.
The microwave surrounds heating, refers to that the perisporium interval of microwave tube is equipped with multiple microwave generators, and particulate protein is starched
Carry out microwave and surround heating.The microwave surrounds heating, is to use microwave frequency microwave power 15-30W's is micro- for 2450MHz
Wave producer, each heat time is depending on containing tank size.
Microwave is ceramic material or polyvinyl resin material；Microwave diameter is about 2 times of the fading depth of microwave half, so,
Microwave penetration microwave tube wall is acted on particulate protein slurry, is ensured the uniform encirclement heating from perisporium, is ensured in particulate protein slurry
From the inside to the outside, the from top to bottom uniformity of Temperature Distribution.Preferably, the microwave tube uses ceramic material, diameter 1400mm,
Length is 3000mm.
The effect microminiaturization ultrasonic wave, diameter 10-40mm ultrasonic probes stretch into ultraphonic pipe apart from the depths of tube hub 1/3,
With 20KHz-40KHz, the w/cm of field strength 70~1002, ultrasonication 3-10 minutes.Between ultrasonic probe is on ultraphonic pipe length direction
Every arrangement.
Preferably, constant temperature, high pressure are kept in enzymolysis process, constant temperature is to act on microwave duration t by interval time t1, to protect
The enzyme digestion reaction temperature of particulate protein slurry is held at 40~65 DEG C；The high pressure, it is so that particulate protein slurry is pressed by 1-5MPa
Pretend use.
The protease be alkali protease caused by microbial fermentation or the protease be selected from alkali protease and
The complex enzyme of flavor protease, wherein, alkali protease addition is that raw material shrimp weighs 0.5-1.0wt%, flavor protease addition
For raw material shrimp weight 0.05-0.1wt%.
Embodiment 2
As shown in Figure 2,3, step 4）In, the preset body is film 5.2, and the upper surface of film 5.2 has been spaced a plurality of
Isolate fin 5.3, the isolation fin 5.3 is in the sine curve interval of same period.The upper and lower surface of film 5.2 is uniformly glued
Dispersible carrier 5.4 is had, such as silicon ribozyme particle, the concentration of immobilized enzyme is determined with bicinchoninic acid method, reaches 2-3 μ g/cm2, it is real
Trampling proves that the immobilized enzyme has good activity.Film 5.2 is wound into the preset body coiling body of certain diameter around the outer peripheral face of axle 5.1
5, the preset body coiling body 5 is fixed in the immobilized enzyme reactor 1 of tubular.Because the preset body coiling body 5 is with tighter
The interval gathered, isolate and a sinusoidal sinusoidal channel is provided between fin 5.3 and lower films, the sinusoidal channel is from preset body
The one end open of coiling body 5, is exported from the other end, and sinusoidal channel extends the chance of protease contacted protein macromolecular chain, is
Decentralized enzymolysis contact provides good condition.
It is weight than 50-60% that the de- bitter enzymolysis protein slurry is concentrated in vacuo into moisture, produces the sea rich in small peptide
Produce small oppossum shrimp and digest full shrimp slurry.
Step 4）Above-mentioned enzyme digestion reaction during, enzymolysis can be accelerated by stretching into new preset body coiling body 5, increase small peptide
Yield.
The present embodiment 2 is to disperse curing technology using enzyme, increases ratio table of the enzyme with the contact of small corpuscular protein activation slurry
Area, high temperature enzyme deactivation step is eliminated, cost is reduced for enzymolysis production.
After testing, the small oppossum shrimp of marine products obtained by the present invention digests full shrimp slurry, and crude protein content is 20~35 wt%, small peptide
Content is 15~20 wt%, 3~6wt% of crude fat content, and ash content is 3~8 wt%, 2~6wt% of lysine, methionine 1~
3wt%, water content are 40~60wt%.Wherein, molecular weight below 1000Da protein hydrolysate proportion for 40~
90%。
A kind of small oppossum shrimp of marine products rich in small peptide digests application of the full shrimp slurry in aquaculture, with accounting for fish meal gross weight extremely
The small oppossum shrimp of marine products described in few 10wt% digests full shrimp slurry and substitutes fish meal feeding cultured fishes.
