CN108060197A - The method for integrated extraction of multiple biological activities ingredient in a kind of sea eel - Google Patents
The method for integrated extraction of multiple biological activities ingredient in a kind of sea eel Download PDFInfo
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- XOAAWQZATWQOTB-UHFFFAOYSA-N taurine Chemical compound NCCS(O)(=O)=O XOAAWQZATWQOTB-UHFFFAOYSA-N 0.000 description 2
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- RUUHDEGJEGHQKL-UHFFFAOYSA-M 2-hydroxypropyl(trimethyl)azanium;chloride Chemical compound [Cl-].CC(O)C[N+](C)(C)C RUUHDEGJEGHQKL-UHFFFAOYSA-M 0.000 description 1
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- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 1
- 239000004472 Lysine Substances 0.000 description 1
- 235000005035 Panax pseudoginseng ssp. pseudoginseng Nutrition 0.000 description 1
- 235000003140 Panax quinquefolius Nutrition 0.000 description 1
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- 125000001301 ethoxy group Chemical group [H]C([H])([H])C([H])([H])O* 0.000 description 1
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- 229920000159 gelatin Polymers 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000008434 ginseng Nutrition 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 229910052740 iodine Inorganic materials 0.000 description 1
- 239000011630 iodine Substances 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
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- 235000015112 vegetable and seed oil Nutrition 0.000 description 1
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- 239000002699 waste material Substances 0.000 description 1
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Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/06—Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/20—Animal feeding-stuffs from material of animal origin
- A23K10/22—Animal feeding-stuffs from material of animal origin from fish
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/158—Fatty acids; Fats; Products containing oils or fats
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/78—Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin or cold insoluble globulin [CIG]
-
- 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
- C11B1/025—Pretreatment by enzymes or microorganisms, living or dead
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- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Zoology (AREA)
- Polymers & Plastics (AREA)
- Wood Science & Technology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Molecular Biology (AREA)
- Biochemistry (AREA)
- Genetics & Genomics (AREA)
- Microbiology (AREA)
- Animal Husbandry (AREA)
- General Health & Medical Sciences (AREA)
- Food Science & Technology (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Biotechnology (AREA)
- Biophysics (AREA)
- Physiology (AREA)
- Biomedical Technology (AREA)
- Marine Sciences & Fisheries (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- General Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Gastroenterology & Hepatology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Toxicology (AREA)
- Meat, Egg Or Seafood Products (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention discloses a kind of method for integrated extraction of multiple biological activities ingredient in sea eel, comprise the following steps：S1 substrates pre-process：The protein content gone in sea eel meat content and sea eel meat in intestines sea eel, eel head and eel bone is measured respectively；Intestines is gone to clean, rub sea eel, and/or eel head and eel bone are cleaned, rubbed, the particle for removing intestines sea eel, eel head and eel bone is diluted with pure water respectively, the protein content gone in intestines sea eel feed liquid, eel head feed liquid and eel bone feed liquid made is respectively 4.5% the 9.0% of its feed liquid total weight, puts into reaction kettle；S2 digests to obtain first time enzymolysis product for the first time；S3 carries out bone fluid phase separation to first time enzymolysis product；S4 extracts to obtain sea eel meat oligopeptide from liquid phase；S5 extracts bone meal, bone collagen oligopeptide and oil from bone；The oil merging post processing that slightly carries in S3 and S5 is obtained refined eel oil or feed rank fish oil by S6.The present invention can make full use of sea eel resource, avoid the wasting of resources.
Description
Technical field
The present invention relates to biochemistry enzymolysis extraction field, more particularly, to multiple biological activities in a kind of sea eel into
The method for integrated extraction divided.
Background technology
Common eel is commonly called as river eel, also known as white eel, blue or green eel, Bai Qiu, snake fish etc., belongs to Osteichthyes sea eel mesh, Anguillidae, common eel
Belong to, be distributed widely in global tropical, subtropical zone and Temperate Region in China, be distributed mainly on the coastal state of Pacific Northwest.The world
Upper common eel shares 19 kinds (including subspecies), and China has described 8 kinds of common eels.Sea eel be a kind of meat flavour it is delicious, it is full of nutrition and
The first-class edible fishes to nourish the body have the laudatory title of " ginseng in water ".
