CN111742979A - Fugu rubripes biological preservative and preparation method and application thereof - Google Patents
Fugu rubripes biological preservative and preparation method and application thereof Download PDFInfo
- Publication number
- CN111742979A CN111742979A CN202010658784.2A CN202010658784A CN111742979A CN 111742979 A CN111742979 A CN 111742979A CN 202010658784 A CN202010658784 A CN 202010658784A CN 111742979 A CN111742979 A CN 111742979A
- Authority
- CN
- China
- Prior art keywords
- takifugu rubripes
- frozen
- protease
- preparation
- rubripes
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 241001441722 Takifugu rubripes Species 0.000 title claims abstract description 64
- 239000003755 preservative agent Substances 0.000 title claims abstract description 27
- 230000002335 preservative effect Effects 0.000 title claims abstract description 27
- 238000002360 preparation method Methods 0.000 title claims abstract description 15
- 241000251468 Actinopterygii Species 0.000 claims abstract description 48
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 46
- 239000004365 Protease Substances 0.000 claims abstract description 27
- 238000000034 method Methods 0.000 claims abstract description 21
- 108091005804 Peptidases Proteins 0.000 claims abstract description 19
- 239000000047 product Substances 0.000 claims abstract description 19
- 238000001816 cooling Methods 0.000 claims abstract description 14
- 150000001875 compounds Chemical class 0.000 claims abstract description 11
- 238000010411 cooking Methods 0.000 claims abstract description 11
- 235000013372 meat Nutrition 0.000 claims abstract description 11
- 238000004537 pulping Methods 0.000 claims abstract description 7
- 238000009210 therapy by ultrasound Methods 0.000 claims abstract description 5
- 238000004140 cleaning Methods 0.000 claims abstract description 3
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 claims abstract 5
- 235000019419 proteases Nutrition 0.000 claims description 17
- 238000003860 storage Methods 0.000 claims description 16
- 241000276707 Tilapia Species 0.000 claims description 12
- 108090000145 Bacillolysin Proteins 0.000 claims description 11
- 108091005658 Basic proteases Proteins 0.000 claims description 11
- 102000035092 Neutral proteases Human genes 0.000 claims description 11
- 108091005507 Neutral proteases Proteins 0.000 claims description 11
- 239000006228 supernatant Substances 0.000 claims description 10
- 108090000526 Papain Proteins 0.000 claims description 8
- 235000019834 papain Nutrition 0.000 claims description 8
- 229940055729 papain Drugs 0.000 claims description 8
- 238000007789 sealing Methods 0.000 claims description 6
- 210000001367 artery Anatomy 0.000 claims description 5
- 238000009461 vacuum packaging Methods 0.000 claims description 5
- 210000001835 viscera Anatomy 0.000 claims description 5
- 238000004321 preservation Methods 0.000 claims description 4
- 241000194108 Bacillus licheniformis Species 0.000 claims description 3
- 244000063299 Bacillus subtilis Species 0.000 claims description 3
- 235000014469 Bacillus subtilis Nutrition 0.000 claims description 3
- 102000005158 Subtilisins Human genes 0.000 claims description 3
- 108010056079 Subtilisins Proteins 0.000 claims description 3
- 238000010257 thawing Methods 0.000 claims description 3
- 230000007071 enzymatic hydrolysis Effects 0.000 claims description 2
- 238000006047 enzymatic hydrolysis reaction Methods 0.000 claims description 2
- 230000001953 sensory effect Effects 0.000 abstract description 9
- 238000011156 evaluation Methods 0.000 abstract description 6
- 239000011248 coating agent Substances 0.000 abstract description 5
- 238000000576 coating method Methods 0.000 abstract description 5
- 230000002255 enzymatic effect Effects 0.000 abstract 1
- 239000000413 hydrolysate Substances 0.000 abstract 1
- 239000007788 liquid Substances 0.000 description 15
- 102000035195 Peptidases Human genes 0.000 description 14
- 102000004169 proteins and genes Human genes 0.000 description 13
- 108090000623 proteins and genes Proteins 0.000 description 13
- 238000007710 freezing Methods 0.000 description 10
- 108090000765 processed proteins & peptides Proteins 0.000 description 9
- 239000000243 solution Substances 0.000 description 9
- VLTRZXGMWDSKGL-UHFFFAOYSA-N perchloric acid Chemical compound OCl(=O)(=O)=O VLTRZXGMWDSKGL-UHFFFAOYSA-N 0.000 description 8
- 241001627955 Tetraodon lineatus Species 0.000 description 7
- 230000008014 freezing Effects 0.000 description 7
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 6
