CN111647275A - Preparation method of edible composite membrane of fish myofibrillar protein - Google Patents
Preparation method of edible composite membrane of fish myofibrillar protein Download PDFInfo
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Abstract
The invention discloses a preparation method of an edible composite membrane of fish myofibrillar protein, which comprises the following steps: extracting myofibrillar protein of grass carp; dissolving chitosan in an acetic acid solution to obtain a chitosan solution with the mass concentration of 1-4%, stirring until the chitosan solution is clear, and filtering insoluble substances; adding distilled water into grass carp myofibrillar protein to ensure that the mass concentration of the grass carp myofibrillar protein is 2% -4%, homogenizing for 1-3 min, adjusting the pH value to 3-4, centrifuging for 10-12 min, and collecting supernatant; adding glycerol into the supernatant, stirring for 30-40 min, uniformly mixing, mixing with a chitosan solution according to a volume ratio of 4-5: 3.5-4, adding a rosemary extract with a mass concentration of 0.05-0.2%, and uniformly mixing to obtain a mixture solution for preparing the composite membrane; and baking the mixture solution in an oven to obtain the fish myofibrillar protein edible composite membrane. The method can improve the mechanical property, water resistance, antibacterial property and oxidation resistance of the composite film.
Description
Technical Field
The invention relates to the technical field of biological materials. More specifically, the invention relates to a preparation method of an edible composite membrane of fish myofibrillar protein.
Background
During the past decades, the production and use of chemically synthesized plastic films for food packaging has grown rapidly. Since synthetic plastics are difficult to degrade, serious environmental problems result. Currently, researchers are seeking biodegradable materials that reduce the environmental problems associated with food packaging. Currently, research is mainly focused on biopolymer materials as raw materials for food packaging and preservation. Since biopolymers have properties of preventing water loss, fragrance loss, oxygen permeation, edibility and biodegradability, etc., biopolymers can be used instead of synthetic packaging materials.
Residues (by-products) of fish processing, which can amount to 70% of the initial weight of fish, are regarded as high-quality raw materials with low commercial value, are mostly unused, cause ecological hazards and economic losses, and thus, it is considered that these by-products are prepared into biodegradable fish protein films. The mechanical properties of single Fish Myofibrillar Protein (FMP) membranes are poor due to the presence of disulfide bonds, which makes them impractical for most applications. To increase the flexibility of such films, hydrophilic plasticizers must be added to reduce protein-protein interactions to reduce film brittleness. However, the addition of the plasticizer reduces the tensile strength and water vapor barrier properties of the film. At present, the modification research of the myofibrillar protein membrane at home and abroad is usually focused on a single modification mode of compounding myofibrillar protein and a single natural high molecular compound, and the research report on the compounding of the myofibrillar protein and a plurality of blending agents is very few.
Disclosure of Invention
An object of the present invention is to solve at least the above problems and/or disadvantages and to provide at least the advantages described hereinafter.
The invention also aims to provide a preparation method of the fish myofibrillar protein edible composite film, which can improve the mechanical property, the water resistance, the antibacterial property and the oxidation resistance of the composite film.
To achieve these objects and other advantages in accordance with the purpose of the invention, there is provided a method for preparing an edible composite film of fish myofibrillar proteins, comprising the steps of:
step one, extracting myofibrillar protein of grass carp;
step two, preparing a chitosan solution: dissolving chitosan in an acetic acid solution to obtain a chitosan solution with the mass concentration of 1-4%, stirring until the chitosan solution is clear, and filtering insoluble substances;
step three, preparing a composite membrane: adding distilled water into grass carp myofibrillar protein to ensure that the mass concentration of the grass carp myofibrillar protein is 2% -4%, homogenizing the grass carp myofibrillar protein for 1-3 min under the condition that the rotating speed is 8000-10000 r/min, adjusting the pH value to 3-4, centrifuging the grass carp myofibrillar protein for 10-12 min at room temperature by using a centrifugal force of 3000-3500 g, and collecting supernatant; adding glycerol into the collected supernatant, stirring for 30-40 min to uniformly mix the glycerol with the collected supernatant, then mixing the glycerol with the prepared chitosan solution according to the volume ratio of 4-5: 3.5-4, adding a rosemary extract with the mass concentration of 0.05-0.2%, and uniformly mixing to obtain a mixture solution for preparing the composite membrane; and (3) drying the mixture solution in an oven at 38-42 ℃ for 15-18 h to obtain the fish myofibrillar protein edible composite membrane.
