CN104789630A - Bluefin tuna collagen hydrolyzate and preparation method thereof - Google Patents
Bluefin tuna collagen hydrolyzate and preparation method thereof Download PDFInfo
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Abstract
The invention relates to a bluefin tuna collagen hydrolyzate and a preparation method thereof. The method comprises the following steps: cleaning bluefin tuna bones, and immersing in an acidic solution to remove inorganic calcium salts; immersing in a sodium chloride solution to remove non-collagen impurity proteins; carrying out degreasing, fishy smell removal and enzymolysis on the treated fish bones, inactivating the enzyme at high temperature, and carrying out hydrothermal extraction to obtain a bluefin tuna collagen hydrolyzate solution; and drying to obtain the collagen hydrolyzate powder. The preparation technique is scientific and reasonable, and fully utilizes the unemployed fish bones in aquatic product processing as the raw material to develop the collagen hydrolyzate with potential market value; the composite enzyme degreasing, fishy smell removal and enzymolysis process is adopted while regulating the enzymolysis conditions of the composite enzyme, thereby greatly shortening the extraction time of the collagen hydrolyzate, preparing the tuna collagen hydrolyzate with different molecular weights and with the advantages of high gel strength, high oxidation resistance activity, high safety, no toxic or side effect and the like, enhancing the production efficiency and lowering the energy consumption.
Description
Technical field
The present invention relates to a kind of fish osso-albumin hydrolyzate and its production and use, particularly relate to a kind of bluefin tuna fish bone collagen hydrolyzate and its production and use.
Background technology
Collagen protein is ubiquitous a kind of macro-molecular protein in animal body, and it is mainly present in the reticular tissue of animal.Collagen protein is widely used in the fields such as food, medicine, organizational project, makeup because it has good biocompatibility, biological activity and biodegradability.Collagen hydrolysate is that collagen protein is hydrolyzed and the hydrolyzate obtained under certain condition.Compared with collagen protein, the molecular weight of collagen hydrolysate is low, water-soluble better, is easily absorbed by the body.Meanwhile, the hydrolyzate of collagen protein is also provided with more biological activity, as: oxidation-resistance, delays senility, and replenishes the calcium and promotes that skeleton development improves immunizing power, promote skin cells regeneration etc.
Current collagen hydrolysate is mainly derived from the animal such as pig, ox.But the problem such as immunogenicity, religious belief may be there is in these collagen hydrolysates.Along with the development of domestic fish processing industry, tankage such as fish head, fish-skin, fish scale, fish-bone, fin etc. that aquatic products processing enterprise produces increase year by year.But the utilization of aquatic products processing tankage is also insufficient, causes the wasting of resources.The collagen hydrolysate deriving from fish endangers without disease and does not have religion to limit.
The traditional technology that collagen hydrolysate extracts can be divided three classes, i.e. acid system, alkaline process and enzyme process.Its ultimate principle is all the characteristic according to collagen protein, changes the external environment of protein, collagen protein is separated from other protein.Acid system and alkaline process are extracted under acidity and alkaline environment by collagen protein, and acid system is relatively more extensive, and alkaline process report is less.Biological enzyme utilizes the gentle high benefit of enzyme reaction to be extracted from different raw materials by collagen protein under certain environmental conditions.Along with the development of Enzymes Industry and constantly widening of Application Areas, zymin output increases rapidly, and market competition advantage constantly increases.
In the selection of enzyme, main selection has papoid, Sumizyme MP, neutral protease.Papoid applied range is many mainly due to its action site, cleavage site, as Methionin, arginic carboxyl terminal; The action site of Sumizyme MP is then main take carboxyl side as the amino acid whose peptide bond of hydrophobic aromatic race; The action site of neutral protease is the leucine of carboxyl side, the peptide bond of phenylalanine.End contains the oxidation-resistance that Histidine and leucic aminoacid sequence can significantly improve protein polypeptide to have investigator to think; When the hydrophobic amino acid in peptide chain, aromatic nucleus aminoacids content height, its oxidation-resistance has significance.Therefore in this patent, in an embodiment, adopted Sumizyme MP to prepare to have the tuna osso-albumin hydrolyzate of high antioxygenic property.
