CN108034487B - Enzymolysis and electric field composite treatment method for tuna leftovers - Google Patents
Enzymolysis and electric field composite treatment method for tuna leftovers Download PDFInfo
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- 230000005684 electric field Effects 0.000 title claims abstract description 68
- 238000000034 method Methods 0.000 title claims abstract description 45
- 235000021190 leftovers Nutrition 0.000 title claims abstract description 35
- 239000002131 composite material Substances 0.000 title claims abstract description 9
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 claims abstract description 118
- 239000007788 liquid Substances 0.000 claims abstract description 81
- 229910002092 carbon dioxide Inorganic materials 0.000 claims abstract description 59
- 239000001569 carbon dioxide Substances 0.000 claims abstract description 58
- 239000002002 slurry Substances 0.000 claims abstract description 53
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims abstract description 34
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 33
- 239000011780 sodium chloride Substances 0.000 claims abstract description 25
- 108090000145 Bacillolysin Proteins 0.000 claims abstract description 24
- 102000035092 Neutral proteases Human genes 0.000 claims abstract description 24
- 108091005507 Neutral proteases Proteins 0.000 claims abstract description 24
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-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
- 238000010438 heat treatment Methods 0.000 claims abstract description 9
- 238000000926 separation method Methods 0.000 claims abstract description 9
- 238000005406 washing Methods 0.000 claims abstract description 9
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 56
- 239000007789 gas Substances 0.000 claims description 56
- 239000012267 brine Substances 0.000 claims description 36
- HPALAKNZSZLMCH-UHFFFAOYSA-M sodium;chloride;hydrate Chemical compound O.[Na+].[Cl-] HPALAKNZSZLMCH-UHFFFAOYSA-M 0.000 claims description 36
- 238000009210 therapy by ultrasound Methods 0.000 claims description 29
- 229910052757 nitrogen Inorganic materials 0.000 claims description 28
- 230000008569 process Effects 0.000 claims description 24
- 108090000526 Papain Proteins 0.000 claims description 16
- 239000004365 Protease Substances 0.000 claims description 16
- 108090000787 Subtilisin Proteins 0.000 claims description 16
- 102000004142 Trypsin Human genes 0.000 claims description 16
- 108090000631 Trypsin Proteins 0.000 claims description 16
- 235000019834 papain Nutrition 0.000 claims description 16
- 229940055729 papain Drugs 0.000 claims description 16
- 238000003756 stirring Methods 0.000 claims description 16
- 239000012588 trypsin Substances 0.000 claims description 16
- 230000002255 enzymatic effect Effects 0.000 claims description 11
- 239000000413 hydrolysate Substances 0.000 claims description 11
- 239000000203 mixture Substances 0.000 claims description 8
- 239000000463 material Substances 0.000 claims description 3
- 230000007071 enzymatic hydrolysis Effects 0.000 claims 2
- 238000006047 enzymatic hydrolysis reaction Methods 0.000 claims 2
- 235000021323 fish oil Nutrition 0.000 abstract description 28
- 238000000605 extraction Methods 0.000 abstract description 5
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 30
- 241000251468 Actinopterygii Species 0.000 description 10
- 239000006227 byproduct Substances 0.000 description 3
- 238000012545 processing Methods 0.000 description 3
- 239000002699 waste material Substances 0.000 description 3
- 235000020777 polyunsaturated fatty acids Nutrition 0.000 description 2
- 206010061218 Inflammation Diseases 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 208000007536 Thrombosis Diseases 0.000 description 1
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 210000000988 bone and bone Anatomy 0.000 description 1
- 230000004641 brain development Effects 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 229940013317 fish oils Drugs 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 239000004519 grease Substances 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000000199 molecular distillation Methods 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 239000003921 oil Substances 0.000 description 1
- 235000019198 oils Nutrition 0.000 description 1
- 230000007180 physiological regulation Effects 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000007127 saponification reaction Methods 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
- 210000001835 viscera Anatomy 0.000 description 1
Classifications
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- 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
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- 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 an enzymolysis and electric field composite treatment method of tuna leftovers, which comprises the following steps: step 1) washing and crushing tuna leftovers by using clear water; preparing saline water with mass concentration of 0.3-0.5g/mL by using sodium chloride, heating the saline water to 25-30 ℃, and adding the saline water into the slurry, wherein the mass ratio of the saline water to the slurry is 1: 5-8; step 2) adding pepsin into the pulp for primary enzymolysis; step 3) continuously introducing carbon dioxide gas into the primary enzymolysis liquid, placing the primary enzymolysis liquid in a high-voltage pulse electric field for treatment, and continuously introducing the carbon dioxide gas, wherein the electric field intensity of the high-voltage pulse electric field is 55-60 kV/cm; step 4), adding neutral protease for secondary enzymolysis; step 5), adding hydrochloric acid, and continuously adding absolute ethyl alcohol; and 6) carrying out centrifugal separation. The invention improves the extraction rate of the fish oil and the content of EPA and DHA in the fish oil.
