CN108041402A - A kind of preparation method of complex biological preservative - Google Patents
A kind of preparation method of complex biological preservative Download PDFInfo
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- CN108041402A CN108041402A CN201711316324.6A CN201711316324A CN108041402A CN 108041402 A CN108041402 A CN 108041402A CN 201711316324 A CN201711316324 A CN 201711316324A CN 108041402 A CN108041402 A CN 108041402A
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- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
- A23L3/34—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals
- A23L3/3454—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals in the form of liquids or solids
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Abstract
The invention belongs to food antiseptic preservation fields, and in particular to a kind of preparation method of complex biological preservative.The present invention by from the fermented bean curd that egg white is impregnated separation and Extraction lysozyme be used as main body, it is green, safe and pollution-free, lysozyme is as a kind of native protein, it can be digested and absorb as nutriment in stomach and intestine, it is non-toxic to human body, it will not remain in vivo, be a kind of very high antistaling agent of security.It enhances the acidproof heat-proof performance of lysozyme, expands scope of restraining fungi, both there is inhibitory action to gram-positive bacteria, also generate inhibitory action to Gram-negative bacteria object to a part by the modification to lysozyme.In addition lysozyme and EDTA, streptococcus lactis peptide, natamycin, several preservative mixed fermentations of chitin oligo saccharide are improved the activity of enzyme, the greatly freshness date of extension food and aquatic products by the present invention, and many convenience are brought to store, transporting etc..
Description
Technical field
The invention belongs to food antiseptic preservation fields, and in particular to a kind of preparation method of complex biological preservative.
Background technology
With economy and social progress, food industry has become one of three big pillars of national economy, and in food
There are many reason for maximum harm is exactly the putrid and deteriorated of food, causes food apoilage in industry, but bacterium, mould, ferment
The invasion and attack of the microorganisms such as female bacterium are the main reason for causing food spoilage.Microbial contamination is to influence the principal element of food security
One of, the effect of preservative is killing or the growth and breeding for inhibiting microorganism, to prevent that the biology of tissue infection and substance is rotten
It loses.Food production, in the process of circulation because microorganism pollution can caused by it is putrid and deteriorated, the addition of preservative is necessary.
Preservative refers to prevent or slow down the putrid and deteriorated substance of organic matter, be added in food and medicine and cosmetics anti-
Rotten agent can inhibit the growth of the microorganisms such as fungi, bacterium and mould, achieve the effect that extend the shelf life.
The preservative that China is applied to food mainly has three kinds of chemical preservative, natural antiseptic agent and compound preservative, but
All there are the Railway Project below this,（1）Chemical method is artificial synthesized：What is largely used in the food industry both at home and abroad at present is anti-
Rotten agent is still what artificial chemistry method synthesized, and raw material, auxiliary material and the additive used in process of production contains in itself or wherein
Impurity human health may all be damaged；（2）Security is poor：These chemical preservatives are not much that nature is deposited naturally
Substance, may to health there is potential chronic toxicity or three-induced effect（It is mutagenesis, teratogenesis, carcinogenic）；
（3）Scope of restraining fungi is narrow：Either chemical preservative or natural antiseptic agent all there are antimicrobial spectrum it is narrow the problem of, such as at present in market
On the Nisi that occupies an important position（Streptococcus lactis peptide）Only have inhibitory action to gram-positive bacteria, and to Gram-negative fungus without
Inhibitory action；（4）Natural antiseptic agent species is few：The natural microbial preservative that uses of China's approval at present only have natamycin and
Nisin etc. a few, kind is very single；（5）The preservative scope of application is small：Most preservatives it is antibacterial require it is applicable
Environment is acid or neutral, as Nisin only has bacteriostasis in acid condition.All these problems are the current world
Various countries' food industry problem urgently to be resolved hurrily.
Therefore it provides a kind of freshness that can guarantee food, the life safe, antiseptic property is excellent, bacteriostasis is strong
Object preservative, be one very it is necessary to thing.
