CN108034487A - The enzymolysis and electric field compounding method of tuna leftover bits and pieces - Google Patents
The enzymolysis and electric field compounding method of tuna leftover bits and pieces Download PDFInfo
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- CN108034487A CN108034487A CN201711412870.XA CN201711412870A CN108034487A CN 108034487 A CN108034487 A CN 108034487A CN 201711412870 A CN201711412870 A CN 201711412870A CN 108034487 A CN108034487 A CN 108034487A
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- enzymolysis
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- carbon dioxide
- pieces
- brine
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- 238000000034 method Methods 0.000 title claims abstract description 41
- 230000005684 electric field Effects 0.000 title claims abstract description 29
- 238000013329 compounding Methods 0.000 title claims abstract description 17
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 claims abstract description 133
- 239000007788 liquid Substances 0.000 claims abstract description 85
- 239000002002 slurry Substances 0.000 claims abstract description 81
- 229910002092 carbon dioxide Inorganic materials 0.000 claims abstract description 66
- 239000001569 carbon dioxide Substances 0.000 claims abstract description 65
- 239000012267 brine Substances 0.000 claims abstract description 50
- HPALAKNZSZLMCH-UHFFFAOYSA-M sodium;chloride;hydrate Chemical compound O.[Na+].[Cl-] HPALAKNZSZLMCH-UHFFFAOYSA-M 0.000 claims abstract description 50
- 101000693530 Staphylococcus aureus Staphylokinase Proteins 0.000 claims abstract description 24
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims abstract description 18
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims abstract description 18
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 18
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 17
- 102000057297 Pepsin A Human genes 0.000 claims abstract description 17
- 108090000284 Pepsin A Proteins 0.000 claims abstract description 17
- 229940111202 pepsin Drugs 0.000 claims abstract description 17
- 239000011780 sodium chloride Substances 0.000 claims abstract description 9
- 238000012545 processing Methods 0.000 claims description 64
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 56
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 32
- 229910052757 nitrogen Inorganic materials 0.000 claims description 28
- 238000002525 ultrasonication Methods 0.000 claims description 21
- 239000004365 Protease Substances 0.000 claims description 17
- 102000005158 Subtilisins Human genes 0.000 claims description 16
- 108010056079 Subtilisins Proteins 0.000 claims description 16
- 108090000526 Papain Proteins 0.000 claims description 15
- 235000019834 papain Nutrition 0.000 claims description 15
- 229940055729 papain Drugs 0.000 claims description 15
- 238000005119 centrifugation Methods 0.000 claims description 13
- 102000004190 Enzymes Human genes 0.000 claims description 10
- 108090000790 Enzymes Proteins 0.000 claims description 10
- 229940088598 enzyme Drugs 0.000 claims description 10
- 239000012263 liquid product Substances 0.000 claims description 6
- 239000000243 solution Substances 0.000 claims description 5
- 239000007789 gas Substances 0.000 claims description 3
- 102000035195 Peptidases Human genes 0.000 claims description 2
- 108091005804 Peptidases Proteins 0.000 claims description 2
- 235000019419 proteases Nutrition 0.000 claims description 2
- 238000003756 stirring Methods 0.000 claims description 2
- GWEVSGVZZGPLCZ-UHFFFAOYSA-N Titan oxide Chemical compound O=[Ti]=O GWEVSGVZZGPLCZ-UHFFFAOYSA-N 0.000 claims 2
- 229910052799 carbon Inorganic materials 0.000 claims 2
- 241000219112 Cucumis Species 0.000 claims 1
- 235000015510 Cucumis melo subsp melo Nutrition 0.000 claims 1
- 241000790917 Dioxys <bee> Species 0.000 claims 1
- FJJCIZWZNKZHII-UHFFFAOYSA-N [4,6-bis(cyanoamino)-1,3,5-triazin-2-yl]cyanamide Chemical compound N#CNC1=NC(NC#N)=NC(NC#N)=N1 FJJCIZWZNKZHII-UHFFFAOYSA-N 0.000 claims 1
- 239000004408 titanium dioxide Substances 0.000 claims 1
- 239000002023 wood Substances 0.000 claims 1
- 235000021323 fish oil Nutrition 0.000 abstract description 29
- 238000011084 recovery Methods 0.000 abstract description 5
- 241000251468 Actinopterygii Species 0.000 description 8
- 238000000926 separation method Methods 0.000 description 5
- 239000000463 material Substances 0.000 description 4
- 238000002604 ultrasonography Methods 0.000 description 4
- 230000008901 benefit Effects 0.000 description 2
- 239000006227 byproduct Substances 0.000 description 2
- 235000019198 oils Nutrition 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 239000002699 waste material Substances 0.000 description 2
- 239000002253 acid Substances 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 230000002785 anti-thrombosis Effects 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 239000003146 anticoagulant agent Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 210000004556 brain Anatomy 0.000 description 1
