CN111937952A - Freezing preservative based on fish scale protein zymolyte and preparation method and application thereof - Google Patents
Freezing preservative based on fish scale protein zymolyte and preparation method and application thereof Download PDFInfo
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- CN111937952A CN111937952A CN202010785570.1A CN202010785570A CN111937952A CN 111937952 A CN111937952 A CN 111937952A CN 202010785570 A CN202010785570 A CN 202010785570A CN 111937952 A CN111937952 A CN 111937952A
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23B—PRESERVING, e.g. BY CANNING, MEAT, FISH, EGGS, FRUIT, VEGETABLES, EDIBLE SEEDS; CHEMICAL RIPENING OF FRUIT OR VEGETABLES; THE PRESERVED, RIPENED, OR CANNED PRODUCTS
- A23B4/00—General methods for preserving meat, sausages, fish or fish products
- A23B4/06—Freezing; Subsequent thawing; Cooling
- A23B4/08—Freezing; Subsequent thawing; Cooling with addition of chemicals or treatment with chemicals before or during cooling, e.g. in the form of an ice coating or frozen block
- A23B4/09—Freezing; Subsequent thawing; Cooling with addition of chemicals or treatment with chemicals before or during cooling, e.g. in the form of an ice coating or frozen block with direct contact between the food and the chemical, e.g. liquid N2, at cryogenic temperature
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23J1/00—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites
- A23J1/04—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from fish or other sea animals
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23J1/00—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites
- A23J1/10—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from hair, feathers, horn, skins, leather, bones, or the like
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23J3/00—Working-up of proteins for foodstuffs
- A23J3/30—Working-up of proteins for foodstuffs by hydrolysis
- A23J3/32—Working-up of proteins for foodstuffs by hydrolysis using chemical agents
- A23J3/34—Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes
- A23J3/341—Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes of animal proteins
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- Health & Medical Sciences (AREA)
- Marine Sciences & Fisheries (AREA)
- Wood Science & Technology (AREA)
- Meat, Egg Or Seafood Products (AREA)
Abstract
The invention relates to the technical field of aquatic food preservation, in particular to a fish scale protein zymolyte-based freezing preservative, a preparation method and application thereof. The frozen fresh-keeping agent is obtained by carrying out enzymolysis on fish scales by using compound protease, wherein the compound protease comprises alkaline protease, trypsin, neutral protease and flavourzyme. The freezing preservative provided by the invention has the functions of oxidation resistance and freezing resistance, and can be applied to the fish slices plated with ice coatings, so that the quality of the product can be kept at a better level after the fish slices are frozen for 9 months, and the sensory quality of the product can be kept over 92% of fresh fish meat, and the freezing preservative is obviously superior to the fish slices plated with ice coatings or tea polyphenols by pure water.
Description
Technical Field
The invention relates to the technical field of aquatic food preservation, in particular to a fish scale protein zymolyte-based freezing preservative, a preparation method and application thereof.
Background
The frozen storage is used as a common aquatic product fresh-keeping mode, more than 90% of water in the aquatic product is frozen, and the enzyme activity and the microbial growth are almost completely inhibited, so that the aquatic product can be preserved for a long time. However, the aquatic products will have the phenomena of quality reduction, drying loss, protein denaturation, fat oxidation and the like in the process of frozen storage. The ice-plated clothes are used as an effective storage means for reducing the dry consumption and protecting aquatic products, and are widely applied to the freezing storage of the aquatic products. The ice-coated clothes can effectively slow down the deterioration of the texture during the frozen storage period, but the color change, the protein oxidation, the water retention and other phenomena usually occur in the simple ice-coated aquatic products along with the prolonging of the storage time. In addition, as people's health consciousness is gradually increased, people begin to pay attention to the problem of residual components of the preservative, so the development of the chemical preservation method is limited. The natural biological preservative is more popular because of the safety, health, good water solubility and small influence on the quality of aquatic products, and can keep the better flavor of the aquatic products. At present, no report of using fish scale zymolyte as a natural preservative for fish fillets and application thereof exists.
Disclosure of Invention
On the basis of the research on the existing ice coating preservation of aquatic products, the invention combines the characteristics of fish scales which are the products of fish bodies, utilizes the fish scales as raw materials, applies a biological enzymolysis technology to prepare the freezing preservative with the functions of oxidation resistance and freezing resistance, applies the freezing preservative to the fish slices coated with the ice coatings, keeps the product quality at a better level after the fish slices are frozen and stored for 9 months, keeps the sensory quality of more than 92 percent of fresh fish meat, and is obviously superior to the fish slices coated with the ice coatings or tea polyphenols by pure water.