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 6
- ZKHQWZAMYRWXGA-UHFFFAOYSA-N Adenosine triphosphate Natural products C1=NC=2C(N)=NC=NC=2N1C1OC(COP(O)(=O)OP(O)(=O)OP(O)(O)=O)C(O)C1O ZKHQWZAMYRWXGA-UHFFFAOYSA-N 0.000 description 4
- GRSZFWQUAKGDAV-UHFFFAOYSA-N Inosinic acid Natural products OC1C(O)C(COP(O)(O)=O)OC1N1C(NC=NC2=O)=C2N=C1 GRSZFWQUAKGDAV-UHFFFAOYSA-N 0.000 description 4
- 241001441723 Takifugu Species 0.000 description 4
- 125000002915 carbonyl group Chemical group [*:2]C([*:1])=O 0.000 description 4
- 238000006243 chemical reaction Methods 0.000 description 4
- 230000000052 comparative effect Effects 0.000 description 4
- 238000005520 cutting process Methods 0.000 description 4
- FDGQSTZJBFJUBT-UHFFFAOYSA-N hypoxanthine Chemical group O=C1NC=NC2=C1NC=N2 FDGQSTZJBFJUBT-UHFFFAOYSA-N 0.000 description 4
- 235000013902 inosinic acid Nutrition 0.000 description 4
- 238000007747 plating Methods 0.000 description 4
- 230000008569 process Effects 0.000 description 4
- 239000002002 slurry Substances 0.000 description 4
- XTWYTFMLZFPYCI-KQYNXXCUSA-N 5'-adenylphosphoric acid Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](COP(O)(=O)OP(O)(O)=O)[C@@H](O)[C@H]1O XTWYTFMLZFPYCI-KQYNXXCUSA-N 0.000 description 3
- XTWYTFMLZFPYCI-UHFFFAOYSA-N Adenosine diphosphate Natural products C1=NC=2C(N)=NC=NC=2N1C1OC(COP(O)(=O)OP(O)(O)=O)C(O)C1O XTWYTFMLZFPYCI-UHFFFAOYSA-N 0.000 description 3
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 3
- 239000004698 Polyethylene Substances 0.000 description 3
- 244000269722 Thea sinensis Species 0.000 description 3
- UDMBCSSLTHHNCD-KQYNXXCUSA-N adenosine 5'-monophosphate Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](COP(O)(O)=O)[C@@H](O)[C@H]1O UDMBCSSLTHHNCD-KQYNXXCUSA-N 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- 239000003651 drinking water Substances 0.000 description 3
- 235000020188 drinking water Nutrition 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 239000000706 filtrate Substances 0.000 description 3
- 235000013305 food Nutrition 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 229910052757 nitrogen Inorganic materials 0.000 description 3
- -1 polyethylene Polymers 0.000 description 3
- 229920000573 polyethylene Polymers 0.000 description 3
- 150000008442 polyphenolic compounds Chemical class 0.000 description 3
- 235000013824 polyphenols Nutrition 0.000 description 3
- 238000005057 refrigeration Methods 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 238000005303 weighing Methods 0.000 description 3
- AUHDWARTFSKSAC-HEIFUQTGSA-N (2S,3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)-2-(6-oxo-1H-purin-9-yl)oxolane-2-carboxylic acid Chemical compound [C@]1([C@H](O)[C@H](O)[C@@H](CO)O1)(N1C=NC=2C(O)=NC=NC12)C(=O)O AUHDWARTFSKSAC-HEIFUQTGSA-N 0.000 description 2
- HORQAOAYAYGIBM-UHFFFAOYSA-N 2,4-dinitrophenylhydrazine Chemical compound NNC1=CC=C([N+]([O-])=O)C=C1[N+]([O-])=O HORQAOAYAYGIBM-UHFFFAOYSA-N 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- UGQMRVRMYYASKQ-UHFFFAOYSA-N Hypoxanthine nucleoside Natural products OC1C(O)C(CO)OC1N1C(NC=NC2=O)=C2N=C1 UGQMRVRMYYASKQ-UHFFFAOYSA-N 0.000 description 2
- 229930010555 Inosine Natural products 0.000 description 2
- UGQMRVRMYYASKQ-KQYNXXCUSA-N Inosine Chemical group O[C@@H]1[C@H](O)[C@@H](CO)O[C@H]1N1C2=NC=NC(O)=C2N=C1 UGQMRVRMYYASKQ-KQYNXXCUSA-N 0.000 description 2
- 241000269799 Perca fluviatilis Species 0.000 description 2
- 238000002835 absorbance Methods 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 230000006866 deterioration Effects 0.000 description 2
- 239000012153 distilled water Substances 0.000 description 2
- 238000001035 drying Methods 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- 229960003786 inosine Drugs 0.000 description 2
- 229940028843 inosinic acid Drugs 0.000 description 2
- 239000004245 inosinic acid Substances 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000003647 oxidation Effects 0.000 description 2
- 238000007254 oxidation reaction Methods 0.000 description 2
- 239000002244 precipitate Substances 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 230000009467 reduction Effects 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- ZKHQWZAMYRWXGA-KQYNXXCUSA-J ATP(4-) Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](COP([O-])(=O)OP([O-])(=O)OP([O-])([O-])=O)[C@@H](O)[C@H]1O ZKHQWZAMYRWXGA-KQYNXXCUSA-J 0.000 description 1
- 241000251464 Coelacanthiformes Species 0.000 description 1
