CN105506043A - Sea eel small molecular peptide extracted from sea eel bones and extraction method thereof - Google Patents
Sea eel small molecular peptide extracted from sea eel bones and extraction method thereof Download PDFInfo
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Abstract
The invention relates to a method for extracting sea eel small molecular active peptide from sea eel bones. The method comprises the following steps of 1, sea eel bone processing, 2, low-temperature smashing, 3, cell dissolution processing, 4, biological enzymolysis processing and 5 postprocessing; the ratio of the obtained sea eel small molecular peptide with the molecular weight being 0-480 d is 88.5% or more; the sea eel small molecular peptide is applicable to a peptide for diagnosis, a peptide vaccine, an antibacterial activity peptide, a cell factor simulation peptide, a cell factor simulation peptide, an antiviral peptide or an anti-tumor peptide and a peptide health care product.
Description
Technical field
The present invention relates to the technology extracting peptide with fish-bone, be specifically related to a kind of method extracting small-molecular peptides from eel bone.
Background technology
The collagen protein extracted from animal such as yak bone has certain limitation in specific absorption, and along with the risk brought mad cow disease, make this comparatively traditional mode not approve by people, and people more and more approve extract corresponding nutritive substance from fish.
Current extraction mainly extracts collagen protein from fish scale, fish-skin, such as extract from tilapia fish scale, extract from deep-sea fish skin such as cod fish-skin, but these extract product molecular weight distribution extensively not easily absorb, the existence of the outstanding problems such as the specific absorption that macromole causes is low, what prevent modern to pursue greatly is of high nutritive value, safety, the realization of the new health care theory such as good absorption.
On the other hand, traditional collagen protein or protein peptide extracting liquid volume are comparatively large, and the production cycle of product is longer, and enzymolysis solution removal of impurities is not thorough, affect collagen protein quality, gained finished product often has bitter off-flavors, and mouthfeel is poor, if filtration rank is inadequate, product water dissolubility will be poor, direct impact is digested and assimilated, and the Resolving probiems effect high to the saltiness of fish extract is undesirable, affects products taste.
Although along with technical progress, people use the advantage of membrane separation technique, improve quality product to a certain extent, also fundamentally do not obtain small active peptides.
Summary of the invention
The present invention, for effectively to solve the problem, provides a kind of brand-new preparation method, take eel bone as raw material, obtain being easy to absorb by innovation means, mouthfeel is good, is of high nutritive value, the eel small active peptides of safety, and the method extraction efficiency is high, technique simply uses.
The present invention adopts the biological enzymolysis of improvement to modify method of double crossing, preparation eel small active peptides, concrete technical scheme is, a kind of method extracting eel small active peptides from eel bone, comprise the steps: (1) eel bone process: cleaned up by fresh eel bone, broken, in digester, employing temperature is the temperature of 120-150 DEG C, the pressure boiling 6-10h of 0.13-0.15Mpa, and bone slag is discharged, obtain bone soup, after quick-frozen, form frozen bone soup oil and freeze block, for subsequent use in the freezer being stored in-18 DEG C; (2) pulverize at low temperature process: adopt pulverizer to freeze block at low temperature to described frozen bone soup oil and carry out pulverize at low temperature, pulverizing temperature controls between 0-4 DEG C, described frozen bone soup oil is frozen block pulverizing and freezes block particle for particle diameter at the frozen bone soup oil of 0.02-0.1 millimeter; (3) cell stripping process: adopt the lower 4 DEG C of oxygen-free waters of normal pressure to freeze block particle to the frozen bone soup oil pulverized and carry out infiltration water treatment, the water saturation time is 6-8 hour, and described oxygen-free water is as the criterion to flood raw material completely; Then fast deep is freezing with thawing process, wherein the Deep-Frozen treatment time is 1-2h, and freezing temp is subzero 22-25 DEG C, and then thawing, namely freeze block particle with the lower 4 DEG C of water-baths of normal pressure to the frozen bone soup oil being in Deep-Frozen and carry out quick ice-melt, the ice-melt treatment time is 3-5h; Freezing with thawing treating processes 2-3 time of fast deep described in repetition, cell wall structure is destroyed, active substance stripping, forms eel oligopeptides bone soup; (4) biological enzymolysis process: described eel oligopeptides bone soup is imported in hydrolytic decomposition pot, add prozyme, described prozyme preparation method is as follows, first careless genus bacillus, bifid Helicobacter pylori, lactobacillus rhamnosus, photosynthetic bacteria, Bacillus licheniformis are placed in triangular flask and cultivate, the temperature of cultivating is 28-33 DEG C, under 200-250rpm/min, cultivate 25-35h, make thalline or spore concentration reach 1 × 10