- 229910002090 carbon oxide Inorganic materials 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 239000002781 deodorant agent Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 235000019197 fats Nutrition 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 239000004519 grease Substances 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 210000000496 pancreas Anatomy 0.000 description 1
- 230000007180 physiological regulation Effects 0.000 description 1
- 235000020777 polyunsaturated fatty acids Nutrition 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 238000007127 saponification reaction Methods 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 235000021081 unsaturated fats Nutrition 0.000 description 1
- 210000002700 urine Anatomy 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
- C11B1/00—Production of fats or fatty oils from raw materials
- C11B1/02—Pretreatment
- C11B1/025—Pretreatment by enzymes or microorganisms, living or dead
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
- C11B1/00—Production of fats or fatty oils from raw materials
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
- C11B1/00—Production of fats or fatty oils from raw materials
- C11B1/02—Pretreatment
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
- C11B3/00—Refining fats or fatty oils
- C11B3/001—Refining fats or fatty oils by a combination of two or more of the means hereafter
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
- C11B3/00—Refining fats or fatty oils
- C11B3/10—Refining fats or fatty oils by adsorption
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
- C11B3/00—Refining fats or fatty oils
- C11B3/16—Refining fats or fatty oils by mechanical means
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- Organic Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Microbiology (AREA)
- Mechanical Engineering (AREA)
- Biochemistry (AREA)
- Meat, Egg Or Seafood Products (AREA)
- Enzymes And Modification Thereof (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The invention discloses the enzymolysis and electric field compounding method of a kind of tuna leftover bits and pieces, including：Step 1) is cleaned up tuna leftover bits and pieces with clear water, is smashed；With the brine that sodium chloride configuration quality concentration is 0.3 0.5g/mL, brine is heated to 25 30 DEG C, is added into slurry, the mass ratio of brine and slurry is 1:5‑8；Step 2) adds pepsin into slurry and carries out primary enzymolysis；Step 3) is continually fed into carbon dioxide into primary enzymolysis liquid, is placed in high-pressure pulse electric and is handled, and continues to be passed through carbon dioxide, the electric field strength of high-pressure pulse electric is 55 60kV/cm；Step 4) adds neutral proteinase and carries out secondary enzymolysis；Step 5) adds hydrochloric acid, continuously adds absolute ethyl alcohol；Step 6) centrifuges.The present invention improves the recovery rate to fish oil, improves the content of EPA and DHA in fish oil.
Description
Technical field
The present invention relates to the enzymolysis and electric field compounding method of agriculture field, more particularly to a kind of tuna leftover bits and pieces.
Background technology
Fish is one of important aquatic resources in China, its yield occupies first place in the world.But many fish process are only limited to
In the utilization to fish body muscle, and it is less to the processing and utilization of the byproducts such as fish guts, fish-bone, fish head, generally as discarded object
Processing.The integral level of China's comprehensive utilization fish pomace is not also high, often the by-product containing higher protein and fat content
How effectively product do not obtain rationally effective performance, this not only causes the waste of resource, but also polluted environment, so profit
With these resources, turn waste into wealth, will be a very important research topic.Contain abundant how unsaturated fat in marine products fish oil
Fat acid, its main component are EPA and DHA, and polyunsaturated fatty acid has anti-inflammatory, antithrombotic, antitumor and promotion brain hair
Educate, strengthen a variety of physiological regulation functions such as memory.The domestic fish oil of extraction at present is mostly using saponification method, molecularly distilled, urine
The single methods such as plain inclusion method, CO2 methods extract fish oil, in terms of purity and oil yield all there are it is respective the shortcomings that；In addition it is international
On the fish oil deodorant problem that processes also be not well solved.Therefore, these fish pomaces how are made full use of, most
The grease for therefrom extracting high quality of big degree, the added value for improving fish processing, reduction environmental pollution become current important
Problem.
The content of the invention
It is an object of the invention to solve at least the above, and provide the advantages of at least will be described later.