- UDMBCSSLTHHNCD-UHFFFAOYSA-N Coenzym Q(11) Natural products C1=NC=2C(N)=NC=NC=2N1C1OC(COP(O)(O)=O)C(O)C1O UDMBCSSLTHHNCD-UHFFFAOYSA-N 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- LNQVTSROQXJCDD-UHFFFAOYSA-N adenosine monophosphate Natural products C1=NC=2C(N)=NC=NC=2N1C1OC(CO)C(OP(O)(O)=O)C1O LNQVTSROQXJCDD-UHFFFAOYSA-N 0.000 description 1
- 230000003064 anti-oxidating effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008033 biological extinction Effects 0.000 description 1
- 230000000740 bleeding effect Effects 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- KGBXLFKZBHKPEV-UHFFFAOYSA-N boric acid Chemical compound OB(O)O KGBXLFKZBHKPEV-UHFFFAOYSA-N 0.000 description 1
- 239000004327 boric acid Substances 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 230000036425 denaturation Effects 0.000 description 1
- 238000004925 denaturation Methods 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000007599 discharging Methods 0.000 description 1
- 238000004821 distillation Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 229940088598 enzyme Drugs 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 235000019634 flavors Nutrition 0.000 description 1
- 238000009920 food preservation Methods 0.000 description 1
- 238000000227 grinding Methods 0.000 description 1
- 229960000789 guanidine hydrochloride Drugs 0.000 description 1
- PJJJBBJSCAKJQF-UHFFFAOYSA-N guanidinium chloride Chemical compound [Cl-].NC(N)=[NH2+] PJJJBBJSCAKJQF-UHFFFAOYSA-N 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- XKVWLLRDBHAWBL-UHFFFAOYSA-N imperatorin Natural products CC(=CCOc1c2OCCc2cc3C=CC(=O)Oc13)C XKVWLLRDBHAWBL-UHFFFAOYSA-N 0.000 description 1
- 239000000395 magnesium oxide Substances 0.000 description 1
- CPLXHLVBOLITMK-UHFFFAOYSA-N magnesium oxide Inorganic materials [Mg]=O CPLXHLVBOLITMK-UHFFFAOYSA-N 0.000 description 1
- AXZKOIWUVFPNLO-UHFFFAOYSA-N magnesium;oxygen(2-) Chemical compound [O-2].[Mg+2] AXZKOIWUVFPNLO-UHFFFAOYSA-N 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 1
- 229960000907 methylthioninium chloride Drugs 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 239000004570 mortar (masonry) Substances 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 238000004806 packaging method and process Methods 0.000 description 1
- 239000008055 phosphate buffer solution Substances 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000006318 protein oxidation Effects 0.000 description 1
- 239000012460 protein solution Substances 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 239000000523 sample Substances 0.000 description 1
- 239000012488 sample solution Substances 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000012086 standard solution Substances 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 238000004448 titration Methods 0.000 description 1
- YNJBWRMUSHSURL-UHFFFAOYSA-N trichloroacetic acid Chemical compound OC(=O)C(Cl)(Cl)Cl YNJBWRMUSHSURL-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- 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
- A23B4/14—Preserving with chemicals not covered by groups A23B4/02 or A23B4/12
- A23B4/18—Preserving with chemicals not covered by groups A23B4/02 or A23B4/12 in the form of liquids or solids
- A23B4/20—Organic compounds; Microorganisms; Enzymes
-
- 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
-
- 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
- A23B4/14—Preserving with chemicals not covered by groups A23B4/02 or A23B4/12
- A23B4/18—Preserving with chemicals not covered by groups A23B4/02 or A23B4/12 in the form of liquids or solids
- A23B4/20—Organic compounds; Microorganisms; Enzymes
- A23B4/22—Microorganisms; Enzymes; Antibiotics
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
Abstract
The invention relates to a takifugu rubripes biological preservative and a preparation method and application thereof, wherein the method comprises the following steps: step 1), unfreezing frozen fish skin, cleaning, pulping, adding water, and cooking at 100-121 ℃; and 2), cooling to 65-75 ℃, carrying out ultrasonic treatment, and then adding compound protease for enzymolysis to obtain the product. The biological preservative for the fish skin enzymatic hydrolysate is applied to the takifugu rubripes plated with the ice coating, the quality of the takifugu rubripes meat is kept at a good level after being frozen and stored for 12 months, and the takifugu rubripes meat is kept more than 94% of the fresh takifugu rubripes meat through sensory evaluation and is obviously superior to the takifugu rubripes plated with the ice coating by pure water.