Considered based on aquaculture cost, will preferably account for the small oppossum shrimp of marine products described in the 25-40wt% of shrimp med gross weight and digest full shrimp slurry
Substitute shrimp med and continuously feed shrimps in culture, the small oppossum shrimp of marine products for adding 25%-30wt% digests full shrimp slurry continuously to feed South America right in vain
Shrimp three months, its average daily gain reach 0.42-0.66g, improve 15% than adding the feed efficiency of control group of shrimp med, compare city
The feed efficiency of the similar hydrolysate in face improves 7%, while enhances the immunologic function of cultivation shrimp, reduces its disease incident.
Experimental data is as follows：
Experiment is experimental subjects from same maternal Penaeus Vannmei, and experiment uses natural light, is cultivated in breeding barrel, cultivation sea
Water is deep sea water, is higher than 7mg/L, salinity using flowing water culture pattern, flow velocity 5L/min, 15 ± 0.5 DEG C of water temperature, dissolved oxygen
32 ‰, PH7.5~8.0.Before experiment starts, stop eating 24h, selects uniform in size, physically fit and disease-free body surface South America right in vain
Shrimp juvenile prawn.Fish is divided into 15 breeding barrels at random, 3 treatment groups, each handles 5 repetitions, every barrel of 25 tails are artificial daily full
Food feeds twice 6:30 and 16:30, after feeding 30 minutes, to the residual bait gauge number in each bucket, according to the flat of every 100 residual baits
Equal weight calculates the residual bait weight do not ingested.It is added in following ratio in common fish meal and is fed：
Control group：Shrimp med 30%；A groups： FH（Similar hydrolysate on the market）30%, B group：The small oppossum shrimp enzyme of marine products that embodiment 1 obtains
Bitter full shrimp slurry 30% is freed, culture experiment amounts to 90 days, the feed intake to fish, feed rate respectively during raising（Weightening/feed）, it is dead
The rate of dying is recorded, as a result as shown in table 1.
The full shrimp slurry of the small oppossum shrimp debittering by enzymatic hydrolysis of the marine products of table 1 substitutes the fish meal cultivation Penaeus Vannmei table of comparisons
| Daily ration processing | Control | A groups | B groups |
| Initial weight, g | 80 | 80 | 80 |
| End weight, g | 110 | 125 | 140 |
| Feed intake, g | 125 | 140 | 155 |
| Feed efficiency, weightening/feed | 0.24 | 0.32 | 0.39 |
| Death rate % | 3.3 | 2.1 | 0.6 |
As seen from Table 1, the small full shrimp slurry of oppossum shrimp debittering by enzymatic hydrolysis of marine products for adding 30wt% continuously feeds Penaeus Vannmei three months, its
Average daily gain reaches 0.66g, improves 15% than adding the feed efficiency of control group B groups of shrimp med, hydrolysate more similar than market
A groups feed efficiency improve 7%, while enhance cultivation fish immunologic function, than add shrimp med control group B groups the death rate
81% is reduced, the A groups death rate of hydrolysate more similar than market reduces by 71%.
The small oppossum shrimp enzymolysis of marine products prepares the preparation method of the full shrimp slurry rich in small peptide, and enzymolysis efficiency is high, production cost
It is low, shrimp med can be substituted and be used for aquaculture, can be as the forage protein source of the economic feed of aquaculture.
The present invention takes full advantage of the marine organisms money of China's abundant using the full fish of the small oppossum shrimp of marine products of low value as raw material
Source；The pulpous state small peptide product of high value is prepared using zymolysis technique, energy consumption is low and technological process is short, remain the nutrition of full fish into
Point；Transition for China's fish meal processing industry product opens new approach.