Preferably, the preparation method of the edible composite membrane for fish myofibrillar protein comprises the following specific steps of:
a. taking white meat on a grass carp body before corpse is corpse, processing the white meat into minced fillet, dissolving the minced fillet by using a low-salt phosphate buffer solution, wherein the volume ratio of the low-salt phosphate buffer solution to the minced fillet is 3-5: 1, and homogenizing for 1-3 min under the condition that the rotating speed is 8000-10000 r/min;
b. standing for 10-20 min, centrifuging at the rotating speed of 3000-;
c. dissolving the precipitate obtained in the step b in a high-salt buffer solution, wherein the volume ratio of the precipitate to the high-salt buffer solution is 1: 3-5, homogenizing for 1-2 min under the condition that the rotating speed is 8000-10000 r/min, and carrying out static extraction for 20-24 h;
d. centrifuging the mixture solution obtained in the step c for 10-15 min at the rotating speed of 1000-14000 r/min, taking supernatant, precipitating the supernatant for 25-35 min by using condensed water, wherein the volume ratio of the condensed water to the supernatant is 8-12: 1, centrifuging for 10-12 min at the rotating speed of 8000-12000 r/min, and taking precipitate, namely grass carp myofibrillar protein;
wherein, the steps are all carried out at the temperature of 1-5 ℃.
Preferably, the preparation method of the edible composite membrane of fish myofibrillar protein, wherein the step of dissolving chitosan in acetic acid solution to obtain chitosan solution with mass concentration of 1% -4%, comprises the following steps:
adding 1ml of glacial acetic acid into 99ml of deionized water to prepare an acetic acid solution with the mass concentration of 1%, and then dissolving 1-4 g of chitosan into the 1% acetic acid solution to obtain a chitosan solution with the mass concentration of 1% -4%.
Preferably, in the third step, 1mol/L hydrochloric acid solution is used to adjust the pH value of the grass carp myofibrillar protein solution to 3-4.
Preferably, the preparation method of the edible composite membrane of fish myofibrillar protein comprises the following steps of drying the mixture solution in an oven at 38-42 ℃ for 15-18 h to obtain the edible composite membrane of fish myofibrillar protein, and specifically comprises the following steps: 40ml of the mixture solution was poured into a 13cm disposable petri dish and then placed in an oven for baking.
Preferably, in the preparation method of the edible composite membrane of the fish myofibrillar protein, the content of the extracted grass carp myofibrillar protein is measured by a biuret method.
The invention at least comprises the following beneficial effects: firstly, compounding grass carp myofibrillar protein and chitosan can increase the mechanical property of the composite membrane, and simultaneously solve the problem of poor water resistance caused by a single grass carp myofibrillar protein membrane; secondly, no chemical synthetic preservative is added in the processing method of the composite membrane, the composite membrane is natural and environment-friendly, and chitosan can play a certain antibacterial role; the glycerol with hydrophilicity is used as a plasticizer, so that the interaction between proteins is reduced, and the brittleness of the composite membrane is reduced; thirdly, after the composite film is formed, the film is in a certain faint yellow color due to Maillard reaction and the color action of the chitosan solution, and has good light resistance; fourthly, the oxidation resistance of the composite membrane is also improved by adding the rosemary extract in the preparation process.
Additional advantages, objects, and features of the invention will be set forth in part in the description which follows and in part will become apparent to those having ordinary skill in the art upon examination of the following or may be learned from practice of the invention.
Drawings
FIG. 1 is a schematic flow chart illustrating a method for preparing an edible composite membrane of fish myofibrillar proteins according to an embodiment of the present invention;
FIG. 2 is a graph showing the results of mechanical property measurements of products obtained in examples 1 to 5;
FIG. 3 is a graph showing the results of antioxidant assay of the products obtained in examples 1 to 5.
