LU501124B1 - Recombinant Minced Fillet Product With High Gelation Properties and Preparation Method Thereof - Google Patents

Recombinant Minced Fillet Product With High Gelation Properties and Preparation Method Thereof Download PDF

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LU501124B1
LU501124B1 LU501124A LU501124A LU501124B1 LU 501124 B1 LU501124 B1 LU 501124B1 LU 501124 A LU501124 A LU 501124A LU 501124 A LU501124 A LU 501124A LU 501124 B1 LU501124 B1 LU 501124B1
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minced fillet
recombinant
fish oil
vitamin
ethyl ester
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LU501124A
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German (de)
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Lu Zhai
Xiaoxia Liu
Renyao Jin
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Univ Zhejiang Gongshang
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    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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    • A23B4/00—General methods for preserving meat, sausages, fish or fish products
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
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    • A23L29/045—Organic compounds containing nitrogen as heteroatom
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
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    • A23L3/349—Organic compounds containing oxygen with singly-bound oxygen
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • 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
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Abstract

The present invention provides a recombinant minced fillet product with a high gelation property and a preparation method thereof. The recombinant minced fillet product is processed by frozen minced fillet, salt, fish oil, vitamin E, casein peptide. The minced fillet product prepared by the method of the present invention has high gel strength, high water holding capacity and good flavor; and the EPA and DHA contained therein have high content and are long-term stable.

Description

Recombinant Minced Fillet Product with High Gelation Properties and Preparation Method Thereof Technical Field The present invention belongs to the field of aquatic food processing, and particularly relates to a recombinant minced fillet product with a high gelation property and a preparation method thereof.
Background Minced fillet is a kind of concentrated myofibrillar protein produced by performing meat separation, rinsing, dewatering, refined filtration and other manufacturing procedures on a fish body.
To improve the storage stability and quality properties of frozen minced fillet, a large number of DHA, EPA and other functional lipid substances in fish body are removed in rinsing process, leading to a massive loss of functionality in the produced minced fillet products.
Further, lipids are essential to maintain the texture and rheological properties of meat paste products, produce a peculiar flavor and to increase high nutritive value.
The lack of lipids in meat paste will produce an uncomfortable sense of touch just like in contact with rubber.
Therefore, exogenous fat/oil is always used as a raw material to be backfilled into minced fillet, thus improving the quality of the minced fillet product.
T minced fillet products have always been an important protein supplement for the elderly due to their high protein and elasticity.
Its research and development have also been developed from the nutritional properties of protein, and the development of increasing functional lipids in the minced fillet products has been slow.
Further, vegetable oil, fat and the like are usually added to the conventional minced fillet product to enhance the gel strength of the minced fillet product.
In recent years, the latest developments in health awareness have increased the demand for functional seafood.
Major functional ingredients in w-3 polyunsaturated fatty acid are Eicosapentaenoic Acid (abbreviated for EPA) and Docosahexaenoic Acid (abbreviated for DHA). EPA is known as the "blood vessel scavenger" and has a degrading effect on cholesterol and blood lipids, and can reduce the prevalence of diseases such as Alzheimer's and arteriosclerosis.
DHA not only plays an indispensable role in human health, but also plays an important role in the development of the human brain and visual system, and enjoys the reputation of "brain gold". w-3 polyunsaturated fatty acids (EPA and DHA) in natural conditions mainly exist in deep sea fish oil in a form of triglyceride, but its content is only 30% around and thus cannot satisfy consumer's demands.
Meanwhile, EPA and DHA contain unsaturated bonds and thus, are very unstable and easily decomposed.
Therefore, it is difficult to achieve functional requirements of the minced fillet product.
The existing recombinant minced fillet products usually have the problems such as, low gel strength, low water holding capacity, poor gel texture properties and poor flavor.
Moreover, EPA and DHA added in minced fillet gel are easily decomposed and thus, hardly accepted by consumers.
Therefore, there is an urgent need to find a recombinant minced fillet product with high gel strength, high water holding capacity, good flavor, and long-term stability and rich in EPA and DHA.
Summary To solve the above problems, the present invention provides a recombinant minced fillet product with a high gelation property.
By adding refined ethyl ester-typed fish oil to frozen minced fillet, and adding vitamin E and casein peptides at the same time, the prepared minced fillet gel has high gel strength, water retention, good flavor, and is rich in EPA and DHA, and the content of EPA and DHA is stable for a long time.
On the one hand, the present invention provides a recombinant minced fillet product, including minced fillet and ethyl ester-typed fish oil.
The preparation method of the ethyl ester-typed fish oil of the present invention includes: firstly refining crude oil, including degumming, deacidification, decolorization, and deodorization; and then transesterifying the refined fish oil with ethanol to obtain the ethyl ester-typed fish oil.
