CN109043114A - A kind of preparation process and its application of mussel extract - Google Patents
A kind of preparation process and its application of mussel extract Download PDFInfo
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- CN109043114A CN109043114A CN201810666201.3A CN201810666201A CN109043114A CN 109043114 A CN109043114 A CN 109043114A CN 201810666201 A CN201810666201 A CN 201810666201A CN 109043114 A CN109043114 A CN 109043114A
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- FBOZXECLQNJBKD-UHFFFAOYSA-N methotrexate Chemical compound C=1N=C2N=C(N)N=C(N)C2=NC=1CN(C)C1=CC=C(C(=O)NC(CCC(O)=O)C(O)=O)C=C1 FBOZXECLQNJBKD-UHFFFAOYSA-N 0.000 description 1
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Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23J1/00—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites
- A23J1/04—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from fish or other sea animals
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23J3/00—Working-up of proteins for foodstuffs
- A23J3/30—Working-up of proteins for foodstuffs by hydrolysis
- A23J3/32—Working-up of proteins for foodstuffs by hydrolysis using chemical agents
- A23J3/34—Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes
- A23J3/341—Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes of animal proteins
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/17—Amino acids, peptides or proteins
- A23L33/18—Peptides; Protein hydrolysates
-
- 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
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Food Science & Technology (AREA)
- Organic Chemistry (AREA)
- Polymers & Plastics (AREA)
- Biochemistry (AREA)
- Molecular Biology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Wood Science & Technology (AREA)
- General Chemical & Material Sciences (AREA)
- Nutrition Science (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Marine Sciences & Fisheries (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Mycology (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Peptides Or Proteins (AREA)
Abstract
It is main by operations such as deodorization grease removal, cleaning, denaturation, enzymatic hydrolysis, concentration and dryings the present invention relates to a kind of preparation process of mussel extract and its application, including step S1~S9.A perfect mussel extract preparation process is provided, this by-product is utilized to mussel shell；Gained mussel extract has the characteristics that sterile, high activity and easily storage；Feeding trial is carried out by rat, the left back sufficient Bones and joints swelling situation of each group rat is detected by MRI scan, feeding mussel peptide rat bone arthroedema and swelling are all alleviated as the result is shown；In the acute RA of complete Freund's adjuvant induction, mussel peptide can prevent the state of an illness from further deteriorating by its anti-inflammatory symptom for alleviating RA with consumer edema effect；In collagen-induced chronic RA, mussel peptide can reach certain preventive effect by its anti-inflammatory acts on consumer edema；Do not observe that adverse reaction caused by traditional treatment RA drug occurs in rat during long-term feeding mussel peptide.
Description
Technical field
The present invention relates to the preparation process of biological products preparation and application field more particularly to a kind of mussel extract and its
Using.
Background technique
In drug research field, terrestrial organism and mineral are always the weight of emphasis and many new drugs that scholars study
Source is wanted, but with the continuous development of drug research, the exploitation of terrestrial resource is further difficult, and sight is focused on marine organisms
It has been trend of the times.Research shows that the extract of many marine organisms have anti-inflammatory, antitumor, antiviral, antibacterial, decompression and
The effects of anticoagulant is the utilizable important medicine source of the mankind.
Ancient book is recorded: mussel taste is salty, warm-natured, nontoxic, enters liver and kidney channel.It can tonifying kidney and benefiting sperm, tune liver blood-nourishing, the goiter that disappears, menstruation regulating
The effect of blood, blood pressure lowering；For the thin thin, dizziness of consumptive disease, night sweat, impotence, pain in the loins, in blood, metrorrhagia and metrostaxis, leukorrhagia, goiter, hernia lump in the abdomen etc.
Disease.Mussel is as one of the marine organisms eaten extensively, and abundance is harmless nontoxic, contained rich in nutrition content, nutriture value
Value is higher than general shellfish and fish, shrimp, meat etc., to boosting metabolism, guarantees that the movable nutrition supply of brain and body has
Positive effect, person " marine egg ".Epidemiological survey is the results show that New Zealand Maoris is local due to long-term consumption
The disease incidence of a kind of Perna viridis, inflammation related disease is significantly reduced than other areas.Clinical research also indicates that, New Zealand's emerald
The fat-soluble extract of mussel respectively reaches 60% and 50% to the curative effect of rheumatic arthritis and osteoarthritis, mussel dry powder with
Arthralgia caused by fat-soluble extract is loose to rheumatic arthritis and sclerotin and arthroncus have good relaxation effect.This
Outside, there are also decreasing heart rates for mussel extract, reduce blood pressure, and increase myocardial blood flow and platelet aggregation-against and anti-atherogenic is hard
The effects of change.Primary Study discovery mussel extract can be absorbed by organisms utilization well, greatly improve body selenium level and resist
Oxidase active mitigates damage of the active oxygen radical to body, protects large biological molecule, while making the growth of transplanted tumor
It is suppressed.Animal experiments show that thick-shell mussel fat-soluble extract can significantly inhibit adjuvant type pass under 100mg/kg dosage
The scorching pedal swelling with Type Ⅱ collagen rats with arthritis of section, reduces arthritis index, improves the pathological symptom of ankle-joint, simultaneously also
Can prevent rat because of body weight increase caused by arthroncus slowly and spleen enlargement phenomenon.