It is a still further object of the present invention to provide the enzymolysis and electric field compounding method of a kind of tuna leftover bits and pieces, bag
Include：
Step 1) is cleaned up tuna leftover bits and pieces with clear water, is smashed, and is added equivalent to the tuna leftover bits and pieces
0.5-2 times of water is mixed into slurry, and the slurry is stirred evenly；
The brine for being 0.3-0.5g/mL with sodium chloride configuration quality concentration, 25-30 DEG C is heated to by brine, by brine plus
Enter into the slurry, the mass ratio of the brine and the slurry is 1:5-8；The slurry for adding brine is held
Stir continuously, the slurry is uniformly mixed with brine；Stand for standby use；
Pepsin is added in the slurry that step 2) is prepared to the step 1) and carries out primary enzymolysis, primary enzymolysis
Temperature is 30-35 DEG C, pH value 2-3, when the primary enzymolysis time is 9-10 small, wherein, the addition of the pepsin is institute
The 2-2.5% for the slurry quality that step 1) is prepared is stated, obtains primary enzymolysis liquid；
The primary enzymolysis liquid obtained by the step 2) processing is placed in a container by step 3), to the primary enzymolysis
Carbon dioxide is continually fed into liquid, the time for being passed through carbon dioxide is 1-2h, and the flow of carbon dioxide is 4-
The container, is placed in high-pressure pulse electric is handled afterwards by 5m/s, in the high-pressure pulse electric processing procedure, after
Continuous to be passed through carbon dioxide, the flow of carbon dioxide is 1-2m/s, after the high-pressure pulse electric processing terminates,
Also continue to be passed through carbon dioxide, the flow of carbon dioxide is protected with the flow in the high-pressure pulse electric processing procedure
Hold unanimously, the electric field strength of the high-pressure pulse electric is 55-60kV/cm, frequency 100-150Hz, pulse width 20-25
μ s, processing time 5-10s；
The feed liquid that the step 3) processing obtains is placed 30-60min by step 4), and it is secondary to add neutral proteinase progress
Enzymolysis, secondary enzymolysis temperature are 40-45 DEG C, pH value 8-9, and the addition of the neutral proteinase is handled for the step 3)
The 2-3% of the feed liquid quality arrived, when the secondary enzymolysis time is 3-4 small, obtains secondary enzymolysis liquid；
Step 5) adds the hydrochloric acid that mass concentration is 2-3% into the secondary enzymolysis liquid, by the secondary enzymolysis liquid
PH is adjusted to 4-5, continuously adds absolute ethyl alcohol, and addition of the absolute ethyl alcohol in the secondary enzymolysis liquid is described secondary
The 65-70% of enzymolysis liquid volume, when standing 20-23 is small；
The feed liquid that the step 5) is prepared is placed in centrifugation apparatus and is centrifuged by step 6), and centrifuge RPMs are
8000-9000r/min, the time of centrifugal treating is 26-33min.
Preferably, in the enzymolysis and electric field compounding method of the tuna leftover bits and pieces, in the step 2),
During primary enzymolysis, while nitrogen is continually fed into the slurry, the flow of nitrogen is 1.5-2.5m/s, is carried out at the same time
Ultrasonication, the supersonic frequency of the ultrasonication is 800-950kHz.
Preferably, in the enzymolysis and electric field compounding method of the tuna leftover bits and pieces, in the step 4),
During secondary enzymolysis, while nitrogen is continually fed into the slurry, the flow of nitrogen is 3.5-5.5m/s, is carried out at the same time
Ultrasonication, the supersonic frequency of the ultrasonication is 650-700kHz.
Preferably, in the enzymolysis and electric field compounding method of the tuna leftover bits and pieces, in the step 4), institute
State neutral proteinase be by trypsase, papain and subtilopeptidase A, the trypsase, papain and
The mass ratio of subtilopeptidase A is 4-5:1-2:0.5-1.
Preferably, in the enzymolysis and electric field compounding method of the tuna leftover bits and pieces, in the step 4), institute
The mass ratio for stating trypsase, papain and subtilopeptidase A is 4:1:0.5.
Preferably, in the enzymolysis and electric field compounding method of the tuna leftover bits and pieces, in the step 3), to
Carbon dioxide is continually fed into the primary enzymolysis liquid, the time for being passed through carbon dioxide is 1h, carbon dioxide
Flow be 4m/s, in the high-pressure pulse electric processing procedure, continue to be passed through carbon dioxide, carbon dioxide
Flow is 1m/s.
The present invention includes at least following beneficial effect：
The enzymolysis and electric field compounding method of a kind of tuna leftover bits and pieces provided by the invention, including：Step 1) is with clearly
Water cleans up tuna leftover bits and pieces, smashes, and adds the water mixing pulp equivalent to 0.5-2 times of the tuna leftover bits and pieces
Thing, the slurry is stirred evenly；The brine for being 0.3-0.5g/mL with sodium chloride configuration quality concentration, brine is heated to
25-30 DEG C, brine is added into the slurry, the mass ratio of the brine and the slurry is 1:5-8；To adding salt
The slurry of water is constantly stirred, and the slurry is uniformly mixed with brine；Stand for standby use；Step 2) is to the step
1) pepsin is added in the slurry being prepared and carries out primary enzymolysis, primary enzymolysis temperature is 30-35 DEG C, pH value 2-3,
When the primary enzymolysis time is 9-10 small, wherein, the addition of the pepsin is the slurry that the step 1) is prepared
The 2-2.5% of quality, obtains primary enzymolysis liquid；The primary enzymolysis liquid obtained by the step 2) processing is placed in one by step 3)
In container, carbon dioxide is continually fed into the primary enzymolysis liquid, the time for being passed through carbon dioxide is 1-2h, two
The flow of carbon oxide gas is 4-5m/s, the container is placed in high-pressure pulse electric is handled afterwards, in the high pressure
In impulse electric field processing procedure, continue to be passed through carbon dioxide, the flow of carbon dioxide is 1-2m/s, in the high pressure
After impulse electric field processing terminates, also continue to be passed through carbon dioxide, the flow of carbon dioxide and the high-voltage pulse
Flow during electric field treatment is consistent, and the electric field strength of the high-pressure pulse electric is 55-60kV/cm, and frequency is
100-150Hz, pulse width are 20-25 μ s, processing time 5-10s；The feed liquid that step 4) obtains the step 3) processing
30-60min is placed, neutral proteinase is added and carries out secondary enzymolysis, secondary enzymolysis temperature is 40-45 DEG C, pH value 8-9, institute
The addition for stating neutral proteinase handles the 2-3% of obtained feed liquid quality for the step 3), and the secondary enzymolysis time is small for 3-4
When, obtain secondary enzymolysis liquid；Step 5) adds the hydrochloric acid that mass concentration is 2-3% into the secondary enzymolysis liquid, by described two
The pH of secondary enzymolysis liquid is adjusted to 4-5, continuously adds absolute ethyl alcohol, addition of the absolute ethyl alcohol in the secondary enzymolysis liquid
The 65-70% accumulated for the secondary enzymolysis liquid, when standing 20-23 is small；The feed liquid that the step 5) is prepared step 6)
It is placed in centrifugation apparatus and is centrifuged, centrifuge RPMs 8000-9000r/min, the time of centrifugal treating is 26-33min.