Description
Technical Field
The invention relates to the field of agriculture, in particular to an enzymolysis and electric field composite treatment method for tuna leftovers.
Background
Fish is one of the important aquatic resources in China, and the yield of the fish is the first place in the world. However, many fish processing processes are limited to the utilization of fish muscle, and the processing and utilization of byproducts such as fish viscera, fish bones, fish heads and the like are less, and the byproducts are generally disposed as waste. The overall level of comprehensive utilization of fish leftovers in China is not high, and byproducts containing higher protein and fat content are not reasonably and effectively utilized, so that not only is the waste of resources caused, but also the environment is polluted, and how to effectively utilize the resources and change waste into valuables is an important research subject. The marine fish oil contains rich polyunsaturated fatty acids, the main components of which are EPA and DHA, and the polyunsaturated fatty acids have various physiological regulation functions of resisting inflammation, resisting thrombus, resisting tumor, promoting brain development, enhancing memory and the like. At present, the fish oil extracted at home is mostly extracted by a single method such as a saponification method, a molecular distillation method, a urea inclusion method, a CO2 method and the like, and the respective defects exist in the aspects of purity and oil yield; in addition, the problem of fishy smell removal of the internationally processed fish oil is not well solved. Therefore, how to fully utilize the fish leftovers and extract high-quality grease from the fish leftovers to the maximum extent, improve the added value of fish processing and reduce environmental pollution is an important issue at present.
Disclosure of Invention
An object of the present invention is to solve at least the above problems and to provide at least the advantages described later.
The invention also aims to provide an enzymolysis and electric field composite treatment method of tuna leftovers, which comprises the following steps:
step 1) washing tuna leftovers with clear water, smashing, adding water which is 0.5-2 times of the tuna leftovers to mix into slurry, and uniformly stirring the slurry;
preparing saline water with the mass concentration of 0.3-0.5g/mL by using sodium chloride, heating the saline water to 25-30 ℃, and adding the saline water into the slurry, wherein the mass ratio of the saline water to the slurry is 1: 5-8; continuously stirring the slurry added with the brine to uniformly mix the slurry with the brine; standing for later use;
step 2) adding pepsin into the pulp prepared in the step 1) for primary enzymolysis, wherein the primary enzymolysis temperature is 30-35 ℃, the pH value is 2-3, and the primary enzymolysis time is 9-10 hours, and the adding amount of the pepsin is 2-2.5% of the mass of the pulp prepared in the step 1) to obtain primary enzymolysis liquid;
step 3) placing the primary enzymolysis liquid obtained by the treatment in the step 2) into a container, continuously introducing carbon dioxide gas into the primary enzymolysis liquid for 1-2h, wherein the flow rate of the carbon dioxide gas is 4-5m/s, then placing the container into a high-voltage pulse electric field for treatment, continuously introducing the carbon dioxide gas during the treatment of the high-voltage pulse electric field, wherein the flow rate of the carbon dioxide gas is 1-2m/s, continuously introducing the carbon dioxide gas after the treatment of the high-voltage pulse electric field is finished, the flow rate of the carbon dioxide gas is consistent with the flow rate during the treatment of the high-voltage pulse electric field, the electric field strength of the high-voltage pulse electric field is 55-60kV/cm, the frequency is 100-150Hz, and the pulse width is 20-25 μ s, the treatment time is 5-10 s;
step 4) placing the feed liquid obtained by the treatment of the step 3) for 30-60min, adding neutral protease for secondary enzymolysis, wherein the secondary enzymolysis temperature is 40-45 ℃, the pH value is 8-9, the adding amount of the neutral protease is 2-3% of the mass of the feed liquid obtained by the treatment of the step 3), and the secondary enzymolysis time is 3-4 hours, so as to obtain secondary enzymolysis liquid;
step 5) adding hydrochloric acid with the mass concentration of 2-3% into the secondary enzymolysis liquid, adjusting the pH of the secondary enzymolysis liquid to 4-5, continuously adding absolute ethyl alcohol, wherein the addition amount of the absolute ethyl alcohol in the secondary enzymolysis liquid is 65-70% of the volume of the secondary enzymolysis liquid, and standing for 20-23 hours;
and step 6) placing the material liquid prepared in the step 5) into a centrifugal device for centrifugal separation, wherein the centrifugal rotation number is 8000-9000r/min, and the centrifugal treatment time is 26-33 min.