Claims (10)
1. the method that a kind of de- bitter enzymolysis of small oppossum shrimp of marine products prepares the full shrimp slurry rich in small peptide, it is characterised in that including following step
Suddenly：
1）Granulating, it will be freeze-dried to water content in 50wt%-70wt%, be crushed to using the small oppossum shrimp of fresh marine products as raw material shrimp
Protein body of the full-size in below 3mm；
2）Micronized, according to protein body and water quality than 1:0.2 adds 0-3 DEG C of pure water, ultra high shear in protein body
Protein particle slurry more than to 150 mesh, it is 1 to add pure water to raw material shrimp and water quality ratio:0.5~5；
3）Microminiaturization, protein particle slurry carry out microwave and surround heating, and 90-100 DEG C of boiling boils 10-20min, be cooled to microminiaturization temperature
55 DEG C~75 DEG C, ultrasonic probe stretches into protein particle slurry and acts on microminiaturization ultrasonic wave so that the protein particle of protein particle slurry
Diameter is reduced to below micron order or micron order, and small corpuscular protein activation slurry is made；
4）Enzymolysis, slurry progress microwave encirclement is activated to small corpuscular protein and is heated to 40 DEG C~65 DEG C of enzyme digestion reaction temperature, by raw material
The protease that fish weighs 0.01~2wt% is fixed on dispersible carrier surface, then dispersible carrier is uniformly fixed on to multiple preset body surfaces
Face, microwave, ultrasound, small corpuscular protein activation slurry are immersed in preset body surface face enzyme digestion reaction cycle alternation and applied 10-20min/ times,
Total 20~60min of enzyme digestion reaction time, obtain enzymolysis protein slurry；
5）Selective Separation, enzymolysis protein is starched and carries out multi-stage booster filtering, Selective Separation goes out molecule from enzymolysis protein slurry
The oligopeptide in 1000-4000Da is measured, residue enzymolysis protein milk is de- bitter enzymolysis protein slurry；The de- bitter enzymolysis protein slurry is true
It is 40-60wt% that sky, which is concentrated into moisture, produces the small oppossum shrimp of marine products rich in small peptide and digests full shrimp slurry；
Step 1）-5）In remaining oxygen<Carried out in 5% nitrogen environmental protection.
2. the method that the de- bitter enzymolysis of the small oppossum shrimp of marine products as claimed in claim 1 prepares the full shrimp slurry rich in small peptide, it is characterised in that
The preset body is polyethylene board, and dispersible carrier is fixed on multiple preset body surface faces, refers to the dispersible carrier even adhesion
To the upper and lower surface of thin polyethylene board, finally by multiple thin polyethylene board compartments every assembling multiple concentric circles annular compositions
Flase floor.
3. the method that the de- bitter enzymolysis of the small oppossum shrimp of marine products as claimed in claim 1 prepares the full shrimp slurry rich in small peptide, it is characterised in that
The preset body is film（5.2）, the film（5.2）Upper surface has been spaced a plurality of isolation fin（5.3）, the isolation
Fin（5.3）In the sine curve of same period, the film（5.2）Upper and lower surface even adhesion has dispersible carrier（5.4), so
Rear film（5.2）Around axle（5.1）Outer peripheral face is wound into the preset body coiling body of certain diameter（5）.
4. the method that the de- bitter enzymolysis of the small oppossum shrimp of the marine products as described in Claims 2 or 3 prepares the full shrimp slurry rich in small peptide, its feature exist
In the microwave surrounds heating, refers to that tubular container perisporium interval is equipped with multiple microwave generators, protein particle slurry is carried out micro-
Ripple surrounds heating.
