CN108576693A - It is a kind of to improve immune and anti-oxidation function anchovy enzymolysis object and preparation method thereof - Google Patents
It is a kind of to improve immune and anti-oxidation function anchovy enzymolysis object and preparation method thereof Download PDFInfo
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Abstract
The present invention relates to a kind of preparation methods improving immune and anti-oxidation function anchovy enzymolysis object.Steps are as follows：Anchovy slurries are first prepared, then prepare anchovy enzymolysis liquid, anchovy fermentation enzymolysis liquid is then prepared, the anchovy enzymolysis liquid that ferments finally is obtained into anchovy enzymolysis object by being concentrated in vacuo, drying.Immune and anti-oxidation function anchovy enzymolysis object is improved the present invention also provides a kind of, such as above-mentioned preparation method is prepared.The present invention is simple using the synchronous anchovy enzymolysis object preparation method for combining biofermentation technique exploitation of enzymolysis, and product keeps preferable functional characteristic, is easier to realize industrialization production.The anchovy enzymolysis object of exploitation, which has, improves immunity and anti-oxidation function, and anchovy has obtained complete utilization in production process, does not generate any by-product.The present invention utilizes the anchovy protein liquid of specific frequency ultrasonic technology processing, can be obviously improved sensibility of the anchovy albumen to enzyme, reduce the usage amount of enzyme.
Description
Technical field
Immune and anti-oxidation function anchovy enzymolysis object and preparation method thereof is improved the present invention relates to a kind of, belongs to animal food
Object manufacture field.
Background technology
Every physiological function of animal immune system and animal is closely bound up, for maintaining animal health, resisting all kinds of diseases
Disease, guarantee animal items metabolism are smoothed out all significant.Immunity of organisms is improved, enhancing body's immunity is to prevent
Various diseases occur with the key point of rehabilitation.Modern nutriology research finds that animal ingestion protein is made through gastral enzymolysis
With rear, digested and assimilated in the form of small peptide mostly, the ratio very little absorbed in the form of free amino acid.Because small peptide with it is free
Amino acid compared to being more convenient for transporting by intestinal wall, peptide specific ionization amino acid digestion faster, absorb it is more.Peptide matters trophism
Another new knowledge to be protein can generate some biologically active peptides with special physiological regulatory function in enzymolysis process.
Immunomodulatory peptides can be by adjusting Immune Organs of Body（Tissue）Growth and development, immunocyte activity with function, antibody produce
Raw, cell factor secretion adjusts body's immunity with expression and the release of immune related signaling molecules etc., is ensureing
Animal（People）Play an important roll in health.
At present about the anti-oxidant of protein hydrolysate, ACE inhibition and the patent application of function of blood sugar reduction in country's patent
It is more but seldom about the enzymolysis product patent application for improving immune function.Especially not only with raising immunity but also with anti-
The enzymolysis product report of oxidative function is seldom.
Anchovy be it is a kind of living in the small fishes of temperate zone ocean at the middle and upper levels, be distributed widely in China Bohai and Yellow Seas and
The East Sea is the food organisms of other economic fish, and anchovy is currently main low value fish in ocean, and yield is larger, and resource does not have
It is effectively utilized.At present using anchovy as raw material, the patent text for the enzymolysis product for improving immune and anti-oxidation function is prepared
It offers and correlative study report yet there are no.Therefore, it is necessary to propose effective technical solution, solve the above problems.
Invention content
The present invention in view of the above shortcomings of the prior art, provides a kind of anchovy enzymolysis object for improving and being immunized with anti-oxidation function
And preparation method thereof.The anchovy enzymolysis object has preferable raising immune and anti-oxidation function.