Description
Technical Field
The invention relates to the technical field of aquatic food preservation, in particular to a takifugu rubripes biological preservative and a preparation method and application thereof.
Background
Fugu rubripes belongs to the Osteichthyes, Fugu orders, Fugu suborder, Fugu families and the genus Fugu. The takifugu rubripes is popular with consumers due to the characteristics of high nutrition and high quality, and has great market potential. At present, the globefish resources in China are rich, with the market opening, the demand of the globefish in China is increased, the storage, transportation and preservation of the fresh globefish are very important, the control of the freshness of the globefish in the storage and transportation process has important significance for the development of the globefish storage and transportation industry in the future, and the further expansion and development of the industry are facilitated.
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 plated ice clothes can effectively slow down the deterioration of the texture during the frozen storage period, but the simply plated ice clothes aquatic products usually have the phenomena of color change, protein oxidation, obvious quality reduction and the like 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 natural preservative for takifugu rubripes and no report of application of the natural preservative are available.
Disclosure of Invention
In order to solve the technical problems, the embodiment of the invention provides a takifugu rubripes biological preservative and a preparation method and application thereof. The invention provides a method for storing and preserving red-fin eastern globefish meat by freezing and coating an ice coating, which is based on a great deal of research, combines the characteristics of the red-fin eastern globefish, utilizes the fish skin as a by-product of processing as a raw material, and applies a biological enzymolysis technology to prepare fish skin enzymolysis product protein peptide with the functions of antioxidation and freeze resistance.
The invention provides a preparation method of a takifugu rubripes biological preservative, which comprises the following steps:
step 1), unfreezing frozen fish skin, cleaning, pulping, adding water, and cooking at 100-121 ℃;
and 2), cooling to 65-75 ℃, carrying out ultrasonic treatment, and then adding compound protease for enzymolysis to obtain the product.
According to some preferred embodiments of the invention, the frozen skin is frozen tilapia skin. The inventor takes specific fish skin as a raw material, and unexpectedly develops a fish skin protein peptide liquid preservative with good anti-freezing function on takifugu rubripes through a large number of experiments and a complete set of preparation method.
According to some preferred embodiments of the present invention, in the step 1), the thawing temperature is 10 to 12 ℃; and/or the amount of the added water is 3.5-4.5 times of the weight of the fish skin; and/or the cooking time is 60-120 min.
According to some preferred embodiments of the present invention, in the step 2), the frequency of the ultrasonic treatment is 40 to 60kH, and the time is 15 to 25 min.
According to some preferred embodiments of the present invention, in the step 2), the compound protease is added in an amount of 0.1 to 0.3% by weight of the frozen fish skin; and/or the enzymolysis temperature is 50-60 ℃, and the time is 0.5-1.0 h.
According to some preferred embodiments of the present invention, the compound protease is composed of alkaline protease, neutral protease and papain, and preferably, the mass ratio of the alkaline protease, the neutral protease and the papain is 1-2: 1: 1-2; and/or the alkaline protease is Alcalase 2.4L and is derived from bacillus licheniformis; the neutral protease is protease derived from bacillus subtilis. According to some preferred embodiments of the present invention, the enzymatic hydrolysis further comprises the following steps: cooling to room temperature, centrifuging for 3-8 min under the condition of 4000-5000 g, collecting supernatant, adjusting the protein content to 1.5-2%, and cooling to 0-4 ℃.