RA is a kind of systemic autoimmune disease, clinically with chronic, symmetry, peripheral, panarthritis venereal disease
Become its main feature, can behave as affected joints pain, swelling and function reduction.Currently, the main therapeutic agent of RA has
Non-steroidal anti-inflammatory drugs (NSAIDs) acts on antirheumatic drug (SAARDs), glucocorticoid etc. slowly, but since drug therapy is bad
The limitation of reaction leads to not effectively promote.
Therefore, the drug that prevention and treatment RA is prepared using mussel extract is particularly important, but does not have correlation also at present
Research, and the preparation process of mussel extract is also not perfect.
Summary of the invention
The purpose of the invention is to overcome the deficiencies of the prior art and provide a kind of preparation process of mussel extract,
Solve the problems, such as that mussel extract preparation process is incomplete.
The present invention is achieved through the following technical solutions:
A kind of preparation process of mussel extract, by weight, comprising the following steps:
S1: taking in 1 part of Mussel Powder addition, 5~50 parts of sodium hydroxide solutions, is ultrasonically treated after 20~30min in constant temperature vibration
Immersion treatment 5~for 24 hours is vibrated in water-bath；
S2: clear water is carried out repeatedly to immersion treatment gains in S1 and is rinsed, is drained；
S3: into S2,5~50 parts of clear water are added in gains, and 20~30min of magnetic agitation carries out colloid mill grinding；
S4: it will be ground in S3 after gains boil 0.5~4h, and be cooled to 40~70 DEG C；
S5: into S4,0.001~0.05 part of alkali protease, 40~70 DEG C of 3~5h of enzymatic hydrolysis are added in gains；
S6: being warming up to 95 DEG C of enzyme deactivation 10min for gains are digested in S5, be cooled to room temperature, 10~20min of centrifugal treating,
Collect centrifugate；
S7: into S6,0.01~5 part of filter aid is added in centrifugate, carries out plate-frame filtering after 20~30min of magnetic agitation, is received
Collect filtrate；
S8: the filtrate in S7 is concentrated into 20~50%, obtains concentrate；
S9: concentrate in S8 is spray-dried up to product.
Further, it is 50 DEG C that temperature is ultrasonically treated in S1, processing power 600w.
Further, in S3 colloid mill grinding the following steps are included:
S3-1: colloid mill gap is adjusted to 3mm, is ground gains are stirred in S3 3 times；
S3-2: colloid mill gap is adjusted to 1mm, gains in S3-1 are ground 2 times；
S3-3: colloid mill gap is adjusted to 0.5mm, gains in S3-2 are ground 1 time.
Further, by weight, alkali protease include 1 part of alkali protease produced by bacillus pumilus 209,
1 part of alkali protease and 1 part of alkali protease produced by Bacillus licheniformis2709 produced by bacillus pumilus 289.
Further, centrifugal rotational speed is 5000r/min in S6.
Further, filter aid includes 1 part of perlite, 1 part of diatomite and 1 part of mussel shell powder.Wherein, perlite, diatom
Soil and mussel shell powder diameter are 100~200 mesh.
Further, condensing mode is concentrated using scraper plate in S8, and after scraper plate concentration, concentrate is through 0.2~0.5 μm of aperture
Filter membrane processing.
The Mussel Powder that the present invention uses, the micro- Huang of color, no mildew, without obvious stink, water content is 5~8%, and hydroxide is molten
Liquid quality solubility is 0.25~4%；Vibration is impregnated in S1 is carried out using constant temperature vibration water-bath, adjusts revolving speed 50r/min, temperature
25℃；Clear water described in S2 and S3 is deionized water；Magnetic agitation rotating speed is 300~500r/min.
Further, mussel extract prepared by the present invention, for making the functional food of prevention rheumatoid arthritis
Product.
Further, effective edible dosage of mussel extract is 25~1000mg/kg in functional food.
Further, the main component of mussel extract is mussel peptide.
Compared with prior art, the present invention has following usefulness:
1) provide a perfect mussel extract preparation process: step is simply easy to realization of industrialization, using hydrogen-oxygen
Change sodium to handle raw material, the stink contained in raw material can be removed, while the lipid in raw material is removed by saponification
Ingredient, wherein ultrasonic treatment improves the efficiency of deodorization and degreasing；
2) to mussel shell, this by-product is utilized, meanwhile, make filter aid using mussel shell powder and (completely or partially replaces
Generation) also reduce the cost of preparation process；
3) gained mussel extract has the characteristics that sterile, high activity and easily storage；
4) Bones and joints swelling and inflammatory reaction caused by RA can be effectively relieved in the mussel extract provided in vivo, to RA
There is certain prevention effect.
Figure of description
Bones and joints Magnetic resonance imaging when the rheumatoid rat of Fig. 1 adjuvant induction takes mussel extract 2 weeks；
Before Fig. 2 collagen-induced rheumatoid rat modeling, modeling 11 days, 21 days when Bones and joints Magnetic resonance imaging.
In Fig. 1: A1 Normal group ♀；A2MTX treatment group ♀；A3 natural recovering group ♀；A4 mussel peptide treatment group ♀；
B1 Normal group ♂；B2MTX treatment group ♂；B3 natural recovering group ♂；B4 mussel peptide treatment group ♂；
In Fig. 2: each group female mice Bones and joints NMR imaging before Aa modeling；Each group hero mouse Bones and joints NMR imaging before Bb modeling；
Each group female mice Bones and joints NMR imaging at A modeling 11 days；Each group hero mouse Bones and joints NMR imaging at B modeling 11 days；
Each group female mice Bones and joints NMR imaging at a modeling 21 days；Each group hero mouse Bones and joints NMR imaging at b modeling 21 days；
1 negative control group；2 low dose therapy groups；3 middle dosage treatment groups；4 high-dose therapy groups.