The technical solution that the present invention solves above-mentioned technical problem is as follows：
A kind of preparation method improving immune and anti-oxidation function anchovy enzymolysis object, steps are as follows：
（1）Prepare anchovy slurries：It selects and freezes fresh anchovy, the water of 1.0~2.0 times of anchovy total amount is added in 121-131 DEG C of condition
Lower processing 0.5-1 hours, is then adjusted to 75-85 DEG C by temperature, using supersonic generator through ultrasonication 15-25 points
Clock obtains anchovy slurries；
（2）Prepare anchovy enzymolysis liquid：Regulating step（1）Protein content is 7-10% in the obtained anchovy slurries；It adds
The compound protease of protein wt 0.1-0.2% is digested, and 0.5~1.5h of enzyme digestion reaction is carried out under the conditions of 50~60 DEG C,
Obtain anchovy enzymolysis liquid；
（3）Prepare anchovy fermentation enzymolysis liquid：By step（2）Obtained anchovy enzymolysis liquid temperature is adjusted to 45-50 DEG C, and anchovy is added
Enzymolysis liquid weight 0.5-1% glucose adds anchovy enzymolysis liquid weight 0.1-0.2% bulgaria lactobacillus fermentation liquid；In 40-
It ferments 8-12 hours under the conditions of 45 DEG C, obtains anchovy fermentation enzymolysis liquid；
（4）By step（3）Obtained anchovy fermentation enzymolysis liquid obtains anchovy enzymolysis object by being concentrated in vacuo, drying.
Preferably, the ultrasonication frequency is 40-60kH.
Preferably, it is 1~2 that the compound protease, which is mass ratio,：1 alkali protease and neutral proteinase composition.
Immune and anti-oxidation function anchovy enzymolysis object is improved the present invention also provides a kind of, prepared by such as above-mentioned preparation method
It obtains.
Advantageous effect：
（1）The present invention is simple using the synchronous anchovy enzymolysis object preparation method for combining biofermentation technique exploitation of enzymolysis, and product is protected
Preferable functional characteristic is held, is easier to realize industrialization production.
（2）The anchovy enzymolysis object of exploitation, which has, improves immunity and anti-oxidation function, and anchovy has obtained entirely in production process
It utilizes, does not generate any by-product.
（3）The present invention utilizes the anchovy protein liquid of specific frequency ultrasonic technology processing, can be obviously improved anchovy albumen
To the sensibility of enzyme, the usage amount of enzyme is reduced.
（4）The product safety of exploitation, the present invention use numerous food grade compound protease（Alkali protease, neutral protein
Enzyme）And lactobacillus bulgaricus, the anchovy enzymolysis object through in a mild condition, being digested by appropriateness and biofermentation obtains.
（5）The anchovy enzymolysis object that the present invention develops has good flavor, can be widely used in animal feed, improves dynamic
The immunocompetence of object.
（6）The ratio that the present invention develops peptide of the anchovy enzymolysis object middle-molecular-weihydroxyethyl less than 1500 is 90% or more.
Specific implementation mode
The present invention is directed to the material characteristic of Optimization of Low Value Fish anchovy, by largely testing, develops a kind of with preferably raising
Immune and anti-oxidation function anchovy enzymolysis object and preparation method.By to pretreatments such as anchovy high pressure, high-frequency ultrasonics, passing through
Fermentation combines zymolysis technique, has antifatigue and anti-oxidation function anchovy enzymolysis with what dry technology produced in conjunction with being concentrated in vacuo
Object establishes the preparation method of a set of anchovy enzymolysis object being simple and efficient.
For a better understanding of the present invention, it is further elaborated below by specific embodiment.
Embodiment 1
A kind of preparation method improving immune and anti-oxidation function anchovy enzymolysis object, steps are as follows：
It selects and freezes fresh anchovy, cleaned with the clean water for meeting sanitary standard for drinking water, anchovy total amount 1.0 is added after anchovy cleaning
Water again is handled 0.5 hour under the conditions of 131 DEG C, temperature is then adjusted to 75 DEG C, using supersonic generator through ultrasonic wave
（Frequency is 40kH）Processing 25 minutes, obtains anchovy slurries.
It is 7% by concentrating or being diluted with water protein content in adjusting anchovy slurries, the compound of protein by weight 0.2% is added
Protease is digested（Alkali protease, neutral proteinase composition, the mass ratio between them are 2：1）, under the conditions of 50 DEG C
Enzyme digestion reaction 1.5h is carried out, anchovy enzymolysis liquid is obtained.
