CN112592517A - Preparation method of high-temperature-resistant lysozyme composite preservative film for food - Google Patents
Preparation method of high-temperature-resistant lysozyme composite preservative film for food Download PDFInfo
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- CN112592517A CN112592517A CN202011477337.3A CN202011477337A CN112592517A CN 112592517 A CN112592517 A CN 112592517A CN 202011477337 A CN202011477337 A CN 202011477337A CN 112592517 A CN112592517 A CN 112592517A
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J5/00—Manufacture of articles or shaped materials containing macromolecular substances
- C08J5/18—Manufacture of films or sheets
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B65—CONVEYING; PACKING; STORING; HANDLING THIN OR FILAMENTARY MATERIAL
- B65D—CONTAINERS FOR STORAGE OR TRANSPORT OF ARTICLES OR MATERIALS, e.g. BAGS, BARRELS, BOTTLES, BOXES, CANS, CARTONS, CRATES, DRUMS, JARS, TANKS, HOPPERS, FORWARDING CONTAINERS; ACCESSORIES, CLOSURES, OR FITTINGS THEREFOR; PACKAGING ELEMENTS; PACKAGES
- B65D65/00—Wrappers or flexible covers; Packaging materials of special type or form
- B65D65/38—Packaging materials of special type or form
- B65D65/42—Applications of coated or impregnated materials
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J2305/00—Characterised by the use of polysaccharides or of their derivatives not provided for in groups C08J2301/00 or C08J2303/00
- C08J2305/08—Chitin; Chondroitin sulfate; Hyaluronic acid; Derivatives thereof
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J2489/00—Characterised by the use of proteins; Derivatives thereof
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- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08K—Use of inorganic or non-macromolecular organic substances as compounding ingredients
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- C08K13/02—Organic and inorganic ingredients
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- C08K—Use of inorganic or non-macromolecular organic substances as compounding ingredients
- C08K5/00—Use of organic ingredients
- C08K5/0008—Organic ingredients according to more than one of the "one dot" groups of C08K5/01 - C08K5/59
- C08K5/0058—Biocides
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08K—Use of inorganic or non-macromolecular organic substances as compounding ingredients
- C08K5/00—Use of organic ingredients
- C08K5/04—Oxygen-containing compounds
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- C08K5/053—Polyhydroxylic alcohols
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Abstract
The invention belongs to the technical field of food preservation, and particularly discloses a preparation method of a high-temperature resistant lysozyme composite preservative film for food, which solves the defects of short food shelf life, damage to human body and high cost in the prior art, and adopts the following technical scheme: the preservative film takes chitosan as a film forming substrate, and is formed into a composite film by adding glycerol, montmorillonite, Tween-80, high temperature resistant lysozyme and holy ginger essential oil. The antibacterial composite preservative film prepared by the preparation method has strong antibacterial capacity, can be used for packaging food or coated on the surface of food to form a coating after being subjected to casting film forming, can effectively inhibit food spoilage caused by microbial pollution and oxidation, prolongs the shelf life of the food, particularly has good inhibition effect on dominant bacterial communities in crayfish food, has good mechanical properties, and can be widely applied to the fields of food preservation, food packaging and the like.
Description
Technical Field
The invention relates to the technical field of food preservation, in particular to a preparation method of a high-temperature resistant lysozyme composite preservative film for food.
Background
With the continuous improvement of living standard, consumers also pay more and more attention to the food safety problem and pursue higher quality food more and more. The upgrading of consumption terminals and circulation supervision forces the upgrading of crawfish breeding management technology, the water body for breeding crawfish is changed from traditional sewage into clear water, in order to ensure the cleanness of high-density breeding water body, farmers usually use water purifying agent in the breeding water body, wherein the most common environment-friendly and safe water purifying agent is bacillus subtilis preparation, but the control technology also causes the content of bacillus subtilis on the body surface of crawfish. The spores have the characteristic of strong stress resistance, are difficult to kill in the processing process, the nutrition of the shrimp meat is rich and balanced, and the bacillus can grow and propagate quickly as long as the environmental temperature is proper, so that the food is rotten. Therefore, the crayfish products must be subjected to cold chain logistics in the whole process from production to sale, the cold chain logistics is high in cost and unstable, and the crayfish is rotten and deteriorated no matter which cold chain link has problems. The deteriorated food is not only a waste of resources, but also a damage to human body.