Protein content is 13% or so (fresh weight) in sea eel, crude fat difference (5.1%- when quality is different
24.0%).Amino acid classes are complete, eight kinds must amino acid account for total amino acid ratio up to 39%, meet the high-quality eggs of FAO/WHO
The requirement in white matter source, wherein lysine account for the 9% of total amino acid content, and contain higher taurine.
In recent years, sea eel industry is fast-developing, and China has become the highest state of common eel aquaculture industry degree in the world
One of family, year sea eel yield about 1.6 × 105Ton accounts for more than the 2/3 of the world, and the gross output value surpasses 10,000,000,000, and export processing amount surpasses 100,000
Ton, exports goods and earns foreign currency 900,000,000 dollars, roast eel accounts for product water outlet and earns foreign exchange first in year.But substantial amounts of roast eel generates enormous amount
Eel head and by-products, the present invention such as eel bone determine, it is 92% left to account for eel head total weight for the sea eel meat in eel head
Protein content is 14.5% or so in the right side, wherein meat, and the sea eel meat in eel bone accounts for 78.5% or so of eel bone total weight,
Therefore protein content 11.5% or so wherein in meat, are badly in need of a kind of integrated extraction work of multiple biological activities ingredient in sea eel
Skill makes sea eel resource be fully used, and avoids wasting.
The content of the invention
It is an object of the invention to overcome defect existing in the prior art, multiple biological activities ingredient in a kind of sea eel is provided
Method for integrated extraction, by the integrated processing to eel head and eel bone, fully extract sea eel or roast eel by-product eel head,
Sea eel meat oligopeptide, sea eel ossein oligopeptide, biological calcium powder and eel oil in eel bone, improve the utilization rate of sea eel, keep away
Exempt from the wasting of resources.
To achieve the above object, technical scheme is as follows：
The method for integrated extraction of multiple biological activities ingredient in a kind of sea eel, which is characterized in that comprise the following steps：
Step S1 substrates pre-process：It measures respectively and goes the sea eel meat content in intestines sea eel, eel head and eel bone and its eel
Protein content in the flesh of fish；Intestines gone to clean, rub sea eel and/or clean eel head and eel bone, rubbed, respectively to going
Intestines sea eel, eel head and eel bone particle are diluted with pure water, so as to get remove intestines sea eel feed liquid, eel head feed liquid and sea eel aggregate
Protein content in liquid is respectively the 4.5%-9.0% of its feed liquid total weight, obtains substrate feed liquid, puts into reaction kettle.The step
The middle content for determining protein in the percentage composition of contained meat and its meat in sea eel, eel head and eel bone respectively, accomplish with
Tot Prot for standard precisely feed intake, also, protein content be feed liquid total weight 4.5%-9.0% when, in all water
In the optimal concentration of substrate for solving enzyme, avoid concentration too low and increase post processing cost.
Step S2 is digested for the first time：Substrate feed liquid is warming up to 40 DEG C -55 DEG C, with CaO tune pH to 7.5-8.5, adds in matter
Measure the compound protease of the 1.0%-5.0% of Tot Prot in the substrate feed liquid obtained for step S1, stirring enzymolysis 3h-6h, enzyme
After solution, with HCl solution flavouring liquid pH to 5.0-7.0,70 DEG C of -90 DEG C of progress destroy the enzyme treatments are warming up to, obtains and digests for the first time
Product.In this step, sea eel meat is directly digested, is not required to boiling, remained the activity of constitutive enzyme in sea eel meat, reduce enzyme
Additive amount, reduce production cost, meanwhile, avoid influencing mouthfeel due to thermophilic digestion, improve product quality.On the other hand, enzyme
Solution time 3h-6h is the optimal enzymolysis time of enzyme used, and enzymolysis time is too short, and enzymolysis is insufficient, and enzymolysis time is long, is easily reduced
Productivity.
Preferably, the compound protease in digesting for the first time includes at least As1.398 neutral proteinases, 2709 basic proteins
Enzyme, trypsase and bromelain；Wherein, the composition of compound protease and its mass percent are：Enzymatic activity for 1 ×
105The As1.398 neutral proteinases 20%-60% of U/g, enzymatic activity are 2 × 105The 2709 alkali protease 20%-60% of U/g,
Enzymatic activity is 1 × 105The trypsase 10%-40% of U/g, enzymatic activity are 6 × 105The bromelain 5%-30% of U/g.It is multiple
The composition of hop protein enzyme is optimised, improves enzymolysis efficiency, and it is more than 80% to make product yield, molecular weight product<1000Da's is oligomeric
Peptide accounts for total protein ratio more than 95%.
