CN105002246A - Fish polypeptide with uniform molecular weight distribution, and preparation method thereof - Google Patents
Fish polypeptide with uniform molecular weight distribution, and preparation method thereof Download PDFInfo
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Abstract
The invention provides a preparation method of a fish polypeptide with uniform molecular weight distribution. The preparation method comprises following steps: fish skin is subjected to pretreatment, and multistage tandem enzymolysis for a period of time; a gelatin crude product is obtained via enzyme inactivation; and the fish polypeptide is obtained via extraction; wherein multistage tandem enzymolysis is designed to be 4 to 6 stages, temperature of each stage of enzymolysis is controlled to be 25 to 55 DEG C, pH value is controlled to be 6 to 9, temperature is increased gradually from the first stage to the next stages, and pH value is reduced gradually. Molecular weight distribution of the fish polypeptide is uniform; application range is wide; the fish polypeptide can be used for preparing cosmetics, drugs, and health food; and absorption effect by human body is excellent.
Description
Technical field
The present invention relates to manufacturing of gelatin manufacture field, in particular to fish polypeptide that a kind of molecular weight distribution is homogeneous and preparation method thereof.
Background technology
Gelatin is the water-soluble polypeptide mixture that collagen hydro obtains, polypeptide molecular weight all has distribution in several thousand to tens0000 scopes, outward appearance is translucent thin slice, particulate state or flaxen powder, gelatin has much special physics and chemical property, as jelly power, affinity, high dispersion, low viscosity characteristics, dispersion stabilization, retentiveness, coating property, toughness and reversibility etc., all there is application in many fields such as food, makeup, medicine, papermaking, imagings.
Gelatin in the market mainly with pigskin, pig bone and ox bone for raw material, the frequent people of making of mad cow disease and foot and mouth disease tend to the gelatin using other animal-origin gradually, and the gelatin that the impact of religion also makes Some Animals originate in addition is difficult to extensive popularization.In recent years, it is increasing with fish-skin to be that the research of gelatin prepared by raw material, its major cause is that the raw material of fish-skin is more stable, gelatin and pig, the Ox horn Glue character of preparation are similar, and about fish disease report fewer, therefore those skilled in the art more and more pay close attention to fish-skin be raw material this respect research.
In prior art, the production method of gelatin mainly comprises alkaline process, acid system, salt and alkali method and enzyme process etc.Acid system and alkaline process are generally used because its technique is simple, with low cost, but acid system and alkaline process can produce certain destruction to the molecular structure of final collagen product, especially alkaline process is larger to tropocollagen molecule destructiveness, and cause range of molecular weight distributions too wide, salt and alkali method is more rare.Acid process gelatin is also called A type gelatin, iso-electric point 7.0-9.0; Alkaline process gelatin also known as Type B gelatin, between iso-electric point 4.7-5.0, it is generally acknowledged the plasticity-of A type gelatin and elasticity better, and the hardness of Type B gelatin is better.Although be industrially sometimes used alone A type gelatin or Type B gelatin, most of pharmagel is the mixture of above two type gelatin, and gelatin common on market has different grades, variable grain size and different molecular weight equal-specification.Alkaline process, acid system all can be used for bringing up again from pigskin, pig bone and ox bone getting gelatin, but extraction process there is some difference, acid system is used for extracting gelatin from pigskin, and alkaline process is used for extracting gelatin from bone.
Acid system and alkaline process are generally used because its technique is simple, with low cost, but acid system and alkaline process can produce certain destruction to the molecular structure of final collagen product, especially alkaline process is larger to tropocollagen molecule destructiveness, cause range of molecular weight distributions too wide, even produce carcinogenic substance, if therefore certain harm can be had to human body as edible Gelatinum oxhide.
Enzyme process is the production method of a kind of advanced person, and with proteolytic enzyme directionally hydrolyzing collagen fabric, gained degrade proteins molecular weight meets human body optimal absorption rate, is method leading, the most safe and reliable in the world in recent years.Its product stable, molecular size is homogeneous, maintain the biological activity that the finished product are very high, and edible safety is relieved, but the enzyme process generally adopted is owing to can cause the random degenerate of collagen protein in production process, molecular weight distribution is wider, thus affects the performance of the fish polypeptide that final enzymolysis obtains.