5. the method that the de- bitter enzymolysis of the small oppossum shrimp of the marine products as described in Claims 2 or 3 prepares the full shrimp slurry rich in small peptide, its feature exist
In the effect microminiaturization ultrasonic wave, diameter 10-40mm ultrasonic probes stretch into tubular container apart from the depths of tube hub 1/3, multiple
Ultrasonic probe arranged for interval on tubular container length direction, with frequency 20-40kHz, field strength 70-100w/cm2, ultrasonication
3-10 minutes.
6. the method that the de- bitter enzymolysis of the small oppossum shrimp of marine products as claimed in claim 1 prepares the full shrimp slurry rich in small peptide, it is characterised in that
The protease is fixed on dispersible carrier surface, prepares Fe3O4Magnetic nanoparticle, it is hydride modified to arrive Fe3O4Magnetic Nano
Grain surface obtains micro- magnetic core, and protease covalent coupling to micro- magnetic core surface is obtained into scattered silicon ribozyme particle.
Used 7. prepared by a kind of de- bitter enzymolysis of small oppossum shrimp of the marine products as described in claim 1-6 in method of the full shrimp slurry rich in small peptide
Alternating action device, it is characterised in that the microwave, ultrasound, small corpuscular protein activation slurry be immersed in preset body digest it is anti-
Answering cycle alternation to apply is realized by cycle alternation acting device, and the enzyme that the alternating action device includes head and the tail connection is consolidated
Surely reaction tube is changed（1）, microwave tube（2）With ultrasonic wave duct（3）, the flase floor or the preset body coiling body（5）It is fixed on enzyme
Immobilized reactant pipe（1）In, slurries are full of the alternating action device with certain pressure, and slurries are every in alternating action device
In alternating action device circulation primary after the certain pause time；During intermittent time, slurries receive microwave action in microwave tube, surpass
Slurries receive ul-trasonic irradiation, enzyme immobilization reaction tube in sonic tube（1）Middle generation enzyme digestion reaction.
Used 8. prepared by a kind of de- bitter enzymolysis of small oppossum shrimp of the marine products as described in claim 1-6 in the method for the full shrimp slurry rich in small peptide
Multi-stage booster filter, it is characterised in that including back taper annular canister（1）, the back taper annular canister（1）Entrance, centre and
Outlet is respectively fixed with first order filter membrane（2）, second level filter membrane（3）With third level filter membrane（4）, internal cone wall, which is provided with, to recoil
Mouthful（10,11）, back taper annular canister（1）Slided on internal cone wall axis and be provided with tapered plug（5）, tapered plug（5）Reach blocks the recoil mouth,
Tapered plug（5）Move afterwards and decontrol the recoil mouth.
9. the method that a kind of de- bitter enzymolysis of small oppossum shrimp of marine products as described in claim 1-6 is any prepares the full shrimp slurry rich in small peptide obtains
The small oppossum shrimp of the marine products rich in small peptide arrived digests full shrimp slurry, it is characterised in that the small oppossum shrimp of marine products digests full shrimp slurry and is in a liquid state
The small peptide content of the small molecule of pulpous state, wherein amino acid residue below 10 is 15~20 wt%.
10. a kind of small oppossum shrimp of marine products as claimed in claim 9 rich in small peptide digests application of the full shrimp slurry in aquaculture, its
It is characterised by, digesting full shrimp slurry replacement shrimp med with the small oppossum shrimp of the marine products for accounting for fish meal gross weight at least 10wt%, continuously feeding is supported
Grow shrimps.
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| CN109170987A (en) * | 2018-11-16 | 2019-01-11 | 无棣县兴亚生物科技有限公司 | A kind of digested tankage production equipment and process |
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Denomination of invention: A method for preparing whole shrimp slurry by Debittering and enzymatic hydrolysis of marine small bran shrimp and its application Effective date of registration: 20220408 Granted publication date: 20201027 Pledgee: Agricultural Bank of China Limited by Share Ltd. Yuhuan branch Pledgor: Yuhuan Wufeng steamed and degreased fish meal factory Registration number: Y2022330000476 |
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