In conclusion, the preservation and preservation of crayfish products are still difficult at present, and relevant theories and technologies need to be studied more deeply. At present, common methods for preserving and preserving freshness include low-temperature refrigeration, heating sterilization, vacuum packaging, preservative addition and the like, wherein the preservative mainly comprises chitosan, tea polyphenol, nisin, potassium sorbate and the like. The prior common preservation method has the following defects: the low-temperature refrigeration cost is high and unstable, the taste and flavor of meat are influenced by high-temperature sterilization, the bacteriostatic spectrum of the existing fresh-keeping agent is not wide enough, or the existing fresh-keeping agent has potential harm to human health. The novel preservative film prepared by using the high-temperature resistant lysozyme, the chitosan and the holy ginger essential oil as main materials has strong heat resistance and flexible use, and can be added at any time of crayfish processing. Most of non-spore bacteria can be removed by heat treatment in the processing process of the crayfish food, the residual active microorganisms in the finished product are mainly spores of bacillus, lysozyme has the function of inhibiting the germination and the propagation of the spores, and the ginger essential oil and the chitosan have broad-spectrum antibacterial activity. The three are combined for use, so that the preservation period of the crayfish food can exceed 3 months, and is prolonged by at least 2 months compared with the traditional preservation technology.
Disclosure of Invention
The invention aims to solve the defects of short shelf life of food, damage to human body and high cost in the prior art, and provides a preparation method of a high-temperature resistant lysozyme composite preservative film for food.
A preparation method of a high-temperature resistant lysozyme composite preservative film for food comprises the following steps:
the method comprises the following steps: preparing 1.5% chitosan acetic acid solution, adding glycerol, montmorillonite, Tween-80, high temperature resistant lysozyme and Langling white ginger essential oil, centrifuging, performing ultrasonic water bath, and circulating for multiple times to obtain composite membrane solution A;
step two: pouring the composite membrane solution A into a template, drying for 46-50h at 30-60 ℃, tearing the membrane, keeping for 12-48h at 20-30 ℃ under 50-60% humidity, softening, and finally washing to obtain the composite preservative membrane.
Preferably, in the first step, the preparation process of the acetic acid solution of 1.5% chitosan comprises: adding chitosan into 1% acetic acid solution, and magnetically stirring at room temperature for 24 hr to dissolve completely to obtain 1.5% acetic acid solution of chitosan.
Preferably, in the first step, the mass of the glycerol is 30% of that of the chitosan, the mass of the added montmorillonite is 2.5% of that of the chitosan, the mass of the added Tween-80 is 0.2% of the total volume, the volume of the added holy ginger essential oil is 2% of the total volume, and the mass of the added high-temperature resistant lysozyme is 0.03% of the total volume.
Preferably, in the first step, the centrifugal speed is 15000rpm/min, each time is 5 minutes, and the ultrasonic water bath is carried out for 15 minutes in an ultrasonic cleaner with the power of 360W, wherein the ultrasonic aim is to remove air bubbles in the composite preservative film.
Preferably, in the first step, the centrifugation and the ultrasonic water bath are in one cycle, the glycerol and the montmorillonite are added before the cycle, and the Tween-80, the high-temperature resistant lysozyme and the holy basil ginger essential oil are added before the third cycle.
Preferably, the composite preservative film is applied to food preservation and food packaging.
Compared with the prior art, the invention has the beneficial effects that: the chitosan solution is prepared by using an acetic acid solution, a certain amount of glycerol, montmorillonite, Tween-80, high-temperature-resistant lysozyme and the holy basil ginger essential oil are added, the contained montmorillonite can enhance the mechanical property of the preservative film, the lysozyme has the function of inhibiting spore germination and reproduction, the holy basil ginger essential oil and chitosan have broad-spectrum antibacterial activity, the content of volatile oil contained in the holy basil ginger is highest, the holy basil ginger is poured into a template and then dried to form a film, and humidity treatment is carried out, so that the composite preservative film is obtained. The antibacterial composite preservative film prepared by the preparation method has strong antibacterial capacity, can be used for packaging food or coated on the surface of food to form a coating after being subjected to casting film forming, can effectively inhibit food spoilage caused by microbial pollution and oxidation, prolongs the shelf life of the food, particularly has good inhibition effect on dominant bacterial communities in crayfish food, has good mechanical properties, and can be widely applied to the fields of food preservation, food packaging and the like.