The invention also provides a takifugu rubripes biological preservative prepared by the preparation method
The invention further provides the application of the biological preservative obtained by the preparation method or the biological preservative in the frozen ice coating storage and preservation of the meat of the takifugu rubripes.
According to some preferred embodiments of the invention, fresh and live takifugu rubripes is subjected to blood bleeding by cutting off the artery behind the head of the fish body, then the viscera of the takifugu rubripes are removed, the takifugu rubripes is rinsed with water and drained, the takifugu rubripes is frozen at the temperature of-35-45 ℃ for 3-5 hours, the frozen takifugu rubripes is immersed in the biological preservative for 10-15 s, the content of ice coat is controlled to be 15-20%, and the takifugu rubripes plated with the ice coat is obtained; then the takifugu rubripes plated with the ice coat is put into a sealing bag and is frozen and stored at the temperature of-18 to-23 ℃ in a vacuum packaging mode.
The invention has the beneficial effects that: (1) the invention develops the natural anti-freezing preservative for the takifugu rubripes by utilizing the fish skin to carry out specific biological enzymolysis treatment; (2) pulping the fish skin, then treating at the temperature of 100-121 ℃, and directly performing enzymolysis by using protease, wherein the total usage amount of the protease is only 0.1-0.3% of the weight of the fish skin. No chemical substances are added in the enzymolysis process; (3) the developed product is safe, and the invention adopts a plurality of food-grade compound proteases (alkaline protease, neutral protease and papain), and obtains the fish skin protein peptide with specific functions, which is 100 percent of the fish skin protein peptide, through moderate enzymolysis under mild conditions; (4) the fish skin protein peptide developed by the invention has the advantages of simple production process, no need of separation and purification, low production cost and better anti-freezing function. (5) The fish skin protein peptide liquid developed by the invention is used for storage and fresh keeping of the takifugu rubripes, has a remarkable fresh-keeping effect, and the sensory quality of the product is kept above 94% after the product is stored for 12 months. (6) The protein peptide liquid developed by the invention can be widely used as a natural preservative for aquatic food.
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: the alkaline protease is Alcalase 2.4L and is derived from Bacillus licheniformis; the neutral protease is protease derived from Bacillus subtilis.
Example 1
The embodiment provides a fresh-keeping method of takifugu rubripes, which comprises the following steps:
1) 50 g of frozen fish skin is selected, unfreezing is carried out at the temperature of 10-12 ℃, the fish skin is cleaned by clean water meeting the sanitary standard of drinking water, the fish skin is pulped by a pulping machine after being cleaned, water with the weight 3.5 times of that of the fish skin is added, and the fish skin is boiled for 120 minutes at the temperature of 110 ℃. Then, the temperature was adjusted to 65 ℃ and the fish skin slurry was obtained by treating the fish skin slurry with ultrasonic waves (frequency of 60kH) for 15 to 25 minutes using an ultrasonic generator.
2) Adding compound protease into the fish skin (1) according to the weight percentage of 0.1% of the weight of the fish skin for enzymolysis, wherein the protease (alkaline protease, neutral protease and papain) consists of 1: 1: 2) performing enzymolysis at 50 deg.C for 0.5h, cooling to room temperature, centrifuging at 5000g for 5min, collecting supernatant, adjusting protein content to 1.5%, and cooling to 0-4 deg.C.
3) Cutting off the posterior artery of the head of the fresh and live takifugu rubripes to bleed, removing internal organs, rinsing with water, draining, freezing the liquid at the temperature of 40 ℃ below zero for 4 hours, and immersing the frozen takifugu rubripes in the liquid at the temperature of 0 ℃ in the step (2) for 15 seconds to obtain the takifugu rubripes plated with the ice coat, wherein the content of the ice coat is 18%.
4) The takifugu rubripes plated with the ice coat is put into a polyethylene sealing bag, and is subjected to vacuum packaging and frozen storage in a refrigeration house at the temperature of between 18 ℃ below zero and 23 ℃ below zero. Freezing for 12 months.