Step S3 bones-oil-liquid phase separation：First time enzymolysis product is isolated by aperture for 10-30 mesh rotary screens
Obtained liquid is separated fuel-displaced and liquid phase by bone and liquid by tube centrifuge.
Step S4 liquid phases extract to obtain sea eel meat oligopeptide：The step S3 liquid phases isolated are decolourized, defishying, so
After filter out impurity and obtain refined filtration liquid, by the concentration of refined filtration liquid, dry sea eel meat oligopeptide.
Specifically, the process of the decoloration and defishying is：In the liquid phase isolated in step S3 add in flocculant into
Row flocculation, adjusts the temperature to 60 DEG C -70 DEG C, adds in powdered carbon while stirring and carries out decoloration deodorization, stirring and adsorbing 30min-60min,
Afterwards, de- charcoal obtains destainer, destainer is adjusted temperature to 60 DEG C -70 DEG C, it is the substrate feed liquid that step S1 is obtained to add in quality
The beta-cyclodextrin of middle Tot Prot 1.0%-15.0% stirs 10min-40min, carries out defishying, deodorant liquid is obtained, by deodorant
Liquid further crosses activated-charcoal column and carries out deodorant decoloration deodorization processing, obtains column liquid.
Specifically, column liquid is taken off into charcoal by ceramic membrane or cottonseed cake flame filter press and obtains refined filtration liquid.
Specifically, it is by the process of the concentration of refined filtration liquid, dry sea eel meat oligopeptide：Refined filtration liquid first is by retaining aperture
The organic film of 50Da-100Da carries out first time concentration, and making solid content, secondly, concentrate is steamed again by depressurizing up to 17%-18%
Hair is concentrated into solid content 40%-55%, then, concentrate is spray-dried to obtain the oligomeric Gly-His-Lys of sea eel meat.
Step S5 bones extract to obtain bone meal, bone collagen oligopeptide and oil：After being cleaned to the step S3 bones isolated
It is put into digester, by solid-liquid ratio 1:1-5 adds in pure water and carries out boiling, isolates bone slag, collagen liquid and oil；The bone
Slag is through rinsing, being dehydrated and dry and obtaining dry bone slag, and dry bone ground-slag is broken, sieve to obtain bone meal；The collagen liquid is put into enzymatic vessel,
The mass percent of collagen in collagen liquid is adjusted as 4.5%-9.0%, adjustment temperature is to 40 DEG C -55 DEG C, with CaO tune
Whole pH to 7.5-8.0 adds in compound protease of the quality for the 1.0%-5.0% of collagen quality in collagen liquid, into
Row second digests, stirring enzymolysis 3h-6h, after enzymolysis, adjusts pH to 5.0-7.0, is warming up to 70 DEG C -90 DEG C, at enzyme deactivation
20min-40min is managed, obtains second of enzymolysis product, then second of enzymolysis product filters out impurity and obtain through decoloration, defishying
Refined filtration liquid, smart concentrating filter liquor and be spray-dried solid the oligomeric Gly-His-Lys of bone collagen；The oil is isolated with step S3
Oil merging.
Specifically, the feed liquid after boiling is first separated it is fuel-displaced, then by remaining feed liquid squeeze into 30-40 mesh rotary screen carry out slag
Liquid separates, and obtains bone slag and collagen liquid respectively, and with pure water rinse bone slag, washing lotion is merged into collagen liquid, to merging
Collagen liquid afterwards separates the collagen liquid and grease that must be further purified, grease herein with tubular type or disk centrifugal separator
It is incorporated into the oil that step S3 is isolated.
Preferably, bone slag is dehydrated with tripod pendulum type batch centrifugal, dewatered bone slag is put into 100 DEG C of -120 DEG C of bakings of drying chamber
It is dry, dry bone slag is obtained, dry bone slag smashes it through -200 mesh sieve of 150 mesh, obtains bone meal.
Preferably, the compound protease in second of enzymolysis includes at least As1.398 neutral proteinases, trypsase
And papain；Wherein, the composition of compound protease and with its mass percent it is：Enzymatic activity is 1 × 105U/g's
As1.398 neutral proteinases 30%-60%, enzymatic activity are 1 × 105The trypsase 15%-40% of U/g, enzymatic activity for 6 ×
105The papain 5%-30% of U/g.