Specifically, the invention provides the following technical scheme:
the invention provides a fish scale protein zymolyte-based freezing preservative which is obtained by carrying out enzymolysis on fish scales by using compound protease, wherein the compound protease comprises alkaline protease, trypsin, neutral protease and flavourzyme.
Preferably, in the frozen fresh-keeping agent, the addition amount of the compound protease is 0.05-0.5% by mass of the dry matter of the fish scales, and preferably 0.1-0.3% by mass of the dry matter of the fish scales.
Preferably, in the frozen fresh-keeping agent, in the compound protease, the mass ratio of the alkaline protease, the trypsin, the neutral protease and the flavourzyme is 0.8-1.2: 1-2: 1-2: 1.
preferably, in the frozen preservative, the fish scales are the scales of tilapia or grass carp.
The invention also provides a preparation method of the frozen preservative, which comprises the following steps:
(1) mixing fish scales with water to obtain a fish scale-water mixture;
(2) sequentially carrying out heat treatment, vibration screen filtration and ultrasonic treatment on the fish scale-water mixture to obtain slurry;
(3) and carrying out enzymolysis on the slurry by using the compound protease to obtain the compound protease.
Preferably, in the preparation method, in the step (1), the amount of water is 400-500 wt% based on the mass of the dry matter of the fish scales;
and/or in the step (2), the temperature of the heat treatment is 115-121 ℃;
and/or in the step (2), the mesh number of the vibrating screen is 30-60 meshes;
and/or in the step (2), the frequency of ultrasonic treatment is 80-100 Kh, the time of ultrasonic treatment is 10-15 min, and the temperature of ultrasonic treatment is 75-85 ℃.
Preferably, in the above preparation method, in step (3), the enzymatic hydrolysis includes the following steps:
adding the compound protease into the slurry, performing enzymolysis for 0.5-1.0 h at 45-55 ℃, and then performing heat preservation for 15-20 min at 90-95 ℃.
Preferably, in the preparation method, in the step (3), after the enzymolysis, the method further comprises the steps of filtering and adjusting the obtained filtrate into a suspension with a protein content of 1.5-2 wt%.
The invention also provides the application of the freezing preservative or the freezing preservative prepared by the preparation method in freezing, ice coating, storing and refreshing of fish.
Preferably, in the application, the fish meat is frozen to the temperature of-18 to-23 ℃ at the temperature of-35 to-40 ℃, then the frozen fish meat is immersed in the freezing preservative for 10 to 15 seconds, the ice coating content is controlled to be 15 to 20 percent, the fish meat plated with the ice coating is obtained, and the fish meat is frozen and stored after being vacuum packaged.
The invention has the following beneficial effects:
(1) the frozen preservative based on the fish scale protein zymolyte is not added with any chemical substance, and can be widely used as a natural preservative for aquatic food;
(2) the frozen preservative based on the fish scale protein zymolyte adopts a plurality of food-grade compound proteases (alkaline protease, trypsin, neutral protease and flavourzyme), and obtains fish scale protein peptides with specific functions, namely 100% of fish scale protein peptides, through moderate enzymolysis under mild conditions;
(3) the frozen preservative based on the fish scale protein zymolyte has simple production process, low production cost and better anti-freezing and anti-oxidation functions, and fish scales do not need deliming treatment and separation and purification;
(4) the fish scale protein zymolyte-based freezing preservative is used for storage and preservation of fish slices, has a remarkable preservation effect, and keeps the sensory quality of the product to be more than 92 percent after the product is stored for 9 months.
Detailed Description
The following examples are intended to illustrate the invention but are not intended to limit the scope of the invention. The examples do not show the specific techniques or conditions, according to the technical or conditions described in the literature in the field, or according to the product specifications.
In the present invention, the instruments and the like used are conventional products which are purchased from regular vendors, not indicated by manufacturers. The process is conventional unless otherwise specified, and the starting materials are commercially available from the open literature. In the following examples, alkaline protease, neutral protease and gustatory protease were obtained from Novozymes and trypsin was obtained from Sigma.
