CN103601802A - Method for promoting fishbone hydrolysis utilization - Google Patents
Method for promoting fishbone hydrolysis utilization Download PDFInfo
- Publication number
- CN103601802A CN103601802A CN201310582156.0A CN201310582156A CN103601802A CN 103601802 A CN103601802 A CN 103601802A CN 201310582156 A CN201310582156 A CN 201310582156A CN 103601802 A CN103601802 A CN 103601802A
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- fish
- bone
- hydrolysis
- protein
- fishbone
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/78—Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin, cold insoluble globulin [CIG]
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/06—Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products
Abstract
Hydrolysis utilization methods of fishbone mainly comprise an acid method, an alkali method, a water-extraction method and an enzyme method. According to the invention, ultrasonic action is further adopted, so that fishbone hydrolysis utilization rate is improved and extraction time is shortened.
Description
Technical field
The method that fish-bone hydrolysis utilizes is main: acid system, alkaline process and enzyme process, the present invention further uses ul-trasonic irradiation, improves fish-bone hydrolysis utilization ratio, has shortened extraction time, makes it obtain better utilised in fields such as food, makeup, medicine and healthcare products.
Background technology
The skin of aquatic animal, bone, fin etc. are containing abundant collagen protein, and are the sources of safe collagen protein.
China is the country of output of aquatic products maximum in the world, is also that unique cultivation amount surpasses the country of amount of fishing in the world, and especially freshwater aquaculture industry development is very fast, and fresh-water fishes output accounts for 60% left and right of national fishery products ultimate production.The fast development of aquaculture will drive the development of fish processing industry, and at present, freshwater product processing is the focus that people pay close attention to always.This also just means that freshwater fish that cultured output is higher is if the ratio of the processed goods such as silver carp, grass carp, tilapia is by increasing.Along with the development of domestic fish processing industry, the tankage that aquatic products processing enterprise produces are if fish head, fish-skin, fish scale, fish-bone, fin etc. are also by increase year after year.The level of current domestic comprehensive utilization of aquatic products is lower, because freshwater fish edible portion ratio is low, the tankage after flesh of fish processing and other secondary material are not fully recycled, or sell as low value feed, fertilizer, or arbitrarily discharge, will cause waste resource contaminate environment.Therefore, make full use of aquatic products processing tankage, to improve overall economic efficiency, become the problem of fresh-water fishes processing enterprise growing interest.The output of fish-bone is maximum, is the emphasis of research.
From fish-bone, extract collagen protein and it furtherd investigate quite general abroad.Collagen protein (Collagen) belongs to scleroprotein class, it is the main component of human body skin, account for 70%～80% of skin dry weight, with insoluble fiber form, be present in all multicellular animals bodies, it is the class protein that in-vivo content is maximum, there is height anti-tensile ability, very important to the formation of animal and human's body skin, blood vessel, bone, muscles and bones, cartilage, in zooblast, play the part of the role of reticular tissue.Collagen protein in food is all generally macromolecular structure (molecular weight is 5 * 104～10 * 104Da), and human body can not effectively absorb and utilize; Only have micromolecular collagen (molecular weight <3000Da), could effectively be absorbed by human body; Some are containing the food of colloid, as Ungula Bovis seu Bubali muscle, pig's feet, chicken wings, cock skin, fish-skin and cartilage, all contain collagen protein, but due to causes such as mad cow disease, foot and mouth disease, the terrestrial animal of take is prepared collagen protein as raw material and is very limited, raw material turns to take hydrocoles as main, and composition structure and the human body of fish are the most approaching, is the collagen protein source that tissue institute identification the most absorbs.Collagen protein is rich content in corium, bone and the scale of fish, its molecule has unique " triple helical " structure (triple helix configuration being twisted into mutually by 3 α-2 peptide chains, each peptide chain is approximately containing 1200 amino acid, comprising unique oxyproline), have now found that 18～19 kinds, the textural difference of various is mainly that the primary structure of polypeptide chain is the difference of amino-acid sequence.Isin glue collagen belongs to type i collagen, and molecular weight is 30 * 104Da, and the long 280nm of fiber, diameter 115nm are slender type needle-like rigid bodies, is about the α-2 peptide chain formation spirane structure of 10 * 104Da by 3 molecular weight.