The present invention carries out the slurry prepared by tuna leftover bits and pieces primary enzymolysis, high-pressure pulse electric is handled, secondary
Enzymolysis, on the one hand improves the recovery rate to fish oil, on the other hand reduces the destruction to EPA and DHA, it is final to improve the two
Content in fish oil；And ultrasonication is used at the same time during primary enzymolysis and secondary enzymolysis, to improve enzymolysis
Efficiency, finally improve the recovery rate of fish oil.
The present invention improves the recovery rate to fish oil, improves the content of EPA and DHA in fish oil.
Further advantage, target and the feature of the present invention embodies part by following explanation, and part will also be by this
The research and practice of invention and be understood by the person skilled in the art.
Embodiment
The present invention is described in further detail below, to make those skilled in the art being capable of evidence with reference to specification word
To implement.
It should be appreciated that such as " having ", "comprising" and " comprising " term used herein do not allot one or more
The presence or addition of a other elements or its combination.
Embodiment 1
Step 1) is cleaned up tuna leftover bits and pieces with clear water, is smashed, and is added equivalent to the tuna leftover bits and pieces 0.5
Water again is mixed into slurry, and the slurry is stirred evenly；
The brine for being 0.3g/mL with sodium chloride configuration quality concentration, is heated to 25 DEG C by brine, brine is added to described
In slurry, the mass ratio of the brine and the slurry is 1:5；The slurry for adding brine is constantly stirred,
The slurry is set to be uniformly mixed with brine；Stand for standby use；
Pepsin is added in the slurry that step 2) is prepared to the step 1) and carries out primary enzymolysis, primary enzymolysis
Temperature is 30 DEG C, pH value 2, when the primary enzymolysis time is 9 small, wherein, the addition of the pepsin is the step 1)
The 2% of the slurry quality being prepared, during primary enzymolysis, is carried out at the same time ultrasonication, the ultrasonication
Supersonic frequency be 800kHz, while nitrogen is continually fed into the slurry, the flow of nitrogen is 1.5m/s, is obtained once
Enzymolysis liquid；
The primary enzymolysis liquid obtained by the step 2) processing is placed in a container by step 3), to the primary enzymolysis
Carbon dioxide is continually fed into liquid, the time for being passed through carbon dioxide is 1h, and the flow of carbon dioxide is 4m/s,
The container is placed in high-pressure pulse electric afterwards and is handled, in the high-pressure pulse electric processing procedure, continues to lead to
Enter carbon dioxide, the flow of carbon dioxide is 1m/s, after the high-pressure pulse electric processing terminates, is also continued to
Carbon dioxide is passed through, the flow of carbon dioxide keeps one with the flow in the high-pressure pulse electric processing procedure
Cause, the electric field strength of the high-pressure pulse electric is 55kV/cm, and frequency 100Hz, pulse width is 20 μ s, and processing time is
5s；
The feed liquid that the step 3) processing obtains is placed 30min by step 4), is added neutral proteinase and is carried out secondary enzyme
Solution, secondary enzymolysis temperature are 40 DEG C, and pH value 8, the addition of the neutral proteinase handles obtained material for the step 3)
The 2% of liquid quality, when the secondary enzymolysis time is 3 small, during secondary enzymolysis, is carried out at the same time ultrasonication, the ultrasound
The supersonic frequency of ripple processing is 650kHz, while nitrogen is continually fed into the slurry, and the flow of nitrogen is 3.5m/s, is obtained
To secondary enzymolysis liquid；Wherein, the neutral proteinase be by trypsase, papain and subtilopeptidase A, it is described
The mass ratio of trypsase, papain and subtilopeptidase A is 4:1:0.5；
Step 5) adds the hydrochloric acid that mass concentration is 2% into the secondary enzymolysis liquid, by the pH of the secondary enzymolysis liquid
Adjust to 4-5, continuously add absolute ethyl alcohol, addition of the absolute ethyl alcohol in the secondary enzymolysis liquid is the secondary enzyme
The 65% of liquid product is solved, when standing 20 is small；
The feed liquid that the step 5) is prepared is placed in centrifugation apparatus and is centrifuged by step 6), and centrifuge RPMs are
8000r/min, the time of centrifugal treating is 26min；
Step 7) is handled in obtained fish oil with the step 6) and adds activated carbon, and the addition of the activated carbon is described
The 3% of the fish oil quality that step 6) processing obtains；Again using centrifugation apparatus by Activated carbon separation, so as to obtain fish oil.