In some examples, the specific steps of the method for preparing ethyl ester-typed fish oil are: 100 g of fish oil is mixed with 30 g of absolute ethanol, 0.6 g of NaOH is added as a catalyst, and the final yield of ethyl ester-typed fish oil is 92%. Further, the ethyl ester-typed fish oil used in the present invention is extracted from marine fish waste.
In some examples, the minced fillet is FF-grade frozen minced fillet.
The inventor has proved through a large number of studies that adding ethyl ester-typed fish oil to frozen minced fillet can not only improve the gelation properties of minced fillet products, but also significantly improve the texture characteristics of minced fillet gel without affecting the activity of endogenous TG enzymes. The activity of the enzyme will not affect the content of myosin heavy chain (MHC), actin and tropomyosin, so that the prepared minced fillet gel has a better flavor.
The ethyl ester-typed fish oil needs to be refined and purified, and the EPA and DHA in the ethyl ester-typed fish oil can be further enriched through the refining and purification process, thereby preparing the minced fillet gel rich in EPA and DHA.
Further, the recombinant minced fillet product also includes vitamin E and casein peptide. Vitamin E is a fat-soluble vitamin and an antioxidant.
Casein peptides have strong antioxidant capacity; and casein peptides have low molecular weight and are easy to absorb.
Studies have shown that the further addition of vitamin E and casein peptides to recombinant minced fillet products can not only make the prepared minced fillet gel more stable, maintain better texture characteristics and water retention, but also keep the high content of EPA and DHA in the minced fillet gel stable for a long time, not easy to decompose, and make the minced fillet gel have a longer shelf life.
Further, the ethyl ester-typed fish oil is 0.8 wt%-1.6 wt% of the minced fillet, and the EPA content in the ethyl ester-typed fish oil is 14-15%.
Further, the vitamin E is 0.5 wt%-1 wt% of the minced fillet, and the casein peptide is 0.5 wt%-1 wt% of the minced fillet. Further, salt is also included, and the salt is 2.0 wt% of the minced fillet.
On the other hand, the present invention provides a preparation method of a recombinant minced fillet product, mainly including adding ethyl ester-typed fish oil, vitamin E and casein peptide to frozen minced fillet.
Further, the ethyl ester-typed fish oil is 0.8 wt%-1.6 wt% of the minced fillet, the vitamin E is 0.5 wt%-1 wt% of the minced fillet, and the casein peptide is 0.5 wt%-1 wt% of the minced fillet.
Further, the minced fillet is FF-grade frozen minced fillet.
Further, the preparation method of a recombinant minced fillet product includes the following steps: (1). taking frozen minced fillet, performing semi-thawing and cutting into pieces, and air chopping for 3-5 min in a food conditioning machine with a revolving speed of 3000 r/min; (2). adding 2.0 wt% salt of the frozen minced fillet, and performing salt chopping for 3-5 min with a revolving speed of 4500 r/min; (3). adding 0.8 wt%-1.6 wt% ethyl ester-typed fish oil of the frozen minced fillet, 0.5 wt%-1 wt% vitamin E and 0.5 wt%-1 wt% casein peptide and performing continuous chopping and blending for 3-5 min with a revolving speed of 4500 r/min; (4). gelating the minced fillet sol obtained in the step (3) at 45°C for 30-60 min, and performing heating and curing for 20-30 min at 90°C; and (5). immediately placing the minced fillet gel obtained in the step (4) to ice water and cooling to a center temperature < 10°C, thus obtaining a recombinant minced fillet product.
Further, the semi-thawing is thawing at 0-10°C to a central temperature of -4~0°C.
Further, during the chopping process, the temperature of the material should be kept <10°C.
In another aspect, the present invention provides the use of ethyl ester-typed fish oil for preparing the above-mentioned recombinant minced fillet product, and the ethyl ester-typed fish oil can improve the gelation characteristics and functional properties of the minced fillet product.
In another aspect, the present invention provides the use of a mixture for the preparation of the recombinant minced fillet product as mentioned above, the mixture includes vitamin E and casein peptide, and the vitamin E and the casein peptide can improve the stability of EPA and DHA in the recombinant minced fillet product.
The beneficial effects of the present invention are as follows: (1) the method of adding ethyl ester-typed fish oil is used to enhance the strength of minced fillet gel and improve the texture characteristics, improve the taste, and enhance the functional characteristics of the minced fillet products;
(2) the present invention renders the minced fillet gel more stable by adding vitamin E and casein peptide, thus keeping better texture properties and water holding capacity.
Meanwhile, the present invention can further keep the high-content EPA and DHA in the minced fillet gel stable for a long time and to be not decomposed easily, thus achieving a 5 longer shelf life of the minced fillet gel.