Detailed Description
The present invention is further described in detail below with reference to the attached drawings so that those skilled in the art can implement the invention by referring to the description text.
It will be understood that terms such as "having," "including," and "comprising," as used herein, do not preclude the presence or addition of one or more other elements or groups thereof.
As shown in fig. 1, an embodiment of the present invention provides a method for preparing an edible composite membrane of fish myofibrillar proteins, comprising the following steps:
step one, extracting myofibrillar protein of grass carp;
step two, preparing a chitosan solution: dissolving chitosan in an acetic acid solution to obtain a chitosan solution with the mass concentration of 1-4%, stirring until the chitosan solution is clear, and filtering insoluble substances;
step three, preparing a composite membrane: adding distilled water into grass carp myofibrillar protein to ensure that the mass concentration of the grass carp myofibrillar protein is 2% -4%, homogenizing the grass carp myofibrillar protein for 1-3 min under the condition that the rotating speed is 8000-10000 r/min, adjusting the pH value to 3-4, centrifuging the grass carp myofibrillar protein for 10-12 min at room temperature by using a centrifugal force of 3000-3500 g, and collecting supernatant; adding glycerol into the collected supernatant, stirring for 30-40 min to uniformly mix the glycerol with the collected supernatant, then mixing the glycerol with the prepared chitosan solution according to the volume ratio of 4-5: 3.5-4, adding a rosemary extract with the mass concentration of 0.05-0.2%, and uniformly mixing to obtain a mixture solution for preparing the composite membrane; and (3) drying the mixture solution in an oven at 38-42 ℃ for 15-18 h to obtain the fish myofibrillar protein edible composite membrane.
Among them, the volume ratio of the supernatant mixed with glycerol to the prepared chitosan solution may be preferably 4.3:3.7, the concentration of the chitosan solution is preferably 2%.
According to the embodiment of the invention, the myofibrillar protein of the grass carp and the chitosan are compounded, so that the mechanical property of the composite membrane can be increased, and the problem of poor water resistance caused by a single membrane of the myofibrillar protein of the grass carp is solved; the processing method of the prepared composite membrane does not add any chemically synthesized preservative, is natural and environment-friendly, and simultaneously, the chitosan can play a certain antibacterial role; the glycerol with hydrophilicity is used as a plasticizer, so that the interaction between proteins is reduced, and the brittleness of the composite membrane is reduced; after the composite film is formed, the film is in a certain faint yellow color due to the Maillard reaction and the color action of a chitosan solution, and has good light resistance; the addition of rosemary extract in the preparation process also increases the oxidation resistance of the composite membrane.
In one embodiment, the preparation method of the edible composite membrane for fish myofibrillar protein comprises the following specific steps of:
a. taking white meat on a grass carp body before corpse is corpse, processing the white meat into minced fillet, dissolving the minced fillet by using a low-salt phosphate buffer solution, wherein the volume ratio of the low-salt phosphate buffer solution to the minced fillet is 3-5: 1, and homogenizing for 1-3 min under the condition that the rotating speed is 8000-10000 r/min;
b. standing for 10-20 min, centrifuging at the rotating speed of 3000-;
c. dissolving the precipitate obtained in the step b in a high-salt buffer solution, wherein the volume ratio of the precipitate to the high-salt buffer solution is 1: 3-5, homogenizing for 1-2 min under the condition that the rotating speed is 8000-10000 r/min, and carrying out static extraction for 20-24 h;
d. centrifuging the mixture solution obtained in the step c for 10-15 min at the rotating speed of 1000-14000 r/min, taking supernatant, precipitating the supernatant for 25-35 min by using condensed water, wherein the volume ratio of the condensed water to the supernatant is 8-12: 1, centrifuging for 10-12 min at the rotating speed of 8000-12000 r/min, and taking precipitate, namely grass carp myofibrillar protein;
wherein, the steps are all carried out at the temperature of 1-5 ℃.