The content of the invention
The technical problems to be solved by the invention：Larger, the antimicrobial spectrum for preservative Side effect currently on the market
It is narrow, the problem of fresh keeping time is short, provide a kind of preparation method of complex biological preservative.
In order to solve the above technical problems, the present invention is using technical solution as described below：
A kind of preparation method of composite biological preservative, which is characterized in that the preparation method includes the following steps：
（1）The bean curd after egg white immersion is taken to be put at -2 ~ 3 DEG C, the bean curd after egg white immersion is made to be cooled to 4 ~ 5 DEG C, Xiang Leng
But exchanger resin is added in bean curd after, keeps the temperature and is stirred continuously, stands to being layered, collects lower sediment, adds in distillation water washing
Lower sediment, it is dry, dry resin is obtained, the ammonium sulfate that mass fraction is 10% is added in into dry resin, stirring elutes,
Filtering, takes filtrate, obtains eluent, eluent is placed in 4 ~ 5 DEG C of waters bath with thermostatic control, and stirring while adds in acetone, must elute
Liquid mixed solution centrifuges eluent mixed solution, collects precipitation；
（2）Above-mentioned precipitation is added in modification liquid, is placed in water bath with thermostatic control, is stirred to react, is centrifuged, is taken supernatant, must change
Property matrix, take Sephadex G-25 columns on modified matrix, with mass fraction be 12% pH 7.4 ± 0.2 phosphate buffer
Elution, collects active modification lysozyme, and freeze-drying obtains modified lysozyme powder；
（3）Above-mentioned modified lysozyme powder is added in fermented nutritive liquid to ferment, collects fermentate, adds in auxiliary anticorrosion agent,
It is uniformly mixed, stands, freeze-drying obtains complex biological preservative.
The step（1）The pretreatment mode of middle exchanger resin is：By exchanger resin under 23 ~ 32 DEG C of temperature conditionss, add
Enter absolute ethyl alcohol sealing and impregnate 8h, the mass ratio of exchanger resin and absolute ethyl alcohol is 2：5, resin after impregnating is taken out, adds in and impregnates
The distillation water washing of 4 times of resin quality afterwards removes the ethyl alcohol in resin, obtains primary resin, is then 5% with mass fraction
The mass ratio of NaOH solution immersion 8h, primary resin and NaOH solution is 2：6, primary resin is taken out, adds in primary resin quality 3
Distilled water again washs resin to neutrality, obtains second resin, finally impregnates second resin 8h with the HCl that mass fraction is 5%,
The mass ratio of second resin and HCl solution is 1：5, second resin is taken out, adds in the distilled water of 5 times of second resin quality by two level
Resin is washed to neutrality, you can.
The step（1）The mass ratio of middle bean curd and exchanger resin is 5：3, the mass ratio of resin and ammonium sulfate is 2：
7, the mass ratio of eluent and acetone is 1：2.
The step（2）The preparation of middle lysozyme modification liquid：It is 2 in mass ratio：3：6, forulic acid, caffeic acid is taken to be dissolved in
Mass fraction is in 15% NaOH solution, and the HCl solution for being 1mol/L with mass fraction adjusts pH to 7.2 ± 0.2, is mixed
Solution is closed, EDAC and deionized water are added in into mixed solution, the mass ratio of mixed solution, EDAC and deionized water is 3：4：7,
1 ~ 2h is stood, obtains modification liquid.
The step（2）The mass ratio of middle precipitation and lyase modification liquid is 1：1.
The step（3）Middle auxiliary anticorrosion agent：In mass ratio 2：1：2 mix EDTA, streptococcus lactis peptide, chitin oligo saccharide
It is even.
The step（3）The mass ratio of middle modified lysozyme powder and fermented nutritive liquid is 3：5, fermentation condition is 20 ~ 35
DEG C, the time is 24 ~ 36 h, fermentate and auxiliary anticorrosion agent in mass ratio 5：2 are uniformly mixed.