Brief Description of the Drawings FIG. 1 is a schematic diagram of the results of SDS-PAGE analysis of recombinant minced fillet product provided by Example 5 of the present invention.
Detailed Description of the Embodiments The present invention will be further described in detail below in conjunction with the embodiments.
It should be pointed out that the following embodiments are intended to facilitate the understanding of the present invention and do not have any limiting effect on it.
The reagents used in this embodiment are all known products, which are obtained by purchasing commercially available products.
Example 1 Influences of Different Kinds of Fish Oil on Prepared Recombinant Minced Fillet Products The preparation method of the recombinant minced fillet product provided in this example is as follows: 1). frozen minced fillet was taken and semi-thawed to a center temperature of -4 to OC, and then cut into pieces, and air chopped for 5 min in a food conditioning machine with a revolving speed of 1500 r/min; 2). 2.0 wt% salt of the frozen minced fillet was added for salt chopping for 5 min with a revolving speed of 2000 r/min; 3). 1.0 wt% fish oil of the frozen minced fillet, 0.5 wt% vitamin E and 0.5 wt% casein peptide were then added, and continuously chopped and blended for 3 min with a revolving speed of 2000 r/min; 4). the minced fillet sol obtained in the step 3) was gelated at 45°C for 30 min, and heated and cured for 20 min at 90°C; and 5). the minced fillet gel obtained in the step 4) was immediately placed to ice water and cooled to a center temperature<10°C, thus obtaining a recombinant minced fillet product.
The material temperature need to be kept <10C during the chopping and blending process, and after being cooled, the minced fillet gel was determined directly.
This experiment was divided into four groups.
For the fish oil in the step 3), ethyl ester-typed fish oil (Zhejiang Xingye Food Co., Ltd.) is used in the first group: triglyceride-typed fish oil (Zhejiang Xingye Food Co., Ltd.) is used in the second group; phospholipid-typed fish oil (Zhejiang Xingye Food Co., Ltd.) is used in the third group, and the fourth group is used as a blank control.
The added casein peptides are added in the form of casein peptide powder.
The preparation method of casein peptide powder: prepare the casein powder into protein dispersion with purified water, adjust the pH of the protein dispersion to between 7-9; heat and keep the temperature at 40°C, and then add neutral protease to the protein dispersion, stir it evenly and enzymatically hydrolyze for 2 h.
The amount of the added neutral protease is 0.2% of the substrate content.
After enzymatic hydrolysis is completed, perform enzyme inactivation at high temperature; centrifuge the enzymatic hydrolysate after enzyme inactivation to remove impurities, keep the centrifugal speed of 12000 r/min, the feeding speed of the peristaltic pump is 1.5 L/min, collect the clear liquid, start the membrane filtration equipment, and pass the clear liquid through the microporous membrane filter, the thickness of the filter membrane is 90 um, the particle size of the filter is 4um, the operating pressure is 0.01 MPa, thus to ensure that the filtrate is clear and transparent, and the filter residue is removed; the filtrate after centrifugal filtration is passed through an ion exchange column and an activated carbon column for desalination and decolorization; the exchanged solution is concentrated in vacuum in sequence, at a concentration temperature of 65°C and a vacuum degree of -0.05 Mpa, to obtain a peptide concentrate, with a relative density controlled at 1.03; and then spray-dried to obtain casein peptide powder.
The four groups of prepared recombinant minced fillet products were subjected to detection and analysis, and the influences of adding different types of fish oil on the hardness, elasticity, cohesiveness, tackiness, chewiness, gel strength, water holding capacity, hydrophobic bond, disulfide bond, and EPA and DHA contents of the prepared recombinant minced fillet products are investigated separately.