Although the collagen hydro physical performance deriving from fish is good, be widely used, but the sight of people does not aim at the tuna bone collagen hydrolyzate being prepared different molecular weight by controlled enzymatic hydrolysis condition, and probes into Gel strength and the anti-oxidant activity of the osso-albumin hydrolyzate of different molecular weight.Tuna bone is as the tankage of the tuna course of processing, and economic worth is utilized fully, and tuna is as a kind of high-quality marine economy fish, and market demand is huge, and the tuna bone quantity of generation is large.CN 103421871 A discloses a kind of preparation technology of tuna collagen hydrolysate, comprise the following steps: A: after raw material tuna bone is carried out soaking and washing, first after using pulverizer pulverization process, enter whizzer continuously centrifuged after mixing with 4 ~ 8 times of volume water and remove supernatant in 30 minutes, vinegar decalcification process 30 ~ 50 minutes, recentrifuge; B: extract: utilize biological enzymolysis tank to carry out enzymolysis processing, reaction conditions is 37 ~ 65 DEG C, pH6.5 ~ 8.0,4 ~ 8 hours reaction times; C: shaping and oven dry: enzymolysis product 30KD ultrafiltration membrane treatment, permeation parts concentrates through nanofiltration membrane, and spraying dry is tuna osso-albumin hydrolyzate finished product.
In the preparation method of its tuna osso-albumin hydrolyzate, do not have to introduce and remove by the degreasing of regulation and control prozyme the method that raw meat enzymolysis and extraction condition obtains the osso-albumin hydrolysate of different molecular weight, and to the Gel strength of tuna osso-albumin hydrolyzate and the not report of oxidation-resistance.The preparation method of the tuna osso-albumin hydrolyzate that this patent is introduced, making full use of the fish-bone do not utilized in processing of aquatic products is raw material, exploitation has the collagen hydrolysate that potential market is worth, and adopt prozyme degreasing except the method for raw meat enzymolysis, greatly shorten the extraction time of collagen hydrolysate, obtain high performance collagen hydrolysate, enhance productivity, reduce energy consumption.
Bluefin tuna is the famous and precious kind in tuna.The high-strength albumen of bluefin tuna low fat, swimming speed reaches 72 kilometers per hour the soonest, generally just 2 ~ 3 kilometers, it can bear the activity of greatest differences like this, and a large amount of free radicals that energy autophage is produced by high-speed motion, illustrate that fish-bone has the ability of very high intensity and very strong scavenging free radicals.Because bluefin tuna has above characteristic, therefore we invest sight Gel strength and the antioxidant property of bluefin tuna osso-albumin hydrolyzate.
Summary of the invention
The object of the present invention is to provide a kind of preparation method of bluefin tuna osso-albumin hydrolyzate, this preparation method can regulate and control the molecular weight of hydrolyzate, and craft science is reasonable, easy handling.
For solving the problems of the technologies described above, the present invention adopts following technical scheme:
A preparation method for bluefin tuna osso-albumin hydrolyzate, comprises the steps:
(1) decalcification process: the fish-bone of bluefin tuna is cleaned up and is placed in acidic solution and soaks;
(2) prozyme degreasing and destinking enzymolysis and extraction: by fish-bone lipase, yeast and protease hydrolyzed after process, go out enzyme, hot water extraction, obtains bluefin tuna osso-albumin hydrolyzate solution;
(3) purifying drying step: hydrolyzate solution is filtered, obtains bluefin tuna osso-albumin hydrolysate powder finished product through lyophilize.
Further, also comprise between step (1) and (2) and remove foreign protein step: after decalcification fish-bone being placed in sodium chloride solution immersion, repeatedly clean with deionized water.
Wherein, the acidic solution in decalcification process is 0.1 ~ 0.2wt% hydrochloric acid soln or sulphuric acid soln, and soak time is 7.5 ~ 15h, and every 1.5h changes an acid solution.