Detailed Description
The present invention will be further illustrated with reference to the following specific examples.
1. The invention provides a preparation method of a high-temperature resistant lysozyme composite preservative film for food, which comprises the following steps: diluting 1mL of glacial acetic acid to a constant volume of 100mL to obtain a 1% glacial acetic acid aqueous solution; adding 1.5g of chitosan into the glacial acetic acid aqueous solution, and magnetically stirring at room temperature for 24 hours to completely dissolve the chitosan to obtain the glacial acetic acid aqueous solution of chitosan; adding 0.45g of glycerol and 0.0375g of montmorillonite into the glacial acetic acid aqueous solution of the chitosan, placing the mixture in a centrifuge with the speed of 15000rpm for centrifugation for 5min, then carrying out water bath in an ultrasonic water bath kettle with the power of 360W for 15min, wherein the centrifugation and the ultrasonic water bath form one cycle, adding 2mL of the white ginger essential oil, 30mg of high-temperature resistant lysozyme and 0.4g of Tween-80 after two cycles, and then carrying out the third cycle to obtain the composite preservative film solution. Pouring the obtained composite membrane solution into a template, drying for 48h at 50 ℃, tearing the membrane, keeping for 30h at 25 ℃ under 55% humidity, softening, and finally washing to obtain the composite preservative membrane, wherein most of non-spore bacteria can be removed by heat treatment in the processing process, the residual active microorganisms in the finished product are mainly spores of bacillus, lysozyme has the function of inhibiting germination and propagation of the spores, and the ginger essential oil and the chitosan have broad-spectrum bacteriostatic activity. The three are combined for use, so that the preservation period of the crayfish food can exceed 3 months, and is prolonged by at least 2 months compared with the traditional preservation technology.
2. Application of composite preservative film in preservation of ready-to-eat crayfishes
(1) Preparation of instant crayfish
Selecting raw materials: selecting raw material shrimps entering a factory in time, removing dead shrimps, old shell shrimps and all impurities, and then grading according to large, medium and small specifications;
secondly, primary processing: soaking crayfish in 15% saline water for 3 hr to discharge dirt, removing head shell, cleaning meat, cleaning, and refrigerating at 4 + -1 deg.C;
③ rinsing water: rinsing the primarily processed crayfish for 0.5h with running water, draining, removing heads of the crayfish, and leaving tails of the crayfish for later use;
fourthly, soaking in soup: adding flavoring agent into water, mixing, heating to boil, keeping boiling for 0.5 hr, cooling to 4 + -1 deg.C, soaking tail of crayfish in the decoction for 2 hr, and draining the decoction;
cooking: heating the soup to a preset temperature, putting the soup-soaked crayfish tails into the soup, boiling the crayfish tails at the temperature of 90 ℃, and respectively heating the crayfish tails for 10min to obtain the instant crayfish tails.
(2) Instant crayfish wrapped by composite preservative film
Wrapping the tail of the instant crayfish with a prepared composite preservative film, then placing the wrapped tail into a tray, and packaging the wrapped tail with a PE film; the control group was directly placed in a tray without freshness-retaining treatment, and vacuum-packed. And (3) storing the packaged instant crayfish tails at normal temperature, sampling at 7 th, 14 th, 21 th, 28 th, 35 th, 42 th, 49 th and 56 th days, performing sensory evaluation, determining the total number of bacterial colonies, the pH value, the content of volatile basic nitrogen, the thiobarbituric acid value and the like, setting three parallels, and repeating each batch of experiments twice to detect the fresh-keeping effect of the composite preservative film on the instant crayfish.