14individual/liter (CFU/L), obtained starter, then in starter, nutrient substance is added, the weight controlling nutrient substance is the 1/5-1/8 of described starter weight, described nutrient substance is wort, namely described prozyme is obtained, add described prozyme according to the weight ratio of eel oligopeptides bone soup and prozyme 1000-1200: 1 and carry out biological enzymolysis process, in described biological enzymolysis process, keep pH in hydrolytic decomposition pot to be 6.8-7.0, described hydrolysis temperature is 60 DEG C, described enzymolysis time is 13-21 DEG C, obtains elementary eel small active peptides; (5) aftertreatment: go out described elementary eel small active peptides enzyme, and be rapidly heated to 180-185 DEG C and keep 20-25s, then 50-60 DEG C/s is cooled to 60 DEG C and makes enzyme deactivation; Then supercentrifuge is adopted by enzymolysis solution with the centrifugation 3-5min of 60000-80000r/min, upper liquid and middle level liquid are merged in input dilution tank body, i.e. enzymolysis parting liquid, subnatant is separated, then adopt hot barrier film cryoconcentration technology to concentrate, thickening temperature is 45-50 DEG C, and vacuum tightness is 0.4-2.0MPa, centrifugal spray drying tower inner drying concentrated solution being transported to 200-220 DEG C becomes powder, namely obtains described eel small active peptides.
Preferably, adopt in step (1) temperature to be the temperature of 130 DEG C, the pressure boiling 7h of 0.14Mpa, bone slag is discharged, and obtains bone soup, after quick-frozen, forms frozen bone soup oil and freezes block, for subsequent use in the freezer being stored in-18 DEG C; Step 2) in, pulverize temperature and control at 2 DEG C, described frozen bone soup oil is frozen block and pulverizes as particle diameter freezes block particle at the frozen bone soup oil of 0.05 millimeter.
Preferably, cell stripping process in step (3): adopt the lower 4 DEG C of oxygen-free waters of normal pressure to freeze block particle to the frozen bone soup oil pulverized and carry out infiltration water treatment, the water saturation time is 7 hours, and described oxygen-free water is as the criterion to flood raw material completely; Then fast deep is freezing with thawing process, wherein the Deep-Frozen treatment time is 2h, and freezing temp is subzero 25 DEG C, and then thawing, namely freeze block particle with the lower 4 DEG C of water-baths of normal pressure to the frozen bone soup oil being in Deep-Frozen and carry out quick ice-melt, the ice-melt treatment time is 3h; Freezing with thawing treating processes 3 times of fast deep described in repetition, cell wall structure is destroyed, active substance stripping, forms eel oligopeptides bone soup.
Preferably, step is cultivated in (4) in triangular flask, and the temperature of cultivation is 30 DEG C, under 200rpm/min, cultivate 28h, makes thalline or spore concentration reach 1 × 10
14individual/liter (CFU/L), add described prozyme according to the weight ratio of eel oligopeptides bone soup and prozyme 1000: 1 and carry out biological enzymolysis process, in described biological enzymolysis process, keep pH in hydrolytic decomposition pot to be 7.0, described hydrolysis temperature is 60 DEG C, described enzymolysis time is 21 DEG C, obtains elementary eel small active peptides.
Preferably, aftertreatment in step (5): go out described elementary eel small active peptides enzyme, and be rapidly heated to 180 DEG C and keep 25s, then 60 DEG C/s is cooled to 60 DEG C and makes enzyme deactivation; Then supercentrifuge is adopted by enzymolysis solution with the centrifugation 4-5min of 60000-70000r/min, upper liquid and middle level liquid are merged in input dilution tank body, i.e. enzymolysis parting liquid, subnatant is separated, then adopt hot barrier film cryoconcentration technology to concentrate, thickening temperature is 45 DEG C, and vacuum tightness is 2.0MPa, centrifugal spray drying tower inner drying concentrated solution being transported to 200 DEG C becomes powder, namely obtains described eel small active peptides.More preferably supercentrifuge by enzymolysis solution with the centrifugation 4min of 60000r/min.