Specific embodiment
In order to make the objectives, technical solutions, and advantages of the present invention clearer, with reference to the accompanying drawings and embodiments, right
The present invention is further elaborated.It should be appreciated that the specific embodiments described herein are merely illustrative of the present invention, and
It is not used in the restriction present invention.
Embodiment 1
A kind of preparation process of mussel extract, by weight, comprising the following steps:
S1: taking in 100g parts of Mussel Powder (water content 5%) 500g parts of addition sodium hydroxide (0.25%) solution, ultrasound (place
Manage power 600w, 50 DEG C for the treatment of temperature) processing 20min after in constant temperature vibration water-bath vibrate immersion treatment (25 DEG C of temperature,
50r/min)5h；
S2: deionized water is carried out repeatedly to gains in S1 and is rinsed, is drained；
S3: into S2,500g deionized water is added in gains, and magnetic agitation (revolving speed 300r/min) 20min carries out colloid
Mill grinding；
S4: it will be ground in S3 after gains boil 0.5h, and be cooled to 40 DEG C；
S5: into S4, the alkali protease of 0.1g, 40 DEG C of enzymatic hydrolysis 3h are added in gains；
S6: 95 DEG C of enzyme deactivation 10min are warming up to by gains are digested in S5, is cooled to room temperature (20 DEG C), is centrifuged (revolving speed
10min 5000r/min) is handled, centrifugate is collected；
S7: into S6,1g filter aid, the laggard andante frame mistake of magnetic agitation (revolving speed 300r/min) 20min is added in centrifugate
Filtrate is collected in filter；
S8: being concentrated into 20% for filtrate in S7, obtains concentrate, and concentrate is handled through the filter membrane in 0.2 μm of aperture；
S9: concentrate in S8 is spray-dried up to product 85g, product yield 85%.
Preferably, in S3 colloid mill grinding the following steps are included:
S3-1: colloid mill gap is adjusted to 3mm, is ground gains are stirred in S3 3 times；
S3-2: colloid mill gap is adjusted to 1mm, gains in S3-1 are ground 2 times；
S3-3: colloid mill gap is adjusted to 0.5mm, gains in S3-2 are ground 1 time.
Preferably, filter aid includes 0.25g perlite, 0.25g diatomite and 0.5g mussel shell powder.Wherein, perlite, silicon
Diatomaceous earth and mussel shell powder diameter are 100 mesh.
Preferably, condensing mode is concentrated using scraper plate in S8.
Embodiment 2
A kind of preparation process of mussel extract, by weight, comprising the following steps:
S1: taking in 100g parts of Mussel Powder (water content 5%) 1500g parts of addition sodium hydroxide solution (0.25%), ultrasound (place
Manage power 600w, 50 DEG C for the treatment of temperature) processing 20min after in constant temperature vibration water-bath vibrate immersion treatment (25 DEG C of temperature,
50r/min)8h；
S2: deionized water is carried out repeatedly to gains in S1 and is rinsed, is drained；
S3: into S2,1500g deionized water is added in gains, and magnetic agitation (revolving speed 300r/min) 20min carries out colloid
Mill grinding；
S4: it will be ground in S3 after gains boil 1.5h, and be cooled to 50 DEG C；
S5: into S4, the alkali protease of 0.5g, 50 DEG C of enzymatic hydrolysis 3.5h are added in gains；
S6: 95 DEG C of enzyme deactivation 10min are warming up to by gains are digested in S5, is cooled to room temperature (20 DEG C), is centrifuged (revolving speed
10min 5000r/min) is handled, centrifugate is collected；
S7: into S6,10g filter aid, the laggard andante frame mistake of magnetic agitation (revolving speed 300r/min) 20min is added in centrifugate
Filtrate is collected in filter；
S8: being concentrated into 30% for filtrate in S7, obtains concentrate, and concentrate is handled through the filter membrane in 0.25 μm of aperture；
S9: concentrate in S8 is spray-dried up to product 84.5g, product yield 84.5%.
Preferably, in S3 colloid mill grinding the following steps are included:
S3-1: colloid mill gap is adjusted to 3mm, is ground gains are stirred in S3 3 times；
S3-2: colloid mill gap is adjusted to 1mm, gains in S3-1 are ground 2 times；
S3-3: colloid mill gap is adjusted to 0.5mm, gains in S3-2 are ground 1 time.
Preferably, filter aid includes 2.5g perlite, 2.5g diatomite and 5g mussel shell powder.Wherein, perlite, diatomite
It is 100 mesh with mussel shell powder diameter.
Preferably, condensing mode is concentrated using scraper plate in S8.