Wherein, the testing methods of hardness, elasticity, cohesiveness, tackiness and chewiness are all tested by texture analyzer, and the parameter settings are consistent with the gel strength. The principle is detailed as follows: this test uses the TMS-Pro texture analyzer produced by FTC in the United States, and the measurement method uses texture profile analysis (TPA). Sample processing: cut the sample into a cylinder with a length of 20 mm and a diameter of 45 mm. The texture parameters used in the TPA test are defined as: Hardness refers to the peak pressure when the sample is compressed for the first time; Springiness is the quotient of the measurement time of the second compression and the time of the first measurement; Cohesiveness is the quotient of the working area of the second compression divided by the working area of the first time. Chewiness is used for solid products, and the calculation formula is HardnessxSpringinessxCohesiveness. The gel strength test method is: cut the sample to be tested into a cylinder with a thickness of 20 mm, equilibrate for 30 minutes at room temperature, then measure the texture characteristics by TPA method, and use a P/5S spherical probe. Calculate the gel strength according to the formula: gel strength (g.cm) = rupture strength (g) x depression depth (cm). The method for detecting water holding capacity is as follows: accurately weigh 3.0 g (W1) minced fillet sample, wrap it with 3 layers of filter paper, centrifuge at 5000 g in a 50 ml centrifuge tube for 15 min, centrifuge at 4°C, and weigh again (W2). Calculate water holding capacity (WHC) according to the formula: WHC(%)=W2/W1x100%. The detection method of hydrophobic bond and disulfide bond is as follows: accurately weigh 2.0 g minced fillet sample and mix with 10 ml different chemical force destroying reagents to homogenize, stir at 4°C for 60 min, centrifuge, and determine the protein concentration in the supernatant by bradford method. Chemical destruction reagents are:
0.05 M NaCl (SA), 0.6 M NaCl (SB), 0.6 M NaCl+1.5 M urea (SC), 0.6 M NaCI+8 M urea (SD), 0.6 M NaCl+8 M urea+0.05 M B-mercapto ethanol (SE). The chemical force is expressed by the difference in supernatant protein concentration between groups: ionic bond is the difference between SB and SA, hydrogen bond is the difference between SC and SB, hydrophobic force is the difference between SD and SC, and disulfide bond is the difference between SE and SD. The detection method of EPA and DHA is as follows: accurately weigh 2.0 g minced fillet gel sample, add 12.0 mL lipid extract (dichloromethane: anhydrous methanol=2:1), ultrasonically extract for 20 min, add 5.0 mL pure water, centrifuge and take the lower layer of nitrogen blowing to obtain crude oil. Methyl esterification method: add 5 mL 0.5 M KOH-methanol solution to the crude oil, water bath at 65°C for 30 min, shake until the oil droplets disappear and cool to room temperature. Add 2.0 mL of 14% boron trifluoride methanol solution, and after 5 minutes of water bath at 65°C, ultrasonic extraction is performed for 10 minutes. Accurately add 2.0 mL of n-hexane, shake and rinse with 2.0 mL of saturated NaCl, centrifuge and take the upper layer to dehydrate with anhydrous sodium sulfate, and filter with 0.22 um for testing. Detected by gas chromatography, gas chromatographic conditions: HP-INNOWAX capillary column (30 mX0.25 mm, 0.15 pm); heating program: initial temperature 50°C for 2 min, then 4°C/min to 250°C, keep for 15 min; the temperature of the injection port is 250°C, the split ratio is 40:1, the injection volume is 1 pL, and the carrier gas flow rate is 0.65 mL/min. The results are shown in Table 1: Table 1. Influences of different kinds of fish oil on the prepared recombinant minced fillet products Fourt First h group Second group Third group group Test (Ethyl (Triglyceride-typ (Phospholipid-typ (Blan item\Group ester-typ ed fish oil) ed fish oil) k ed fish contro oil) 1) Elasticity
11.89 10.38 10.83 12.06 mm Cohesivene
0.39 0.37 0.34 0.33 ss
U501124 | me | [me |”
502.91 389.64 372.99 g/cm 0 Water holding 82.35 77.97 77.26 78.14 capacity %
FAIRE
9.38 6.58 5.90 5.53 c bond g/L om [om [on [em [ee
0.62 0.39 0.39 0.36 bond g/L It can be seen from Table 1 that compared with the blank control in the fourth group, the ethyl ester-typed fish oil is added to significantly improve the hardness, elasticity, cohesiveness, chewiness of the recombinant minced fillet products prepared and other texture properties of the gel, and further promote the gel strength and water holding capacity as well as chemical action force of the gel, and to bring rich EPA, DHA and other functional nutrient substances for the minced fillet. During the formation of minced fillet gel, with the significant decrease of hydrogen bonds, the hydrophobic force is the main force for the minced fillet gel structure, and the hydrophobic bonds and disulfide bonds are the main force for maintaining the minced fillet gel network structure. lonic bonds are usually formed between the oppositely charged amino acid residues. The addition of ethyl ester-typed fish oil provides a hydrophobic environment, which leads to the unfolding of protein molecules and exposes the buried amino acid residues, thereby increasing protein structure and protein electrostatic interaction. At the same time, the addition of ethyl ester-typed fish oil causes increased exposure of the hydrophobic side chains of minced fillet protein and a significant increase in hydrophobic interactions. The possible reason for the increase in disulfide bonds is that the hydrophobic environment provided by the ethyl ester-typed fish oil exposes the sulfhydryl groups inside the protein, which is beneficial to the formation of disulfide bond crosslinks.