Example 2
The embodiment provides a fresh-keeping method of takifugu rubripes, which comprises the following steps:
1) 500 g of frozen tilapia skin is selected, unfreezing is carried out at the temperature of 10-12 ℃, the tilapia skin is cleaned by clean water meeting the sanitary standard of drinking water, the tilapia skin is pulped into pulp by a pulping machine after being cleaned, water with the weight 4.0 times of the tilapia skin is added, and the tilapia skin is boiled for 60 minutes at the temperature of 121 ℃. Then, the temperature was adjusted to 75 ℃ and the mixture was treated with ultrasonic waves (frequency: 40kH) for 25 minutes using an ultrasonic generator to obtain a fish skin slurry.
2) Adding compound protease into the fish skin in the step (1) according to the weight percentage of 0.2% of the weight of the fish skin for enzymolysis, wherein the protease (alkaline protease, neutral protease and papain) comprises the following components in a mass ratio of 2: 1: 1) carrying out enzymolysis reaction at 60 ℃ for 1.0h, cooling to room temperature, centrifuging for 8 minutes at 4000g, collecting supernatant, adjusting protein content to 2%, and cooling to 0-4 ℃;
3) cutting off the posterior artery of the head of the fresh and alive fugu rubripes to bleed, removing internal organs, rinsing with water, draining, freezing at-40 ℃ for 5 hours, immersing the frozen fugu rubripes in the liquid at 0 ℃ in the step (2) for 12s, and controlling the content of the ice coat to be 15-18% to obtain the fugu rubripes plated with the ice coat.
4) The takifugu rubripes plated with the ice coat is put into a polyethylene sealing bag, and is subjected to vacuum packaging and frozen in a refrigeration house at the temperature of between 18 ℃ below zero and 23 ℃ below zero for 12 months.
Example 3
The embodiment provides a fresh-keeping method of takifugu rubripes, which comprises the following steps:
1) 1000 g of frozen tilapia skin is selected, unfreezing is carried out at the temperature of 10-12 ℃, the tilapia skin is cleaned by clean water meeting the sanitary standard of drinking water, the tilapia skin is pulped into pulp by a pulping machine after being cleaned, water with the weight 4.5 times of the tilapia skin is added, and the tilapia skin is boiled for 120 minutes at the temperature of 100 ℃. Then, the temperature was adjusted to 75 ℃ and the mixture was treated with ultrasonic waves (frequency: 60kH) for 15 minutes using an ultrasonic generator to obtain a fish skin slurry.
2) Adding compound protease into the fish skin (1) according to the weight percentage of 0.3% of the weight of the fish skin for enzymolysis, wherein the protease (alkaline protease, neutral protease and papain) consists of 1: 1: 1) carrying out enzymolysis reaction at 55 ℃ for 1.0h, cooling to room temperature, centrifuging for 4 minutes at 5000g, collecting supernatant, adjusting protein content to 2%, and cooling to 0-4 ℃;
3) cutting off the posterior artery of the head of the fresh and live takifugu rubripes to bleed, removing internal organs, rinsing with water, draining, freezing at-40 ℃ for 5 hours, immersing the frozen takifugu rubripes in the liquid at 0 ℃ in the step (2) for 15s, and controlling the content of the ice coat to be 20% to obtain the takifugu rubripes plated with the ice coat.
4) The takifugu rubripes plated with the ice coat is put into a polyethylene sealing bag, and is subjected to vacuum packaging and frozen in a refrigeration house at the temperature of between 18 ℃ below zero and 23 ℃ below zero for 12 months.
Comparative example 1
The comparative example provides a method for plating takifugu rubripes plated with tea polyphenol liquid ice coat, which is the same as that of example 2 (takifugu rubripes plated with fish skin protein peptide liquid ice coat), except that the plating liquid is 2% tea polyphenol liquid.
Comparative example 2
This comparative example provides a method for plating takifugu rubripes plated with pure water ice coat, which is the same as that of example 2 (takifugu rubripes plated with fish skin protein peptide liquid ice coat), except that the plating liquid is pure water.
Experimental example 1 determination of Fugu rubripes quality
1. Measurement of cooking loss ratio
Thawing Fugu rubripes, weighing fish meat of certain weight, packaging into fresh-keeping bag, discharging air, sealing, boiling in water bath at 85 deg.C for 20min, taking out, cooling at room temperature for 10min, drying with filter paper, weighing again, and calculating the loss rate of cooking. The cooking loss (%) (weight of fish before cooking-weight of fish after cooking)/weight of fish before cooking x 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 the formula of A × D/(C × B), wherein A represents the absorbance, B represents the concentration of the liquid protein to be detected (mg/mL), C represents the molar absorption coefficient, the value is 13600mol-1 cm-1. L, and D is the dilution factor, 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.