Embodiment 2
Step 1) is cleaned up tuna leftover bits and pieces with clear water, is smashed, and is added equivalent to 2 times of the tuna leftover bits and pieces
Water be mixed into slurry, the slurry is stirred evenly；
The brine for being 0.5g/mL with sodium chloride configuration quality concentration, is heated to 30 DEG C by brine, brine is added to described
In slurry, the mass ratio of the brine and the slurry is 1:8；The slurry for adding brine is constantly stirred,
The slurry is set to be uniformly mixed with brine；Stand for standby use；
Pepsin is added in the slurry that step 2) is prepared to the step 1) and carries out primary enzymolysis, primary enzymolysis
Temperature is 35 DEG C, pH value 3, when the primary enzymolysis time is 10 small, wherein, the addition of the pepsin is the step 1)
The 2.5% of the slurry quality being prepared, during primary enzymolysis, is carried out at the same time ultrasonication, at the ultrasonic wave
The supersonic frequency of reason is 950kHz, while nitrogen is continually fed into the slurry, and the flow of nitrogen is 2.5m/s, obtains one
Secondary enzymolysis liquid；
The primary enzymolysis liquid obtained by the step 2) processing is placed in a container by step 3), to the primary enzymolysis
Carbon dioxide is continually fed into liquid, the time for being passed through carbon dioxide is 2h, and the flow of carbon dioxide is 5m/s,
The container is placed in high-pressure pulse electric afterwards and is handled, in the high-pressure pulse electric processing procedure, continues to lead to
Enter carbon dioxide, the flow of carbon dioxide is 2m/s, after the high-pressure pulse electric processing terminates, is also continued to
Carbon dioxide is passed through, the flow of carbon dioxide keeps one with the flow in the high-pressure pulse electric processing procedure
Cause, the electric field strength of the high-pressure pulse electric is 60kV/cm, and frequency 150Hz, pulse width is 25 μ s, and processing time is
10s；
The feed liquid that the step 3) processing obtains is placed 60min by step 4), is added neutral proteinase and is carried out secondary enzyme
Solution, secondary enzymolysis temperature are 45 DEG C, and pH value 9, the addition of the neutral proteinase handles obtained material for the step 3)
The 3% of liquid quality, when the secondary enzymolysis time is 4 small, during secondary enzymolysis, is carried out at the same time ultrasonication, the ultrasound
The supersonic frequency of ripple processing is 700kHz, while nitrogen is continually fed into the slurry, and the flow of nitrogen obtains for 5.5m/s
To secondary enzymolysis liquid；Wherein, the neutral proteinase be by trypsase, papain and subtilopeptidase A, it is described
The mass ratio of trypsase, papain and subtilopeptidase A is 5:2:1；
Step 5) adds the hydrochloric acid that mass concentration is 3% into the secondary enzymolysis liquid, by the pH of the secondary enzymolysis liquid
Adjust to 4-5, continuously add absolute ethyl alcohol, addition of the absolute ethyl alcohol in the secondary enzymolysis liquid is the secondary enzyme
The 70% of liquid product is solved, when standing 23 is small；
The feed liquid that the step 5) is prepared is placed in centrifugation apparatus and is centrifuged by step 6), and centrifuge RPMs are
9000r/min, the time of centrifugal treating is 33min；
Step 7) is handled in obtained fish oil with the step 6) and adds activated carbon, and the addition of the activated carbon is described
The 4% of the fish oil quality that step 6) processing obtains；Again using centrifugation apparatus by Activated carbon separation, so as to obtain fish oil.