Anchovy enzymolysis liquid temperature is adjusted to 45 DEG C, 1% glucose of anchovy enzymolysis liquid weight is added, adds fish enzymolysis liquid
(viable count is 1-5 × 10 to 0.1% bulgaria lactobacillus fermentation liquid of weight12A/g)；It ferments 12 hours, obtains under the conditions of 45 DEG C
To anchovy fermentation enzymolysis liquid.
Anchovy is fermented enzymolysis liquid, by being concentrated in vacuo, drying, is improved immune and anti-oxidation function anchovy enzymolysis
Object.
Embodiment 2
A kind of preparation method improving immune and anti-oxidation function anchovy enzymolysis object, steps are as follows：
It selects and freezes fresh anchovy, cleaned with the clean water for meeting sanitary standard for drinking water, anchovy total amount 2.0 is added after anchovy cleaning
Water again is handled 1 hour under the conditions of 121 DEG C, temperature is then adjusted to 85 DEG C, using supersonic generator through ultrasonic wave（Frequently
Rate is 50kH）Processing 15 minutes, obtains anchovy slurries.
It is 8% by concentrating or being diluted with water protein content in adjusting anchovy slurries, the compound of protein by weight 0.2% is added
Protease is digested（Alkali protease, neutral proteinase composition, the mass ratio between them are 2：1）, under the conditions of 55 DEG C
Enzyme digestion reaction 0.5h is carried out, anchovy enzymolysis liquid is obtained.
Anchovy enzymolysis liquid temperature is adjusted to 50 DEG C, 0.5% glucose of anchovy enzymolysis liquid weight is added, adds fish enzymolysis
(viable count is 1-5 × 10 to 0.2% bulgaria lactobacillus fermentation liquid of liquid weight12A/g)；It ferments 8 hours, obtains under the conditions of 45 DEG C
To anchovy fermentation enzymolysis liquid.
Anchovy is fermented enzymolysis liquid, by being concentrated in vacuo, drying, is improved immune and anti-oxidation function anchovy enzymolysis
Object.
Embodiment 3
A kind of preparation method improving immune and anti-oxidation function anchovy enzymolysis object, steps are as follows：
It selects and freezes fresh anchovy, cleaned with the clean water for meeting sanitary standard for drinking water, anchovy total amount 1.5 is added after anchovy cleaning
Water again is handled 1 hour under the conditions of 125 DEG C, temperature is then adjusted to 80 DEG C, using supersonic generator through ultrasonic wave（Frequently
Rate is 50kH）Processing 20 minutes, obtains anchovy slurries.
It is 10% by concentrating or being diluted with water protein content in adjusting anchovy slurries, answering for protein by weight 0.15% is added
Hop protein enzyme is digested（Alkali protease, neutral proteinase composition, the mass ratio between them are 1：1）, in 60 DEG C of conditions
Lower progress enzyme digestion reaction 1.0h, obtains anchovy enzymolysis liquid.
Anchovy enzymolysis liquid temperature is adjusted to 50 DEG C, 1% glucose of anchovy enzymolysis liquid weight is added, adds fish enzymolysis liquid
(viable count is 1-5 × 10 to 0.1% bulgaria lactobacillus fermentation liquid of weight12A/g)；It ferments 10 hours, obtains under the conditions of 45 DEG C
To anchovy fermentation enzymolysis liquid.
Anchovy is fermented enzymolysis liquid, by being concentrated in vacuo, drying, is improved immune and anti-oxidation function anchovy enzymolysis
Object.
Test example 1
The measurement of anchovy enzymolysis object delay fatigue function of the present invention is tested
Test specimen：Sample 1,2,3 is respectively anchovy enzymolysis object prepared by embodiment 1,2,3.