Fish-bone collagen protein all can be hydrolyzed by trypsinase and Proteinase K, and hydrolysate molecular size range is different, complicated component.In fish-bone collagen protein, glycine content is the abundantest, is L-Ala, L-glutamic acid and arginine successively.
Fish-bone can be prepared antibacterial peptide.Protein, under enzymic hydrolysis effect, can produce the peptide section with certain functional property, and these polypeptide molecular weights is relatively little; And the common rearrangement with important structure in enzymolysis dissociation process, stand out in the hydrophobic region that causes some to be originally embedded in protein molecule inside.In addition, polypeptide has new nutrition, function and biological nature.Research shows, biologically active peptides regulating gastrointestinal movement, regulate immunity system, hypertension, antibacterial, antithrombotic, antiviral, anticancer, remove free radical and promote the aspects such as mineral element absorption to play an important role.Research for many years shows that antibacterial peptide has broad-spectrum bactericidal action, and relative molecular weight is less, has the advantages such as thermally-stabilised, good water solubility.Up to the present have been found that more than the 600 kind of Endogenous antimicrobial polypeptide being produced by bacterium, plant and animal.Fish-bone is the fine source of antibacterial peptide.
Fish-bone can be prepared Hydrolyzed protein liquid.In fish-bone, protein content is about 15%, and biological value is high, however bone collagen in the non-digestible bone of human body.Enzymolysis can be hydrolyzed into the bone collagen in bone in polypeptide and L-amino acid, greatly improves its nutritive value and functional performance.It is main that enzyme solution adopts single enzymolysis and combinative enzyme hydrolysis, but protein has produced bitter taste after above-mentioned incomplete hydrolysis, and hydrolyzed solution need be through desalting treatment, thereby has limited the application of protein hydrolystate in food and medicine.Adopt substep mode of action hydrolysis fish-bone albumen, expectation improves hydrolysis effect, obtains low without bitter taste, salts contg, can to directly apply to food and medicine field fish-bone hydrolysis prods.
The method that fish-bone hydrolysis utilizes is main: acid system, alkaline process and enzyme process, in actual extracting process, often mutually combine between Different Extraction Method, to obtain good extraction effect meanwhile.Its ultimate principle is all by fish-bone hydrolysis, or collagen protein is separated from other oroteins, or bone is further hydrolyzed, and obtains Hydrolyzed protein liquid or antimicrobial polypeptide, for food Medicines and Health Product etc.
Summary of the invention
The method that fish-bone hydrolysis utilizes is main: acid system, alkaline process, water extraction and enzyme process, the present invention further uses ul-trasonic irradiation, improves fish-bone hydrolysis utilization ratio, has shortened extraction time.
Embodiment
Embodiment mono-
Fish-bone extracts collagen protein
Control group:
Raw material pre-treatment: it is stand-by that-20 ℃ of refrigerators are used or be placed in to raw material fish after getting its vertebrae and thoroughly removing the muscle adhering to immediately.
The extraction of fish-bone collagen protein and purifying: after fish-bone fully minces, with the 0.1mol/L NaOH solution of 20 times of its quality, at 4 ℃, soak 4h, to remove noncollagen protein composition and to prevent the impact of endogenous protease on collagen protein.Fish-bone distilled water repetitive scrubbing, the 0.5mol/L EDTA solution (pH7.4) that fully drains 5 times of its quality of rear use soaks 5d at 4 ℃, changes 1 EDTA solution every day to play decalcification, and uses distilled water repetitive scrubbing.After draining, add 10% aqueous isopropanol of 20 times to soak 1d at 4 ℃, to remove fat.Through distilled water repetitive scrubbing, after draining, add again the 0.5mol/L acetic acid solution of equivalent and soak 3d at 4 ℃.Through 4 ℃, after the centrifugal 30min of 20000 * g, obtain supernatant liquor and be thick collagen protein.Get appropriate thick collagen protein add NaCl to solution final concentration be 0.9mol/L, separate out flocks simultaneously, 4 ℃, the centrifugal 15min of 12000 * g obtains throw out and is the collagen protein after saltouing.Stand-by in rearmounted-25 ℃ of refrigerators of lyophilize.