Embodiment 3
Step 1) is cleaned up tuna leftover bits and pieces with clear water, is smashed, and is added and is got a foothold 2 times equivalent to the tuna
Water is mixed into slurry, and the slurry is stirred evenly；
The brine for being 0.3g/mL with sodium chloride configuration quality concentration, is heated to 25 DEG C by brine, brine is added to described
In slurry, the mass ratio of the brine and the slurry is 1:6；The slurry for adding brine is constantly stirred,
The slurry is set to be uniformly mixed with brine；Stand for standby use；
Pepsin is added in the slurry that step 2) is prepared to the step 1) and carries out primary enzymolysis, primary enzymolysis
Temperature is 35 DEG C, pH value 3, when the primary enzymolysis time is 10 small, wherein, the addition of the pepsin is the step 1)
The 2.5% of the slurry quality being prepared, during primary enzymolysis, is carried out at the same time ultrasonication, at the ultrasonic wave
The supersonic frequency of reason is 950kHz, while nitrogen is continually fed into the slurry, and the flow of nitrogen is 1.5m/s, obtains one
Secondary enzymolysis liquid；
The primary enzymolysis liquid obtained by the step 2) processing is placed in a container by step 3), to the primary enzymolysis
Carbon dioxide is continually fed into liquid, the time for being passed through carbon dioxide is 2h, and the flow of carbon dioxide is 4m/s,
The container is placed in high-pressure pulse electric afterwards and is handled, in the high-pressure pulse electric processing procedure, continues to lead to
Enter carbon dioxide, the flow of carbon dioxide is 1m/s, after the high-pressure pulse electric processing terminates, is also continued to
Carbon dioxide is passed through, the flow of carbon dioxide keeps one with the flow in the high-pressure pulse electric processing procedure
Cause, the electric field strength of the high-pressure pulse electric is 56kV/cm, and frequency 120Hz, pulse width is 22 μ s, and processing time is
8s；
The feed liquid that the step 3) processing obtains is placed 30min by step 4), is added neutral proteinase and is carried out secondary enzyme
Solution, secondary enzymolysis temperature are 43 DEG C, and pH value 8, the addition of the neutral proteinase handles obtained material for the step 3)
The 2% of liquid quality, when the secondary enzymolysis time is 3 small, during secondary enzymolysis, is carried out at the same time ultrasonication, the ultrasound
The supersonic frequency of ripple processing is 650kHz, while nitrogen is continually fed into the slurry, and the flow of nitrogen is 4m/s, is obtained
Secondary enzymolysis liquid；Wherein, the neutral proteinase is by trypsase, papain and subtilopeptidase A, the pancreas
The mass ratio of protease, papain and subtilopeptidase A is 5:2:0.5；
Step 5) adds the hydrochloric acid that mass concentration is 3% into the secondary enzymolysis liquid, by the pH of the secondary enzymolysis liquid
Adjust to 4-5, continuously add absolute ethyl alcohol, addition of the absolute ethyl alcohol in the secondary enzymolysis liquid is the secondary enzyme
The 70% of liquid product is solved, when standing 23 is small；
The feed liquid that the step 5) is prepared is placed in centrifugation apparatus and is centrifuged by step 6), and centrifuge RPMs are
8500r/min, the time of centrifugal treating is 26min；
Step 7) is handled in obtained fish oil with the step 6) and adds activated carbon, and the addition of the activated carbon is described
The 3% of the fish oil quality that step 6) processing obtains；Again using centrifugation apparatus by Activated carbon separation, so as to obtain fish oil.
Embodiment 4
Step 1) is cleaned up tuna leftover bits and pieces with clear water, is smashed, and is added equivalent to 2 times of the tuna leftover bits and pieces
Water be mixed into slurry, the slurry is stirred evenly；
The brine for being 0.3g/mL with sodium chloride configuration quality concentration, is heated to 25 DEG C by brine, brine is added to described
In slurry, the mass ratio of the brine and the slurry is 1:5；The slurry for adding brine is constantly stirred,
The slurry is set to be uniformly mixed with brine；Stand for standby use；
Pepsin is added in the slurry that step 2) is prepared to the step 1) and carries out primary enzymolysis, primary enzymolysis
Temperature is 35 DEG C, pH value 3, when the primary enzymolysis time is 10 small, wherein, the addition of the pepsin is the step 1)
The 2.5% of the slurry quality being prepared, during primary enzymolysis, is carried out at the same time ultrasonication, at the ultrasonic wave
The supersonic frequency of reason is 900kHz, while nitrogen is continually fed into the slurry, and the flow of nitrogen is 2m/s, is obtained once
Enzymolysis liquid；
The primary enzymolysis liquid obtained by the step 2) processing is placed in a container by step 3), to the primary enzymolysis
Carbon dioxide is continually fed into liquid, the time for being passed through carbon dioxide is 1h, and the flow of carbon dioxide is 4m/s,
The container is placed in high-pressure pulse electric afterwards and is handled, in the high-pressure pulse electric processing procedure, continues to lead to
Enter carbon dioxide, the flow of carbon dioxide is 1m/s, after the high-pressure pulse electric processing terminates, is also continued to
Carbon dioxide is passed through, the flow of carbon dioxide keeps one with the flow in the high-pressure pulse electric processing procedure
Cause, the electric field strength of the high-pressure pulse electric is 55kV/cm, and frequency 130Hz, pulse width is 20 μ s, and processing time is
8s；
The feed liquid that the step 3) processing obtains is placed 40min by step 4), is added neutral proteinase and is carried out secondary enzyme
Solution, secondary enzymolysis temperature are 43 DEG C, and pH value 8, the addition of the neutral proteinase handles obtained material for the step 3)
The 2% of liquid quality, when the secondary enzymolysis time is 3 small, during secondary enzymolysis, is carried out at the same time ultrasonication, the ultrasound
The supersonic frequency of ripple processing is 680kHz, while nitrogen is continually fed into the slurry, and the flow of nitrogen is 3.5m/s, is obtained
To secondary enzymolysis liquid；Wherein, the neutral proteinase be by trypsase, papain and subtilopeptidase A, it is described
The mass ratio of trypsase, papain and subtilopeptidase A is 4:1:1；
Step 5) adds the hydrochloric acid that mass concentration is 3% into the secondary enzymolysis liquid, by the pH of the secondary enzymolysis liquid
Adjust to 4-5, continuously add absolute ethyl alcohol, addition of the absolute ethyl alcohol in the secondary enzymolysis liquid is the secondary enzyme
The 70% of liquid product is solved, when standing 23 is small；
The feed liquid that the step 5) is prepared is placed in centrifugation apparatus and is centrifuged by step 6), and centrifuge RPMs are
9000r/min, the time of centrifugal treating is 33min；
Step 7) is handled in obtained fish oil with the step 6) and adds activated carbon, and the addition of the activated carbon is described
The 4% of the fish oil quality that step 6) processing obtains；Again using centrifugation apparatus by Activated carbon separation, so as to obtain fish oil.