Compared with other methods, advantageous effects are the present invention：
The present invention by from the fermented bean curd that egg white is impregnated separation and Extraction lysozyme be used as main body, it is green, safe and pollution-free,
Lysozyme can be digested and be absorbed as nutriment in stomach and intestine as a kind of native protein, non-toxic to human body, also not
It can remain in vivo, be a kind of very high antistaling agent of security.The present invention enhances lysozyme by the modification to lysozyme
Acidproof heat-proof performance expands scope of restraining fungi, both there is inhibitory action to gram-positive bacteria, also to a part of Gram-negative bacteria
Object generates inhibitory action.The present invention mixes lysozyme and EDTA, streptococcus lactis peptide, natamycin, several preservatives of chitin oligo saccharide
Close fermentation, improve the activity of enzyme, greatly extend the freshness date of food and aquatic products, for store, transport etc. bring it is all multi-party
Just.
Specific embodiment
Bean curd choose in：Shanghai Qingmei Green Food Co., Ltd.
Raw material：Take new fresh bean curd and egg white, in mass ratio 5：7, new fresh bean curd is placed in 2 ~ 3h of immersion in egg white, is obtained
Bean curd after egg white immersion.
Exchanger resin pre-processes：By resin under 23 ~ 32 DEG C of temperature conditionss, add in absolute ethyl alcohol sealing and impregnate 8h, resin
Mass ratio with absolute ethyl alcohol is 2：5, resin after impregnating is taken out, adds in the distillation water washing of 4 times of resin quality after impregnating, is removed
Ethyl alcohol in resin obtains primary resin, then impregnates 8h, primary resin and NaOH solution with the NaOH solution that mass fraction is 5%
Mass ratio be 2：6, primary resin is taken out, the distilled water for adding in 3 times of primary resin quality washs resin to neutrality, obtains two level
Resin finally impregnates second resin 8h with the HCl that mass fraction is 5%, and the mass ratio of second resin and HCl solution is 1：5, it takes
Go out second resin, the distilled water for adding in 5 times of second resin quality washs second resin to neutrality, obtains exchanger resin.
The preparation of lysozyme modification liquid：It is forulic acid, coffee in 15% solution that forulic acid, caffeic acid is taken, which to be dissolved in mass fraction,
Coffee acid and the mass ratio of NaOH solution are 2：3：6, the HCl solution for being 1mol/L with mass fraction adjusts pH to 7.2 ± 0.2, obtains
To mixed solution, EDAC and deionized water are added in into mixed solution, the mass ratio of mixed solution, EDAC and deionized water is 3：
4：7,1 ~ 2h is stood, obtains modification liquid.
The preparation of fermented nutritive liquid：According to the mass fraction, 4 ~ 7 parts of beef extract, 12 ~ 15 parts of peptone, starch 13 ~ 16 are taken
Part, 5 ~ 6 parts of NaCl, 15 ~ 17 parts of glucose, KH2PO41.2 ~ 2.5 parts, MgSO4·7H20.8 ~ 1.6 part of O, vitamin 5 ~ 10
Part, 1000 parts of water, 7.2 ± 0.1,121 DEG C of sterilizing 20min of pH value.