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
5 takifugu rubripes were arbitrarily selected, and sensory evaluation groups (15 persons) trained specially evaluated the fish sensory quality comprehensively according to table 1, and the average value was finally obtained. 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 Fugu rubripes sensory evaluation criteria
The results of comparing the main quality index of different treated fish meat after frozen storage at-18 to-23 ℃ for 12 months in this experimental example are shown in Table 2.
TABLE 2 comparison of the main quality index of differently processed fish meat after frozen storage at-18 to-23 deg.C for 12 months
The above results show that the takifugu rubripes biological preservative prepared in examples 1-3 of the present invention and the takifugu rubripes coated with the takifugu rubripes biological preservative prepared in example 2 of the present invention have a sensory evaluation of 23.7 minutes after being frozen and stored for 12 months, the sensory quality is maintained at 94% or more, the cooking loss rate is less than 12%, the disulfide bond content is less than 3.4mol/105g, the carbonyl content is less than 1.5nmol/mg protein, and the volatile basic nitrogen is less than 8mg/100g, which is significantly better than that of the pure water icing group and the non-icing group, and is also better than that of the 2% tea polyphenol liquid icing group. The fish skin biological preservative developed by the invention can obviously prolong the storage life of the takifugu rubripes and has obvious effects on oxidation resistance and freezing resistance.
Although the invention has been described in detail hereinabove with respect to a general description and specific embodiments thereof, it will be apparent to those skilled in the art that modifications or improvements may 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 preparation method of a takifugu rubripes biological preservative is characterized by comprising the following steps:
step 1), unfreezing frozen fish skin, cleaning, pulping, adding water, and cooking at 100-121 ℃;
and 2), cooling to 65-75 ℃, carrying out ultrasonic treatment, and then adding compound protease for enzymolysis to obtain the product.
2. The method of claim 1, wherein the frozen skin is frozen tilapia skin.
3. The preparation method according to claim 1, wherein in the step 1), the thawing temperature is 10-12 ℃; and/or the amount of the added water is 3.5-4.5 times of the weight of the fish skin; and/or the cooking time is 60-120 min.
4. The method according to any one of claims 1 to 3, wherein in the step 2), the ultrasonic treatment is performed at a frequency of 40 to 60kH for a time of 15 to 25 min.
5. The preparation method according to any one of claims 1 to 4, wherein in the step 2), the compound protease is added in an amount of 0.1 to 0.3% by weight of the frozen fish skin; and/or the enzymolysis temperature is 50-60 ℃, and the time is 0.5-1.0 h.
6. The preparation method according to claim 5, wherein the compound protease is composed of alkaline protease, neutral protease and papain, and preferably, the mass ratio of the alkaline protease to the neutral protease to the papain is 1-2: 1: 1-2; and/or the alkaline protease is Alcalase 2.4L and is derived from bacillus licheniformis; the neutral protease is protease derived from bacillus subtilis.
7. The method of claim 5, further comprising the following steps after the enzymatic hydrolysis: cooling to room temperature, centrifuging for 3-8 min under the condition of 4000-5000 g, collecting supernatant, adjusting the protein content to 1.5-2%, and cooling to 0-4 ℃.
8. A takifugu rubripes biological preservative characterized in that the takifugu rubripes biological preservative obtained by the preparation method according to any one of claims 1 to 7 is used.
9. The use of the biological preservative obtained by the preparation method according to any one of claims 1 to 7 or the biological preservative according to claim 8 in the frozen coated ice storage and preservation of takifugu rubripes meat.