In one embodiment, the method for preparing the edible fish myofibrillar protein composite film, wherein the step of dissolving chitosan in an acetic acid solution to obtain a chitosan solution with a mass concentration of 1% -4%, specifically comprises the following steps:
adding 1ml of glacial acetic acid into 99ml of deionized water to prepare an acetic acid solution with the mass concentration of 1%, and then dissolving 1-4 g of chitosan into the 1% acetic acid solution to obtain a chitosan solution with the mass concentration of 1% -4%.
In one embodiment, in the third step, a hydrochloric acid solution of 1mol/L is used to adjust the pH value of the grass carp myofibrillar protein solution to 3-4.
In one specific embodiment, the preparation method of the edible fish myofibrillar protein composite film comprises the following steps of drying the mixture solution in an oven at 38-42 ℃ for 15-18 hours to obtain the edible fish myofibrillar protein composite film, and specifically comprises the following steps: 40ml of the mixture solution was poured into a 13cm disposable petri dish and then placed in an oven for baking.
In one embodiment, in the preparation method of the edible composite membrane of fish myofibrillar protein, the extracted grass carp myofibrillar protein is subjected to content measurement by using a biuret method.
Specific examples will be given below for illustration:
example 1
S10, extracting myofibrillar protein of grass carp: s11, taking white meat on the grass carp before corpse stupefaction, removing blood and impurities, processing the white meat into minced fillet by using a food conditioner, dissolving the minced fillet by using a low-salt phosphate buffer solution, wherein the volume ratio of the low-salt phosphate buffer solution to the minced fillet is 3:1, and homogenizing the minced fillet for 1min at the rotating speed of 8000 r/min; s12, standing for 10min, centrifuging for 8min at the rotating speed of 3000r/min, repeating for 3 times, and taking the precipitate; s13, dissolving the precipitate obtained in the step S12 in a high-salt buffer solution, wherein the volume ratio of the precipitate to the high-salt buffer solution is 1: 3, homogenizing for 1min at the rotating speed of 8000r/min, and carrying out static extraction for 20 h; s14, centrifuging the mixture solution obtained in the step S13 for 10min at the rotation speed of 1000r/min, taking supernatant, precipitating the supernatant for 25min by using condensed water, wherein the volume ratio of the condensed water to the supernatant is 8:1, centrifuging for 10min at the rotation speed of 8000r/min, and taking precipitate, namely grass carp myofibrillar protein; wherein, the steps are all carried out at the temperature of 1 ℃, and the content of the extracted grass carp myofibrillar protein is measured by a biuret method.
S20, preparation of chitosan solution: adding 1ml of glacial acetic acid into 99ml of deionized water to prepare an acetic acid solution with the mass concentration of 1%, then dissolving 1g of chitosan in the 1% acetic acid solution to obtain a chitosan solution with the mass concentration of 1%, stirring until the chitosan solution is clear, and filtering insoluble substances;
s30, preparation of a composite membrane: adding distilled water into grass carp myofibrillar protein to enable the mass concentration of the grass carp myofibrillar protein to be 2%, homogenizing the grass carp myofibrillar protein for 1min under the condition that the rotating speed is 8000r/min, adjusting the pH value of the grass carp myofibrillar protein solution to 3 by using 1mol/L hydrochloric acid solution, centrifuging the grass carp myofibrillar protein solution for 10min at the room temperature by using a centrifugal force of 3000g, and collecting supernatant; adding glycerol into the collected supernatant, stirring for 30min to uniformly mix the glycerol with the collected supernatant, then mixing the glycerol with the prepared chitosan solution according to the volume ratio of 4:3.5, adding 0.05% of rosemary extract, and uniformly mixing to obtain a mixture solution for preparing the composite membrane; pouring 40ml of the mixture solution into a disposable culture dish of 13cm, then placing the culture dish into an oven for baking, and baking for 15 hours at the temperature of 38 ℃ to obtain the fish myofibrillar protein edible composite membrane.