Embodiment 3
A kind of preparation process of mussel extract, by weight, comprising the following steps:
S1: taking in 100g parts of Mussel Powder (water content 6.5%) 250g parts of addition sodium hydroxide solution (2%), ultrasound (processing
Power 600w, 50 DEG C for the treatment of temperature) processing 25min after in constant temperature vibration water-bath vibrate immersion treatment (25 DEG C of temperature, 50r/
min)12h；
S2: deionized water is carried out repeatedly to gains in S1 and is rinsed, is drained；
S3: into S2,2500g deionized water is added in gains, and magnetic agitation (revolving speed 300r/min) 20min carries out colloid
Mill grinding；
S4: it will be ground in S3 after gains boil 2h, and be cooled to 60 DEG C；
S5: into S4, the alkali protease of 1g, 60 DEG C of enzymatic hydrolysis 4h are added in gains；
S6: 95 DEG C of enzyme deactivation 10min are warming up to by gains are digested in S5, is cooled to room temperature (20 DEG C), is centrifuged (revolving speed
15min 5000r/min) is handled, centrifugate is collected；
S7: into S6,100g filter aid, the laggard andante frame mistake of magnetic agitation (revolving speed 300r/min) 20min is added in centrifugate
Filtrate is collected in filter；
S8: being concentrated into 40% for filtrate in S7, obtains concentrate, and concentrate is handled through the filter membrane in 0.3 μm of aperture；
S9: concentrate in S8 is spray-dried up to product 87g, product yield 87%.
Preferably, in S3 colloid mill grinding the following steps are included:
S3-1: colloid mill gap is adjusted to 3mm, is ground gains are stirred in S3 3 times；
S3-2: colloid mill gap is adjusted to 1mm, gains in S3-1 are ground 2 times；
S3-3: colloid mill gap is adjusted to 0.5mm, gains in S3-2 are ground 1 time.
Preferably, filter aid includes 25g perlite, 25g diatomite and 100g mussel shell powder.Wherein, perlite, diatomite
It is 150 mesh with mussel shell powder diameter.
Preferably, condensing mode is concentrated using scraper plate in S8.
Embodiment 4
A kind of preparation process of mussel extract, by weight, comprising the following steps:
S1: taking in 100g parts of Mussel Powder (water content 7.3%) 3000g parts of addition sodium hydroxide solution (3%), ultrasound (place
Manage power 600w, 50 DEG C for the treatment of temperature) processing 25min after in constant temperature vibration water-bath vibrate immersion treatment (25 DEG C of temperature,
50r/min)16h；
S2: deionized water is carried out repeatedly to gains in S1 and is rinsed, is drained；
S3: into S2,3000g deionized water is added in gains, and magnetic agitation (revolving speed 300r/min) 25min carries out colloid
Mill grinding；
S4: it will be ground in S3 after gains boil 2.5h, and be cooled to 70 DEG C；
S5: into S4, the alkali protease of 2.5g, 70 DEG C of enzymatic hydrolysis 4h are added in gains；
S6: 95 DEG C of enzyme deactivation 10min are warming up to by gains are digested in S5, is cooled to room temperature (20 DEG C), is centrifuged (revolving speed
15min 5000r/min) is handled, centrifugate is collected；
S7: into S6,200g filter aid, the laggard andante frame mistake of magnetic agitation (revolving speed 300r/min) 25min is added in centrifugate
Filtrate is collected in filter；
S8: being concentrated into 50% for filtrate in S7, obtains concentrate, and concentrate is handled through the filter membrane in 0.35 μm of aperture；
S9: concentrate in S8 is spray-dried up to product 85.5g, product yield 85.5%.
Preferably, in S3 colloid mill grinding the following steps are included:
S3-1: colloid mill gap is adjusted to 3mm, is ground gains are stirred in S3 3 times；
S3-2: colloid mill gap is adjusted to 1mm, gains in S3-1 are ground 2 times；
S3-3: colloid mill gap is adjusted to 0.5mm, gains in S3-2 are ground 1 time.
Preferably, filter aid includes 50g perlite, 50g diatomite and 100g mussel shell powder.Wherein, perlite, diatomite
It is 150 mesh with mussel shell powder diameter.
Preferably, condensing mode is concentrated using scraper plate in S8.
Embodiment 5
A kind of preparation process of mussel extract, by weight, comprising the following steps:
S1: taking in 100g parts of Mussel Powder (water content 7.3%) 3500g parts of addition sodium hydroxide solution (4%), ultrasound (place
Manage power 600w, 50 DEG C for the treatment of temperature) processing 30min after in constant temperature vibration water-bath vibrate immersion treatment (25 DEG C of temperature,
50r/min)19h；
S2: deionized water is carried out repeatedly to gains in S1 and is rinsed, is drained；
S3: into S2,4000g deionized water is added in gains, and magnetic agitation (revolving speed 300r/min) 25min carries out colloid
Mill grinding；
S4: it will be ground in S3 after gains boil 3h, and be cooled to 70 DEG C；
S5: into S4, the alkali protease of 5g, 70 DEG C of enzymatic hydrolysis 4.5h are added in gains；
S6: 95 DEG C of enzyme deactivation 10min are warming up to by gains are digested in S5, is cooled to room temperature (20 DEG C), is centrifuged (revolving speed
15min 5000r/min) is handled, centrifugate is collected；
S7: into S6,300g filter aid, the laggard andante frame mistake of magnetic agitation (revolving speed 300r/min) 25min is added in centrifugate
Filtrate is collected in filter；
S8: being concentrated into 50% for filtrate in S7, obtains concentrate, and concentrate is handled through the filter membrane in 0.45 μm of aperture；
S9: concentrate in S8 is spray-dried up to product 87.8g, product yield 87.8%.
Preferably, in S3 colloid mill grinding the following steps are included:
S3-1: colloid mill gap is adjusted to 3mm, is ground gains are stirred in S3 3 times；
S3-2: colloid mill gap is adjusted to 1mm, gains in S3-1 are ground 2 times；
S3-3: colloid mill gap is adjusted to 0.5mm, gains in S3-2 are ground 1 time.