Preferably, in the method for combined treatment of enzymolysis and electric field of tuna leftovers, in the step 2), nitrogen is continuously introduced into the slurry at the same time in the process of one enzymolysis, the flow rate of the nitrogen is 1.5-2.5m/s, and ultrasonic treatment is performed at the same time, wherein the ultrasonic frequency of the ultrasonic treatment is 800-950 kHz.
Preferably, in the method for combined treatment of enzymolysis and electric field of tuna leftovers, in the step 4), nitrogen is continuously introduced into the slurry at the same time in the secondary enzymolysis process, the flow rate of the nitrogen is 3.5-5.5m/s, and ultrasonic treatment is performed at the same time, wherein the ultrasonic frequency of the ultrasonic treatment is 650-700 kHz.
Preferably, in the method for the enzymolysis and electric field composite treatment of the tuna leftovers, in the step 4), the neutral protease is trypsin, papain and subtilisin, and the mass ratio of the trypsin to the papain to the subtilisin is 4-5:1-2: 0.5-1.
Preferably, in the enzymolysis and electric field composite treatment method of the tuna leftovers, in the step 4), the mass ratio of the trypsin to the papain to the subtilisin is 4:1: 0.5.
Preferably, in the method for the enzymolysis and electric field combined treatment of the tuna leftovers, in the step 3), carbon dioxide gas is continuously introduced into the primary enzymolysis liquid, the time for introducing the carbon dioxide gas is 1 hour, the flow rate of the carbon dioxide gas is 4m/s, and the carbon dioxide gas is continuously introduced in the high-voltage pulse electric field treatment process, wherein the flow rate of the carbon dioxide gas is 1 m/s.
The invention at least comprises the following beneficial effects:
the invention provides an enzymolysis and electric field composite treatment method of tuna leftovers, which comprises the following steps: step 1) washing tuna leftovers with clear water, smashing, adding water which is 0.5-2 times of the tuna leftovers to mix into slurry, and uniformly stirring the slurry; preparing saline water with the mass concentration of 0.3-0.5g/mL by using sodium chloride, heating the saline water to 25-30 ℃, and adding the saline water into the slurry, wherein the mass ratio of the saline water to the slurry is 1: 5-8; continuously stirring the slurry added with the brine to uniformly mix the slurry with the brine; standing for later use; step 2) adding pepsin into the pulp prepared in the step 1) for primary enzymolysis, wherein the primary enzymolysis temperature is 30-35 ℃, the pH value is 2-3, and the primary enzymolysis time is 9-10 hours, and the adding amount of the pepsin is 2-2.5% of the mass of the pulp prepared in the step 1) to obtain primary enzymolysis liquid; step 3) placing the primary enzymolysis liquid obtained by the treatment in the step 2) into a container, continuously introducing carbon dioxide gas into the primary enzymolysis liquid for 1-2h, wherein the flow rate of the carbon dioxide gas is 4-5m/s, then placing the container into a high-voltage pulse electric field for treatment, continuously introducing the carbon dioxide gas during the treatment of the high-voltage pulse electric field, wherein the flow rate of the carbon dioxide gas is 1-2m/s, continuously introducing the carbon dioxide gas after the treatment of the high-voltage pulse electric field is finished, the flow rate of the carbon dioxide gas is consistent with the flow rate during the treatment of the high-voltage pulse electric field, the electric field strength of the high-voltage pulse electric field is 55-60kV/cm, the frequency is 100-150Hz, and the pulse width is 20-25 μ s, the treatment time is 5-10 s; step 4) placing the feed liquid obtained by the treatment of the step 3) for 30-60min, adding neutral protease for secondary enzymolysis, wherein the secondary enzymolysis temperature is 40-45 ℃, the pH value is 8-9, the adding amount of the neutral protease is 2-3% of the mass of the feed liquid obtained by the treatment of the step 3), and the secondary enzymolysis time is 3-4 hours, so as to obtain secondary enzymolysis liquid; step 5) adding hydrochloric acid with the mass concentration of 2-3% into the secondary enzymolysis liquid, adjusting the pH of the secondary enzymolysis liquid to 4-5, continuously adding absolute ethyl alcohol, wherein the addition amount of the absolute ethyl alcohol in the secondary enzymolysis liquid is 65-70% of the volume of the secondary enzymolysis liquid, and standing for 20-23 hours; and step 6) placing the material liquid prepared in the step 5) into a centrifugal device for centrifugal separation, wherein the centrifugal rotation number is 8000-9000r/min, and the centrifugal treatment time is 26-33 min.