Embodiment 5
Step 1) is cleaned up tuna leftover bits and pieces with clear water, is smashed, and is added equivalent to 2 times of the tuna leftover bits and pieces
Water be mixed into slurry, the slurry is stirred evenly；
The brine for being 0.5g/mL with sodium chloride configuration quality concentration, is heated to 30 DEG C by brine, brine is added to described
In slurry, the mass ratio of the brine and the slurry is 1:8；The slurry for adding brine is constantly stirred,
The slurry is set to be uniformly mixed with brine；Stand for standby use；
Pepsin is added in the slurry that step 2) is prepared to the step 1) and carries out primary enzymolysis, primary enzymolysis
Temperature is 35 DEG C, pH value 3, when the primary enzymolysis time is 10 small, wherein, the addition of the pepsin is the step 1)
The 2.5% of the slurry quality being prepared, during primary enzymolysis, is carried out at the same time ultrasonication, at the ultrasonic wave
The supersonic frequency of reason is 870kHz, while nitrogen is continually fed into the slurry, and the flow of nitrogen is 2.5m/s, obtains one
Secondary enzymolysis liquid；
The primary enzymolysis liquid obtained by the step 2) processing is placed in a container by step 3), to the primary enzymolysis
Carbon dioxide is continually fed into liquid, the time for being passed through carbon dioxide is 2h, and the flow of carbon dioxide is 4.5m/
The container, is placed in high-pressure pulse electric is handled afterwards by s, in the high-pressure pulse electric processing procedure, continues
Be passed through carbon dioxide, the flow of carbon dioxide is 1m/s, after the high-pressure pulse electric processing terminates, also after
Continuous to be passed through carbon dioxide, the flow of carbon dioxide keeps one with the flow in the high-pressure pulse electric processing procedure
Cause, the electric field strength of the high-pressure pulse electric is 55kV/cm, and frequency 100Hz, pulse width is 20 μ s, and processing time is
5s；
The feed liquid that the step 3) processing obtains is placed 30-60min by step 4), and it is secondary to add neutral proteinase progress
Enzymolysis, secondary enzymolysis temperature are 45 DEG C, pH value 9, and the addition step 3) processing of the neutral proteinase obtains
The 3% of feed liquid quality, during secondary enzymolysis, is carried out at the same time ultrasonication, the supersonic frequency of the ultrasonication is
700kHz, when the secondary enzymolysis time is 4 small, while is continually fed into nitrogen into the slurry, and the flow of nitrogen is 3.5m/s,
Obtain secondary enzymolysis liquid；Wherein, the neutral proteinase is by trypsase, papain and subtilopeptidase A, institute
The mass ratio for stating trypsase, papain and subtilopeptidase A is 5:2:1；
Step 5) adds the hydrochloric acid that mass concentration is 3% into the secondary enzymolysis liquid, by the pH of the secondary enzymolysis liquid
Adjust to 4-5, continuously add absolute ethyl alcohol, addition of the absolute ethyl alcohol in the secondary enzymolysis liquid is the secondary enzyme
The 70% of liquid product is solved, when standing 23 is small；
The feed liquid that the step 5) is prepared is placed in centrifugation apparatus and is centrifuged by step 6), and centrifuge RPMs are
8800r/min, the time of centrifugal treating is 26min；
Step 7) is handled in obtained fish oil with the step 6) and adds activated carbon, and the addition of the activated carbon is described
The 3% of the fish oil quality that step 6) processing obtains；Again using centrifugation apparatus by Activated carbon separation, so as to obtain fish oil.
Compliance test result：
Tuna oil is extracted using the method for embodiment 1 to embodiment 5.Wherein, each embodiment processing
1000g tuna leftover bits and pieces.The index for the fish oil that embodiment 1 is extracted to embodiment 5 is shown in Table 1.The present invention is improved to golden rifle
The recovery rate of fish fish oil, improves the content of EPA and DHA in fish oil.