In view of this, special proposition the present invention.
In prior art, the production technique that alkaline process prepares fishskin gelatin is as follows:
(1) raw material of different varieties of classifying needs to be separated; Identical raw material, different tissues, as skin, squama, fish glue etc., needs separately; Different raw materials, identical organizing also will separate.
(2) raw material that rinsing is fresh, as long as can drop into subsequent processing with clear water rinsed clean; Drying raw material clear water processes after soaking fully water suction again; Salted raw material could use after needing immersion desalination.Rinse cycle need change water, prevents bacterial reproduction, affects raw materials quality.
(3) degreasing raw material needs to carry out skimming treatment.Degreasing method generally uses chemical method degreasing, by the calcium salt in tissue, makes soft texture, and colloid expands, in order to carrying glue.Using during acid treatment to prevent precipitation from producing, and changes liquid in time.
(4) liming liming is the very important ring of Feedstock treating, raw material under the effect of alkali, the tissue of the raw material that can not only loosen, and can dissolve and remove the organism that those affect colloid amount, as soluble proteins, pigment etc.
(5) in and raw material deliming, in and, washing raw material through milk of lime soak after, washing and adjust pH.
(6) endure glue glue raw material together with water altogether heat and the process that changes gelatin into be called and endure glue, enduring glue is critical process in manufacturing of gelatin.Collagen is transformed into gelatin, and water molecules entered the space of tropocollagen molecule interchain before this, then makes some splitting of chain of tropocollagen molecule triple helix structure, thus becomes water-soluble gelatin.Accelerate the carrying out of this process.But the hydrolytic action of collagen is not just stop after being transformed into gelatin, but is carrying out with keeping, until generate amino acid.Therefore, must control temperature and time when enduring glue, to obtain high-quality gelatin.Endure light glue at about 60 DEG C, with filter cotton, gac, diatomite etc. as flocculating aids, with plate-and-frame filter press filter, obtain clarification glue.Glue is separated with whizzer again, further except impurity such as degreases.From raw material, extract gelatin, adopt shunting to endure the method for glue.Namely first in glue pot, put into a certain amount of hot water, and drop in pot by processed good raw material, one side slowly heats up, and the raw material after expansion is heated and shrinks gradually and enter out a large amount of water, finally makes the whole raw material of water yield energy submergence added.Then at a certain temperature, collagen is just constantly hydrolyzed into gelatin and water-soluble, like this through a few hours, when reaching finite concentration, is released by light glue in pot, Here it is first glue.And then hot water is added in pot, comparatively front for temperature raising 5-10 DEG C continued to endure glue, this is second, and carry out repeatedly successively, temperature also correspondingly raises gradually, can boil, all to be endured out by the collagen in raw material for last one.Glue process is endured in impact following several factor: the temperature of enduring glue should be low unsuitable high, and temperature is high by impelling collagen hydro to become the speed of gelatin protein to accelerate, and also will speed up the secondary hydrolysis speed of gelatin simultaneously, causes colloid impure.Senior gelatin, endures glue temperature no more than 70 DEG C, because the secondary hydrolysis speed of more than 70 DEG C gelatin will significantly be accelerated; When enduring glue, pH value should be best near the iso-electric point of gelatin protein.At this moment collagen hydro becomes the speed of gelatin and gelatin secondary hydrolysis speed the slowest all, and the glue endured out is best in quality; The time of enduring glue should shortly should not be grown, and the time is longer, and secondary hydrolysis product increases, and makes colloid impure, Quality Down.General control is at 3-8h.
The technique of enzyme process fishskin gelatin is as follows: the fish-skin-pre-treatment-adjustment pH-enzymolysis-survey degree of hydrolysis-enzyme that goes out-hydrolyzed solution-vacuum filtration (gac does flocculating aids)-get gelatin slightly carries product.