Described prozyme degreasing and destinking enzymolysis and extraction refers to: mixed with 1:9 ~ 20 by weight proportion of deionized water by fish-bone, pH is regulated to be 2 ~ 10, lipase is added according to concentration (quality of the quality/fish-bone of lipase) 0.3 ~ 0.5wt%, yeast is added according to concentration (quality of the quality/fish-bone of yeast) 2.0 ~ 3.0wt%, proteolytic enzyme is added according to concentration (quality of the quality/fish-bone of proteolytic enzyme) 0.5 ~ 2.0wt%, hydrolysis temperature 40 ~ 55 DEG C, enzymolysis 6 ~ 10 hours; Enzymolysis solution is heated to 90 ~ 95 DEG C of enzyme 10 ~ 15min that go out, hot water extraction osso-albumin hydrolyzate.
The concentration of described sodium chloride solution is 3 ~ 5wt%, and soaking the fish-bone time is 10 ~ 12h.
Another object of the present invention is to provide a kind of bluefin tuna osso-albumin hydrolyzate, this collagen hydrolysate safe without toxic side effect, and has stronger Gel strength and antioxygenation.
This bluefin tuna osso-albumin hydrolyzate has above-mentioned preparation method to obtain, and the weight average molecular weight range of this collagen hydrolysate is at 1 ~ 30KDa.When weight-average molecular weight is 10 ~ 30KDa, molecular weight distribution index D (M
w/ M
n) scope is when being 2.33 ~ 3.61, this collagen hydrolysate and Gel strength better, its Gel strength is 200 ~ 270Bloom g.When weight-average molecular weight is 1 ~ 3KDa, molecular weight distribution index D (M
w/ M
n) scope is when being 1.12-1.31, this collagen hydrolysate oxidation-resistance is better, and Scavenging action to hydroxyl free radical is 60 ~ 80%, and oxygen radical removing rate is 50 ~ 65%.
Compared to existing technology, the present invention has technique effect useful as follows:
The present invention selects lipase, yeast, proteolytic enzyme and as complex enzyme zymohydrolysis, the collagen hydrolysate of different molecular weight is discharged largely by regulation and control complex enzyme hydrolysis condition and greatly shortens the extraction time of collagen hydrolysate, obtain high performance collagen hydrolysate, enhance productivity, reduce energy consumption; The collagen hydrolysate of the different molecular weight that bluefin tuna fish-bone enzymolysis obtains, Gel strength is high, and safely, have no side effect, antioxygenation is remarkable.Making full use of the fish-bone do not utilized in processing of aquatic products is raw material, and exploitation has the collagen hydrolysate that potential market is worth.
Embodiment
Below in conjunction with embodiment, the present invention will be further described, but protection scope of the present invention is not limited to this.
The invention provides a kind of preparation method of collagen hydrolysate, it is as follows that it comprises step: decalcification removes foreign protein treatment step, prozyme degreasing and destinking enzymolysis and extraction step, purifying drying step.
Fish-bone, except in foreign protein step, fully drains rear with adding 0.1 ~ 0.2wt% acid soak, 5 ~ 24h by decalcification of the present invention, is preferably 7.5 ~ 15h, every 1.5h and changes an acid solution, remove the calcium salt in fish-bone; Working concentration is 2 ~ 8wt%, and preferably the sodium chloride solution immersion fish-bone of 3 ~ 5wt% removes foreign protein, and soak time is 8 ~ 14 hours, preferably 10 ~ 12 hours, then uses washed with de-ionized water fish-bone, removes sodium-chlor.