Test method：
1.1 experimental animal feedings and grouping
80 cleaning grade Kunming mouses, in clean Animal Lab. adaptability raise one week, 8~10/cage, sub-cage rearing,
Bedding and padding dried sanitary is kept, is changed clothes every three days primary.It sweeps on time daily, carries out the work such as sterilization, ensure clean animal
Laboratory neat and tidy.25 ± 2 DEG C of room temperature, relative humidity 50 ± 5%, optical illumination inner room is divided into 12 h, normal with mouse standard feed
Rule raising, ad lib and drinking-water, record weekly a mouse weight.After one week laundering period, 80 mouse are divided at random
Be 4 groups, i.e. blank group, 1 group of sample, 2 groups of sample, 3 groups of sample.Naive mice feeds the physiological saline of equivalent, other 3 groups
Dosage be set as 160 mg/kg/d, every morning 9:00-10:It is fed during 30, after feeding 20 days, detects given the test agent
Immune effect.
1.1.1 Immune Organs Index
It plucks eyeball method and takes blood in case detecting immunoglobulin, above-mentioned mouse is dissected after taking blood, wins thymus gland, spleen, is used
Filter paper sucks the bloodstain on surface, and electronic balance is used in combination to weigh the quality of mouse spleen, thymus gland, seeks Thymus and spleen index K values:K=
Organ weight/weight, every group of 20 mouse.
1.1.2 serum IgG, IgM, IgA assay：IgG, IgM, IgA concentration are detected using ELISA kit.
Table 1 show the influence table of anchovy enzymolysis object and control group of the present invention to mouse weight.
Table 2 show influence table of the anchovy enzymolysis object of the present invention to mouse immune shoot formation and antibody level of serum.
The influence of 1 anchovy enzymolysis object of the present invention of table and control group to mouse weight（g）
| Sample | Before experiment | First week | Second week | Third week | 4th week |
| Blank group | 20.92±0.96 | 32.68±1.07 | 36.78±1.18 | 38.88±1.49 | 39.81±1.03 |
| Sample 1 | 20.68±0.83 | 33.55±1.34 | 37.39±1.22 | 39.45±1.78 | 39.98±1.29 |
| Sample 2 | 20.67±0.86 | 34.23±1.09 | 37.88±1.29 | 39.05±1.62 | 40.24±1.18 |
| Sample 3 | 20.74±0.69 | 34.07±1.48 | 38.18±1.02 | 39.03±1.39 | 40.33±1.25 |
Influence of the 2 anchovy enzymolysis object of the present invention of table to mouse immune shoot formation and antibody level of serum
| Sample | Index and spleen index (%) | Thymus index (%) | IgG（g/L） | IgM（g/L） | IgA（g/L） |
| Blank group | 4.78±0.16 | 1.76±0.09 | 2.79±0.08 | 1.29±0.06 | 1.16±0.02 |
| Sample 1 | 5.47±0.09 | 2.28±0.06 | 2.92±0.06 | 1.68±0.09 | 1.40±0.05 |
| Sample 2 | 5.52±0.14 | 2.30±0.07 | 2.95±0.08 | 1.70±0.07 | 1.39±0.04 |
| Sample 3 | 5.58±0.22 | 2.31±0.05 | 2.94±0.05 | 1.73±0.08 | 1.42±0.07 |
As can be seen from Table 1 and Table 2, fish anchovy enzymolysis object is to the no significant impact of the growth of mouse, but anchovy enzymolysis object energy
Enough significantly improve the Immune Organs Index of mouse（Index and spleen index, thymus index）And the antibody level of serum of mouse（IgG、IgM、
IgA）.
Test example 2
The measurement of anchovy enzymolysis object antioxidant activity of the present invention is tested
Test specimen：The anchovy enzymolysis object that respectively prepared by embodiment 1, embodiment 2, embodiment 3 of sample 1,2,3.
Test method：
（1）Scavenging ability of DPPH free radical：1.5 mL of antioxidation active peptides of 20 μ g/mL is taken, 99.5 % ethyl alcohol, 1.5 mL is added
It is mixed with 0.02 % DPPH ethanol solutions, 0.675 mL, vibrates mixing, 30 min of water-bath is protected from light at room temperature, then in 517 nm
Lower detection architecture light absorption value.Light absorption value is lower, and the scavenging ability of DPPH free radical of system is stronger.