To compare enzyme process and alkaline process fishskin gelatin, can find:
(1) homogeneity problem: the molecular weight of gelatin polypeptides all has distribution from several thousand to tens0000, the molecular weight ranges of enzyme process gelatin is lower than the molecular weight ranges of alkaline process gelatin and acid process gelatin in theory, but in preparation of gelatin with enzyme process, substrate molecule weight range is comparatively large, causes the molecular weight ranges of the finished product to be difficult to homogeneous.The gelatin how preparing range of molecular weight distributions relative narrower is the common problem generally preparing gelatin existence at present.
(2) Product Safety: acid system and alkaline process can produce certain destruction to the molecular structure of final collagen product, especially alkaline process is larger to tropocollagen molecule destructiveness, cause producing carcinogenic substance, so generally it goes without doing production food grade collagen protein, and the security of enzyme process gelatin is higher.
(3) production cost: the production cost of gelatin mainly comprises the consumption such as chemical reagent such as water, electricity, soda acid, also needs to add zymin etc. in enzyme process glue process.Comparatively speaking, the temperature in enzyme process glue process is lower, and the consumption of electricity is less.
(4) production cycle: the cycle of Production by Enzymes gelatin was at about 7 days, and alkaline process and acid system prepare the production cycle of gelatin at about 9 days.
(5) technique degree of cleaning: the initial stage of enzyme process adhesive-preparing technology needs to consume a certain amount of bronsted lowry acids and bases bronsted lowry etc., the later stage consumption of enzymolysis reaction reagent is lower, and acid system and enzyme process need a large amount of reagent in production technique, the discharge of waste water is the principal element affecting environment.
Therefore, all there are certain relative merits in various adhesive-preparing technology, and generally speaking, enzyme process adhesive-preparing technology is better than chemical method, is also that fish-skin prepares the focus in gelatin research in recent years.
Although enzyme process fishskin gelatin method belongs to a kind of method optimum in prior art, but the usual way of enzyme process isinglass is generally designed to single-stage enzymolysis, namely under the condition that the temperature in enzymatic vessel and pH value are all determined, proteolytic enzyme is selected to carry out enzymolysis to substrate, such operation due to temperature and pH value all constant, it is also unique that the kind of proteolytic enzyme is chosen, in long-time enzymolysis process, collagen protein can be easy to degrade, the substrate that molecular weight ranges cannot be provided homogeneous for enzyme digestion reaction, also hydrolysis result and enzymolysis product yield can be affected, enzymolysis product is made to be the mixture of the different multiple peptide of molecular size range, mass percentage content shared by tripeptides is only about 10%, residue is tetrapeptide to icosapeptide not etc., the molecular weight of gelatin polypeptides all has distribution from several thousand to tens0000, be made into heath food in the future, medicine or makeup are not easily absorbed by the body, affect product effect, and then affect the promotion efficiency of product.
In order to solve the technical problem of above appearance, enzymolysis process is designed to plural serial stage enzymolysis form by the present invention, each rank is according to the Denaturing of collagen protein and the problem fully taking into account the degraded suppressing collagen protein, choose suitable temperature, pH value, also has the kind of enzyme, the processing parameter indexs such as the consumption of enzyme, by the optimization of these technic indexs, make collagen protein can be dissolved in acid or alkaline solution, but do not degrade, for enzymic degradation reacts the substrate providing molecular weight homogeneous.Utilize the change of this dissolving properties of collagen protein to devise pH and thermograde multiple tank adverse current, finally realize collagen protein and dissolve but do not degrade.Not only increase the yield of enzymolysis product, after testing in fish polypeptide product the mass percentage content of tripeptides more than 70%, molecular weight control is between 700-1000, just because of the molecular weight ranges controlling substrate just can make the molecular weight distribution of the finished product homogeneous.
Wherein, pretreated step, mainly for realizing degreasing and deliming, comprises and fish-skin is passed through the step scaled, clean, soak and pulverize, also can carry out pre-treatment with reference to the operation steps of the enzyme process gelatin of prior art.