The method carries out primary enzymolysis, high-voltage pulse electric field treatment and secondary enzymolysis on the slurry prepared from the tuna leftovers, so that on one hand, the extraction rate of the fish oil is improved, on the other hand, the damage to EPA and DHA is reduced, and the final content of the EPA and the DHA in the fish oil is improved; and ultrasonic treatment is adopted in the processes of primary enzymolysis and secondary enzymolysis simultaneously so as to improve the enzymolysis efficiency and finally improve the extraction rate of the fish oil.
The invention improves the extraction rate of the fish oil and the content of EPA and DHA in the fish oil.
Additional advantages, objects, and features of the invention will be set forth in part in the description which follows and in part will become apparent to those having ordinary skill in the art upon examination of the following or may be learned from practice of the invention.
Detailed Description
The present invention is described in further detail below to enable those skilled in the art to practice the invention with reference to the description.
It will be understood that terms such as "having," "including," and "comprising," as used herein, do not preclude the presence or addition of one or more other elements or groups thereof.
Example 1
Step 1) washing tuna leftovers with clear water, smashing, adding water which is 0.5 times of the tuna leftovers into the smashed tuna leftovers to mix into slurry, and uniformly stirring the slurry;
preparing brine with the mass concentration of 0.3g/mL by using sodium chloride, heating the brine to 25 ℃, and adding the brine into the slurry, wherein the mass ratio of the brine to the slurry is 1: 5; continuously stirring the slurry added with the brine to uniformly mix the slurry with the brine; standing for later use;
step 2) adding pepsin into the pulp obtained in the step 1) for primary enzymolysis, wherein the primary enzymolysis temperature is 30 ℃, the pH value is 2, and the primary enzymolysis time is 9 hours, wherein the adding amount of the pepsin is 2% of the mass of the pulp obtained in the step 1), and in the primary enzymolysis process, ultrasonic treatment is simultaneously carried out, the ultrasonic frequency of the ultrasonic treatment is 800kHz, nitrogen is continuously introduced into the pulp, and the flow of the nitrogen is 1.5m/s, so that primary enzymolysis liquid is obtained;
step 3) placing the primary enzymatic hydrolysate obtained by the treatment in the step 2) into a container, continuously introducing carbon dioxide gas into the primary enzymatic hydrolysate for 1h, wherein the flow rate of the carbon dioxide gas is 4m/s, then placing the container into a high-voltage pulse electric field for treatment, continuously introducing the carbon dioxide gas during the treatment process of the high-voltage pulse electric field, wherein the flow rate of the carbon dioxide gas is 1m/s, and after the treatment of the high-voltage pulse electric field is finished, the carbon dioxide gas is also continuously introduced, the flow rate of the carbon dioxide gas is consistent with the flow rate during the treatment process of the high-voltage pulse electric field, the electric field strength of the high-voltage pulse electric field is 55kV/cm, the frequency is 100Hz, the pulse width is 20 mu s, and the treatment time is 5 s;
step 4) placing the feed liquid obtained by the treatment in the step 3) for 30min, adding neutral protease for secondary enzymolysis, wherein the temperature of the secondary enzymolysis is 40 ℃, the pH value is 8, the adding amount of the neutral protease is 2% of the mass of the feed liquid obtained by the treatment in the step 3), the time of the secondary enzymolysis is 3 hours, and in the process of the secondary enzymolysis, ultrasonic treatment is simultaneously carried out, the ultrasonic frequency of the ultrasonic treatment is 650kHz, nitrogen is continuously introduced into the pulp, and the flow of the nitrogen is 3.5m/s, so that secondary enzymolysis liquid is obtained; the neutral protease is composed of trypsin, papain and subtilisin, and the mass ratio of the trypsin to the papain to the subtilisin is 4:1: 0.5;
step 5) adding hydrochloric acid with the mass concentration of 2% into the secondary enzymolysis liquid, adjusting the pH of the secondary enzymolysis liquid to 4-5, continuously adding absolute ethyl alcohol, wherein the addition amount of the absolute ethyl alcohol in the secondary enzymolysis liquid is 65% of the volume of the secondary enzymolysis liquid, and standing for 20 hours;
step 6) placing the feed liquid prepared in the step 5) into a centrifugal device for centrifugal separation, wherein the centrifugal revolution is 8000r/min, and the centrifugal treatment time is 26 min;
step 7) adding activated carbon into the fish oil obtained by the treatment in the step 6), wherein the adding amount of the activated carbon is 3% of the mass of the fish oil obtained by the treatment in the step 6); and separating the activated carbon by adopting centrifugal equipment to obtain the fish oil.