Degreasing and destinking of the present invention extracts in complex enzyme hydrolysis step, decalcification fish-bone is drained, part by weight 1:5 ~ 20 of fish-bone and deionized water, preferred 1:9 ~ 20, pH is regulated to be 2 ~ 10, be 0.1 ~ 0.6wt% according to concentration (quality of the quality/fish-bone of lipase), be preferably 0.3 ~ 0.5wt% and add lipase, according to concentration (quality of the quality/fish-bone of yeast is the same) 1.0 ~ 5.0wt%, be preferably 2.0 ~ 3.0wt% and add yeast, be 0.25 ~ 3.0wt% according to concentration (quality of the quality/fish-bone of Sumizyme MP), be preferably 0.5 ~ 2.0wt% and add a kind of proteolytic enzyme, hydrolysis temperature is adjusted to 30 ~ 60 DEG C, be preferably 40 ~ 55 DEG C, enzymolysis 5 ~ 12 hours, be preferably 6 ~ 10 hours, enzymolysis solution is heated to 90 ~ 95 DEG C of enzyme 10 ~ 15min that go out.Hot water extraction osso-albumin hydrolyzate also filters.
The present invention regulates the alkaline matter of pH to be not particularly limited, and is placed in its aqueous solution display alkalescence.Be preferably inorganic alkaline.The example of mineral alkali includes but not limited to sodium hydroxide, potassium hydroxide, lithium hydroxide or calcium hydroxide.
The present invention regulates the acidic substance of pH to be not particularly limited, and is placed in its aqueous solution display acidity.Be preferably organic acid or mineral acid.Organic acids is as acetic acid, citric acid.Acidic substance of the present invention are more preferably mineral acid.The example of mineral acid includes but not limited to hydrochloric acid, sulfuric acid, phosphoric acid or nitric acid.
Remove fat enzyme used in the present invention to be not particularly limited.Lipase is alkaline lipase, neutral lipase and acid lipase enzyme such as.
The proteolytic enzyme that in the present invention, enzymolysis is used is not particularly limited.Proteases is as stomach en-, trypsinase, aspartic protease, Sumizyme MP, neutral protease and papoid.In a concrete enforcement of the present invention, proteolytic enzyme is Sumizyme MP.
The collagen hydrolysate that the present invention extracts from bluefin tuna fish-bone, a kind of tuna fish bone collagen hydrolyzate of gained, is characterized in that the weight average molecular weight range of this collagen hydrolysate is at 1 ~ 30KDa.When weight-average molecular weight is 10 ~ 30KDa, molecular weight distribution index D (M
w/ M
n) scope is when being 2.33 ~ 3.61, the film-forming properties of this collagen hydrolysate and Gel strength better, its Gel strength is 200 ~ 270Bloom g.Weight average molecular weight range is 1 ~ 3KDa, molecular weight distribution index D (M
w/ M
n) scope is when being 1.12-1.31, it has good oxidation-resistance, and Scavenging action to hydroxyl free radical is 60 ~ 80%, and oxygen radical removing rate is 50 ~ 60%, high compared with the oxidation-resistance of the collagen hydrolysate extracted in other fish-bones (Scavenging action to hydroxyl free radical 9.42%, oxygen radical removing rate is 42.5%; The oxidation-resistance of self-control bluefin tuna osso-albumin hydrolyzate and other fish bone collagen hydrolyzates measures all uses same standard, that is: Scavenging action to hydroxyl free radical is the spectrophotometry reacting hydroxyl radical free radical and the removing thereof produced according to Fenton, and oxygen radical removing rate adopts assay NBT photoreduction to measure).
Embodiment 1
For bluefin tuna fish-bone for relatively small molecular weight collagen hydrolysate prepared by raw material, carry out following processing step operation successively:
Step 1: thaw bluefin tuna fish-bone, with cold water cleaning, under room temperature, soak 7.5h by fish-bone 0.1wt% hydrochloric acid soln, every 1.5h changes an acid solution, removes calcium salt; Be that the sodium-chlor of 4wt% soaks fish-bone 12 hours again with concentration, remove foreign protein (foreign protein is the albumen of non-collagen);
Step 2: fish-bone is cleaned up, drains, the part by weight 1:10 of fish-bone and deionized water adds deionized water, pH is regulated to be 8, be that 0.4wt% adds alkaline lipase (Lipase) according to concentration, yeast (aldehyde material of fish-bone be oxidized to acid and then remove smelling removal) is put into according to concentration 2.0wt%, be that 1.5wt% adds a kind of Sumizyme MP (Novo type serine protease) according to concentration, hydrolysis temperature is adjusted to 50 DEG C, enzymolysis 10 hours; Enzymolysis solution is heated to 90 ~ 95 DEG C of enzyme 15min that go out.70 DEG C of hot water extraction collagen hydrolysates;
Step 3: by dry for the collagen hydrolysate liquid freezing obtained, obtain finished product.