The place that should be noted that is, the rank of plural serial stage enzymolysis is set to 4-6 level, then connect between every grade, it is the suitable scope obtained by a large amount of experiments that the temperature of each rank concrete and pH choose, and first level sets to other temperature of next stage according to the trend raised gradually, pH value is then in the trend reduced gradually, because found by a large amount of practical studies, in the basic conditions, the denaturation temperature of collagen protein raises with pH and reduces, under acid washing conditions, the denaturation temperature of collagen protein reduces with pH and reduces, its sex change critical temperature is close to 60 DEG C in neutral conditions, be such as under the condition of 8.0 at pH, the sex change critical temperature of collagen protein is 53 DEG C.Therefore pH and choosing of temperature will match, and according to pH along with the increase of rank reduces gradually, the rule that temperature raises gradually along with the increase of rank carries out gradient selection, the quality of guarantee enzymolysis product.
In order to be improved the quality of products by Optimization Technology index, the rank of plural serial stage enzymolysis is preferably set to 5 grades, and the temperature range of each rank controls between 35-55 DEG C, and pH controls between 7-8.The enzymolysis time altogether of plural serial stage enzymolysis is 10-24h, preferred 15-20h.Further, the consumption of the enzyme that each rank is used, preferably in suitable scope, for being the 0.5-2% through pretreated fish-skin quality, is preferably 1-1.5%.
In addition, choosing of the kind of enzyme is also important, the enzyme used is preferably Sumizyme MP race from bacterial origin, comprises each rank enzyme used and comprises the mixture of one or more in B group enzyme, neutral protease and the aspartic protease that Bacillus subtilus belongs to.Wherein, described neutral protease comprises the mixture of one or more in the proteolytic enzyme of grey strepto-secretion, wooden pawl proteolytic enzyme, ficoin and Traumanase.Described aspartic protease comprises from the oozy proteolytic enzyme of the mixture of one or more in aspergillus niger, mould and brown aspergillus.
Finally, for the manufacture of the raw material mainly fish-skin of isinglass, comprise band squama or the fish-skin of scaling, and band fin or fish-skin not with fin.The kind of fish has: Japanese eel, tuna, extra large meals, sturgeon, shark, cod, catfish, lung fish, carp, Buddhist nun sieve perch, abalone, mackerel (Spanish mackerel, mackerel), east squid, salmon (Sa Menyu, salmon), chopsticks sole (tongue sole) etc.These fish-skins are (comprising fish scale, air bladder, fin, fish-bone) tankage of fish processing industry.
The fish polypeptide prepared by above-mentioned preparation method, molecular weight is between 700-1000, major part is collagen tripeptide, molecular weight distribution is homogeneous, make medicine, heath food and cosmetic effect good, fully by human body is absorbed its nutritive element, the demand of different crowd can be suitable for, can be applicable by great dynamics, very there is commercial application value.
Compared with prior art, beneficial effect of the present invention is:
(1) preparation method of the homogeneous fish polypeptide of a kind of molecular weight distribution is embodiments provided, preparation method is simple, it is low and easy to operate to invest, the molecular weight ranges achieving product fish polypeptide gelatin is controlled, and pollution-free, the abundant environmental protection of three-waste free discharge in short production cycle and production process;
(2) creationary on the basis of preparing isinglass at existing enzyme process single-stage enzymolysis process is designed to plural serial stage enzymolysis process, the kind of the temperature of each rank enzymolysis, pH and enzyme, consumption be the suitable scope by great many of experiments optimization all, the distribution of its component is homogeneous after testing for enzymolysis product, for follow-up the method is promoted, provide can the data of reference, have certain directive significance;
(3) also as in the present invention, narrow molecular-weight is not had and the appearance of the product of the fish polypeptide of homogeneous molecular weight distribution in prior art, the present invention has all started the beginning in preparation method and product performance optimization, in gelatin synthesis manufacture field, still belong to the first, and the equipment to match with preparation technology is common equipment, floor space is little, less investment, whole process simple to operate can realize Automated condtrol, uses manpower and material resources sparingly;
(4) the fish polypeptide that obtains of the preparation method of the embodiment of the present invention, molecular weight is between 700-1000, major part is collagen tripeptide, molecular weight distribution is homogeneous, makes medicine, heath food and cosmetic effect good, can fully by human body is absorbed its nutritive element, edible green health safety, be suitable for the demand of different crowd, can be applicable by great dynamics, very there is commercial application value.