Example 2
S10, extracting myofibrillar protein of grass carp: s11, taking white meat on the grass carp before corpse mumps, removing blood and impurities after red color is not taken, processing the white meat into minced fillet by using a food conditioning machine, and dissolving the minced fillet by using a low-salt phosphate buffer solution, wherein the volume ratio of the low-salt phosphate buffer solution to the minced fillet is 5:1, homogenizing for 3min under the condition that the rotating speed is 10000 r/min; s12, standing for 20min, centrifuging for 12min at the rotating speed of 5000r/min, repeating for 5 times, and taking the precipitate; s13, dissolving the precipitate obtained in the step S12 in a high-salt buffer solution, wherein the volume ratio of the precipitate to the high-salt buffer solution is 1: 5, homogenizing for 2min under the condition that the rotating speed is 10000r/min, and carrying out static extraction for 24 h; s14, centrifuging the mixture solution obtained in the step S13 for 15min at the rotation speed of 14000r/min, taking supernatant, precipitating the supernatant for 35min by using condensed water, wherein the volume ratio of the condensed water to the supernatant is 12:1, centrifuging for 12min at the rotation speed of 12000r/min, and taking precipitate, namely grass carp myofibrillar protein; wherein, the steps are all carried out at the temperature of 5 ℃, and the content of the extracted grass carp myofibrillar protein is measured by a biuret method.
S20, preparation of chitosan solution: adding 1ml of glacial acetic acid into 99ml of deionized water to prepare an acetic acid solution with the mass concentration of 1%, then dissolving 4g of chitosan into the 1% acetic acid solution to obtain a chitosan solution with the mass concentration of 4%, stirring until the chitosan solution is clear, and filtering insoluble substances;
s30, preparation of a composite membrane: adding distilled water into grass carp myofibrillar protein to enable the mass concentration of the grass carp myofibrillar protein to be 4%, homogenizing the grass carp myofibrillar protein for 3min under the condition that the rotating speed is 10000r/min, adjusting the pH value of the grass carp myofibrillar protein solution to 4 by using 1mol/L hydrochloric acid solution, centrifuging the grass carp myofibrillar protein solution for 12min at room temperature by using a centrifugal force of 3500g, and collecting supernatant; adding glycerol into the collected supernatant, stirring for 40min to uniformly mix the glycerol with the collected supernatant, then mixing the glycerol with the prepared chitosan solution according to the volume ratio of 5:4, adding 0.2% of rosemary extract by mass concentration, and uniformly mixing to obtain a mixture solution for preparing the composite membrane; pouring 40ml of the mixture solution into a disposable culture dish of 13cm, then placing the culture dish into an oven for baking, and baking for 18 hours at the temperature of 42 ℃ to obtain the fish myofibrillar protein edible composite membrane.
Example 3
S10, extracting myofibrillar protein of grass carp: s11, taking white meat on the grass carp before corpse stupefaction, removing blood and impurities, processing the white meat into minced fillet by using a food conditioning machine, dissolving the minced fillet by using a low-salt phosphate buffer solution, wherein the volume ratio of the low-salt phosphate buffer solution to the minced fillet is 4:1, and homogenizing the minced fillet for 1min at the rotating speed of 9000 r/min; s12, standing for 15min, centrifuging for 10min at the rotating speed of 4000r/min, repeating for 3 times, and taking the precipitate; s13, dissolving the precipitate obtained in the step S12 in a high-salt buffer solution, wherein the volume ratio of the precipitate to the high-salt buffer solution is 1: 4, homogenizing for 1min at the rotating speed of 9000r/min, and carrying out static extraction for 22 h; s14, centrifuging the mixture solution obtained in the step S13 for 10min at the rotating speed of 12000r/min, taking supernatant, precipitating the supernatant for 30min by using condensed water, wherein the volume ratio of the condensed water to the supernatant is 10:1, centrifuging for 10min at the rotating speed of 10000r/min, and taking precipitate, namely grass carp myofibrillar protein; wherein, the steps are all carried out at the temperature of 4 ℃, and the content of the extracted grass carp myofibrillar protein is measured by a biuret method.