Ultrasonic wave group: 0.5mol/L EDTA solution (pH7.4) soaks in leaching process at 4 ℃, applies 20kHz ultrasonic wave with ultrasonic generator simultaneously, and extraction time shortens to half of control group.
Result: control group extraction yield is 28.3%, and ultrasonic wave group is %, and ultrasonic wave group has obtained better extraction effect, and has shortened extraction time.
Embodiment bis-
Fish-bone is prepared antibacterial peptide
Control group:
The pre-treatment of raw material: the fresh fish-bone → cleaning → homogenate → enzymolysis → enzyme that goes out → centrifugal → supernatant liquor → lyophilize → mensuration protein content
Antibacterial peptide preparation: fish-bone is cleaned up, after pre-treatment, with ether, at 30 ℃, carry out degreasing, freezing standby after the homogenate packing after degreasing.Homogenate after degreasing adds water, stomach en-enzymolysis, and pepsic best enzymolysis process condition is: concentration of substrate 1:4, enzyme addition 2000U/g, enzymolysis time 2.5h, 37 ℃ of hydrolysis temperatures, optimum pH 7.0.The enzyme 10min that goes out at 85 ℃ after certain hour, centrifugal 10min (4000r/min) obtains supernatant liquor, gets a certain amount of supernatant liquor and measures its anti-microbial activity.
Ultrasonic wave group: in enzyme process enzymolysis process, apply 20kHz ultrasonic wave with ultrasonic generator, extraction time shortens to half of control group simultaneously.
Result: control group is-202.7% to colibacillary inhibiting rate, and ultrasonic wave group is-378.3%, and ultrasonic wave group has obtained better extraction effect, and has shortened extraction time.
Embodiment tri-
Fish-bone is prepared protein hydrolyte
Control group:
The pre-treatment of raw material: it is stand-by that-20 ℃ of refrigerators are used or be placed in to raw material fish after getting its vertebrae and thoroughly removing the muscle adhering to immediately.
The preparation of protein hydrolyte: fish-bone is boiling 40min at 120 ℃.After boiling, by solid-liquid ratio 1:4 (W/V), add water making beating (the three-in-one stirrer in HR2864 type Philip), add a certain amount of enzyme, stir evenly, be placed in and under design temperature, be hydrolyzed into setting-up time (regularly stirring).Initial pH8.0, uses 1500U/g Pr Alcalase after 55 ℃ of hydrolysis 2h, then continues hydrolysis 2h with 1500U/g Pr Flavorzyme.Reaction finishes the rear boiling water bath enzyme 10min that goes out, and reaction solution, in the centrifugal 10min of 3000r/min, is removed upper strata fat, collects supernatant liquor, is fish-bone protein hydrolyte, standby.
Ultrasonic wave group: in enzymolysis and extraction process, apply 20kHz ultrasonic wave with ultrasonic generator, extraction time shortens to half of control group simultaneously.
Result: control group extraction yield is 53.5%, and ultrasonic wave group is 71.7%, and ultrasonic wave group has obtained better extraction effect, and has shortened extraction time.
Claims (5)
1. select suitable method hydrolysis fish-bone and utilize.
2. select the method for the fish-bone extraction collagen protein in suitable right 1.
3. select fish-bone in suitable right 1 to prepare the method for antibacterial peptide.
4. the fish-bone in the suitable right 1 of selection is prepared the method for protein hydrolyte.
5. in the method for right 1-4, with ultrasonic generator, apply 20kHz ultrasonic wave to improve fish-bone hydrolysis utilization ratio simultaneously, shortened extraction time.