Summary of the invention
The first object of the present invention is the preparation method of the fish polypeptide providing a kind of molecular weight distribution homogeneous, to solve the technical problem of above-mentioned appearance, there is the possibility that production cost is low, reduce the degraded of stripping collagen protein, the fish peptide molecule weight range prepared is homogeneous, edible safety coefficient is high, the advantage such as pollution-free in short production cycle and production process, abundant environmental protection of three-waste free discharge.
The second object of the present invention is the fish polypeptide providing a kind of molecular weight distribution obtained by above-mentioned preparation method homogeneous, the molecular weight of this fish polypeptide is controlled, there is homogeneous molecular weight distribution, applied widely, inhibition and generation cosmetic, medicine and protective foods can be used for, the advantages such as absorption of human body is effective.
In order to realize above-mentioned purpose of the present invention, spy by the following technical solutions:
The invention provides a kind of method preparing the homogeneous fish polypeptide of molecular weight distribution, comprise the steps: that, by after fish-skin pre-treatment, through plural serial stage enzymolysis for some time, then obtain gelatin crude product after making enzyme deactivation, namely extracting obtains fish polypeptide;
Wherein, the rank of plural serial stage enzymolysis is set to 4-6 level, and the temperature range of each rank controls between 25-55 DEG C, and pH controls between 6-9, and other temperature raises gradually from first level to next stage, and pH value then reduces gradually.
Embodiment
Below in conjunction with embodiment, embodiment of the present invention are described in detail, but it will be understood to those of skill in the art that the following example only for illustration of the present invention, and should not be considered as limiting the scope of the invention.Unreceipted actual conditions person in embodiment, the condition of conveniently conditioned disjunction manufacturers suggestion is carried out.Agents useful for same or the unreceipted production firm person of instrument, be and can buy by commercially available the conventional products obtained.
Embodiment 1
The preparation method of fish polypeptide is as follows:
1) fish-skin of Japanese eel is carried out pre-treatment: clean up after first being scaled by fish-skin, soak with clear water to make it fully absorb water, carry out swelling with re-using acid treatment after saline soak, soak through milk of lime and wash, should be noted during immersion and change liquid in time, to prevent the generation of bacterial reproduction and precipitation, fish skin raw material is pulverized after taking off ester step through above-mentioned deliming, with for subsequent use;
2) mode of action of 4 enzymatic vessel plural serial stages is adopted to the fish-skin enzymolysis after pulverizing to reduce the degraded of the collagen protein of stripping, the temperature of each enzymatic vessel controls between 25-55 DEG C, pH controls between 6-9, and set from the first enzymatic vessel to the temperature of the 4th enzymatic vessel according to the trend raised gradually, pH value then according to the trend reduced gradually, the mixture of one or more in the proteolytic enzyme being chosen for the secretion of grey strepto-of the enzyme in each enzymatic vessel, wooden pawl proteolytic enzyme, ficoin and Traumanase;
3) altogether enzyme heat inactivation is made to obtain gelatin crude product after enzymolysis 10h, then after the step of concentrated, decolouring, dry and sterilizing, obtain finished product, the molecular weight distribution of fish polypeptide finished product is between 700-1000, and the mass percentage content of tripeptides is more than 70%.