A kind of preparation of complex biological preservative, includes the following steps：
The extraction of lysozyme：Bean curd -2 ~ 3 DEG C constant temperature is taken to be cooled to 4 ~ 5 DEG C, exchanger resin, bean curd are added in into bean curd after cooling
Mass ratio with exchanger resin is 9：3,23 ~ 28 DEG C keep the temperature 8h, are stirred continuously in insulating process, then stand 2 ~ 5h and are extremely layered,
Lower sediment is collected, adds in distillation water washing lower sediment, removes foreign protein, it is dry, last dry resin is taken, adds in quality point
Number is 10% ammonium sulfate, and the mass ratio of resin and ammonium sulfate is 2：7, stirring 1 ~ 1.5h of elution, filtering takes filtrate,
Obtain eluent；Eluent is placed in 4 ~ 5 DEG C of waters bath with thermostatic control, stirring while is added dropwise acetone, eluent and acetone
Mass ratio is 1：2, precipitation is collected in centrifugation；
The modification of lysozyme：Above-mentioned precipitation is pressed into mass fraction 1：1 ratio is added in lysozyme modification liquid, and in 30 DEG C of perseverances
18 ~ 24 h are stirred in tepidarium, after reaction, centrifugal filtration takes filtrate, obtains modified matrix；10 ~ 12 parts of modifications are taken by quality
Sephadex G-25 columns on matrix are eluted, flow velocity with the phosphate buffer of mass fraction 50mmol/L pH 7.4 ± 0.2
The modification lysozyme for having lysozyme activity peak in wavelength 280nm is collected in 1ml/min, detection, and freeze-drying obtains modified bacteriolyze
Enzyme powder；
Above-mentioned modified lysozyme powder addition fermented nutritive liquid is fermented, fermentation condition is 20 ~ 35 DEG C, the time for 24 ~
36h, after the completion of fermentation, collection fermentate, in mass ratio 2：1：2 add in EDTA, streptococcus lactis peptide, chitin oligo saccharide as auxiliary anticorrosion
Agent, fermentate and auxiliary anticorrosion agent in mass ratio 5：2 are uniformly mixed, and stand, and freeze-drying obtains complex biological preservative.
Embodiment 1
Bean curd choose in：Shanghai Qingmei Green Food Co., Ltd.
Raw material：Take new fresh bean curd and egg white, in mass ratio 5：7, fermented bean curd is placed in egg white and impregnates 2h.
Exchanger resin pre-processes：By resin under 23 DEG C of temperature conditionss, add in absolute ethyl alcohol sealing and impregnate 8h, resin and nothing
The mass ratio of water-ethanol is 2：5, resin after impregnating is taken out, adds in the distillation water washing of 4 times of resin quality after impregnating, removes resin
In ethyl alcohol, obtain primary resin, then impregnate 8h, the matter of primary resin and NaOH solution with the NaOH solution that mass fraction is 5%
Amount is than being 2：6, primary resin is taken out, the distilled water for adding in 3 times of primary resin quality washs resin to neutrality, obtains two level tree
Fat finally impregnates second resin 8h with the HCl that mass fraction is 5%, and the mass ratio of second resin and HCl solution is 1：5, it takes out
Second resin, the distilled water for adding in 5 times of second resin quality wash second resin to neutrality, obtain exchanger resin.
The preparation of lysozyme modification liquid：It is forulic acid, coffee in 15% solution that forulic acid, caffeic acid is taken, which to be dissolved in mass fraction,
Coffee acid and the mass ratio of NaOH solution are 2：3：6, the HCl solution for being 1mol/L with mass fraction adjusts pH to 7.2 ± 0.2, obtains
To mixed solution, EDAC and deionized water are added in into mixed solution, the mass ratio of mixed solution, EDAC and deionized water is 3：
4：7,1h is stood, obtains modification liquid.
The preparation of fermented nutritive liquid：According to the mass fraction, 4 parts of beef extract, 12 parts of peptone, 13 parts of starch, NaCl 5 are taken
Part, 15 parts of glucose, KH2PO41.2 parts, MgSO4·7H20.8 part of O, 5 parts of vitamin, 1000 parts of water, pH value 7.2 ±
0.1,121 DEG C of sterilizing 20min.