S20, preparation of chitosan solution: adding 1ml of glacial acetic acid into 99ml of deionized water to prepare an acetic acid solution with the mass concentration of 1%, then dissolving 2g of chitosan into the 1% acetic acid solution to obtain a chitosan solution with the mass concentration of 1% -4%, stirring until the chitosan solution is clear, and filtering insoluble substances;
s30, preparation of a composite membrane: adding distilled water into grass carp myofibrillar protein to enable the mass concentration of the grass carp myofibrillar protein to be 2%, homogenizing for 1min under the condition that the rotating speed is 9000r/min, adjusting the pH value of the grass carp myofibrillar protein solution to 3 by using 1mol/L hydrochloric acid solution, centrifuging for 10min at the centrifugal force of 3000g at room temperature, and collecting supernatant; adding glycerol into the collected supernatant, stirring for 30min to uniformly mix the glycerol with the collected supernatant, then mixing the glycerol with the prepared chitosan solution according to the volume ratio of 4.3:3.7, adding 0.10% of rosemary extract by mass concentration, and uniformly mixing to obtain a mixture solution for preparing the composite membrane; pouring 40ml of the mixture solution into a disposable culture dish of 13cm, then placing the culture dish into an oven for baking, and baking for 16 hours at the temperature of 40 ℃ to obtain the fish myofibrillar protein edible composite membrane.
Example 4
S10, extracting myofibrillar protein of grass carp: s11, taking white meat on the grass carp before corpse stupefaction, removing blood and impurities, processing the white meat into minced fillet by using a food conditioning machine, dissolving the minced fillet by using a low-salt phosphate buffer solution, wherein the volume ratio of the low-salt phosphate buffer solution to the minced fillet is 4:1, and homogenizing the minced fillet for 2min at the rotating speed of 9000 r/min; s12, standing for 17min, centrifuging for 9min at the rotating speed of 4000r/min, repeating for 4 times, and taking the precipitate; s13, dissolving the precipitate obtained in the step S12 in a high-salt buffer solution, wherein the volume ratio of the precipitate to the high-salt buffer solution is 1: 4, homogenizing for 2min under the condition that the rotating speed is 85000r/min, and carrying out static extraction for 23 h; s14, centrifuging the mixture solution obtained in the step S13 for 13min at the rotation speed of 13000r/min, taking supernatant, precipitating the supernatant for 30min by using condensed water, wherein the volume ratio of the condensed water to the supernatant is 9:1, centrifuging for 11min at the rotation speed of 11000r/min, and taking precipitate, namely grass carp myofibrillar protein; wherein, the steps are all carried out at the temperature of 3 ℃, and the content of the extracted grass carp myofibrillar protein is measured by a biuret method.
S20, preparation of chitosan solution: adding 1ml of glacial acetic acid into 99ml of deionized water to prepare an acetic acid solution with the mass concentration of 1%, then dissolving 3g of chitosan into the 1% acetic acid solution to obtain a chitosan solution with the mass concentration of 3%, stirring until the chitosan solution is clear, and filtering insoluble substances;
s30, preparation of a composite membrane: adding distilled water into grass carp myofibrillar protein to enable the mass concentration of the grass carp myofibrillar protein to be 3%, homogenizing the grass carp myofibrillar protein for 2min under the condition that the rotating speed is 9500r/min, adjusting the pH value of the grass carp myofibrillar protein solution to 3 by using 1mol/L hydrochloric acid solution, centrifuging the grass carp myofibrillar protein solution for 11min at the room temperature by using a centrifugal force of 3400g, and collecting supernatant; adding glycerol into the collected supernatant, stirring for 35min to uniformly mix, then mixing with the prepared chitosan solution according to the volume ratio of 4:3.5, adding 0.15% of rosemary extract, and uniformly mixing to obtain a mixture solution for preparing the composite membrane; pouring 40ml of the mixture solution into a disposable culture dish of 13cm, then placing the culture dish into an oven for baking, and baking for 17 hours at the temperature of 39 ℃ to obtain the fish myofibrillar protein edible composite membrane.