Collagen hydrolysate powder These parameters method obtained detects, and wherein weight-average molecular weight is 2336Da, molecular weight distribution index D (M
w/ M
n) when being 1.13, collagen hydrolysate is faint yellow, without fishy smell.Scavenging action to hydroxyl free radical is 74.9%, and oxygen radical removing rate is 56.8%.
Note: Scavenging action to hydroxyl free radical is the spectrophotometry reacting hydroxyl radical free radical and the removing thereof produced according to Fenton, Scavenging action to hydroxyl free radical measures, based on the trapping agent of the hydroxyl radical free radical that Whitfield's ointment produces as Fenton reagent reaction, there is the principle of maximum absorption at wavelength 510nm place, make FeSO
4, H
2o
2, Whitfield's ointment-ethanol becomes a reaction system, examination antioxidant is on the impact of capture by salicylic acid hydroxyl radical free radical; Oxygen radical removing rate is the oxyradical adopting assay NBT photoreduction-spectrophotometry, the mensuration of clearance rate, be chain reaction based on mouse thymus cells process, oxyradical can be produced, it has the principle of maximum absorption at wavelength 319nm place, and the content of himself oxidation products can detect with spectrophotometer.(Xu Xiangrong, Wang Wenhua, Li Huabin etc. colorimetric method for determining Fenton reacts the hydroxy radical qiao and application [J] thereof that produce. Progress in Biochemistry and Biophysics. 1999, the few China of 26 (1): 67-69. Korea Spro, Zhu Jingbo, Wang Yanyan etc. assay NBT photoreduction measures the technique study [J] of anti-oxidant activity. and China brewages and analyzes and detect. 2009,207:155-157.).
Embodiment 2
For bluefin tuna fish-bone for larger molecular weight collagen hydrolysate prepared by raw material, carry out following processing step operation successively:
Step 1: thaw bluefin tuna fish-bone, with cold water cleaning, under room temperature, will soak 15h with in 0.2wt% citric acid solution, every 1.5h changes an acid solution, removes calcium salt; Be that the sodium-chlor of 2wt% soaks fish-bone 6 hours again with concentration, remove foreign protein;
Step 2: fish-bone is cleaned up, drains, the part by weight 1:20 of fish-bone and deionized water adds deionized water, pH is regulated to be 7, be that 0.2wt% adds neutral lipase (Lipase) according to concentration, yeast (aldehyde material of fish-bone be oxidized to acid and then remove smelling removal) is put into according to concentration 2.0wt%, be that 1.0wt% adds a kind of papoid (L-Cysteine HCL Anhydrous) according to concentration, hydrolysis temperature is adjusted to 40 DEG C, enzymolysis 8 hours; Enzymolysis solution is heated to 90 ~ 95 DEG C of enzyme 15min that go out; 70 DEG C of hot water extraction collagen hydrolysates.
Step 3: by dry for the collagen hydrolysate liquid freezing obtained, obtain finished product.
Collagen hydrolysate powder These parameters method obtained detects, and wherein weight-average molecular weight is 29586Da, molecular weight distribution index D (M
w/ M
n) when being 3.61, collagen hydrolysate is faint yellow, without fishy smell.Its Gel strength is 265.3Bloom g.
It will be recognized by those skilled in the art, under the prerequisite not departing from protection scope of the present invention, various amendment, change and combination can be carried out to above-mentioned embodiment, and think that this amendment, change and combination are within the scope of originality thought.