Example 2
Step 1) washing tuna leftovers with clear water, smashing, adding water which is 2 times of the tuna leftovers to mix into slurry, and uniformly stirring the slurry;
preparing brine with the mass concentration of 0.5g/mL by using sodium chloride, heating the brine to 30 ℃, and adding the brine into the slurry, wherein the mass ratio of the brine to the slurry is 1: 8; continuously stirring the slurry added with the brine to uniformly mix the slurry with the brine; standing for later use;
step 2) adding pepsin into the pulp obtained in the step 1) for primary enzymolysis, wherein the primary enzymolysis temperature is 35 ℃, the pH value is 3, and the primary enzymolysis time is 10 hours, wherein the adding amount of the pepsin is 2.5% of the mass of the pulp obtained in the step 1), and in the primary enzymolysis process, ultrasonic treatment is simultaneously carried out, the ultrasonic frequency of the ultrasonic treatment is 950kHz, nitrogen is continuously introduced into the pulp, and the flow of the nitrogen is 2.5m/s, so that primary enzymolysis liquid is obtained;
step 3) placing the primary enzymatic hydrolysate obtained by the treatment in the step 2) into a container, continuously introducing carbon dioxide gas into the primary enzymatic hydrolysate for 2 hours, wherein the flow rate of the carbon dioxide gas is 5m/s, then placing the container into a high-voltage pulse electric field for treatment, continuously introducing the carbon dioxide gas during the treatment process of the high-voltage pulse electric field, wherein the flow rate of the carbon dioxide gas is 2m/s, and after the treatment of the high-voltage pulse electric field is finished, the carbon dioxide gas is also continuously introduced, the flow rate of the carbon dioxide gas is consistent with the flow rate during the treatment process of the high-voltage pulse electric field, the electric field strength of the high-voltage pulse electric field is 60kV/cm, the frequency is 150Hz, the pulse width is 25 μ s, and the treatment time is 10 s;
step 4) placing the feed liquid obtained by the treatment in the step 3) for 60min, adding neutral protease for secondary enzymolysis, wherein the temperature of the secondary enzymolysis is 45 ℃, the pH value is 9, the adding amount of the neutral protease is 3% of the mass of the feed liquid obtained by the treatment in the step 3), the time of the secondary enzymolysis is 4 hours, in the process of the secondary enzymolysis, ultrasonic treatment is simultaneously carried out, the ultrasonic frequency of the ultrasonic treatment is 700kHz, nitrogen is continuously introduced into the pulp, and the flow of the nitrogen is 5.5m/s to obtain secondary enzymolysis liquid; the neutral protease is composed of trypsin, papain and subtilisin, and the mass ratio of the trypsin to the papain to the subtilisin is 5:2: 1;
step 5) adding hydrochloric acid with the mass concentration of 3% into the secondary enzymolysis liquid, adjusting the pH of the secondary enzymolysis liquid to 4-5, continuously adding absolute ethyl alcohol, wherein the addition amount of the absolute ethyl alcohol in the secondary enzymolysis liquid is 70% of the volume of the secondary enzymolysis liquid, and standing for 23 hours;
step 6) placing the feed liquid prepared in the step 5) into a centrifugal device for centrifugal separation, wherein the centrifugal revolution is 9000r/min, and the centrifugal treatment time is 33 min;
step 7) adding activated carbon into the fish oil obtained by the treatment in the step 6), wherein the adding amount of the activated carbon is 4% of the mass of the fish oil obtained by the treatment in the step 6); and separating the activated carbon by adopting centrifugal equipment to obtain the fish oil.