Embodiment 2
The preparation method of fish polypeptide is as follows:
1) fish-skin of Japanese eel is carried out pre-treatment: clean up after first being scaled by fish-skin, soak with clear water to make it fully absorb water, carry out swelling with re-using acid treatment after saline soak, soak through milk of lime and wash, should be noted during immersion and change liquid in time, to prevent the generation of bacterial reproduction and precipitation, fish skin raw material is pulverized after taking off ester step through above-mentioned deliming, with for subsequent use;
2) mode of action of 5 enzymatic vessel plural serial stages is adopted to the fish-skin enzymolysis after pulverizing to reduce the degraded of the collagen protein of stripping, the temperature of each enzymatic vessel controls between 35-55 DEG C, pH controls between 7-8, and set from the first enzymatic vessel to the temperature of the 5th enzymatic vessel according to the trend raised gradually, pH value is then according to the trend reduced gradually, enzyme in each enzymatic vessel be chosen for from aspergillus niger, the oozy proteolytic enzyme of the mixture of one or more in mould and brown aspergillus, the consumption of the enzyme in each enzymatic vessel is 0.5% of the fish-skin quality after pulverizing,
3) altogether after enzymolysis 24h, enzyme heat inactivation is made to obtain gelatin crude product again, then irradiate after with the step of sterilizing through concentrated, activated carbon filtration decolouring, drying and Co 60, obtain finished product, the molecular weight distribution of fish polypeptide finished product is between 700-1000, and the mass percentage content of tripeptides is more than 70%.
Embodiment 3
The preparation method of fish polypeptide is as follows:
1) fish-skin of Japanese eel is carried out pre-treatment: clean up after first being scaled by fish-skin, soak with clear water to make it fully absorb water, carry out swelling with re-using acid treatment after saline soak, soak through milk of lime and wash, should be noted during immersion and change liquid in time, to prevent the generation of bacterial reproduction and precipitation, fish skin raw material is pulverized after taking off ester step through above-mentioned deliming, with for subsequent use;
2) mode of action of 6 enzymatic vessel plural serial stages is adopted to the fish-skin enzymolysis after pulverizing to reduce the degraded of the collagen protein of stripping, the temperature of each enzymatic vessel controls between 35-55 DEG C, pH controls between 7-8, and set from the first enzymatic vessel to the temperature of the 6th enzymatic vessel according to the trend raised gradually, pH value is then according to the trend reduced gradually, the B group enzyme being chosen for Bacillus subtilus genus of the enzyme in each enzymatic vessel, the mixture of one or more in neutral protease and aspartic protease, the consumption of the enzyme in each enzymatic vessel is 2% of the fish-skin quality after pulverizing,
3) altogether after enzymolysis 15h, enzyme heat inactivation is made to obtain gelatin crude product again, then irradiate after with the step of sterilizing through concentrated, activated carbon filtration decolouring, drying and Co 60, obtain finished product, the molecular weight distribution of fish polypeptide finished product is between 700-1000, and the mass percentage content of tripeptides is more than 70%.
Embodiment 4
The preparation method of fish polypeptide is as follows:
1) fish-skin of shark is carried out pre-treatment: clean up after first being scaled by fish-skin, soak with clear water to make it fully absorb water, carry out swelling with re-using acid treatment after saline soak, soak through milk of lime and wash, should be noted during immersion and change liquid in time, to prevent the generation of bacterial reproduction and precipitation, fish skin raw material is pulverized after taking off ester step through above-mentioned deliming, with for subsequent use;
2) mode of action of 5 enzymatic vessel plural serial stages is adopted to the fish-skin enzymolysis after pulverizing to reduce the degraded of the collagen protein of stripping, the temperature of each enzymatic vessel controls between 35-55 DEG C, pH controls between 7-8, , and set from the first enzymatic vessel to the temperature of the 5th enzymatic vessel according to the trend raised gradually, pH value is then according to the trend reduced gradually, the B group enzyme being chosen for Bacillus subtilus genus of the enzyme in each enzymatic vessel, the mixture of one or more in neutral protease and aspartic protease, the consumption of the enzyme in each enzymatic vessel is the 1-1.5% of the fish-skin quality after pulverizing,
3) altogether after enzymolysis 20h, enzyme heat inactivation is made to obtain gelatin crude product again, then irradiate after with the step of sterilizing through concentrated, activated carbon filtration decolouring, drying and Co 60, obtain finished product, the molecular weight distribution of fish polypeptide finished product is between 700-1000, and the mass percentage content of tripeptides is more than 70%.