Compared with the triglyceride-typed fish oil in the second group and the phospholipid-typed fish oil in the third group, the ethyl ester-typed fish oil is added to further improve the texture properties of gel, bringing better taste and flavor for the minced fillet gel. Meanwhile, after the addition of the triglyceride-typed fish oil, the prepared minced fillet gel has significantly higher EPA and DHA content.
Therefore, when preparing recombinant minced fillet products with high gel properties, it is most preferable to add ethyl ester-typed fish oil.
Example 2 Influences of Vitamin E and Casein Peptides on Prepared Recombinant Minced Fillet Products The preparation method of the recombinant minced fillet product provided in this example is as follows: 1). frozen minced fillet was taken and semi-thawed to a center temperature of -4 to OC, and then cut into pieces, and air chopped for 5 min in a food conditioning machine with a revolving speed of 1500 r/min; 2). 2.0 wt% salt of the frozen minced fillet was added for salt chopping for 5 min with a revolving speed of 2000 r/min; 3). 1.0 wt% fish oil of the frozen minced fillet, 0.5 wt% vitamin E or 0.5 wt% casein peptide were then added, and continuously chopped and blended for 3 min with a revolving speed of 2000 r/min; 4). the minced fillet sol obtained in the step 3) was gelated at 45°C for 30 min, and heated and cured for 20 min at 90°C; and 5). the minced fillet gel obtained in the step 4) was immediately placed to ice water and cooled to a center temperature<10°C, thus obtaining a recombinant minced fillet product. The material temperature need to be kept <10°C during the chopping and blending process, and after being cooled, the minced fillet gel was determined directly.
This experiment was divided into six groups; in the step 3), no addition of vitamin E or casein peptide was taken in the first group; the addition of vitamin E was taken in the second group; the addition of casein peptide was taken in the third group; the simultaneous addition of vitamin E and casein peptide was taken in the fourth group; vitamin C was used in the fifth group as an antioxidant; and the simultaneous addition of vitamin C and casein peptide was taken in the sixth group.
The six groups of the prepared minced fillet recombinant products were subjected to detection and analysis, and the influences on the hardness, elasticity, cohesiveness, tackiness, chewiness, gel strength, water holding capacity, hydrophobic bond, disulfide bond, EPA and DHA contents, and stability of EPA, DHA contents of the prepared recombinant minced fillet products are investigated separately.
The test method is shown in Example 1, where the stability of EPA and DHA contents is tested by taking the minced fillet recombinant products, adding 0.05% potassium sorbate, and placing it at high temperature of 37°C for 3 days, and then testing the EPA and DHA contents in it.
The results are shown in Table 2: Table 2. Influences of vitamin E and casein peptide on the prepared minced fillet recombinant products Fourt Sixth h First Third group Secon group grou group Fifth (Vitam d (Vitam Test p (Case group in C + group in E + item\Group (Not in (Vitam casei (Vitam casei adde peptid in C) n in C) n d) e) peptid peptid e) e) Hardness
9.58 9.62 9.65 9.77 9.57 9.64
N Elasticity 10.6
10.70 11.26 12.78 10.64 10.86 mm 4
37.3 Chewiness 38.37 38.29 41.64 38.28 38.99 Gel
488. 483.7 494 1 537.7 483.9 483.9 strength 93 4 9 1 5 8 g/cm Water
81.3 holding 81.36 82.29 84.46 81.36 81.40 2 capacity % Hydrophobi
8.44 8.98 8.94 10.87 8.98 c bond g/L Disulfide
0.49 0.50 53.82 0.53 0.50 0.51 bond g/L
15.2 DHA% 15.27 15.27 15.81 15.27 15.27 7 EPA% after
0.89 3.27 1.32 5.43 1.58 2.35 3d DHA%
1.57 8.69 3.78 18.71 4.01 4.72 after 3 d It can be seen from Table 2 that vitamin E and casein peptide have certain influences on the gel texture properties of the prepared recombinant minced fillet products, and have significant effects on the improvement of EPA and DHA content stability. After vitamin E 5 and casein peptide are added in the fourth group, the content of EPA and DHA does not change after the prepared recombinant minced fillet product is put for 3 d at a high temperature of 37°C. When there is no addition, or only vitamin E or casein peptide is added in other groups, the content of EPA and DHA significantly decreases. As can be seen, the simultaneous addition of vitamin E and casein peptide may significantly promote the stability of the content of EPA and DHA, such that the prepared recombinant minced fillet product is more stable. When vitamin C is respectively added or vitamin C and casein peptide are simultaneously added to the fifth group and the sixth group, there is no significant promotion effect on the stability of the content of EPA and DHA, after being put at a high temperature of 37°C for 3 d, there is a certain decline in the content of EPA and DHA. Therefore, the addition of both vitamin E and casein peptides to recombinant minced fillet products can significantly improve the stability of EPA and DHA contents and further improve the gel texture properties of the recombinant minced fillet products. Example 3 Influences of Content of Ethyl Ester-Typed Fish Oil on Prepared Recombinant Minced Fillet Products The method provided in Example 2 was taken to prepare a recombinant minced fillet product in this example, where, the ethyl ester-typed fish oil had a content of 0.5%, 0.8%, 1%, 1.2%, 1.6%, and 1.8% respectively, six groups in total; 0.5% vitamin C and casein peptide were simultaneously added for each group. The influences on the hardness, elasticity, cohesiveness, tackiness, chewiness, gel strength, water holding capacity, hydrophobic bond, disulfide bond, and EPA and DHA contents of the prepared recombinant minced fillet products are investigated. The test method is as described in Example 1, and the test results are shown in Table 3. Table 3. Influences of the content of ethyl ester-typed fish oil on the prepared minced fillet recombinant products Fourt First Secon Fifth Sixth Third h Test group d group group group group item\Group (0.5% group (1.6% (1.8% (1%) (1.2% ) (0.8%) ) ) ) men | am | sm | am | am | aw | aw Elasticity