Example 3
Step 1) washing tuna leftovers with clear water, smashing, adding water which is 2 times of the tuna leftovers to mix into slurry, and uniformly stirring the slurry;
preparing brine with the mass concentration of 0.3g/mL by using sodium chloride, heating the brine to 25 ℃, and adding the brine into the slurry, wherein the mass ratio of the brine to the slurry is 1: 6; continuously stirring the slurry added with the brine to uniformly mix the slurry with the brine; standing for later use;
step 2) adding pepsin into the pulp obtained in the step 1) for primary enzymolysis, wherein the primary enzymolysis temperature is 35 ℃, the pH value is 3, and the primary enzymolysis time is 10 hours, wherein the adding amount of the pepsin is 2.5% of the mass of the pulp obtained in the step 1), and in the primary enzymolysis process, ultrasonic treatment is simultaneously carried out, the ultrasonic frequency of the ultrasonic treatment is 950kHz, nitrogen is continuously introduced into the pulp, and the flow of the nitrogen is 1.5m/s, so that primary enzymolysis liquid is obtained;
step 3) placing the primary enzymatic hydrolysate obtained by the treatment in the step 2) into a container, continuously introducing carbon dioxide gas into the primary enzymatic hydrolysate for 2 hours, wherein the flow rate of the carbon dioxide gas is 4m/s, then placing the container into a high-voltage pulse electric field for treatment, continuously introducing the carbon dioxide gas during the treatment process of the high-voltage pulse electric field, wherein the flow rate of the carbon dioxide gas is 1m/s, and after the treatment of the high-voltage pulse electric field is finished, the carbon dioxide gas is also continuously introduced, the flow rate of the carbon dioxide gas is consistent with the flow rate during the treatment process of the high-voltage pulse electric field, the electric field strength of the high-voltage pulse electric field is 56kV/cm, the frequency is 120Hz, the pulse width is 22 mus, and the treatment time is 8 s;
step 4) placing the feed liquid obtained by the treatment in the step 3) for 30min, adding neutral protease for secondary enzymolysis, wherein the temperature of the secondary enzymolysis is 43 ℃, the pH value is 8, the adding amount of the neutral protease is 2% of the mass of the feed liquid obtained by the treatment in the step 3), the time of the secondary enzymolysis is 3 hours, in the process of the secondary enzymolysis, ultrasonic treatment is simultaneously carried out, the ultrasonic frequency of the ultrasonic treatment is 650kHz, nitrogen is continuously introduced into the pulp, and the flow of the nitrogen is 4m/s, so that secondary enzymolysis liquid is obtained; the neutral protease is composed of trypsin, papain and subtilisin, and the mass ratio of the trypsin to the papain to the subtilisin is 5:2: 0.5;
step 5) adding hydrochloric acid with the mass concentration of 3% into the secondary enzymolysis liquid, adjusting the pH of the secondary enzymolysis liquid to 4-5, continuously adding absolute ethyl alcohol, wherein the addition amount of the absolute ethyl alcohol in the secondary enzymolysis liquid is 70% of the volume of the secondary enzymolysis liquid, and standing for 23 hours;
step 6) placing the feed liquid prepared in the step 5) into a centrifugal device for centrifugal separation, wherein the centrifugal revolution is 8500r/min, and the centrifugal treatment time is 26 min;
step 7) adding activated carbon into the fish oil obtained by the treatment in the step 6), wherein the adding amount of the activated carbon is 3% of the mass of the fish oil obtained by the treatment in the step 6); and separating the activated carbon by adopting centrifugal equipment to obtain the fish oil.
Example 4
Step 1) washing tuna leftovers with clear water, smashing, adding water which is 2 times of the tuna leftovers to mix into slurry, and uniformly stirring the slurry;
preparing brine with the mass concentration of 0.3g/mL by using sodium chloride, heating the brine to 25 ℃, and adding the brine into the slurry, wherein the mass ratio of the brine to the slurry is 1: 5; continuously stirring the slurry added with the brine to uniformly mix the slurry with the brine; standing for later use;
step 2) adding pepsin into the pulp obtained in the step 1) for primary enzymolysis, wherein the primary enzymolysis temperature is 35 ℃, the pH value is 3, and the primary enzymolysis time is 10 hours, wherein the adding amount of the pepsin is 2.5% of the mass of the pulp obtained in the step 1), and in the primary enzymolysis process, ultrasonic treatment is simultaneously carried out, the ultrasonic frequency of the ultrasonic treatment is 900kHz, nitrogen is continuously introduced into the pulp, and the flow of the nitrogen is 2m/s, so that primary enzymolysis liquid is obtained;
step 3) placing the primary enzymatic hydrolysate obtained by the treatment in the step 2) into a container, continuously introducing carbon dioxide gas into the primary enzymatic hydrolysate for 1h, wherein the flow rate of the carbon dioxide gas is 4m/s, then placing the container into a high-voltage pulse electric field for treatment, continuously introducing the carbon dioxide gas during the treatment process of the high-voltage pulse electric field, wherein the flow rate of the carbon dioxide gas is 1m/s, and after the treatment of the high-voltage pulse electric field is finished, the carbon dioxide gas is also continuously introduced, the flow rate of the carbon dioxide gas is consistent with the flow rate during the treatment process of the high-voltage pulse electric field, the electric field intensity of the high-voltage pulse electric field is 55kV/cm, the frequency is 130Hz, the pulse width is 20 mus, and the treatment time is 8 s;
step 4) placing the feed liquid obtained by the treatment in the step 3) for 40min, adding neutral protease for secondary enzymolysis, wherein the temperature of the secondary enzymolysis is 43 ℃, the pH value is 8, the adding amount of the neutral protease is 2% of the mass of the feed liquid obtained by the treatment in the step 3), the time of the secondary enzymolysis is 3 hours, in the process of the secondary enzymolysis, ultrasonic treatment is simultaneously carried out, the ultrasonic frequency of the ultrasonic treatment is 680kHz, nitrogen is continuously introduced into the pulp, and the flow of the nitrogen is 3.5m/s, so that secondary enzymolysis liquid is obtained; the neutral protease is composed of trypsin, papain and subtilisin, and the mass ratio of the trypsin to the papain to the subtilisin is 4:1: 1;
step 5) adding hydrochloric acid with the mass concentration of 3% into the secondary enzymolysis liquid, adjusting the pH of the secondary enzymolysis liquid to 4-5, continuously adding absolute ethyl alcohol, wherein the addition amount of the absolute ethyl alcohol in the secondary enzymolysis liquid is 70% of the volume of the secondary enzymolysis liquid, and standing for 23 hours;
step 6) placing the feed liquid prepared in the step 5) into a centrifugal device for centrifugal separation, wherein the centrifugal revolution is 9000r/min, and the centrifugal treatment time is 33 min;
step 7) adding activated carbon into the fish oil obtained by the treatment in the step 6), wherein the adding amount of the activated carbon is 4% of the mass of the fish oil obtained by the treatment in the step 6); and separating the activated carbon by adopting centrifugal equipment to obtain the fish oil.