(3) Sensory evaluation and determination of instant crayfish after composite preservative film wrapping treatment
And (3) selecting 9 evaluators to score and evaluate the fleshy tissues, the body surfaces and the odor after training, wherein each full score is 3, calculating each average score, and adding all the scores according to the weight of 0.25 to serve as a final score. Specific scoring criteria are given in table 1:
| item | 3 points of | 2 is divided into | 1 minute (1) | 0 point (min) |
| Fleshy tissue | Moderate elasticity and tight connection between meat and shell | The elasticity is reduced, the hardness is increased somewhat or the material becomes soft | Poor elasticity of shrimp meat and loose connection between meat and shell | The shrimp body is sticky and milky white, and the shrimp meat is not shaped during shelling |
| Body surface | Bright red shrimp shell without abnormal black spots | The shrimp shell turns dark red without abnormal black spots | Shrimp shell without solidHas color and luster, and a few black spots appear | Shrimp shell has no inherent color and more black spots |
| Smell(s) | Has normal fragrance | Has light fragrance | Strong odor | Strong peculiar smell |
TABLE 1 sensory Scoring criteria
(4) Determination of the Total number of colonies
According to a pouring plate counting method specified in GB/T4789.2-2010, the method is slightly modified as follows: taking out crayfish at 7 th, 14 th, 21 th, 28 th, 35 th, 42 th, 49 th and 56 th days, performing primary treatment, rapidly cutting into pieces under sterile environment, taking 1g of cut shrimp meat, adding 9ml of 0.1% sterile normal saline, and shaking and mixing uniformly. After 10-fold dilution of the shrimp homogenate, the culture was poured into 1mL of several suitable dilutions. Colony count determination agar was counted using PCA plates and counted after incubation at 37. + -. 1 ℃ for 48 h.
(5) Determination of the pH value
According to the method specified in GB5009.237-2016, the method is slightly modified as follows: taking out crayfish at 7 th, 14 th, 21 th, 28 th, 35 th, 42 th, 49 th and 56 th days, adding 5g of shrimp body muscle mashed by tissue masher into a conical flask, adding 45mL of distilled water cooled after boiling, standing for 30min, filtering, and measuring pH of the filtrate with a precision acidimeter.
(6) Determination of volatile basic Nitrogen (TVB-N) content
According to the automatic Kjeldahl determination instrument method specified in GB5009.237-2016, the details are as follows: taking out crayfish at 7 th, 14 th, 21 th, 28 th, 35 th, 42 th, 49 th and 56 th days, taking 5g of crayfish body, placing in a small beaker, adding 45mL0.6mol/L perchloric acid solution, homogenizing for 2min, centrifuging, and storing the filtrate at 2-6 ℃ for later use. The measurement is carried out by adopting a K9840 Kjeldahl automatic azotometer, and the instrument conditions are set to be 40mL of 30g/L boric acid absorption liquid, 20mL of water addition amount, 20mL of alkali addition amount and 5min of steaming time. Titration was performed with 0.01mol/L hydrochloric acid standard titration solution and a mixed indicator (1 part of 2g/L methyl red ethanol solution mixed with 1 part of 1g/L methylene blue ethanol solution). Tube blank and reagent blank experiments were performed for each batch. The TVB-N value is expressed in mg/100 g.
X=[(V1-V2)×0.01×14×100]/(m×5/100)
X: the content of volatile basic nitrogen in the sample is in mg/100g;
v1: the volume of the hydrochloric acid standard solution is consumed, in mL;
v2: the volume of the hydrochloric acid standard solution is consumed for the reagent blank, and the unit is mL;
m: sample mass in g.
(7) Measurement of Thiobabarbituric acid value and the like
Taking out crayfish at 7 th, 14 th, 21 th, 28 th, 35 th, 42 th, 49 th and 56 th days, mixing 10g of crayfish with 20mL of 7.5% trichloroacetic acid (containing 0.1% EDTA) and stirring for 1h, extracting Malondialdehyde (MDA), filtering to obtain 5mL of supernatant and 0.02mol/L thiobarbituric acid, carrying out water bath at 95 ℃ for 30min, cooling, measuring by a spectrophotometer at a wavelength of 530nm, drawing a standard curve by using the known concentration of MDA, and quantifying the thiobarbituric acid value by using the concentration of MDA, wherein the value is expressed by mu mol/g.