Example 1
(1) 1000 g of fresh tilapia scales are used, clean water meeting the sanitary standard of drinking water is used for cleaning the scales, water which is 5 times of the scales in weight is added after the scales are cleaned, the scales are boiled for 60 minutes at the temperature of 121 ℃, then a 40-mesh vibrating screen is used for filtering, the temperature is adjusted to 75 ℃, and an ultrasonic generator is used for processing for 15 minutes by ultrasonic waves (the frequency is 80kH) to obtain scale slurry;
(2) adding compound protease into the fish scale (1) according to the weight percentage of 0.2% of the weight of the fish scales for enzymolysis, wherein the protease consists of (alkaline protease + trypsin + neutral protease + flavourzyme), and the mass ratio of the protease to the trypsin is 1: 1: 2: 1) the enzymolysis reaction is carried out for 1.0h at 50 ℃, the temperature is adjusted to 95 ℃, and the temperature is kept for 15 minutes. Filtering with gauze, collecting filtrate, adjusting protein content to 2 wt%, and cooling to 0 deg.C;
(3) slaughtering tilapia, removing viscera, taking fillets, rinsing with water, draining, freezing in a refrigeration house at-40 ℃ to-18 ℃, immersing the frozen fillets in the liquid at 0 ℃ in the step (2) for 15s, and controlling the ice coat content to be about 17% to obtain the fillets plated with the ice coats;
(4) the fish fillets plated with the ice coat are filled into a polyethylene sealing bag, are packaged in vacuum, are placed in a refrigeration house at the temperature of minus 40 ℃ and are frozen until the temperature is minus 15 ℃, and are frozen in the refrigeration house at the temperature of minus 23 ℃.
Comparative example 1
Comparative example 1 is the same as the method for coating ice coating on tilapia fillets of example 1, and the difference is only that: the plating liquid is 2 wt% of tea polyphenol liquid.
Comparative example 2
Comparative example 2 is the same as the method for coating ice coating on tilapia fillets of example 1, and the difference is only that: the plating liquid is pure water.
Comparative example 3
Comparative example 3 differs from example 1 only in that: the protease consists of a single alkaline protease.
Example 2
(1) Selecting frozen grass carp scales, cleaning the scales with clean water meeting the sanitary standard of drinking water, adding water with the weight 4 times of the scales after cleaning the scales, boiling for 120 minutes at 121 ℃, filtering with a 30-mesh vibrating screen, adjusting the temperature to 85 ℃, and treating for 10 minutes by ultrasonic waves (the frequency is 80kH) by using an ultrasonic generator to obtain scale slurry;
(2) adding compound protease into the fish scale (1) according to the weight percentage of 0.1% of the weight of the fish scales for enzymolysis, wherein the protease consists of (alkaline protease + trypsin + neutral protease + flavourzyme), and the mass ratio of the protease to the trypsin is 1: 2: 1: 1) carrying out enzymolysis reaction at 55 deg.C for 0.5h, adjusting temperature to 90 deg.C, maintaining for 20min, filtering with gauze, collecting filtrate, adjusting protein content to 2 wt%, and cooling to 0 deg.C;
(3) slaughtering the grass carp, removing internal organs, taking fillets, rinsing with water, draining, freezing in a refrigeration house at the temperature of-35 ℃ to the temperature of-18 ℃, immersing the frozen fillets in the liquid at the temperature of 0 ℃ in the step (2) for 12s, and controlling the content of ice coating to be about 16% to obtain the ice-coated grass carp fillets;
(4) the grass carp fillets plated with the ice coat are filled into a polyethylene sealing bag, are packaged in vacuum, are placed in a refrigeration house with the temperature of minus 35 ℃ and are frozen to the temperature of minus 15 ℃, and are frozen in the refrigeration house with the temperature of minus 23 ℃.
Comparative example 4
Comparative example 4 is the same as the ice coating method of the grass carp fillets of example 2, except that: the plating liquid is 2 wt% of tea polyphenol liquid.
Comparative example 5
Comparative example 5 is the same as the ice coating method of the grass carp fillets of example 2, except that: the plating liquid is pure water.
Experimental example determination of quality of fillets
1. Measurement of cooking loss ratio
After the fish fillets are unfrozen, weighing fish meat with a certain weight, putting the fish meat into a fresh-keeping bag, exhausting air, sealing, boiling in a water bath kettle at 85 ℃ for 20min, taking out, cooling at room temperature for 10min, wiping the fish meat with filter paper, weighing again, and calculating the cooking loss rate.