Claims (8)
1. a preparation method for bluefin tuna osso-albumin hydrolyzate, is characterized in that, comprises the steps:
(1) decalcification process: the fish-bone of bluefin tuna is cleaned up and is placed in acidic solution and soaks;
(2) prozyme degreasing and destinking enzymolysis and extraction: by fish-bone lipase, yeast and protease hydrolyzed after process, go out enzyme, hot water extraction, obtains bluefin tuna osso-albumin hydrolyzate solution;
(3) purifying drying step: hydrolyzate solution is filtered, obtains bluefin tuna osso-albumin hydrolysate powder finished product through lyophilize.
2. preparation method according to claim 1, is characterized in that, also comprises and remove foreign protein step between step (1) and (2): after decalcification fish-bone being placed in sodium chloride solution immersion, repeatedly clean with deionized water.
3. preparation method according to claim 1 and 2, is characterized in that, the acidic solution in decalcification process is 0.1 ~ 0.2wt% hydrochloric acid soln or sulphuric acid soln, and soak time is 7.5 ~ 15h, and every 1.5h changes an acid solution.
4. preparation method according to claim 1 and 2, it is characterized in that, described prozyme degreasing and destinking enzymolysis and extraction refers to: mixed 1:9 ~ 20 by weight proportion with deionized water by fish-bone, pH is regulated to be 2 ~ 10, lipase is added according to concentration (quality of the quality/fish-bone of lipase) 0.3 ~ 0.5wt%, yeast is added according to concentration (quality of the quality/fish-bone of yeast) 2.0 ~ 3.0wt%, proteolytic enzyme is added according to concentration (quality of the quality/fish-bone of proteolytic enzyme) 0.5 ~ 2.0wt%, hydrolysis temperature 40 ~ 55 DEG C, enzymolysis 6 ~ 10 hours; Enzymolysis solution is heated to 90 ~ 95 DEG C of enzyme 10 ~ 15min that go out, hot water extraction osso-albumin hydrolyzate.
5. preparation method according to claim 2, is characterized in that, the concentration of described sodium chloride solution is 3 ~ 5wt%, and soaking the fish-bone time is 10 ~ 12h.
6. a bluefin tuna osso-albumin hydrolyzate, is characterized in that the preparation method's preparation adopted described in claim 1 or 2.
7. bluefin tuna osso-albumin hydrolyzate as claimed in claim 6, is characterized in that, when osso-albumin hydrolyzate weight-average molecular weight is 10 ~ 30KDa, and molecular weight distribution index D (M
w/ M
n) scope is when being 2.33 ~ 3.61, its Gel strength is 200 ~ 270Bloom g.
8. bluefin tuna osso-albumin hydrolyzate as claimed in claim 6, is characterized in that, when osso-albumin hydrolyzate weight-average molecular weight is 1 ~ 3KDa, and molecular weight distribution index D (M
w/ M
n) scope is when being 1.12-1.31, collagen hydrolysate Scavenging action to hydroxyl free radical is 60 ~ 80%, and oxygen radical removing rate is 50 ~ 65%.
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| CN105795315A (en) * | 2016-04-08 | 2016-07-27 | 中国水产科学研究院南海水产研究所 | Deodorizing and degreasing method of bass slices |
| CN106420504A (en) * | 2016-12-01 | 2017-02-22 | 北京化工大学 | Antioxidant skin-care composition containing ocean collagen degradation product and its application |
| CN106755241A (en) * | 2016-12-28 | 2017-05-31 | 北京化工大学 | It is a kind of to promote tuna bone collagen polypeptide of bone cell growth and preparation method thereof |
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| 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 |
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| CN106420504A (en) * | 2016-12-01 | 2017-02-22 | 北京化工大学 | Antioxidant skin-care composition containing ocean collagen degradation product and its application |
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| CN106755241B (en) * | 2016-12-28 | 2021-05-04 | 北京化工大学 | Tuna bone collagen polypeptide for promoting bone cell growth and preparation method thereof |
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