Blank control is to change 1.5 mL of sample solution into deionized water 1.5 mL.
DPPH free radical scavenging abilities %=（（Blank absorbency-sample light absorption value）/ blank absorbency）×100
（2）Reducing power measures：1 mL of antioxidation active peptides of 20 μ g/mL is taken, 0.2 M phosphate buffers are added（pH 6.6）2.5
ML and 1%（Mass fraction）2.5 mL of potassium ferricyanide solution, mixing, then in 50 DEG C of 20 min of heating water bath.It takes out rapid cold
But, it is added 10%（Mass fraction）Trichloroacetic acid（TCA）2.5 mL of solution is uniformly mixed, then centrifuges 10 at 3000 g
min.2.5 mL of supernatant is taken, deionized water 2.5 mL and 1% is added（Mass fraction）0.5 mL of liquor ferri trichloridi, it is fully mixed
It is even, 10 min are reacted at room temperature, and absorbance is measured with 700 nm wavelength.Light absorption value table at the i.e. available 700 nm wavelength of reducing power
Show.
Table 3 show the antioxidant activity tests result table of fish anchovy enzymolysis object of the present invention.
The antioxidant activity tests result of 3 fish anchovy enzymolysis object of the present invention of table
From table 3 it can be seen that anchovy enzymolysis object of the present invention has preferable oxidation resistance, it is clear under conditions of 20 μ g/mL
Except DPPH free radical abilities reach 88% or more；Reducing power reaches 0.865 or more, is a kind of preferable oxidation resistance zymolyte；This hair
The ratio of bright peptide of the exploitation anchovy enzymolysis object middle-molecular-weihydroxyethyl less than 1500 is 90% or more.
The present invention can be summarized with others without prejudice to the concrete form of the spirit or essential characteristics of the present invention.Therefore, nothing
By from the point of view of which point, the embodiment above of the invention can only all be considered the description of the invention and cannot limit this hair
Bright, claims indicate the scope of the present invention, and above-mentioned explanation does not point out the scope of the present invention, therefore, with this
Any change in the comparable meaning and scope of claims of invention is all considered as being included in the claim of the present invention
In the range of book.
Claims (4)
1. a kind of preparation method improving immune and anti-oxidation function anchovy enzymolysis object, which is characterized in that steps are as follows：
（1）Prepare anchovy slurries：It selects and freezes fresh anchovy, the water of 1.0~2.0 times of anchovy total amount is added in 121-131 DEG C of condition
Lower processing 0.5-1 hours, is then adjusted to 75-85 DEG C by temperature, using supersonic generator through ultrasonication 15-25 points
Clock obtains anchovy slurries；
（2）Prepare anchovy enzymolysis liquid：Regulating step（1）Protein content is 7-10% in the obtained anchovy slurries；It adds
The compound protease of protein wt 0.1-0.2% is digested, and 0.5~1.5h of enzyme digestion reaction is carried out under the conditions of 50~60 DEG C,
Obtain anchovy enzymolysis liquid；
（3）Prepare anchovy fermentation enzymolysis liquid：By step（2）Obtained anchovy enzymolysis liquid temperature is adjusted to 45-50 DEG C, and anchovy is added
Enzymolysis liquid weight 0.5-1% glucose adds anchovy enzymolysis liquid weight 0.1-0.2% bulgaria lactobacillus fermentation liquid；In 40-
It ferments 8-12 hours under the conditions of 45 DEG C, obtains anchovy fermentation enzymolysis liquid；
（4）By step（3）Obtained anchovy fermentation enzymolysis liquid obtains anchovy enzymolysis object by being concentrated in vacuo, drying.
2. preparation method as described in claim 1, which is characterized in that the ultrasonication frequency is 40-60kH.
3. preparation method as described in claim 1, which is characterized in that the compound protease is that mass ratio is 1~2：1 alkali
Property protease and neutral proteinase composition.
4. a kind of improving immune and anti-oxidation function anchovy enzymolysis object, which is characterized in that as described in claim any one of 1-3
Preparation method be prepared.
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