Cooking loss (%) - (weight of fish before cooking-weight of fish after cooking)/weight of fish before cooking [. 100 ]
2. The disulfide bond content was determined by the method of Benjakul et al (Food Chemistry,2003,80(4): 535-544).
The disulfide bond content is calculated by A × D/(C × B), wherein A represents absorbance, B represents concentration of liquid protein to be measured (mg/mL), and C represents molar absorption coefficient (13600 mol)-1·cm-1L and D are dilution times, and the experiment is 1.25.
3. And (3) determination of carbonyl content:
the carbonyl content was determined by the DNPH method (Oliver, 1987). An equal volume of 10mM DNPH-HCl solution was added to 1mL of myofibrillar protein solution (2mg/mL) and reacted in the dark for 1 h. After the reaction was completed, 10% (w/v) trichloroacetic acid solution was added to terminate the reaction and centrifuged at 10,000g for 5 min. After discarding the supernatant, the unreacted DNPH in the precipitate was washed three times with ethanol/ethyl acetate (1: 1, v/v) solution. The washed precipitate was dissolved with 3mL of 6M guanidine hydrochloride and incubated in a 37 ℃ water bath for 15 min. After the solution had returned to room temperature, the absorbance of the solution was measured at 370nm (A)370). Molar extinction coefficient (22,000M) Using 2, 4-dinitrophenylhydrazone-1cm-1) The carbonyl content was calculated and expressed as nmol/mg protein.
The carbonyl content was calculated as (nmol/mg protein) ═ OD (370nm) -blank)/(22 × concentration (mg/ml)) × 125 × 10 × 5.
4. Referring to a method for measuring volatile basic nitrogen in SC/T3032-2007 standard aquatic products, a semi-micro distillation method is adopted to measure the volatile basic nitrogen. Weighing 5.00g of minced fish meat, placing in a 100mL beaker, adding 100mL of distilled water, stirring for 30min, filtering with medium-speed filter paper, and collecting the filtrate. 5mL of the filtrate was rapidly mixed with 5mL of 10g/L magnesium oxide suspension in a digestion tube, and the mixture was distilled for 5min using a Kjeldahl apparatus with a conical flask containing 10mL of 20g/L boric acid and 5-6 drops of methyl red-methylene blue mixed indicator as the receiver. The absorption solution was titrated with 0.010mol/L hydrochloric acid standard solution, and bluish purple was used as the end point of the titration. Meanwhile, a blank test was conducted by replacing the filtrate with an equal amount of distilled water.
5. The K value was determined by reference to the method of Fan et al (Food Chemistry,2008,108(1): 148-. Placing 1.00g of minced fish meat in a mortar, adding 2mL of 10% cold perchloric acid, grinding, centrifuging at 6000rpm for 5min, leaving the supernatant for later use, washing the residue with 2mL of 5% cold perchloric acid, centrifuging twice, and combining the supernatants for three times. Adjusting the pH value of the supernatant to 6.40 +/-0.05 by using 10mol/L NaOH solution, 1mol/L NaOH solution and 5% perchloric acid solution, centrifuging at 6000rpm for 5min, washing residues by using perchloric acid with pH of 6.4, combining the supernatants and fixing the volume to 10 mL. The sample solution was filtered through a 0.22 μm aqueous membrane, and then measured by HPLC using COSMOSIL 5C18-PAQ (4.6X 250mm) as a column, phosphate buffer solution of pH 6.8 as a mobile phase, 254nm as a detection wavelength, 50 μ L as a sample amount, and 1mL/min as a flow rate. And calculating the concentration of the ATP-related substance according to the standard curve and the peak area. The K value is calculated as follows:
6. Measurement of sensory evaluation
10 fillets were selected arbitrarily, the fish sensory qualities were scored by a specially trained sensory panel (15 persons) according to table 1, and finally the average was taken. The five index results are added as the final sensory evaluation result, 25 means absolute freshness, and 12 means that more obvious quality deterioration has occurred.
TABLE 1 sensory evaluation criteria for fillets
The results of comparing the main quality indexes of the differently processed fillets after being frozen for 9 months are shown in the table 2, wherein the fresh fillets refer to the fillets which are just slaughtered and not frozen, and the groups without ice coating refer to the fillets without ice coating and being frozen for 9 months.