Preferably, collagen total amount 1.0%-15.0% in collagen liquid is incorporated as in second of enzymolysis product
Beta-cyclodextrin carries out deodorant, obtains deodorant liquid.
Preferably, active powdered carbon of the quality for the 0.5%-1.5% of collagen liquid volume is added in deodorant liquid, 60
30min-60min is stirred at DEG C -70 DEG C, decolorization is carried out, afterwards, takes off charcoal with plate and frame filter press, obtain destainer.
Preferably, filter out impurity with the stud fine filter in 0.2 μm -0.4 μm of aperture and obtain refined filtration liquid.
Preferably, the refined filtration liquid first carries out first time concentration, solid content with molecular cut off 50Da-100Da organic films
Up to 17%-18%；Afterwards, concentrate carries out secondary concentration through being evaporated under reduced pressure again, and solid content reaches 40%-55%；Finally, concentrate
It is spray-dried, obtain the gelatine collagen oligopeptide powder of solid.
Step S6 oil extracts must refine eel oil or feed rank fish oil：The oil and step S6 that step S3 is isolated are isolated
Oil merging, refined eel oil is obtained by edible oil and fat process for refining or feed is obtained by feed rank grease process for refining
Rank fish oil.
The edible oil and fat process for refining includes lipin deacidifying, decoloration, deodorization, degumming, dehydration.
Lipin deacidifying is carried out with aliphatic acid in potassium hydroxide, is decolourized and is deodorized with atlapulgite.
In conclusion advantages of the present invention is：
(1) containing for protein in the percentage composition of contained meat and its meat is determined in sea eel, eel head and eel bone respectively
Amount accomplishes precisely feeding intake using Tot Prot as standard；
(2) protein concentration in substrate feed liquid is determined in 4.5%-9.0%, in the optimal concentration of substrate of all hydrolases
It is interior, it avoids concentration too low and increases post processing cost；
(3) enzymolysis time 3h-6h also in the range of the optimal enzymolysis time of enzyme used, raises labour productivity；
(4) meat on sea eel and head and bone is directly digested without boiling, retains the activity of constitutive enzyme in sea eel meat, Ke Yishi
When the amount for reducing external enzyme, production cost is reduced, avoids influencing mouthfeel due to thermophilic digestion, improve product quality；
(5) compound protease used in is screened through single enzyme, optimum organization, improves enzymolysis efficiency, and product yield is more than 80%,
Molecular weight product<The oligopeptide of 1000Da accounts for total protein ratio more than 95%；
(6) sea eel hull melanin and ferroheme are removed with flocculant, obtains light enzymolysis liquid, improve product color；
(7) refined filtration liquid reduces energy consumption, improves product again through vacuum evaporation after organic film once concentrates
Quality；
(8) product is by decoloration, defishying, and in good taste, raw meat is not bitter；
(9) with CaO flavouring liquid pH, calcium content in oligopeptide is improved, consumer is made to take sea eel oligopeptide simultaneously
With supplement calcium；
(10) by active ingredient sea eel meat oligopeptide, sea eel ossein oligopeptide, eel oil and sea eel in sea eel and head, bone
Bone meal is totally separated, and eel head, bone resource is made to be fully used, the cost of material of these products is reduced, substantially increases
Business Economic Benefit, while accomplish to alleviate environmental pollution without waste discharge in addition to a small amount of waste water of cleaning equipment again.
Description of the drawings
Fig. 1 is the flow diagram of the method for integrated extraction of multiple biological activities ingredient in a kind of sea eel of the invention.
Specific embodiment
Below in conjunction with the accompanying drawings, the specific embodiment of the present invention is described in further detail.
In specific embodiment of the invention below, referring to Fig. 1, Fig. 1 is a variety of lifes in a kind of sea eel of the invention
The flow diagram of the method for integrated extraction of object active ingredient.As shown in Figure 1, multiple biological activities in a kind of sea eel of the present invention
The method for integrated extraction of ingredient, comprises the following steps：
Step S1 measures the albumen in sea eel meat content and its sea eel meat in intestines sea eel, eel head and eel bone respectively
Content；Intestines gone to clean, rub sea eel and/or clean eel head and eel bone, rubbed, respectively to removing intestines sea eel, eel head
Diluted with eel bone particle with pure water, so as to get the protein gone in intestines sea eel feed liquid, eel head feed liquid and eel bone feed liquid
Content is respectively the 4.5%-9.0% of its feed liquid total weight, obtains substrate feed liquid, puts into reaction kettle.