11.81 11.72 11.81 12.74 11.67 11.54 mm
Gel strength 463.2 482.7 500.4 502.2 500.6 496.2 g/cm 7 1 9 9 0 8 Water holding 79.37 79.92 82.29 82.80 82.93 82.58 capacity % oo] on] vem] em] en] um
6.59 7.60 9.08 9.35 9.31 9.32 bond g/L
EEE EE
0.33 0.39 0.58 0.61 0.52 0.44 bond g/L It can be seen from Table 3 that with the increase of ethyl ester-typed fish oil in the addition amount, the hardness, elasticity, cohesiveness, tackiness and chewiness of the minced fillet increase accordingly, being up to the maximum values (P<0.05) when the addition amount is 1.2%. Afterwards, with the further increase of ethyl ester-typed fish oil in the addition amount, there is no obvious change in hardness, elasticity, cohesiveness, tackiness and chewiness of the minced fillet. lonic bond and disulfide bond show a trend of first rising then descending, and being up to the maximum values when the addition amount of the fish oil is 1.2%.
With the increase in the amount of ethyl ester-typed fish oil added, the hydrophobic interaction rises sharply (P<0.05) with the increase of the fish oil in the addition amount; and the hydrogen bond shows a significant decline trend. In overall consideration, the addition amount of the ethyl ester-typed fish oil should be within a range of 0.8%-1.6%, most preferably, 1.2%.
Example 4 Influences of Contents of Vitamin E and Casein Peptide on Prepared Recombinant Minced Fillet Products
The method provided in Example 2 was taken to prepare a recombinant minced fillet product in this example, where, the ethyl ester-typed fish oil had a content of 1.2% respectively, four groups in total; different amount of vitamin E and casein peptide were added for each group. The contents of vitamin E and casein peptides in each group are: the first group contained 0.1% vitamin E and 0.1% casein peptide; the second group contained 0.5% vitamin E and 0.5% casein peptide; the third group contained 1% vitamin E and 1% casein peptide; and the fourth group contained 1.5% vitamin E and 1.5% casein peptide. The influences on the hardness, elasticity, cohesiveness, tackiness, chewiness, gel strength, water holding capacity, hydrophobic bond, disulfide bond, EPA and DHA contents, and stability of EPA, DHA contents of the prepared recombinant minced fillet products and effects of lowering blood lipids after feeding mice are investigated separately. The test method is shown in Example 2, and the results are shown in Table 4. Table 4. Influences of the contents of vitamin E and casein peptide on the prepared minced fillet recombinant products Second Fourth First group Third group group group (0.1% (1.0% (0.5% (1.5% Test Vitamin E Vitamin E Vitamin E Vitamin E item\Group +0.1% +1.0% +0.5% +1.5% casein casein casein casein peptide) peptide) peptide) peptide) Elasticity mm 11.63 13.87 13.77 12.98
39.37 42.21 42.22 41.77 Gel strength 539.71 538.59
527.02 537.05 g/cm Water holding 83.95 85.01 85.07 84.39
U501124 Hydrophobic 11.37 11.23
10.53 11.07 bond g/L Disulfide 0.81 0.82
0.79 bond g/L DHA% 18.75 18.77 18.78 18.76 EPA% after 3
1.66 4.48 4.49 4.41 d DHA% after 3
3.08 15.27 15.30 15.11 d It can be seen from Table 4 that when the addition amount of vitamin E and casein peptide is within the range of 0.5%-1%, the stability of the content of EPA and DHA may be improved obviously, such that the prepared recombinant minced fillet product has very good lipid-lowering effect, and the gel texture properties of the recombinant minced fillet product and the taste may be further improved; when the addition amount of vitamin E and casein peptide is 0.1%, the effect of improving the stability of the content of EPA and DHA decreases obviously; when the addition amount of vitamin E and casein peptide is
1.5%, the effect of improving the stability of the content of EPA and DHA shows a decline trend compared with 1%. Moreover, there is no significant effect in the aspect of improving gel texture properties of the recombinant minced fillet product. Therefore, the addition amount of vitamin E and casein peptide should be within the range of 0.5%-1%, preferably, the addition amount of both vitamin E and casein peptide is 0.5%. Example 5 SDS-PAGE Analysis of Recombinant Minced Fillet Products In the minced fillet, myosin heavy chain (MHC) is the main protein, which plays a decisive role in the formation of minced fillet gel, followed by actin and tropomyosin. The endogenous TG enzyme can catalyze the formation of myosin heavy chain e-(y-glutamyl) lysine non-disulfide covalent bond, which will not be destroyed by SDS and DTT in the electrophoresis system. Therefore, the effect of ethyl ester-typed fish oil on the catalysis of endogenous TG enzymes can be observed by gel electrophoresis. In this example, the recombinant minced fillet product prepared in the second group of Example 4 is used for SDS-PAGE analysis, and compared with the control group without adding ethyl ester-typed fish oil. The result is shown in FIG. 1.