Example 5
S10, extracting myofibrillar protein of grass carp: s11, taking white meat on the grass carp before corpse stupefaction, removing blood and impurities, processing the white meat into minced fillet by using a food conditioning machine, dissolving the minced fillet by using a low-salt phosphate buffer solution, wherein the volume ratio of the low-salt phosphate buffer solution to the minced fillet is 4:1, and homogenizing the minced fillet for 2min at the rotating speed of 9000 r/min; s12, standing for 15min, centrifuging for 10min at the rotating speed of 5000r/min, repeating for 5 times, and taking the precipitate; s13, dissolving the precipitate obtained in the step S12 in a high-salt buffer solution, wherein the volume ratio of the precipitate to the high-salt buffer solution is 1: 4, homogenizing for 2min at the rotating speed of 9000r/min, and carrying out static extraction for 21 h; s14, centrifuging the mixture solution obtained in the step S13 for 10min at the rotation speed of 12000r/min, taking supernatant, precipitating the supernatant for 30min by using condensed water, wherein the volume ratio of the condensed water to the supernatant is 9:1, centrifuging the supernatant for 012min at the rotation speed of 9000r/min, and taking precipitate, namely grass carp myofibrillar protein; wherein, the steps are all carried out at the temperature of 4 ℃, and the content of the extracted grass carp myofibrillar protein is measured by a biuret method.
S20, preparation of chitosan solution: adding 1ml of glacial acetic acid into 99ml of deionized water to prepare an acetic acid solution with the mass concentration of 1%, then dissolving 2g of chitosan into the 1% acetic acid solution to obtain a chitosan solution with the mass concentration of 2%, stirring until the chitosan solution is clear, and filtering insoluble substances;
s30, preparation of a composite membrane: adding distilled water into grass carp myofibrillar protein to enable the mass concentration of the grass carp myofibrillar protein to be 2%, homogenizing for 1min under the condition that the rotating speed is 9000r/min, adjusting the pH value of the grass carp myofibrillar protein solution to 3 by using 1mol/L hydrochloric acid solution, centrifuging for 12min at the room temperature by using a centrifugal force of 3000g, and collecting supernatant; adding glycerol into the collected supernatant, stirring for 40min to uniformly mix the glycerol with the collected supernatant, then mixing the glycerol with the prepared chitosan solution according to the volume ratio of 4:3.5, and uniformly mixing to obtain a mixture solution for preparing the composite membrane; pouring 40ml of the mixture solution into a disposable culture dish of 13cm, then placing the culture dish into an oven for baking, and baking for 17 hours at the temperature of 38 ℃ to obtain the fish myofibrillar protein edible composite membrane.
The products obtained in examples 1, 2, 3, 4 and 5 were subjected to mechanical property measurements. A rectangular sample of 50mm multiplied by 20mm is prepared by a sample preparation knife, the sample is placed in a constant temperature and humidity box with the temperature of 25 ℃ and the relative humidity of 50 percent for balancing for 12h, after the sample is taken out, a texture analyzer is used for measuring the Tensile Strength (TS) and the elongation at break (E), the length of the sample between two chucks is 30mm, the test speed is 2.00mm/s, 3 times of parallel tests are carried out, and the average value is taken (Wang N2015).
Tensile strength: TS ═ F/A
In the formula: TS-tensile strength (MPa) of the sample;
f is the maximum tensile force (N) to which the sample is subjected at break;
a-cross-sectional area of the sample before testing (mm 2).
Elongation at break: e ═ L0/L0%
In the formula: e-elongation at break of the sample;
l0-length of sample before testing, mm;
l-the length of the sample at break, mm.
The test results are shown in fig. 2.
The oxidation resistance measurements were carried out on the products obtained in examples 1, 2, 3, 4 and 5. To a film sample (25mg) was added 3ml of distilled water and stirred vigorously for 3 h. After this, the mixture was centrifuged at 3000g for 10 minutes using a centrifuge. The resulting supernatant was further used for measurement of DPPH free radical scavenging activity and iron reducing antioxidant capacity. To the membrane extract (1.5ml) was added 1.5ml of 2, 2-diphenyl-1-pyridinehydrazide (DPPH) in 95% ethanol, followed by thorough mixing. Thereafter, the mixture was kept in the dark at room temperature for 30 minutes, and then the absorbance was measured at 517nm using a spectrophotometer. DPPH free radical scavenging activity was calculated and expressed as a percentage of DPPH scavenging activity. The test results are shown in fig. 3.