The present invention determine whole sea eel meat content for going intestines sea eel account for it is whole go 94% or so of intestines sea eel gross mass, it is whole
The protein content that item is gone in the sea eel meat of intestines sea eel accounts for the 13.0% of whole sea eel meat content for going intestines sea eel；On eel head
Sea eel meat weight accounts for 92% or so of total eel head weight, and the protein content in sea eel meat on eel head is on eel head
14.5% or so of sea eel meat weight；Sea eel meat weight in eel bone accounts for 78.5% or so of total sea eel bone weight, eel bone
In sea eel meat in protein content be 11.5% or so of the sea eel meat weight in eel bone.
In the present embodiment, eel head 100kg is taken, with crusher in crushing or rubbing, obtains eel head particle.By feed liquid quality
Than=1:1 adds pure water, i.e. 100kg eel heads add 100kg water, stir evenly.By the sea eel meat weight in the above-mentioned eel head measured
Amount accounts for 92% or so of total eel head weight, and the protein content in sea eel meat on eel head is the sea eel meat weight on eel head
Amount 14.5% or so, therefore, the protein-contg quality of 100kg eel heads be 100kg × 92% × 14.5%=13.34kg, bottom
Material liquid includes eel head particle and water, and therefore, the concentration of albumen is 13.34kg/200kg=6.67% in substrate feed liquid.
Step S2 is digested for the first time：Substrate feed liquid is warming up to 40 DEG C -55 DEG C, flavouring liquid pH to 7.5-8.5, adds in quality
For the compound protease of the 1.0%-5.0% of albumen gross mass in substrate feed liquid, stir and digest 3h-6h, after enzymolysis, condiment
Liquid pH to 5.0-7.0 is warming up to 70 DEG C of -90 DEG C of progress destroy the enzyme treatments, obtains first time enzymolysis product.
In the present embodiment, feed liquid is heated to 45 DEG C ± 1 DEG C；With CaO tune pH to 8.0 ± 0.1；Add compound protease
467g (account for albumen total weight 3.5%), compound protease is to be compounded after the screening of single enzyme system, and empirical tests, proportioning are
As1.398 neutral proteinases 1 × 105U/g 20%-60% (w/w), 2709 alkali proteases 2 × 105U/g 20%-60% (w/
W), trypsase 1 × 105U/g 10%-40% (w/w), bromelain 6 × 105U/g 5%-30% (w/w), stirring enzymolysis
4h, after enzymolysis, HCl solution flavouring liquid pH to 6.0 ± 0.1 is warming up to 80 DEG C, 15min enzyme deactivations sterilization.
Step S3 bones-oil-liquid phase separation isolate first time enzymolysis product by aperture for 10-30 mesh rotary screens
Obtained liquid is separated fuel-displaced and liquid phase by bone and liquid by tube centrifuge.
In the present embodiment, by feed liquid by 20 mesh rotary screens by feed liquid and broken osteodiastasis, obtain liquid and broken bone；With appropriate
The broken bone of pure water, isolates bone, and washing lotion is merged into liquid；The liquid separates (16300r/ by tube centrifuge
Min), enzymolysis liquid (liquid phase), grease (oil) and particulate matter are obtained, particulate matter is separated after being washed with appropriate pure water and is incorporated in bone, washing lotion
It is merged into enzymolysis liquid.
Step S4 liquid phases extract to obtain sea eel meat oligopeptide：The step S3 liquid phases isolated are decolourized, defishying, so
After filter out impurity and obtain refined filtration liquid, by the concentration of refined filtration liquid, dry sea eel meat oligopeptide.
In the present embodiment, it is 1.0% (w/v) HACC that concentration is added in enzymolysis liquid (liquid phase that i.e. step S3 is isolated)
(2- hydroxypropyltrimethylammonium chloride chitosan) solution, dosage is 5.0% (v/v) of enzymolysis liquid volume, after fully stirring evenly, enriching
It spends for 3.0% (w/v) CTS (chitosan), dosage is enzymolysis liquid volume 2.0% (v/v), and 10min is stood after fully stirring evenly, and is centrifuged
Separation, obtains faint yellow clarification enzymolysis liquid, pigment and impurity extreme portions are removed.