10. The application of the takifugu rubripes as claimed in claim 9, wherein the takifugu rubripes is cut off the artery at the back of the head of a fish body for bloodletting, then the viscera of the takifugu rubripes are removed, the takifugu rubripes is rinsed with water, drained and frozen at-35-45 ℃ for 3-5 hours, the frozen takifugu rubripes is immersed in the biological preservative for 10-15 s, the content of the ice coat is controlled to be 15-20%, and the takifugu rubripes plated with the ice coat is obtained; then the takifugu rubripes plated with the ice coat is put into a sealing bag and is frozen and stored at the temperature of-18 to-23 ℃ in a vacuum packaging mode.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010658784.2A CN111742979A (en) | 2020-07-09 | 2020-07-09 | Fugu rubripes biological preservative and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010658784.2A CN111742979A (en) | 2020-07-09 | 2020-07-09 | Fugu rubripes biological preservative and preparation method and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN111742979A true CN111742979A (en) | 2020-10-09 |
Family
ID=72711092
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010658784.2A Pending CN111742979A (en) | 2020-07-09 | 2020-07-09 | Fugu rubripes biological preservative and preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111742979A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112493305A (en) * | 2020-12-02 | 2021-03-16 | 上海海洋大学 | Method for preserving fugu obscurus |
| CN113575881A (en) * | 2021-08-03 | 2021-11-02 | 佛山市顺德万汇美食工业研究有限公司 | Method for making raw fish |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102239899A (en) * | 2011-04-27 | 2011-11-16 | 中国农业大学 | Fishskin zymolyte preservative as well as preparation method and preservation method thereof |
| CN107811026A (en) * | 2017-10-30 | 2018-03-20 | 浙江天和水产股份有限公司 | A kind of shrimp bio-preservative and its application |
-
2020
- 2020-07-09 CN CN202010658784.2A patent/CN111742979A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102239899A (en) * | 2011-04-27 | 2011-11-16 | 中国农业大学 | Fishskin zymolyte preservative as well as preparation method and preservation method thereof |
| CN107811026A (en) * | 2017-10-30 | 2018-03-20 | 浙江天和水产股份有限公司 | A kind of shrimp bio-preservative and its application |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112493305A (en) * | 2020-12-02 | 2021-03-16 | 上海海洋大学 | Method for preserving fugu obscurus |
| CN113575881A (en) * | 2021-08-03 | 2021-11-02 | 佛山市顺德万汇美食工业研究有限公司 | Method for making raw fish |
| CN113575881B (en) * | 2021-08-03 | 2023-08-04 | 佛山市顺德万汇美食工业研究有限公司 | Method for making raw fish |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Field et al. | Utilization of glucose oxidase for extending the shelf‐life of fish | |
| US4284653A (en) | Process for handling and processing fish meat | |
| CN104322650A (en) | Coating preservation method of pseudosciaena crocea | |
| CN111742979A (en) | Fugu rubripes biological preservative and preparation method and application thereof | |
| CN107811026B (en) | Shrimp meat biological preservative and application thereof | |
| Hernandez‐Herrero et al. | Influence of raw fish quality on some physicochemical and microbial characteristics as related to ripening of salted anchovies (Engraulis encrasicholus L) | |
| CN107372780A (en) | Fish removes fishy preservative | |
| CN107087767A (en) | A kind of biologic fishy smell removing technology of hair scale roe | |
| FERREIRA et al. | Liquifying cod fish frames under acidic conditions with a fungal enzyme | |
| CN111937952A (en) | Freezing preservative based on fish scale protein zymolyte and preparation method and application thereof | |
| JP3352818B2 (en) | How to treat meat | |
| JPH05103586A (en) | Freezing and freeze-drying denaturation suppressing substance, frozen and freeze-dried product containing the same substance and its production | |
| JP2003310208A (en) | Discoloration preventing agent for food and method for preventing discoloration of food | |
| JP5197100B2 (en) | Method for producing retort shrimp | |
| JPH11266803A (en) | Production of softened product of hard collagen obtained from scale of fish, and its utilization for food | |
| Flick et al. | Changes in nitrogenous compounds in freshwater crayfish (Procambarus clarkii) tail meat stored in ice | |
| JPH06125694A (en) | Freeze-aging of frozen shrimp and processed shrimp food prepared by using the aging method | |
| JP2003250499A (en) | Method for soaking fish body with vinegar and composed liquid to be used for the same | |
| Kingwascharapong et al. | Effect of strong alkaline solutions on yield and characteristics of Pacific white shrimp (Litopenaeus vannamei). | |
| CN111903931A (en) | Pickling agent for large yellow croaker and pickling method thereof | |
| Yetim et al. | The influence of egg white and tumbling on proximate composition and protein fractions of fresh catfish muscle | |
| KR100639236B1 (en) | Inhibiting method of browning of mideodeok meat, mideodeok meat which browning is inhibited, and products using it | |
| FR2821245A1 (en) | A method of coating food products, especially skinless sausages with a binder solution containing sodium alginate and a calcium salt insoluble at neutral pH and activating with an acid, especially vinegar or ascorbic acid | |
| Sikorski et al. | The effect of heat-induced changes in nitrogenous constituents on the properties of seafoods | |
| JP2855562B2 (en) | Frozen fish egg and method for producing the same |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20201009 |