Preferably, filter aid includes 75g perlite, 75g diatomite and 150g mussel shell powder.Wherein, perlite, diatomite
It is 200 mesh with mussel shell powder diameter.
Preferably, condensing mode is concentrated using scraper plate in S8.
Embodiment 6
A kind of preparation process of mussel extract, by weight, comprising the following steps:
S1: taking in 100g parts of Mussel Powder (water content 7.3%) 4000g parts of addition sodium hydroxide solution (4%), ultrasound (place
Manage power 600w, 50 DEG C for the treatment of temperature) processing 30min after in constant temperature vibration water-bath vibrate immersion treatment (25 DEG C of temperature,
50r/min)24h；
S2: deionized water is carried out repeatedly to gains in S1 and is rinsed, is drained；
S3: into S2,5000g deionized water is added in gains, and magnetic agitation (revolving speed 300r/min) 30min carries out colloid
Mill grinding；
S4: it will be ground in S3 after gains boil 4h, and be cooled to 70 DEG C；
S5: into S4, the alkali protease of 1.25g, 70 DEG C of enzymatic hydrolysis 5h are added in gains；
S6: 95 DEG C of enzyme deactivation 10min are warming up to by gains are digested in S5, is cooled to room temperature (20 DEG C), is centrifuged (revolving speed
20min 5000r/min) is handled, centrifugate is collected；
S7: into S6,500g filter aid, the laggard andante frame mistake of magnetic agitation (revolving speed 300r/min) 30min is added in centrifugate
Filtrate is collected in filter；
S8: being concentrated into 50% for filtrate in S7, obtains concentrate, and concentrate is handled through the filter membrane in 0.5 μm of aperture；
S9: concentrate in S8 is spray-dried up to product 88.6g, product yield 88.6%.
Preferably, in S3 colloid mill grinding the following steps are included:
S3-1: colloid mill gap is adjusted to 3mm, is ground gains are stirred in S3 3 times；
S3-2: colloid mill gap is adjusted to 1mm, gains in S3-1 are ground 2 times；
S3-3: colloid mill gap is adjusted to 0.5mm, gains in S3-2 are ground 1 time.
Preferably, filter aid includes 125g perlite, 125g diatomite and 250g mussel shell powder.Wherein, perlite, diatom
Soil and mussel shell powder diameter are 200 mesh.
Preferably, condensing mode is concentrated using scraper plate in S8.
Embodiment 7
A kind of preparation process of mussel extract, by weight, comprising the following steps:
S1: taking in 100g parts of Mussel Powder (water content 3.5%) 350g parts of addition sodium hydroxide solution (0.2%), ultrasound (place
Manage power 600w, 50 DEG C for the treatment of temperature) processing 15min after in constant temperature vibration water-bath vibrate immersion treatment (25 DEG C of temperature,
50r/min)4h；
S2: deionized water is carried out repeatedly to gains in S1 and is rinsed, is drained；
S3: into S2,400g deionized water is added in gains, and magnetic agitation (revolving speed 300r/min) 15min carries out colloid
Mill grinding；
S4: it will be ground in S3 after gains boil 0.45h, and be cooled to 35 DEG C；
S5: into S4, the alkali protease of 0.8g, 35 DEG C of enzymatic hydrolysis 2.5h are added in gains；
S6: 95 DEG C of enzyme deactivation 10min are warming up to by gains are digested in S5, is cooled to room temperature (20 DEG C), is centrifuged (revolving speed
8min 5000r/min) is handled, centrifugate is collected；
S7: into S6,0.8g filter aid, the laggard andante frame mistake of magnetic agitation (revolving speed 300r/min) 15min is added in centrifugate
Filtrate is collected in filter；
S8: being concentrated into 19% for filtrate in S7, obtains concentrate, and concentrate is handled through the filter membrane in 0.2 μm of aperture；
S9: concentrate in S8 is spray-dried up to product 37.2g, product yield 37.2%.
Preferably, in S3 colloid mill grinding the following steps are included:
S3-1: colloid mill gap is adjusted to 3mm, is ground gains are stirred in S3 3 times；
S3-2: colloid mill gap is adjusted to 1mm, gains in S3-1 are ground 2 times；
S3-3: colloid mill gap is adjusted to 0.5mm, gains in S3-2 are ground 1 time.
Preferably, filter aid includes 0.2g perlite, 0.2g diatomite and 0.4g mussel shell powder.Wherein, perlite, diatom
Soil and mussel shell powder diameter are 100 mesh.
Preferably, condensing mode is concentrated using scraper plate in S8.
Embodiment 8
The preparation process for present embodiments providing a kind of mussel extract, in addition to the following contents is different, remaining content is the same as real
Example 7 is applied, the difference is that:
S9: product 35.6g, product yield 35.6% are obtained；
Perlite, diatomite and mussel shell powder diameter are 50 mesh.
Embodiment 9
The preparation process for present embodiments providing a kind of mussel extract, in addition to the following contents is different, remaining content is the same as real
Example 7 is applied, the difference is that:
S8: filtrate is concentrated into 20%, obtains concentrate, and concentrate is handled through the filter membrane in 0.2 μm of aperture；
S9: product 38g, product yield 38% are obtained.