A kind of preparation of complex biological preservative, includes the following steps：
The extraction of lysozyme：Bean curd -2 ~ 3 DEG C constant temperature is taken to be cooled to 4 ~ 5 DEG C, exchanger resin is added in into bean curd after cooling, is kept the temperature
And be stirred continuously, the mass ratio of fermented bean curd and exchanger resin is 9：3,23 DEG C keep the temperature 8h, are stirred continuously in insulating process, Ran Houjing
2h is put to being layered, collects lower sediment, adds in distillation water washing lower sediment, removes foreign protein, it is dry, dry resin is taken, is added
Enter mass fraction for 10% ammonium sulfate, the mass ratio of resin and ammonium sulfate is 2：7, stirring elution 1h, filtering takes
Filtrate obtains eluent；By eluent ice bath, stirring while, is added dropwise acetone, and the mass ratio of eluent and acetone is 1：2,
Precipitation is collected in centrifugation；
The modification of lysozyme：Above-mentioned precipitation is pressed into mass fraction 1：1 ratio is added in lysozyme modification liquid, is obtained lysozyme and is changed
Property mixture, stirs 18h, after reaction, centrifugal filtration takes filter by lysozyme modified mixture in 30 DEG C of waters bath with thermostatic control
Liquid obtains modified matrix；Sephadex G-25 columns on 10 ~ 12 parts of modified matrix are taken by quality, with mass fraction 50mmol/L pH
7.4 ± 0.2 phosphate buffer elution, flow velocity 1ml/min, detection, collect has lysozyme activity peak in wavelength 280nm
Modified lysozyme, freeze-drying obtain modified lysozyme powder；
Above-mentioned modified lysozyme powder addition fermented nutritive liquid is fermented, fermentation condition is 20 DEG C, and the time is fermentation for 24 hours
After finishing, collection fermentate, in mass ratio 2：1：2 addition EDTA, streptococcus lactis peptide, chitin oligo saccharide are uniformly mixed, and are stood, freezing is dry
It is dry, obtain complex biological preservative.
Embodiment 2
Bean curd choose in：Shanghai Qingmei Green Food Co., Ltd.
Raw material：Take new fresh bean curd and egg white, in mass ratio 5：7, bean curd is placed in egg white and impregnates 2.5h.
Exchanger resin pre-processes：By resin under 27 DEG C of temperature conditionss, add in absolute ethyl alcohol sealing and impregnate 8h, resin and nothing
The mass ratio of water-ethanol is 2：5, resin after impregnating is taken out, adds in the distillation water washing of 4 times of resin quality after impregnating, removes resin
In ethyl alcohol, obtain primary resin, then impregnate 8h, the matter of primary resin and NaOH solution with the NaOH solution that mass fraction is 5%
Amount is than being 2：6, primary resin is taken out, the distilled water for adding in 3 times of primary resin quality washs resin to neutrality, obtains two level tree
Fat finally impregnates second resin 8h with the HCl that mass fraction is 5%, and the mass ratio of second resin and HCl solution is 1：5, it takes out
Second resin, the distilled water for adding in 5 times of second resin quality wash second resin to neutrality, obtain exchanger resin.
The preparation of lysozyme modification liquid：It is forulic acid, coffee in 15% solution that forulic acid, caffeic acid is taken, which to be dissolved in mass fraction,
Coffee acid and the mass ratio of NaOH solution are 2：3：6, the HCl solution for being 1mol/L with mass fraction adjusts pH to 7.2 ± 0.2, obtains
To mixed solution, EDAC and deionized water are added in into mixed solution, the mass ratio of mixed solution, EDAC and deionized water is 3：
4：7,1.5h is stood, obtains modification liquid.
The preparation of fermented nutritive liquid：According to the mass fraction, 4 ~ 7 parts of beef extract, 13 parts of peptone, 14 parts of starch, NaCl are taken
5 parts, 17 parts of glucose, KH2PO41.8 parts, MgSO4·7H21.2 parts of O, 7 parts of vitamin, 1000 parts of water, pH value 7.2 ±
0.1,121 DEG C of sterilizing 20min.