Comparative example 1
The preparation method of fish polypeptide is as follows:
1) fish-skin of shark is carried out pretreated step substantially identical with the pre-treatment step of embodiment 4, with for subsequent use;
2) enzymatic vessel is adopted to carry out enzymolysis to the fish-skin after pulverizing, the temperature of enzymatic vessel controls between 35-55 DEG C, pH controls between 7-8, the mixture of one or more be chosen in B group enzyme that Bacillus subtilus belongs to, neutral protease and aspartic protease of the enzyme in enzymatic vessel, the consumption of the enzyme in enzymatic vessel is the 1-1.5% of the fish-skin quality after pulverizing;
3) altogether after enzymolysis 20h, enzyme heat inactivation is made to obtain gelatin crude product again, then irradiate after with the step of sterilizing through concentrated, activated carbon filtration decolouring, drying and Co 60, obtain finished product, the molecular weight distribution of fish polypeptide finished product is between 4000-40000, and the mass percentage content of tripeptides is 10%.
Although illustrate and describe the present invention with specific embodiment, however it will be appreciated that can to make when not deviating from the spirit and scope of the present invention many other change and amendment.Therefore, this means to comprise all such changes and modifications belonged in the scope of the invention in the following claims.
Claims (10)
1. a preparation method for the fish polypeptide that molecular weight distribution is homogeneous, is characterized in that, comprise the steps:
After fish-skin pre-treatment, through plural serial stage enzymolysis for some time, then obtain gelatin crude product after making enzyme deactivation, namely extracting obtains fish polypeptide;
Wherein, the rank of plural serial stage enzymolysis is set to 4-6 level, and the temperature range of each rank enzymolysis controls between 25-55 DEG C, and pH controls between 6-9, and other temperature raises gradually from first level to next stage, and pH value then reduces gradually.
2. the preparation method of the fish polypeptide that molecular weight distribution according to claim 1 is homogeneous, is characterized in that, the rank of plural serial stage enzymolysis is set to 5 grades, and the temperature range of each rank controls between 35-55 DEG C, and pH controls between 7-8.
3. the preparation method of the fish polypeptide that molecular weight distribution according to claim 1 is homogeneous, is characterized in that, the enzymolysis time altogether of plural serial stage enzymolysis is 10-24h.
4. the preparation method of the fish polypeptide that molecular weight distribution according to claim 3 is homogeneous, is characterized in that, the enzymolysis time altogether of plural serial stage enzymolysis is 15-20h.
5. the preparation method of the fish polypeptide that molecular weight distribution according to claim 1 is homogeneous, is characterized in that, during each rank enzymolysis, the quality of enzyme used is the 0.5-2% through pretreated fish-skin quality, is preferably 1-1.5%.
6. the preparation method of the fish polypeptide that molecular weight distribution according to claim 1 is homogeneous, is characterized in that, each rank enzyme used comprises the mixture of one or more in B group enzyme, neutral protease and the aspartic protease that Bacillus subtilus belongs to.
7. the preparation method of the fish polypeptide that molecular weight distribution according to claim 6 is homogeneous, is characterized in that, described neutral protease comprises the mixture of one or more in the proteolytic enzyme of grey strepto-secretion, wooden pawl proteolytic enzyme, ficoin and Traumanase.
8. the preparation method of the fish polypeptide that molecular weight distribution according to claim 6 is homogeneous, is characterized in that, described aspartic protease comprises from the oozy proteolytic enzyme of the mixture of one or more in aspergillus niger, mould and brown aspergillus.
9. the preparation method of the fish polypeptide that molecular weight distribution according to claim 1 is homogeneous, is characterized in that, pretreated step comprises: by the step of fish-skin through scaling, cleaning, soak and pulverizing.
10. the fish polypeptide prepared by the preparation method of the homogeneous fish polypeptide of the molecular weight distribution described in any one of claim 1-9.
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Application publication date: 20151028 |