And then enzymolysis liquid is warming up to 65 DEG C or so, add in powdered activated carbon 1.0% (w/v), stirring deodorant, decoloration
Refined 40min, takes off charcoal with plate and frame filter press and obtains coarse filtration liquid；Coarse filtration liquid is squeezed into reaction kettle, is heated to 65 DEG C or so, with total egg
White matter amount is 100% meter, adds in 1.0% beta-cyclodextrin (w/w), and stir process 30min carries out embedding deodorant and deodorizes.Deodorant
Liquid crosses seed activity charcoal post, and deodorant decoloration again is deodorized；It is the ceramic membrane of 50nm or cottonseed cake plate-frame filtering that column liquid, which is crossed, by aperture
Machine takes off charcoal, obtains refined filtration liquid；Refined filtration liquid carries out first time concentration by the organic film of molecular cut off 50Da, reaches solid content
17%-18%；By a concentrate by being concentrated under reduced pressure into solid content 40% or so, secondary concentration liquid is obtained；Concentrate is sprayed
It is dry, sea eel meat oligopeptide 11.04kg is obtained, yield is 82.3% (percentage for accounting for Tot Prot in eel head meat), and product divides
Son amount<The oligopeptide of 1000Da accounts for total oligopeptide weight ratio as 95.0%.
Step S5 bones extract to obtain bone meal, bone collagen oligopeptide and oil：After being cleaned to the step S3 bones isolated
It is put into digester, by solid-liquid ratio 1:1-5 adds in pure water and carries out boiling, isolates bone slag, collagen liquid and oil；The bone
Slag is through rinsing, being dehydrated and dry and obtaining dry bone slag, and dry bone ground-slag is broken, sieve to obtain bone meal；The collagen liquid is put into reaction kettle,
The mass percent of collagen in collagen liquid is adjusted as 4.5%-9.0%, adjustment temperature is to 40 DEG C -55 DEG C, with CaO tune
Whole pH to 7.5-8.0 adds in compound protease of the quality for the 1.0%-5.0% of collagen quality in collagen liquid, into
Row second digests, stirring enzymolysis 3h-6h, after enzymolysis, adjusts pH to 5.0-7.0, is warming up to 70 DEG C -90 DEG C, at enzyme deactivation
20min-40min is managed, obtains second of enzymolysis product, then second of enzymolysis product filters out impurity and obtain through decoloration, defishying
The oligomeric Gly-His-Lys of bone collagen of refined filtration liquid, smart concentrating filter liquor and dry solid；The oil is incorporated into step S6.
In the present embodiment, it is put into after the step S3 bones isolated are cleaned in digester, by solid-liquid ratio 1:3 plus pure
Water；Steam is opened, pot inner pressure keeps 20bar, maintains 2h, stops heating；When pot inner pressure is down to 0, feed liquid is squeezed into 40
It is separated in mesh rotary screen, obtains collagen liquid and bone slag, bone slag is merged into through appropriate pure water, washing lotion in collagen liquid.
Collagen liquid is separated into de-oiling by tube centrifuge, obtains de-oiling collagen liquid and oil, oil is collected, and is separated with step S3
The oil merging gone out, further to be refined to grease in subsequent technique.
The case collagen liquid of de-oiling is returned into enzymatic vessel, adjusts the mass percent of collagen in collagen liquid
For 7.5%；Feed liquid is warming up to 45 DEG C ± 1 DEG C；With CaO flavouring liquids pH to 7.5-7.7；Adding compound protease 2.5%, (w/w is accounted for
The mass percent of Tot Prot in collagen liquid), 45 DEG C of stirring enzymolysis 4h.It is compounded after the screening of single enzyme system, and experience
Card, its proportioning of compound protease is that enzymatic activity is 1 × 105The As1.398 neutral proteinases 30%-60% of U/g, enzymatic activity 1
×105The trypsase 15%-40% of U/g, enzymatic activity are 6 × 105The papain 5%-30% of U/g.After enzymolysis,
With HCl solution flavouring liquid pH6.0 ± 0.1；Feed liquid is warming up to 75 DEG C, adds 1.0% (w/v) powdered activated carbon, stirring and adsorbing
40min, can enzyme deactivation reach decolorizing effect again；Feed liquid is squeezed into de- charcoal in plate and frame filter press and obtains coarse filtration liquid；Coarse filtration liquid passes through hole
0.2 μm of footpath stud refined filtration, obtains refined filtration liquid；Refined filtration liquid is concentrated with the organic film of molecular cut off 80Da, obtains a concentrate,
Solid content 17%-18%；Concentrate carries out secondary concentration with reduction vaporization thickener again, obtains the two of solid content 45% or so
Secondary concentrate；Secondary concentration liquid is spray-dried to obtain sea eel bone collagen peptide powder 3.23kg, yield 80.75%, product point
Son amount<The oligopeptide of 1000Da accounts for the 95.4% of total oligopeptide weight.