While embodiments of the invention have been disclosed above, it is not intended to be limited to the uses set forth in the specification and examples. It can be applied to all kinds of fields suitable for the present invention. Additional modifications will readily occur to those skilled in the art. It is therefore intended that the invention not be limited to the exact details and illustrations described and illustrated herein, but fall within the scope of the appended claims and equivalents thereof.
Claims (6)
1. The preparation method of the fish myofibrillar protein edible composite membrane is characterized by comprising the following steps:
step one, extracting myofibrillar protein of grass carp;
step two, preparing a chitosan solution: dissolving chitosan in an acetic acid solution to obtain a chitosan solution with the mass concentration of 1-4%, stirring until the chitosan solution is clear, and filtering insoluble substances;
step three, preparing a composite membrane: adding distilled water into grass carp myofibrillar protein to ensure that the mass concentration of the grass carp myofibrillar protein is 2% -4%, homogenizing the grass carp myofibrillar protein for 1-3 min under the condition that the rotating speed is 8000-10000 r/min, adjusting the pH value to 3-4, centrifuging the grass carp myofibrillar protein for 10-12 min at room temperature by using a centrifugal force of 3000-3500 g, and collecting supernatant; adding glycerol into the collected supernatant, stirring for 30-40 min to uniformly mix the glycerol with the collected supernatant, then mixing the glycerol with the prepared chitosan solution according to the volume ratio of 4-5: 3.5-4, adding a rosemary extract with the mass concentration of 0.05-0.2%, and uniformly mixing to obtain a mixture solution for preparing the composite membrane; and (3) drying the mixture solution in an oven at 38-42 ℃ for 15-18 h to obtain the fish myofibrillar protein edible composite membrane.
2. The method for preparing the edible composite film of fish myofibrillar protein according to claim 1, wherein the specific steps for extracting the grass carp myofibrillar protein are as follows:
a. taking white meat on a grass carp body before corpse is corpse, processing the white meat into minced fillet, dissolving the minced fillet by using a low-salt phosphate buffer solution, wherein the volume ratio of the low-salt phosphate buffer solution to the minced fillet is 3-5: 1, and homogenizing for 1-3 min under the condition that the rotating speed is 8000-10000 r/min;
b. standing for 10-20 min, centrifuging at the rotating speed of 3000-;
c. dissolving the precipitate obtained in the step b in a high-salt buffer solution, wherein the volume ratio of the precipitate to the high-salt buffer solution is 1: 3-5, homogenizing for 1-2 min under the condition that the rotating speed is 8000-10000 r/min, and carrying out static extraction for 20-24 h;
d. centrifuging the mixture solution obtained in the step c for 10-15 min at the rotating speed of 1000-14000 r/min, taking supernatant, precipitating the supernatant for 25-35 min by using condensed water, wherein the volume ratio of the condensed water to the supernatant is 8-12: 1, centrifuging for 10-12 min at the rotating speed of 8000-12000 r/min, and taking precipitate, namely grass carp myofibrillar protein;
wherein, the steps are all carried out at the temperature of 1-5 ℃.
3. The method for preparing the edible composite membrane of fish myofibrillar protein according to claim 1, wherein the step of dissolving chitosan in acetic acid solution to obtain chitosan solution with mass concentration of 1-4% specifically comprises the following steps:
adding 1ml of glacial acetic acid into 99ml of deionized water to prepare an acetic acid solution with the mass concentration of 1%, and then dissolving 1-4 g of chitosan into the 1% acetic acid solution to obtain a chitosan solution with the mass concentration of 1% -4%.
4. The method for preparing the edible composite membrane of fish myofibrillar protein according to claim 3, wherein in the third step, 1mol/L hydrochloric acid solution is used to adjust the pH value of the grass carp myofibrillar protein solution to 3-4.
5. The preparation method of the edible fish myofibrillar protein composite film according to claim 1, characterized in that the mixture solution is dried in an oven at 38-42 ℃ for 15-18 h to obtain the edible fish myofibrillar protein composite film, which specifically comprises the following steps: 40ml of the mixture solution was poured into a 13cm disposable petri dish and then placed in an oven for baking.
6. The method for preparing the edible composite film of fish myofibrillar protein according to claim 2, wherein the extracted myofibrillar protein of grass carp is subjected to content measurement by a biuret method.
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