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| CN201310582156.0A CN103601802A (en) | 2013-11-15 | 2013-11-15 | Method for promoting fishbone hydrolysis utilization |
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| CN201310582156.0A CN103601802A (en) | 2013-11-15 | 2013-11-15 | Method for promoting fishbone hydrolysis utilization |
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| CN103601802A true CN103601802A (en) | 2014-02-26 |
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105087729A (en) * | 2015-08-14 | 2015-11-25 | 浙江省海洋开发研究院 | Tuna bone collagen peptide preparation method |
| CN105087728A (en) * | 2015-08-14 | 2015-11-25 | 浙江省海洋开发研究院 | Preparation method of selenium-chelated collagen peptide with tuna bone collagen serving as source |
| CN105087730A (en) * | 2015-08-14 | 2015-11-25 | 浙江省海洋开发研究院 | High-F (Fischer)-value tuna oligopeptide preparation method |
| CN105435307A (en) * | 2015-11-30 | 2016-03-30 | 广西医科大学 | Natural-tissue-derived decellularized and decalcified bone material and preparation method thereof |
| CN105801666A (en) * | 2014-12-30 | 2016-07-27 | 浙江海洋学院 | Preparation method of miichthys miiuy maw protein peptide and application thereof |
| CN106858033A (en) * | 2017-01-10 | 2017-06-20 | 大连工业大学 | A kind of preparation method of deodorant cod SPP1 |
| CN107897632A (en) * | 2017-11-10 | 2018-04-13 | 雷笑天 | A kind of preparation method of high-moisture herbage bundle natural fungicidal agent |
| CN109180808A (en) * | 2018-09-30 | 2019-01-11 | 青岛蓝色康典海洋生物科技有限公司 | A kind of fish scale collagen and its preparation method and application |
| CN110484586A (en) * | 2019-09-26 | 2019-11-22 | 深圳市江牧实业有限公司 | A method of flavor peptides are prepared from Tilapia bone albumen |
-
2013
- 2013-11-15 CN CN201310582156.0A patent/CN103601802A/en active Pending
Cited By (14)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105801666A (en) * | 2014-12-30 | 2016-07-27 | 浙江海洋学院 | Preparation method of miichthys miiuy maw protein peptide and application thereof |
| CN105801666B (en) * | 2014-12-30 | 2021-06-04 | 浙江海洋学院 | Preparation method and application of miichthys miiuy swim bladder protein peptide |
| CN105087729B (en) * | 2015-08-14 | 2019-02-12 | 浙江省海洋开发研究院 | A kind of preparation method of tuna fish bone collagen protein peptides |
| CN105087728A (en) * | 2015-08-14 | 2015-11-25 | 浙江省海洋开发研究院 | Preparation method of selenium-chelated collagen peptide with tuna bone collagen serving as source |
| CN105087730A (en) * | 2015-08-14 | 2015-11-25 | 浙江省海洋开发研究院 | High-F (Fischer)-value tuna oligopeptide preparation method |
| CN105087729A (en) * | 2015-08-14 | 2015-11-25 | 浙江省海洋开发研究院 | Tuna bone collagen peptide preparation method |
| CN105087728B (en) * | 2015-08-14 | 2018-09-14 | 浙江省海洋开发研究院 | A kind of preparation method of tuna fish bone collagen protein sources selenium chelating collagen peptide |
| CN105087730B (en) * | 2015-08-14 | 2018-09-14 | 浙江省海洋开发研究院 | A kind of Preparation method of high-F-value tuna oligopeptide |
| CN105435307A (en) * | 2015-11-30 | 2016-03-30 | 广西医科大学 | Natural-tissue-derived decellularized and decalcified bone material and preparation method thereof |
| CN106858033A (en) * | 2017-01-10 | 2017-06-20 | 大连工业大学 | A kind of preparation method of deodorant cod SPP1 |
| CN107897632A (en) * | 2017-11-10 | 2018-04-13 | 雷笑天 | A kind of preparation method of high-moisture herbage bundle natural fungicidal agent |
| CN109180808A (en) * | 2018-09-30 | 2019-01-11 | 青岛蓝色康典海洋生物科技有限公司 | A kind of fish scale collagen and its preparation method and application |
| CN109180808B (en) * | 2018-09-30 | 2021-12-21 | 青岛蓝色康典海洋生物科技有限公司 | Fish scale collagen and preparation method and application thereof |
| CN110484586A (en) * | 2019-09-26 | 2019-11-22 | 深圳市江牧实业有限公司 | A method of flavor peptides are prepared from Tilapia bone albumen |
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Application publication date: 20140226 |