The bone slag that will be separated is rinsed 2-3 times with pure water, obtains os purum slag；Os purum slag is dehydrated with tripod pendulum type batch centrifugal；It will be de-
Os purum slag after water is placed in drying chamber or baking oven and is dried to water content below 2% for 100 DEG C -120 DEG C；The bone residues ultramicro powder of drying is broken
By 150 mesh, food-grade organism calcium powder is obtained.
Step S6 oil extracts must refine eel oil：The oil merging that the oil and step S6 that step S3 is isolated are isolated, passes through
Edible oil and fat process for refining obtains refined eel oil.
In the present embodiment, grease first measures its acid value and iodine value after merging.It is refined by feed rank and food grade two
Technique carries out, and is carried out with reference to vegetable oil process for refining.
Feed rank fish oil (second-rate crude oil) process for refining is as follows：
Crude fish oil (crude oil) → filtering → depickling → washing → drying → decoloration → modulation (adds 300ppm-500ppm ethoxies
Base quinoline makees antioxidant).
Edible (or health-care oil) (the preferable crude oil of quality) process for refining is as follows：
Crude fish oil (crude oil) → filtering → degumming → depickling → washing → decoloration → winterization → deodorization → modulation (adds
The VE (vitamin E) of the TBHQ (tert-butylhydroquinone) or addition 0.5%-1% of 100ppm-200ppm make antioxidant)
Wherein, lipin deacidifying is carried out with aliphatic acid using in potassium hydroxide, is decolourized and deodorized with atlapulgite.
The foregoing is only a preferred embodiment of the present invention, but protection scope of the present invention be not limited thereto,
Any one skilled in the art in the technical scope disclosed by the present invention, technique according to the invention scheme and its
Inventive concept is subject to equivalent substitution or change, should be covered by the protection scope of the present invention.
Claims (9)
1. the method for integrated extraction of multiple biological activities ingredient in a kind of sea eel, which is characterized in that comprise the following steps：
Step S1 substrates pre-process：The sea eel meat content and its sea eel meat gone in intestines sea eel, eel head and eel bone are measured respectively
In protein content；Intestines gone to clean, rub sea eel and/or clean eel head and eel bone, rubbed, respectively to going intestines eel
The particle of fish, eel head and eel bone is diluted with pure water, so as to get go intestines sea eel feed liquid, eel head feed liquid and eel bone feed liquid
In protein content be respectively its feed liquid total weight 4.5%-9.0%, obtain substrate feed liquid, put into reaction kettle；
Step S2 is digested for the first time：Substrate feed liquid is warming up to 40 DEG C -55 DEG C, with CaO tune pH to 7.5-8.5, adding in quality is
The compound protease of the 1.0%-5.0% of albumen gross mass in the substrate feed liquid that step S1 is obtained, stirring enzymolysis 3h-6h, enzymolysis
After, with HCl solution flavouring liquid pH to 5.0-7.0,70 DEG C of -90 DEG C of progress destroy the enzyme treatments are warming up to, obtain enzymolysis production for the first time
Object；
Step S3 bones-oil-liquid phase separation：First time enzymolysis product is isolated into bone by aperture for 10-30 mesh rotary screens
And liquid, obtained liquid is separated into fuel-displaced and liquid phase by tubular type or disk centrifugal separator；
Step S4 liquid phases extract to obtain sea eel meat oligopeptide：The step S3 liquid phases isolated are decolourized, defishying, then mistake
It filters out impurity and obtains refined filtration liquid, refined filtration liquid is further concentrated, is spray-dried to obtain sea eel meat oligopeptide；
Step S5 bones extract to obtain bone meal, bone collagen oligopeptide and oil：It is put into after being cleaned to the step S3 bones isolated
In digester, by feed liquid mass ratio 1:1-5 adds in pure water and carries out boiling, isolates bone slag, collagen liquid and oil；The bone
Slag is through rinsing, being dehydrated and dry and obtaining dry bone slag, and dry bone ground-slag is broken, sieve to obtain bone meal；The collagen liquid is put into reaction kettle,
The mass percent of collagen in collagen liquid is adjusted as 4.5%-9.0%, adjustment temperature is to 40 DEG C -55 DEG C, with CaO tune