According to the composite preservative film disclosed by the invention, the holy ginger essential oil and the high-temperature resistant lysozyme are added into the chitosan solution to prepare the chitosan-based composite preservative film, and the chitosan-based composite preservative film is used for packaging food through casting film formation or is coated on the surface of the food to form a coating, so that the food spoilage caused by microbial pollution and oxidation can be effectively inhibited, the quality guarantee period of the food is prolonged, particularly, the composite preservative film has a good inhibiting effect on dominant bacterial flora in crayfish food, has good mechanical properties, and can be widely applied to the fields of food preservation, food packaging and the like.
The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be considered to be within the technical scope of the present invention, and the technical solutions and the inventive concepts thereof according to the present invention should be equivalent or changed within the scope of the present invention.
Claims (6)
1. A preparation method of a high-temperature resistant lysozyme composite preservative film for food is characterized by comprising the following steps:
the method comprises the following steps: preparing 1.5% chitosan acetic acid solution, adding glycerol, montmorillonite, Tween-80, high temperature resistant lysozyme and Langling white ginger essential oil, centrifuging, performing ultrasonic water bath, and circulating for multiple times to obtain composite membrane solution A;
step two: pouring the composite membrane solution A into a template, drying for 46-50h at 30-60 ℃, tearing the membrane, keeping for 12-48h at 20-30 ℃ under 50-60% humidity, softening, and finally washing to obtain the composite preservative membrane.
2. The preparation method of the high-temperature resistant lysozyme composite preservative film for food according to claim 1, wherein in the first step, the preparation process of the 1.5% chitosan acetic acid solution comprises the following steps: adding chitosan into 1% acetic acid solution, and magnetically stirring at room temperature for 24 hr to dissolve completely to obtain 1.5% acetic acid solution of chitosan.
3. The method for preparing the composite preservative film for food high temperature resistant lysozyme according to claim 1, wherein in the first step, the mass of glycerol is 30% of that of chitosan, the mass of added montmorillonite is 2.5% of that of chitosan, the mass of added Tween-80 is 0.2% of the total volume, the volume of added Zingiber glaucum essential oil is 2% of the total volume, and the mass of added high temperature resistant lysozyme is 0.03% of the total volume.
4. The method for preparing the high temperature resistant lysozyme composite preservative film for food according to claim 1, wherein in the first step, the centrifugal speed is 15000rpm/min, 5 minutes each time, and the ultrasonic water bath is carried out for 15 minutes in an ultrasonic cleaner with the power of 360W, wherein the ultrasonic aim is to remove air bubbles in the composite preservative film.
5. The preparation method of the high temperature resistant lysozyme composite preservative film for food according to claim 1, wherein in the first step, the centrifugation and the ultrasonic water bath are performed in one cycle, the glycerol and the montmorillonite are added before the cycle, and the Tween-80, the high temperature resistant lysozyme and the essential oil of the holy ginger are added before the third cycle.
6. The use of the composite preservative film prepared by the method of any one of claims 1 to 5 in food preservation and food packaging.
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| CN202011477337.3A CN112592517A (en) | 2020-12-15 | 2020-12-15 | Preparation method of high-temperature-resistant lysozyme composite preservative film for food |
| CN202110894468.XA CN113388169A (en) | 2020-12-15 | 2021-08-05 | Preparation method of high-temperature-resistant lysozyme composite preservative film for food |
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| CN103275358A (en) * | 2013-05-20 | 2013-09-04 | 吉林大学 | Method for preparing chitosan-based composite preservative film or coating |
| CL2015003698A1 (en) * | 2015-12-22 | 2016-06-17 | Univ Santiago Chile | Degradable film for packaging of fruits and vegetables comprising a polymer matrix based on polyolefin, which incorporates an antimicrobial active agent (biocide or fungicide) of essential oil or said essential oil, and also incorporates degrading agents, and microencapsulation process of said Active antimicrobial essential oil agent, and film preparation method. |
| CN105801890A (en) * | 2016-04-14 | 2016-07-27 | 裴俊 | Method for preparing high-toughness konjac glucomannan preservative film |
| CN106750576B (en) * | 2016-12-05 | 2019-05-07 | 深圳职业技术学院 | A kind of Antibacterial composite preservative film and preparation method thereof |
| CN107232375A (en) * | 2017-06-06 | 2017-10-10 | 安徽科技学院 | A kind of preparation method of the white Jiang Longxu oral health sugar in Tongling |
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