Embodiment 10
The preparation process for present embodiments providing a kind of mussel extract, in addition to the following contents is different, remaining content is the same as real
Example 7 is applied, the difference is that:
S5: into S4, the alkali protease of 1g, 40 DEG C of enzymatic hydrolysis 3h are added in gains；
S9: product 36.9g, product yield 36.9% are obtained.
Embodiment 11
The preparation process for present embodiments providing a kind of mussel extract, in addition to the following contents is different, remaining content is the same as real
Example 7 is applied, the difference is that:
S4: it will be ground in S3 after gains boil 0.5h, and be cooled to 50 DEG C；
S9: product 37.7g, product yield 37.7% are obtained.
Embodiment 12
The preparation process for present embodiments providing a kind of mussel extract, in addition to the following contents is different, remaining content is the same as real
Example 7 is applied, the difference is that:
S1: immersion treatment (25 DEG C of temperature, 50r/min) 5h is vibrated in constant temperature vibration water-bath after processing 20min；
S9: product 38.2g, product yield 38.2% are obtained.
Embodiment 13
A kind of preparation process of mussel extract, by weight, comprising the following steps:
S1: taking in 150g parts of Mussel Powder (water content 7.3%) 4500g parts of addition sodium hydroxide solution (4.5%), ultrasound
(processing power 600w, 50 DEG C for the treatment of temperature) vibrates immersion treatment (temperature 25 after handling 35min in constant temperature vibration water-bath
DEG C, 50r/min) 26h；
S2: deionized water is carried out repeatedly to gains in S1 and is rinsed, is drained；
S3: into S2,5500g deionized water is added in gains, and magnetic agitation (revolving speed 300r/min) 35min carries out colloid
Mill grinding；
S4: it will be ground in S3 after gains boil 4.5h, and be cooled to 75 DEG C；
S5: into S4, the alkali protease of 1.8g, 75 DEG C of enzymatic hydrolysis 5.5h are added in gains；
S6: 95 DEG C of enzyme deactivation 10min are warming up to by gains are digested in S5, is cooled to room temperature (20 DEG C), is centrifuged (revolving speed
25min 5000r/min) is handled, centrifugate is collected；
S7: into S6,600g filter aid, the laggard andante frame mistake of magnetic agitation (revolving speed 300r/min) 35min is added in centrifugate
Filter collects filtrate, is ultrasonically treated (processing power 1000w, 50 DEG C for the treatment of temperature) 35min to filtrate；
S8: being concentrated into 40% for filtrate in S7, obtains concentrate, and concentrate is handled through the filter membrane in 0.6 μm of aperture；
S9: concentrate in S8 is spray-dried up to product 78.3g, product yield 52.2%.
Preferably, in S3 colloid mill grinding the following steps are included:
S3-1: colloid mill gap is adjusted to 3mm, is ground gains are stirred in S3 3 times；
S3-2: colloid mill gap is adjusted to 1mm, gains in S3-1 are ground 2 times；
S3-3: colloid mill gap is adjusted to 0.5mm, gains in S3-2 are ground 1 time.
Preferably, filter aid includes 150g perlite, 150g diatomite and 300g mussel shell powder.Wherein, perlite, diatom
Soil and mussel shell powder diameter are 250 mesh.
Preferably, condensing mode is concentrated using scraper plate in S8.
Embodiment 14
The preparation process for present embodiments providing a kind of mussel extract, in addition to the following contents is different, remaining content is the same as real
Example 13 is applied, the difference is that:
S9: product 79.4g, product yield 52.9% are obtained；
Perlite, diatomite and mussel shell powder diameter are 200 mesh.
Embodiment 15
The preparation process for present embodiments providing a kind of mussel extract, in addition to the following contents is different, remaining content is the same as real
Example 13 is applied, the difference is that:
S8: filtrate is concentrated into 30%, obtains concentrate, and concentrate is handled through the filter membrane in 0.5 μm of aperture；
S9: product 78.3g, product yield 52.2% are obtained.
Embodiment 16
The preparation process for present embodiments providing a kind of mussel extract, in addition to the following contents is different, remaining content is the same as real
Example 13 is applied, the difference is that:
S5: into S4, the alkali protease of 1.5g, 40 DEG C of enzymatic hydrolysis 5h are added in gains；
S9: product 80.3g, product yield 53.5% are obtained.
Embodiment 17
The preparation process for present embodiments providing a kind of mussel extract, in addition to the following contents is different, remaining content is the same as real
Example 13 is applied, the difference is that:
S4: it will be ground in S3 after gains boil 4h, and be cooled to 70 DEG C；
S9: product 76.7g, product yield 51.1% are obtained.
Embodiment 18
The preparation process for present embodiments providing a kind of mussel extract, in addition to the following contents is different, remaining content is the same as real
Example 13 is applied, the difference is that:
S1: immersion treatment (25 DEG C of temperature, 50r/min) are vibrated for 24 hours in constant temperature vibration water-bath after processing 30min；
S9: product 75.8g, product yield 50.5% are obtained.