Table 1
Although embodiment of the present invention is disclosed as above, it is not restricted in specification and embodiment listed
With it can be applied to various suitable the field of the invention completely, can be easily for those skilled in the art
Realize other modification, therefore under the universal limited without departing substantially from claim and equivalency range, it is of the invention and unlimited
In specific details.
Claims (6)
1. the enzymolysis and electric field compounding method of a kind of tuna leftover bits and pieces, it is characterised in that including：
Step 1) is cleaned up tuna leftover bits and pieces with clear water, is smashed, and is added equivalent to 0.5-2 times of the tuna leftover bits and pieces
Water be mixed into slurry, the slurry is stirred evenly；
The brine for being 0.3-0.5g/mL with sodium chloride configuration quality concentration, 25-30 DEG C is heated to by brine, by brine add to
In the slurry, the mass ratio of the brine and the slurry is 1:5-8；The slurry for adding brine is carried out constantly
Stirring, makes the slurry be uniformly mixed with brine；Stand for standby use；
Pepsin is added in the slurry that step 2) is prepared to the step 1) and carries out primary enzymolysis, primary enzymolysis temperature
For 30-35 DEG C, pH value 2-3, when the primary enzymolysis time is 9-10 small, wherein, the addition of the pepsin is the step
The 2-2.5% of the rapid slurry quality 1) being prepared, obtains primary enzymolysis liquid；
The primary enzymolysis liquid obtained by the step 2) processing is placed in a container by step 3), into the primary enzymolysis liquid
Carbon dioxide is continually fed into, the time for being passed through carbon dioxide is 1-2h, and the flow of carbon dioxide is 4-5m/s,
The container is placed in high-pressure pulse electric afterwards and is handled, in the high-pressure pulse electric processing procedure, continues to lead to
Enter carbon dioxide, the flow of carbon dioxide is 1-2m/s, after the high-pressure pulse electric processing terminates, also after
Continuous to be passed through carbon dioxide, the flow of carbon dioxide keeps one with the flow in the high-pressure pulse electric processing procedure
Causing, the electric field strength of the high-pressure pulse electric is 55-60kV/cm, and frequency 100-150Hz, pulse width is 20-25 μ s,
Processing time is 5-10s；
The feed liquid that the step 3) processing obtains is placed 30-60min by step 4), is added neutral proteinase and is carried out secondary enzyme
Solution, secondary enzymolysis temperature are 40-45 DEG C, pH value 8-9, and the addition of the neutral proteinase obtains for the step 3) processing
Feed liquid quality 2-3%, the secondary enzymolysis time for 3-4 it is small when, obtain secondary enzymolysis liquid；
Step 5) adds the hydrochloric acid that mass concentration is 2-3% into the secondary enzymolysis liquid, by the pH tune of the secondary enzymolysis liquid
Section continuously adds absolute ethyl alcohol, addition of the absolute ethyl alcohol in the secondary enzymolysis liquid is the secondary enzymolysis to 4-5
The 65-70% of liquid product, when standing 20-23 is small；
The feed liquid that the step 5) is prepared is placed in centrifugation apparatus and is centrifuged by step 6), and centrifuge RPMs are
8000-9000r/min, the time of centrifugal treating is 26-33min.
2. the enzymolysis and electric field compounding method of tuna leftover bits and pieces as claimed in claim 1, it is characterised in that the step
It is rapid 2) in, during primary enzymolysis, while nitrogen is continually fed into the slurry, the flow of nitrogen is 1.5-2.5m/s
Ultrasonication is carried out at the same time, the supersonic frequency of the ultrasonication is 800-950kHz.
3. the enzymolysis and electric field compounding method of tuna leftover bits and pieces as claimed in claim 2, it is characterised in that the step
It is rapid 4) in, during secondary enzymolysis, while nitrogen is continually fed into the slurry, the flow of nitrogen is 3.5-5.5m/
S, is carried out at the same time ultrasonication, the supersonic frequency of the ultrasonication is 650-700kHz.
4. the enzymolysis and electric field compounding method of tuna leftover bits and pieces as claimed in claim 1, it is characterised in that the step
It is rapid 4) in, the neutral proteinase be by trypsase, papain and subtilopeptidase A, the trypsase, wood
The mass ratio of melon protease and subtilopeptidase A is 4-5:1-2:0.5-1.
5. the enzymolysis and electric field compounding method of tuna leftover bits and pieces as claimed in claim 4, it is characterised in that the step
It is rapid 4) in, the mass ratio of the trypsase, papain and subtilopeptidase A is 4:1:0.5.
6. the enzymolysis and electric field compounding method of tuna leftover bits and pieces as claimed in claim 1, it is characterised in that the step
It is rapid 3) in, be continually fed into carbon dioxide into the primary enzymolysis liquid, the time for being passed through carbon dioxide is 1h, dioxy
The flow for changing carbon gas is 4m/s, in the high-pressure pulse electric processing procedure, continues to be passed through carbon dioxide, titanium dioxide
The flow of carbon gas is 1m/s.
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