Whole pH to 7.5-8.0 adds in compound protease of the quality for the 1.0%-5.0% of collagen quality in collagen liquid, into
Row second digests, stirring enzymolysis 3h-6h, after enzymolysis, adjusts pH to 5.0-7.0, is warming up to 70 DEG C -90 DEG C, at enzyme deactivation
20min-40min is managed, obtains second of enzymolysis product, then second of enzymolysis product filters out impurity and obtain through decoloration, defishying
Refined filtration liquid, smart concentrating filter liquor and is spray-dried to obtain the oligomeric Gly-His-Lys of bone collagen；The oil conjunction that the oil is isolated with step S3
And；
Step S6 oil extracts must refine eel oil or feed rank fish oil：The oil that the oil and step S6 that step S3 is isolated are isolated
Merge, refined eel oil is obtained by edible oil and fat process for refining or feed rank is obtained by feed rank grease process for refining
Fish oil.
2. the method for integrated extraction of multiple biological activities ingredient in a kind of sea eel according to claim 1, which is characterized in that
In the step S2, the compound protease in digesting for the first time includes at least As1.398 neutral proteinases, 2709 basic proteins
Enzyme, trypsase and bromelain.
3. the method for integrated extraction of multiple biological activities ingredient in a kind of sea eel according to claim 2, which is characterized in that
The composition of the compound protease and its mass percent are：Enzymatic activity is 1 × 105The As1.398 neutral proteinases of U/g
20%-60%, enzymatic activity are 2 × 105The 2709 alkali protease 20%-60% of U/g, enzymatic activity are 1 × 105The tryptose of U/g
Enzyme 10%-40%, enzymatic activity are 6 × 105The bromelain 5%-30% of U/g.
4. the method for integrated extraction of multiple biological activities ingredient in a kind of sea eel according to claim 1, which is characterized in that
In the step S4 and S5, the decolorization is：Flocculant is added in be flocculated, add in powdered carbon stirring and adsorbing and cross activated carbon
A kind of mode or various ways combination in column.
5. the method for integrated extraction of multiple biological activities ingredient in a kind of sea eel according to claim 1, which is characterized in that
In the step S4 and S5, the defishying carries out defishying to add in beta-cyclodextrin.
6. the method for integrated extraction of multiple biological activities ingredient in a kind of sea eel according to claim 1, which is characterized in that
In the step S4 and S5, the acquisition of the refined filtration liquid is to be taken off using ceramic membrane or cottonseed cake flame filter press after charcoal through stud refined filtration
Device is obtained by filtration.
7. the method for integrated extraction of multiple biological activities ingredient in a kind of sea eel according to claim 1, which is characterized in that
In the step S4 and S5, the process by the concentration of refined filtration liquid, spray drying is：Refined filtration liquid is by retaining aperture 50Da- first
The organic film of 100Da carries out first time concentration, and making solid content, secondly, concentrate is concentrated again by being evaporated under reduced pressure up to 17%-18%
To solid content 40%-50%, then, concentrate is spray-dried to obtain solid powder.
8. the method for integrated extraction of multiple biological activities ingredient in a kind of sea eel according to claim 1, which is characterized in that
In the step S5, described second digest in compound protease include at least As1.398 neutral proteinases, trypsase and
Papain.
9. the method for integrated extraction of multiple biological activities ingredient in a kind of sea eel according to claim 8, which is characterized in that
In the step S5, described second digest in compound protease composition and with its mass percent be：Enzymatic activity for 1 ×
105The As1.398 neutral proteinases 30%-60% of U/g, enzymatic activity are 1 × 105The trypsase 15%-40% of U/g, enzymatic activity
For 6 × 105The papain 5%-30% of U/g.
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| CN116622436A (en) * | 2023-07-25 | 2023-08-22 | 烟台海润嘉生物科技有限公司 | Production method for synchronously extracting tuna fish oil and collagen peptide |
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