Arranging above embodiments 1~18 the results are shown in Table 1:
Table 1: 1~18 result statistical form of embodiment
From table 1 it follows that in Examples 1 to 6, mussel extract product yield with higher (84.5~
88.6%), the product yield of embodiment 7~12 is minimum (35.6~38.2%), and the product yield of embodiment 13~18 is taken second place
(50.5~53.5%)；Component analysis (biuret method, referring to " bone is peaceful is carried out to the ingredient of mussel extract in embodiment 1~18
Polypeptide at analysis and its assay ", Fang Hangjun, Chinese biochemical drug magazine, CHINESE JOURNAL OF
BIOCHEMICAL PHARMACEUTICS, 02 phase in 1999), it is concluded that
In Examples 1 to 6 content of peptides be 91.2~95.7% (implement 1 be 93.4%, embodiment 2 be 94.8%, implementation
3 for 91.4%, to implement 4 be 91.2%, implements 5 is 95.4%, implements 6 is that 95.7%), containing a small amount of polysaccharide and other are miscellaneous
Matter；Content of peptides is lower than 65% in embodiment 7~18, and other impurities are more.
It chooses embodiment 1,3 and 6 gained mussel extracts is uniformly mixed as mussel extract treatment group, made using rat
Feeding trial is carried out for subjects；Rats by intraperitoneal injection dosage 300mg/kg chloraldurate (10% chloraldurate 0.3ml/
It 100g) anaesthetizes, fixed rat, injects 0.1ml complete Freund's adjuvant, two days later, same position in the left back whole plantar fat pad of rat
Setting injection 0.05ml 2.5ug/ml TNF-α, (tumor necrosis factor, it can promote T cell to generate various inflammatory factors, in turn
Promote the generation of inflammatory reaction.) booster immunization, negative control group respectively at same position injection same volume physiological saline,
Start feeding mussel extract normal saline solution, 1 times a week, methopterin (MTX, for anti-folic acid class antimetabolite after one day；
There is very strong immunosuppressive action；Cell of the selectively acting in proliferation prevents immunoblast division growth；Body fluid is exempted from
Epidemic disease and cellular immunity have inhibiting effect；Also there is stronger anti-inflammatory effect；It is mainly used for autoimmune disease such as rheumatoid
Arthritis, systemic loupus erythematosus and dermatomyositis etc..) inject weekly once, continuously after two weeks (administrations are shown in Table 2), each group
Prostrate is in nuclear magnetic resonance apparatus after rat is anaesthetized with isoflurane, and foot Bones and joints left back to rat are scanned that (scanning result is shown in
Attached drawing 1), as a result statistics is shown in Table 3.
Table 2: the rat rheumatoid arthritis dosage of adjuvant induction
From figure 1 it appears that prostrate is in core after anaesthetizing each group rat with isoflurane when taking mussel extract 2 weeks
In magnetic resonance tool, foot Bones and joints left back to rat are scanned, scanning result such as Fig. 1, Normal group female rats figure A1
Bones and joints such as white arrow is signified, and articular cavity is of moderate size, and without obvious hydrops, synovium of joint schemes A2 (MTX without hyperplasia and oedema
Treatment group) compared with scheming A1, joint has slight oedema, but schemes A3 compared with scheming A2, and joint is obviously brighter, prompts joint water
The more serious and arthroedema that swells is more；A4 (mussel extract treatment group) is schemed compared with scheming A3, symptom is obviously alleviated,
And close to MTX treatment group；Equally, it can be seen that this species diversity male big from male rat Bones and joints MR imaging results figure B
It is more obvious with mouse.
It chooses embodiment 1,3 and 6 gained mussel extracts is uniformly mixed as mussel extract treatment group, made using rat
Feeding trial is carried out for subjects；First divide 3 kinds of dosage to rat oral gavage 15 days with mussel extract, then rats by intraperitoneal injection
Dosage 300mg/kg chloraldurate (10% chloraldurate 0.3ml/100g) anesthesia, fixed rat, in the left back whole plantar rouge of rat
Fat pad injects 0.1ml II Collagen Type VI, and after a week, same position injects 0.1ml II Collagen Type VI booster immunization, negative control component
Not in the physiological saline of same position injection same volume.Animal model preparation is whole to use mussel extract stomach-filling, once a day
(administrations are shown in Table 3).
Table 3: collagen-induced rat rheumatoid arthritis dosage
When 1 day before modeling, administration 11 days, administration 21 days, prostrate is in core after each group rat is anaesthetized with isoflurane
In magnetic resonance tool, foot Bones and joints Magnetic resonance imaging scanning left back to each group rat, scanning result such as Fig. 2, Aa, Bb generation respectively
The female mice of 1 day each group and male mouse before table modeling, the female mice of each group and male mouse when A, B respectively represent modeling 11 days, a, b are respectively represented
The female mice of each group and male mouse at modeling 21 days；As shown in Figure 2, each group rat Bones and joints compared with each group before modeling, after modeling
All there is different degrees of oedema and rigid or even mild malformation；When being administered 11 days, each group female mice Bones and joints symptom improves simultaneously
Unobvious, when being administered 21 days, each group is all restored, but compared with scheming a1 (negative control group), figure a2 (low dose therapy group),
Scheme a3 (middle dosage treatment group), figure a4 (high-dose therapy group), improvement degree is relatively obvious, but dose-effect relationship between each treatment group
It is not obvious；For male mouse, when being administered 11 days, figure B2 (low dose therapy group), figure B3 (middle dosage treatment group), figure B4 (high agent
Amount treatment group) Bones and joints swelling is obviously improved compared with scheming B1 (negative control group), and it is the most obvious to scheme B2；When being administered 21 days,
Each group hero mouse Bones and joints symptom difference is not obvious or even negative control group improvement becomes apparent.