It can be seen from FIG. 1 that compared with the control group without addition of ethyl ester-typed fish oil, the color of myosin heavy chain bands in the minced fillet gel with fish oil added has no significant change, indicating that the ethyl ester fish oil does not affect endogenous TG enzymes. The activity of the minced fillet gel mainly affects the strength of the minced fillet gel through chemical forces, which can make the minced fillet gel have a better taste. Example 6 Identification of Volatile Flavor Substances In this example, the recombinant minced fillet product prepared in the second group of Example 4 was used for identification of volatile flavor substances, and the standard library was searched and matched, and the relative percentage content was calculated by the area normalization method. It is identified the volatile flavor components and sensory thresholds of recombinant minced fillet products added with ethyl ester fish oil, vitamin E and casein peptides. The addition amount of ethyl ester fish oil is 0%, 0.4%, 0.8%, and
1.6% respectively. See Table 5 for details. 57, 65, 63, 66 and 70 volatile components were detected in minced fillet gels with different ethyl ester fish oil additions, mainly aldehydes, ketones, alcohols, hydrocarbons and lipids. According to the sensory threshold, the relative odor activity value (ROAV) value is calculated to analyze the volatile flavor substances. In the group without addition of ethyl ester fish oil, although the relative content of decanal is not large, it contributes the most to the overall flavor of minced fillet according to its ultra-low sensory threshold. Therefore, the ROAV of decanal is defined as 100, calculating ROAV of other volatile flavor substances, and the results are shown in Table 5. Similarly, among the minced fillet added with ethyl ester fish oil, ethyl caproate contributes the most to the overall flavor of minced fillet. The ROAV value is defined as 100, and the ROAV of other volatile flavor substances is obtained.
There are 10, 13, 12, 13 and 14 key flavor substances for the 5 graded ethyl ester fish oil additions, including nonanal, n-octanal, hexanal, benzaldehyde, heptaldehyde, 2-ethylfuran and ( Z)-4-heptenal is a common key flavor substance; there are 8, 2, 2, 2, and 3 important flavor components that affect the flavor of minced fillet, among which 2-nonanone is common. Comparative analysis is available without adding ethyl ester fish oil group and adding ethyl ester fish oil group. First, the relative content of esters in the ethyl ester fish oil group is significantly higher than that of the non-adding group. This may be due to the flavor composition of the ethyl fish oil itself. . Secondly, compared with the group without fish oil, the relative content of hexanal, heptanal and 1-octen-3-ol increased significantly. Low-molecular-weight aldehydes, alcohols, ketones, and a small amount of furan, naphthalene and other substances are the main components of the fishy odor of aquatic products, which shows that the fishy odor of the minced fillet gel added with ethyl ester fish oil is aggravated. By sensory smell, when the addition amount is
1.2%, the fishy smell is very insignificant, and the flavor is better. Table 5 Relative content and sensory threshold of volatile flavor components in minced fillet gel Compound Relative content /% Sensory Sensory description 0% 04% 08% 12% 16% threshold (ug/kg) Acetaldehyde 0.03 0.09 009 003 0.03 15 3-Methylbutanal 025 0.04 0.03 0.2 Almond, nutty aroma Hexanal 2.09 047 0.38 0.38 0.33 4.5 Fishy, fruity, leafy aroma Enanthal 0.83 0.16 0.13 0.13 0.11 3 Fishy, nutty aroma (Z)-4-heptenal 088 0.36 0.30 0.33 0.29 10 Vegetable, linseed oil-like Octanal 1.12 0.20 0.19 0.15 0.13 0.5 Fat, orange, honey aroma Benzaldehyde 0.98 0.35 0.36 0.34 0.29 3 Almond, nutty aroma Nonanal 4.32 0.37 0.29 0.20 0.17 1 Fatty, grassy aroma Decanal 0.83 0.1 Sweet, waxy, floral aroma Myristaldehyde 0.38 14 Hexadecaldehyde 071 081 054 042 Glutaraldehyde 003 0.05 0.03 Propionaldehyde 0.05 9.5 Nutty smell 1-heptanol 0.07 3 1-octene-3-ol 1.71 0.22 0.15 0.14 1 Fishy, mushroom aroma, oily 2-ethyl-1-hexanol 0.19 270000