A kind of preparation of complex biological preservative, includes the following steps：
The extraction of lysozyme：Bean curd -2 ~ 3 DEG C constant temperature is taken to be cooled to 4 ~ 5 DEG C, exchanger resin, bean curd are added in into bean curd after cooling
The mass ratio of breast and exchanger resin is 9：3,25 DEG C keep the temperature 8h, are stirred continuously in insulating process, then stand 3h to being layered, collect
Lower sediment adds in distillation water washing precipitation, removes foreign protein, dry, takes dry resin, adds in the sulphur that mass fraction is 10%
The mass ratio of acid ammonium solution, resin and ammonium sulfate is 2：7, stirring elution 1.2h, filtering takes filtrate, obtains eluent；It will wash
De- liquid ice bath, stirring while, are added dropwise acetone, and the mass ratio of eluent and acetone is 1：2, precipitation is collected in centrifugation；
The modification of lysozyme：Above-mentioned precipitation is pressed into mass fraction 1：1 ratio is added in lysozyme modification liquid, and in 30 DEG C of perseverances
22h is stirred in tepidarium, after reaction, centrifugal filtration takes filtrate, obtains modified matrix；It is taken by quality on 11 parts of modified matrix
Sephadex G-25 columns are eluted, flow velocity 1ml/min with the phosphate buffer of mass fraction 50mmol/L pH 7.4 ± 0.2,
Detection.The modification lysozyme that there is lysozyme activity peak in wavelength 280nm is collected, freeze-drying obtains modified lysozyme powder；
Above-mentioned modified lysozyme powder addition fermented nutritive liquid is fermented, fermentation condition is 27 DEG C, and the time is 24 ~ 36h, is sent out
After the completion of ferment, collection fermentate, in mass ratio 5：2：1：2 addition EDTA, streptococcus lactis peptide, chitin oligo saccharide are uniformly mixed, and are stood, cold
It is lyophilized dry, obtain complex biological preservative.
Embodiment 3
Bean curd choose in：Shanghai Qingmei Green Food Co., Ltd.
Raw material：Take new fresh bean curd and egg white, in mass ratio 5：7, fermented bean curd is placed in egg white and impregnates 3h.
Exchanger resin pre-processes：By resin under 32 DEG C of temperature conditionss, add in absolute ethyl alcohol sealing and impregnate 8h, resin and nothing
The mass ratio of water-ethanol is 2：5, resin after impregnating is taken out, adds in the distillation water washing of 4 times of resin quality after impregnating, removes resin
In ethyl alcohol, obtain primary resin, then impregnate 8h, the matter of primary resin and NaOH solution with the NaOH solution that mass fraction is 5%
Amount is than being 2：6, primary resin is taken out, the distilled water for adding in 3 times of primary resin quality washs resin to neutrality, obtains two level tree
Fat finally impregnates second resin 8h with the HCl that mass fraction is 5%, and the mass ratio of second resin and HCl solution is 1：5, it takes out
Second resin, the distilled water for adding in 5 times of second resin quality wash second resin to neutrality, obtain exchanger resin.
The preparation of lysozyme modification liquid：It is forulic acid, coffee in 15% solution that forulic acid, caffeic acid is taken, which to be dissolved in mass fraction,
Coffee acid and the mass ratio of NaOH solution are 2：3：6, the HCl solution for being 1mol/L with mass fraction adjusts pH to 7.2 ± 0.2, obtains
To mixed solution, EDAC and deionized water are added in into mixed solution, the mass ratio of mixed solution, EDAC and deionized water is 3：
4：7,2h is stood, obtains modification liquid.
The preparation of fermented nutritive liquid：According to the mass fraction, 7 parts of beef extract, 15 parts of peptone, 16 parts of starch, NaCl 6 are taken
Part, 17 parts of glucose, KH2PO42.5 parts, MgSO4·7H21.6 parts of O, 10 parts of vitamin, 1000 parts of water, pH value 7.2 ±
0.1,121 DEG C of sterilizing 20min.