TABLE 2 comparison of the main quality index of differently processed fillets after frozen storage for 9 months
The above results show that the frozen fresh-keeping agent for tilapia scales and grass carp scales prepared in example 1 of the present invention and tilapia fillets and grass carp fillets plated with ice coats using the frozen fresh-keeping agent are frozen and stored for 9 months, the sensory evaluation reaches 23.0 minutes, the sensory quality is maintained above 92%, the cooking loss rate is lower than 12%, the disulfide bond content is lower than 3.0mol/105g, the carbonyl content is lower than 1.5nmol/mg protein, and the volatile basic nitrogen is lower than 8mg/100g, which is significantly better than that of pure water ice coat groups and ice coat groups not plated with ice coats, and is also better than that of ice coat groups plated with 2% tea polyphenol liquid. The fish scale freezing preservative for the fish body can obviously prolong the quality of the storage period of tilapia fillets and grass carp fillets, and has obvious effects on oxidation resistance and freezing resistance.
Although the invention has been described in detail hereinabove by way of general description, specific embodiments and experiments, it will be apparent to those skilled in the art that many modifications and improvements can be made thereto based on the invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.
Claims (10)
1. A frozen fresh-keeping agent based on fish scale protein zymolyte is characterized in that the frozen fresh-keeping agent is obtained by carrying out enzymolysis on fish scales through compound protease, and the compound protease comprises alkaline protease, trypsin, neutral protease and flavourzyme.
2. A frozen fresh-keeping agent according to claim 1, characterized in that the addition amount of the compound protease is 0.05-0.5% by mass, preferably 0.1-0.3% by mass, based on the dry matter of the fish scales.
3. A frozen fresh-keeping agent according to claim 1 or 2, characterized in that in the compound protease, the mass ratio of the alkaline protease, the trypsin, the neutral protease and the flavourzyme is 0.8-1.2: 1-2: 1-2: 1.
4. a frozen fresh-keeping agent according to any one of claims 1 to 3, characterized in that the fish scales are the scales of tilapia or grass carp.
5. A method for preparing a frozen fresh-keeping agent according to any one of claims 1 to 4, characterized by comprising the steps of:
(1) mixing fish scales with water to obtain a fish scale-water mixture;
(2) sequentially carrying out heat treatment, vibration screen filtration and ultrasonic treatment on the fish scale-water mixture to obtain slurry;
(3) and carrying out enzymolysis on the slurry by using the compound protease to obtain the compound protease.
6. The preparation method according to claim 5, wherein in the step (1), the water is used in an amount of 400-500 wt% based on the mass of the dry matter of the fish scales;
and/or in the step (2), the temperature of the heat treatment is 115-121 ℃;
and/or in the step (2), the mesh number of the vibrating screen is 30-60 meshes;
and/or in the step (2), the frequency of ultrasonic treatment is 80-100 Kh, the time of ultrasonic treatment is 10-15 min, and the temperature of ultrasonic treatment is 75-85 ℃.
7. The method according to claim 5 or 6, wherein in the step (3), the enzymolysis comprises the following steps:
adding the compound protease into the slurry, performing enzymolysis for 0.5-1.0 h at 45-55 ℃, and then performing heat preservation for 15-20 min at 90-95 ℃.
8. The preparation method according to any one of claims 5 to 7, wherein in the step (3), after the enzymolysis, the method further comprises the steps of filtering and adjusting the obtained filtrate into a suspension with the protein content of 1.5-2 wt%.
9. The application of the frozen fresh-keeping agent according to any one of claims 1 to 4 or the frozen fresh-keeping agent prepared by the preparation method according to any one of claims 5 to 8 in frozen ice-coated storage and fresh keeping of fish meat.
10. The application of the fish meat preservative as claimed in claim 9, wherein the fish meat is frozen at the temperature of-18 to-23 ℃ in an environment of-35 to-40 ℃, then the frozen fish meat is immersed in the frozen preservative for 10 to 15s, the content of ice coating is controlled to be 15 to 20%, and the fish meat plated with the ice coating is obtained and frozen after being vacuum-packed.
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Cited By (1)
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| CN113249424A (en) * | 2021-06-18 | 2021-08-13 | 海南热带海洋学院 | Hydrolysate with anti-freezing activity prepared from carp scales, anti-freezing agent and application of hydrolysate |
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Application publication date: 20201117 |