Octanol 0.18 110 Earthy, metallic smell Phenylethanol 0.14 0.03 750 1-penten-3-ol 0.07 0.05 0.07 0.07 400 Fruit, vegetable aroma 3-octanone 0.25 28 Ether smell 2-nonanone 0.37 0.11 0.12 0.08 0.08 38.9 Fruity, sweet aroma 2-undecone 049 0.14 0.17 0.10 7 Fruity, grassy aroma 2-octanone 0.03 0.02 50 Pelargonic acid 0.16 3000 n-hexadecanoic acid 3.74 10000 Acetic acid 0.05 22000 Sour smell Myristic acid 0.27 0.10 10000 3-methyl-1,4-heptadiene 0.32 0.26 020 022 0.19 Ethylbenzene 0.36 29 Aroma Scent O-xylene 0.57 68.6 Naphthalene 026 010 014 0.16 0.12 21 Ethyl butyrate 0.10 0.10 0.10 4.5 Ethyl valerate 0.23 0.20 1.5 Fruity, fruity ester aroma Ethyl caproate 0.52 0.47 0.47 0.31 1 Ethyl hexadecanoate 16.36 1668 17.29 18.19 Ethyl oleate 10.20 1061 11.96 12.85 Ethyl stearate 6.89 7.22 8.59 9.03 2-ethylfuran 0.21 0.14 0.08 0.12 0.10 2.3 Bean, malt aroma Methoxyphenyl-oxime 1.48 0.64 0.24 0.29 0.25
Although the present invention is disclosed as above, the present invention is not limited to this.
Any person skilled in the art can make various changes and modifications without departing from the spirit and scope of the present invention.
Therefore, the protection scope of the present invention should be subject to the scope defined by the claims.

Claims (2)

Claims
1. A recombinant minced fillet product, comprising minced fillet, ethyl ester-typed fish oil, vitamin E, casein peptide and salt; wherein the ethyl ester-typed fish oil is 0.8 wt%-1.6 wt% of the minced fillet, the vitamin E is 0.5 wt%-1 wt% of the minced fillet, the casein peptide is 0.5 wt%-1 wt% of the minced fillet, and the salt is 2.0 wt% of the minced fillet.
2. À preparation method of a recombinant minced fillet product, comprising the following steps: a. taking frozen minced fillet, performing semi-thawing and cutting into pieces, and air chopping for 3-5 min in a food conditioning machine with a revolving speed of 1500-3000 r/min; b. adding 2.0 wt% salt of the frozen minced fillet, and performing salt chopping for 3-5 min with a revolving speed of 2000-4500 r/min; c. adding 0.8 wt%-1.6 wt% ethyl ester-typed fish oil of the frozen minced fillet, 0.5 wt%-1 wt% vitamin E and 0.5 wt%-1 wt% casein peptide and performing continuous chopping and blending for 3-5 min with a revolving speed of 2000-4500 r/min; d. gelating the minced fillet sol obtained in the step c at 45°C for 30-60 min, and performing heating and curing for 20-30 min at 90°C; and e. immediately placing the minced fillet jelly obtained in the step d to ice water and cooling to a center temperature < 10°C, thus obtaining a recombinant minced fillet product.
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EP0096902B1 (en) * 1982-06-16 1988-09-28 Taiyo Fishery Co., Ltd. Method for the production of protein food products or protein food materials in paste state and method for the production of food products from these materials
US7108873B2 (en) * 1994-04-19 2006-09-19 Applied Elastomerics, Inc. Gelatinous food elastomer compositions and articles
US5456938A (en) * 1993-01-27 1995-10-10 Penwest Foods Co. Cryoprotected surimi product
US7410663B2 (en) * 2002-08-07 2008-08-12 Kao Corporation Oil or fat composition
JP4316943B2 (en) * 2003-06-27 2009-08-19 日油株式会社 Gel food composition
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