Example 5
Step 1) washing tuna leftovers with clear water, smashing, adding water which is 2 times of the tuna leftovers to mix into slurry, and uniformly stirring the slurry;
preparing brine with the mass concentration of 0.5g/mL by using sodium chloride, heating the brine to 30 ℃, and adding the brine into the slurry, wherein the mass ratio of the brine to the slurry is 1: 8; continuously stirring the slurry added with the brine to uniformly mix the slurry with the brine; standing for later use;
step 2) adding pepsin into the pulp obtained in the step 1) for primary enzymolysis, wherein the primary enzymolysis temperature is 35 ℃, the pH value is 3, and the primary enzymolysis time is 10 hours, wherein the adding amount of the pepsin is 2.5% of the mass of the pulp obtained in the step 1), and in the primary enzymolysis process, ultrasonic treatment is simultaneously carried out, the ultrasonic frequency of the ultrasonic treatment is 870kHz, nitrogen is continuously introduced into the pulp, and the flow of the nitrogen is 2.5m/s, so that primary enzymolysis liquid is obtained;
step 3) placing the primary enzymatic hydrolysate obtained by the treatment in the step 2) into a container, continuously introducing carbon dioxide gas into the primary enzymatic hydrolysate for 2 hours, wherein the flow rate of the carbon dioxide gas is 4.5m/s, then placing the container into a high-voltage pulse electric field for treatment, continuously introducing the carbon dioxide gas during the treatment of the high-voltage pulse electric field, wherein the flow rate of the carbon dioxide gas is 1m/s, and after the treatment of the high-voltage pulse electric field is finished, the flow rate of the carbon dioxide gas is consistent with the flow rate during the treatment of the high-voltage pulse electric field, the electric field intensity of the high-voltage pulse electric field is 55kV/cm, the frequency is 100Hz, the pulse width is 20 μ s, and the treatment time is 5 s;
step 4) placing the feed liquid obtained by the treatment in the step 3) for 30-60min, adding neutral protease for secondary enzymolysis, wherein the temperature of the secondary enzymolysis is 45 ℃, the pH value is 9, the adding amount of the neutral protease is 3% of the mass of the feed liquid obtained by the treatment in the step 3), simultaneously performing ultrasonic treatment in the secondary enzymolysis process, the ultrasonic frequency of the ultrasonic treatment is 700kHz, the time of the secondary enzymolysis is 4 hours, and simultaneously continuously introducing nitrogen into the pulp, wherein the flow rate of the nitrogen is 3.5m/s, so as to obtain secondary enzymolysis liquid; the neutral protease is composed of trypsin, papain and subtilisin, and the mass ratio of the trypsin to the papain to the subtilisin is 5:2: 1;
step 5) adding hydrochloric acid with the mass concentration of 3% into the secondary enzymolysis liquid, adjusting the pH of the secondary enzymolysis liquid to 4-5, continuously adding absolute ethyl alcohol, wherein the addition amount of the absolute ethyl alcohol in the secondary enzymolysis liquid is 70% of the volume of the secondary enzymolysis liquid, and standing for 23 hours;
step 6) placing the feed liquid prepared in the step 5) into centrifugal equipment for centrifugal separation, wherein the centrifugal revolution is 8800r/min, and the centrifugal treatment time is 26 min;
step 7) adding activated carbon into the fish oil obtained by the treatment in the step 6), wherein the adding amount of the activated carbon is 3% of the mass of the fish oil obtained by the treatment in the step 6); and separating the activated carbon by adopting centrifugal equipment to obtain the fish oil.