A kind of preparation of complex biological preservative, includes the following steps：
The extraction of lysozyme：Fermented bean curd ice bath is taken to be cooled to 5 DEG C, is stirred evenly, adds in exchanger resin, fermented bean curd and exchanger resin
Mass ratio be 9：3,28 DEG C keep the temperature 8h, are stirred continuously in insulating process, then stand 5h to being layered, collect lower sediment, add
Enter to distill water washing precipitation, remove foreign protein, take the last ammonium sulfate for being filtered dry resin, adding in that mass fraction is 10%, resin
Mass ratio with ammonium sulfate is 2：7, stirring 1 ~ 1.5h of elution, filtering takes filtrate, obtains eluent；By eluent ice bath, stir
It is added dropwise acetone while mixing, the mass ratio of eluent and acetone is 1：2, precipitation is collected in centrifugation；
The modification of lysozyme：Above-mentioned precipitation is pressed into mass fraction 1：1 ratio is added in lysozyme modification liquid, is obtained lysozyme and is changed
Property mixture, stirs 24 h, after reaction, centrifugal filtration takes filter by lysozyme modified mixture in 30 DEG C of waters bath with thermostatic control
Liquid obtains modified matrix；According to the mass fraction, Sephadex G-25 columns on 12 parts of modified matrix are taken, with mass fraction 50mmol/L
The phosphate buffer elution of pH 7.4 ± 0.2, flow velocity 1ml/min, detection, collect has lysozyme activity in wavelength 280nm
The modification lysozyme at peak, freeze-drying, obtains modified lysozyme powder；
Above-mentioned modified lysozyme powder addition fermented nutritive liquid is fermented, fermentation condition is 35 DEG C, time 36h, fermentation
After finishing, fermentate is collected, adds in EDTA, streptococcus lactis peptide, chitin oligo saccharide, fermentate, EDTA, streptococcus lactis peptide, chitin oligo saccharide are pressed
Mass ratio 5：2：1：2 are uniformly mixed, and stand, and freeze-drying obtains complex biological preservative.
Comparative example：The biological preservative of Nantong biochemistry Co., Ltd production
Method：Taking the cherry of 4 parts of 500g, the calculation embodiment and the biology prepared by comparative example for choosing equivalent are prevented as test specimen
Rotten agent, is used it on test specimen, and the sample of used preservative is placed under room temperature and is preserved, and is preserved 15 days, detects various kinds
The specific situation of change of product.
The specific detection case of biological preservative such as table 1
Table 1
Embodiment is that separation and Extraction lysozyme is as main body from the bean curd that egg white is impregnated, and lysozyme is as a kind of native protein
Matter can be digested and absorb as nutriment in stomach and intestine, non-toxic to human body, will not remain in vivo, so this hair
Bright prepared preservative will not do harm to huamn body.
Judge embodiment with the biological preservative prepared by comparative example to gram-positive bacteria by filter paper inhibition zone method
With the size of the antibacterial circle diameter of Gram-negative bacteria, diameter is bigger, and it is better to the fungistatic effect of this strain to show.
The specific detection case of biological preservative fungistatic effect such as table 2
Table 2
For embodiment is compared with comparative example, bacteriostasis property is good, and scope of restraining fungi is big.
From the foregoing, it will be observed that raw material used in composite biological preservative prepared by the present invention is natural material, non-toxic, nothing
Side effect, scope of restraining fungi is big, long to the fresh keeping time of food, is a kind of safe and efficient biological preservative, is worthy to be popularized and makes
With.
Claims (7)
1. a kind of preparation method of composite biological preservative, which is characterized in that the preparation method includes the following steps：
（1）The bean curd after egg white immersion is taken to be put at -2 ~ 3 DEG C, the bean curd after egg white immersion is made to be cooled to 4 ~ 5 DEG C, Xiang Leng
But exchanger resin is added in bean curd after, keeps the temperature and is stirred continuously, stands to being layered, collects lower sediment, adds in distillation water washing
Lower sediment takes precipitation drying, obtains dry resin, adds in the ammonium sulfate that mass fraction is 10%, stirring elution, and filtering takes
Filtrate obtains eluent, and eluent is placed in 4 ~ 5 DEG C of waters bath with thermostatic control, and stirring while adds in acetone, and it is molten to obtain eluent mixing
Liquid centrifuges eluent mixed solution, collects precipitation；
（2）Above-mentioned precipitation is added in modification liquid, is placed in water bath with thermostatic control, is stirred to react, is centrifuged, is taken supernatant, must change
Property matrix, take Sephadex G-25 columns on modified matrix, with mass fraction be 12% pH 7.4 ± 0.2 phosphate buffer
Elution, collects active modification lysozyme, and freeze-drying obtains modified lysozyme powder；
（3）Above-mentioned modified lysozyme powder is added in fermented nutritive liquid to ferment, fermentate is collected, is added in into fermentate auxiliary
Preservative is helped, is uniformly mixed, is stood, freeze-drying obtains complex biological preservative.