In above embodiments, the left back sufficient Bones and joints swelling situation of each group rat is detected by MRI scan, is as a result shown
Show that feeding mussel peptide rat bone arthroedema and swelling are all alleviated；In the acute RA of complete Freund's adjuvant induction, make a gift of
Shellfish peptide can prevent the state of an illness from further deteriorating by its anti-inflammatory symptom for alleviating RA with consumer edema effect；Collagen-induced slow
Property RA in, mussel peptide can reach certain preventive effect by its anti-inflammatory and consumer edema effect；In the mistake of long-term feeding mussel peptide
Do not observe that adverse reaction caused by traditional treatment RA drug occurs in rat in journey.
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, all in essence of the invention
Made any modifications, equivalent replacements, and improvements etc., should all be included in the protection scope of the present invention within mind and principle.
Claims (9)
1. a kind of preparation process of mussel extract, which is characterized in that by weight, comprising the following steps:
S1: taking in 1 part of Mussel Powder addition, 5~50 parts of sodium hydroxide solutions, is ultrasonically treated after 20~30min in constant temperature vibration water-bath
Immersion treatment 5~for 24 hours is vibrated in pot；
S2: clear water is carried out repeatedly to immersion treatment gains in S1 and is rinsed, is drained；
S3: into S2,5~50 parts of clear water are added in gains, and 20~30min of magnetic agitation carries out colloid mill grinding；
S4: it will be ground in S3 after gains boil 0.5~4h, and be cooled to 40~70 DEG C；
S5: into S4,0.001~0.05 part of alkali protease, 40~70 DEG C of 3~5h of enzymatic hydrolysis are added in gains；
S6: being warming up to 95 DEG C of 10~20min of enzyme deactivation for gains are digested in S5, be cooled to room temperature, 10~20min of centrifugal treating,
Collect centrifuged supernatant；
S7: into S6,0.01~5 part of filter aid is added in centrifugate, and plate-frame filtering is carried out after 20~30min of magnetic agitation, collects filter
Liquid；
S8: the filtrate in S7 is concentrated into 20~50%, obtains concentrate；
S9: concentrate in S8 is spray-dried up to product.
2. a kind of preparation process of mussel extract as described in claim 1, which is characterized in that being ultrasonically treated temperature in S1 is
50 DEG C, processing power is 600~1000w.
3. a kind of preparation process of mussel extract as described in claim 1, which is characterized in that colloid mill, which is ground, in S3 includes
Following steps:
S3-1: colloid mill gap is adjusted to 3mm, is ground gains are stirred in S3 3 times；
S3-2: colloid mill gap is adjusted to 1mm, gains in S3-1 are ground 2 times；
S3-3: colloid mill gap is adjusted to 0.5mm, gains in S3-2 are ground 1 time.
4. a kind of preparation process of mussel extract as described in claim 1, which is characterized in that by weight, basic protein
Enzyme includes 1 part of alkali protease produced by bacillus pumilus 209,1 portion of basic protein produced by bacillus pumilus 289
Enzyme and 1 part of alkali protease produced by Bacillus licheniformis2709.
5. a kind of preparation process of mussel extract as described in claim 1, which is characterized in that centrifugal rotational speed is in S6
5000r/min。
6. a kind of preparation process of mussel extract as described in claim 1, which is characterized in that by weight, filter aid packet
Include 1 part of perlite, 1 part of diatomite and 2 parts of mussel shell powder.
7. a kind of preparation process of mussel extract as described in claim 1, which is characterized in that
Condensing mode is concentrated using scraper plate in S8, and after scraper plate concentration, concentrate is handled through the filter membrane in 0.2~0.5 μm of aperture.
8. a kind of application of the mussel extract prepared using claim 1~7, which is characterized in that for making prevention class wind
The functional food of wet arthritis.
9. application as claimed in claim 8, which is characterized in that by weight, mussel extract is edible in functional food
Dosage is 25~1000mg/kg.
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| CN1620887A (en) * | 2004-12-15 | 2005-06-01 | 汤兴然 | Edible fatless animal protein and its preparation method and uses |
| CN103992387A (en) * | 2014-05-29 | 2014-08-20 | 浙江海洋学院 | Mussel cooking liquid active peptide as well as preparation method and application thereof |
| CN104513842A (en) * | 2013-09-26 | 2015-04-15 | 青岛蓝农谷农产品研究开发有限公司 | Production method of odorless moisture-retaining collagen |
| KR20160007744A (en) * | 2014-06-26 | 2016-01-21 | 마산수산업협동조합 | Composition for anti-hangover and anti-obesity containing extract of Mytilus edulis |
| CN106148467A (en) * | 2016-08-31 | 2016-11-23 | 湖北瑞邦生物科技有限公司 | A kind of preparation method of mussel active peptide |
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- 2018-06-22 CN CN201810666201.3A patent/CN109043114A/en active Pending
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1620887A (en) * | 2004-12-15 | 2005-06-01 | 汤兴然 | Edible fatless animal protein and its preparation method and uses |
| CN104513842A (en) * | 2013-09-26 | 2015-04-15 | 青岛蓝农谷农产品研究开发有限公司 | Production method of odorless moisture-retaining collagen |
| CN103992387A (en) * | 2014-05-29 | 2014-08-20 | 浙江海洋学院 | Mussel cooking liquid active peptide as well as preparation method and application thereof |
| KR20160007744A (en) * | 2014-06-26 | 2016-01-21 | 마산수산업협동조합 | Composition for anti-hangover and anti-obesity containing extract of Mytilus edulis |
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