Effect verification:
tuna fish oil was extracted using the methods of examples 1-5. Wherein each example processed 1000g of tuna offal. The indices of the fish oils extracted in examples 1 to 5 are shown in table 1. The invention improves the extraction rate of the tuna fish oil and the content of EPA and DHA in the fish oil.
TABLE 1
While embodiments of the invention have been disclosed above, it is not limited to the applications listed in the description and the embodiments, which are fully applicable in all kinds of fields of application of the invention, and further modifications may readily be effected by those skilled in the art, so that the invention is not limited to the specific details without departing from the general concept defined by the claims and the scope of equivalents.
Claims (3)
1. An enzymolysis and electric field composite treatment method for tuna leftovers is characterized by comprising the following steps:
step 1) washing tuna leftovers with clear water, smashing, adding water which is 0.5-2 times of the tuna leftovers to mix into slurry, and uniformly stirring the slurry;
preparing saline water with the mass concentration of 0.3-0.5g/mL by using sodium chloride, heating the saline water to 25-30 ℃, and adding the saline water into the slurry, wherein the mass ratio of the saline water to the slurry is 1: 5-8; continuously stirring the slurry added with the brine to uniformly mix the slurry with the brine; standing for later use;
step 2) adding pepsin into the pulp obtained in the step 1) for carrying out primary enzymolysis, wherein the primary enzymolysis temperature is 30-35 ℃, the pH value is 2-3, the primary enzymolysis time is 9-10 hours, nitrogen is continuously introduced into the pulp at the same time, the flow of the nitrogen is 1.5-2.5m/s, ultrasonic treatment is carried out at the same time, the ultrasonic frequency of the ultrasonic treatment is 800-950kHz, and the adding amount of the pepsin is 2-2.5% of the mass of the pulp obtained in the step 1), so as to obtain primary enzymolysis liquid;
step 3) placing the primary enzymolysis liquid obtained by the treatment in the step 2) into a container, continuously introducing carbon dioxide gas into the primary enzymolysis liquid for 1-2h, wherein the flow rate of the carbon dioxide gas is 4-5m/s, then placing the container into a high-voltage pulse electric field for treatment, continuously introducing the carbon dioxide gas during the treatment of the high-voltage pulse electric field, wherein the flow rate of the carbon dioxide gas is 1-2m/s, continuously introducing the carbon dioxide gas after the treatment of the high-voltage pulse electric field is finished, the flow rate of the carbon dioxide gas is consistent with the flow rate during the treatment of the high-voltage pulse electric field, the electric field strength of the high-voltage pulse electric field is 55-60kV/cm, the frequency is 100-150Hz, and the pulse width is 20-25 μ s, the treatment time is 5-10 s;
step 4) placing the feed liquid obtained by the treatment of the step 3) for 30-60min, adding neutral protease for secondary enzymolysis, wherein the temperature of the secondary enzymolysis is 40-45 ℃, the pH value is 8-9, simultaneously introducing nitrogen into the pulp continuously, the flow rate of the nitrogen is 3.5-5.5m/s, simultaneously performing ultrasonic treatment, the ultrasonic frequency of the ultrasonic treatment is 650-700kHz, the adding amount of the neutral protease is 2-3% of the mass of the feed liquid obtained by the treatment of the step 3), and the time of the secondary enzymolysis is 3-4 hours, so as to obtain secondary enzymolysis liquid; wherein the neutral protease is composed of trypsin, papain and subtilisin, and the mass ratio of the trypsin to the papain to the subtilisin is 4-5:1-2: 0.5-1;
step 5) adding hydrochloric acid with the mass concentration of 2-3% into the secondary enzymolysis liquid, adjusting the pH of the secondary enzymolysis liquid to 4-5, continuously adding absolute ethyl alcohol, wherein the addition amount of the absolute ethyl alcohol in the secondary enzymolysis liquid is 65-70% of the volume of the secondary enzymolysis liquid, and standing for 20-23 hours;
and step 6) placing the material liquid prepared in the step 5) into a centrifugal device for centrifugal separation, wherein the centrifugal rotation number is 8000-9000r/min, and the centrifugal treatment time is 26-33 min.
2. The method for combined enzymatic hydrolysis and electric field treatment of tuna offal according to claim 1, wherein in the step 4), the mass ratio of the trypsin, the papain and the subtilisin is 4:1: 0.5.
3. The method for combined enzymatic hydrolysis and electric field treatment of tuna leftovers according to claim 1, wherein in the step 3), carbon dioxide gas is continuously introduced into the primary enzymatic hydrolysate for 1 hour, the flow rate of the carbon dioxide gas is 4m/s, and the carbon dioxide gas is continuously introduced during the high-voltage pulsed electric field treatment process, and the flow rate of the carbon dioxide gas is 1 m/s.
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