2. the preparation method of composite biological preservative according to claim 1, which is characterized in that the step（1）Middle friendship
The pretreatment mode for changing resin is：By exchanger resin under 23 ~ 32 DEG C of temperature conditionss, add in absolute ethyl alcohol sealing and impregnate 8h, hand over
The mass ratio for changing resin and absolute ethyl alcohol is 2：5, resin after impregnating is taken out, adds in the distillation washing of 4 times of resin quality after impregnating
Wash, remove resin in ethyl alcohol, obtain primary resin, then with mass fraction be 5% NaOH solution impregnate 8h, primary resin with
The mass ratio of NaOH solution is 2：6, primary resin is taken out, the distilled water for adding in 3 times of primary resin quality washs resin into
Property, second resin is obtained, the mass ratio of second resin 8h, second resin and HCl solution are finally impregnated with the HCl that mass fraction is 5%
For 1：5, second resin is taken out, the distilled water for adding in 5 times of second resin quality washs second resin to neutrality, you can.
3. the preparation method of composite biological preservative according to claim 1, which is characterized in that the step（1）Middle beans
The rotten mass ratio with exchanger resin is 5：3, the mass ratio of resin and ammonium sulfate is 2：7, the mass ratio of eluent and acetone
For 1：2.
4. the preparation method of composite biological preservative according to claim 1, which is characterized in that the step（2）In it is molten
The preparation of bacterium enzyme modification liquid：Forulic acid, caffeic acid is taken to be dissolved in the NaOH solution that mass fraction is 15%, forulic acid, caffeic acid
Mass ratio with NaOH solution is 2：3：6, the HCl solution for being 1mol/L with mass fraction adjusts pH to 7.2 ± 0.2, is mixed
Solution is closed, EDAC and deionized water are added in into mixed solution, the mass ratio of mixed solution, EDAC and deionized water is 3：4：7,
1 ~ 2h is stood, obtains modification liquid.
5. the preparation method of composite biological preservative according to claim 1, which is characterized in that the step（2）In sink
It is 1 to form sediment with the mass ratio of lyase modification liquid：1.
6. the preparation method of composite biological preservative according to claim 1, which is characterized in that the step（3）In it is several
Kind auxiliary anticorrosion agent：In mass ratio 2：1：2 are uniformly mixed EDTA, streptococcus lactis peptide, chitin oligo saccharide.
7. the preparation method of composite biological preservative according to claim 1, which is characterized in that the step（3）In change
Property lysozyme powder and fermented nutritive liquid mass ratio be 3：5, fermentation condition be 20 ~ 35 DEG C, the time be 24 ~ 36 h, fermentate
Mass ratio with auxiliary anticorrosion agent is 5：2.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113475718A (en) * | 2021-05-19 | 2021-10-08 | 滁州欣发食品有限公司 | Processing method of food additive with sterilization function |
| CN115581233A (en) * | 2022-11-11 | 2023-01-10 | 中国农业科学院烟草研究所(中国烟草总公司青州烟草研究所) | Combined use method of tobacco root secretion methyl ferulate and chitosan oligosaccharide and application thereof |
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2017
- 2017-12-12 CN CN201711316324.6A patent/CN108041402A/en not_active Withdrawn
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113475718A (en) * | 2021-05-19 | 2021-10-08 | 滁州欣发食品有限公司 | Processing method of food additive with sterilization function |
| CN115581233A (en) * | 2022-11-11 | 2023-01-10 | 中国农业科学院烟草研究所(中国烟草总公司青州烟草研究所) | Combined use method of tobacco root secretion methyl ferulate and chitosan oligosaccharide and application thereof |
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