The described eel small active peptides that the present invention also provides aforesaid method to prepare, the molecular weight of described eel small-molecular peptides is that the daltonian accounting of 0-480 is no less than 88.2%, the molecular weight of concrete described eel small-molecular peptides is the daltonian accounting of 0-480 is 88.61%, the daltonian accounting of 480-1000 is 9.43%, being greater than 1000 daltonian accountings is 1.96%, and color and luster is faint yellow.
The present invention also provides the application of the above-mentioned eel small-molecular peptides stated; Described eel small-molecular peptides is applicable to diagnosis peptide, peptide vaccine, antibacterial active peptide, cytokine simulating peptide, antiviral peptide or natineoplaston, and peptide health product.
Positively effect of the present invention:
First the eel micromolecule active polypeptide prepared of the present invention is pure, be better than the polypeptide prepared in prior art, eel micromolecule active polypeptide content prepared by the present invention is close to 100%, the molecular weight of described eel small-molecular peptides is that the daltonian accounting of 0-480 is no less than 88.2%, the molecular weight of concrete described eel small-molecular peptides is the daltonian accounting of 0-480 is 88.61%, the daltonian accounting of 480-1000 is 9.43%, and being greater than 1000 daltonian accountings is 1.96%, and color and luster is faint yellow.
Secondly adopt bioprotein enzymolysis technology that eel bone feed degradation is become collagen helical peptide; Adopt ultra micro film and cryoconcentration technology separation eel small active peptides, product has stability, oxidation-resistance, easily preserves, easily be absorbed by the body, the eel small active peptides powder obtained both can be washed by water can, capsule can be made again, people are made to obtain health-care effect in daily life process, existing pharmaceutical use, nutritious again.Adopt biotechnology, comprehensively employ cell stripping process, biological enzymolysis process and separation and purification, be condensed into peptide biological health-care product, for eel bone provides new approach.Eel small-molecular peptides prepared by the present invention, can be applied to peptide medicament and peptide class healthcare products, and range of application is relatively more extensive, such as, for diagnosis peptide, peptide vaccine, antibacterial active peptide, cytokine simulating peptide, antiviral peptide and/or natineoplaston etc.Eel small-molecular peptides prepared by the present invention meets the use needs of growing medicine and healthcare products, can prepare use on a large scale safely.
Embodiment
Below in conjunction with embodiment, the present invention is described in further details.
Embodiment 1
A kind of method extracting eel small active peptides from eel bone, comprise the steps: (1) eel bone process: cleaned up by fresh eel bone, fragmentation, in digester, employing temperature is the temperature of 120 DEG C, the pressure boiling 10h of 0.13Mpa, bone slag is discharged, and obtains bone soup, after quick-frozen, form frozen bone soup oil and freeze block, for subsequent use in the freezer being stored in-18 DEG C; (2) pulverize at low temperature process: adopt pulverizer to freeze block at low temperature to described frozen bone soup oil and carry out pulverize at low temperature, pulverizing temperature controls at 4 DEG C, described frozen bone soup oil is frozen block and pulverizes as particle diameter freezes block particle at the frozen bone soup oil of 0.02 millimeter; (3) cell stripping process: adopt the lower 4 DEG C of oxygen-free waters of normal pressure to freeze block particle to the frozen bone soup oil pulverized and carry out infiltration water treatment, the water saturation time is 8 hours, and described oxygen-free water is as the criterion to flood raw material completely; Then fast deep is freezing with thawing process, wherein the Deep-Frozen treatment time is 2h, and freezing temp is subzero 22 DEG C, and then thawing, namely freeze block particle with the lower 4 DEG C of water-baths of normal pressure to the frozen bone soup oil being in Deep-Frozen and carry out quick ice-melt, the ice-melt treatment time is 3h; Freezing with thawing treating processes 2 times of fast deep described in repetition, cell wall structure is destroyed, active substance stripping, forms eel oligopeptides bone soup; (4) biological enzymolysis process: described eel oligopeptides bone soup is imported in hydrolytic decomposition pot, add prozyme, described prozyme preparation method is as follows, first careless genus bacillus, bifid Helicobacter pylori, lactobacillus rhamnosus, photosynthetic bacteria, Bacillus licheniformis are placed in triangular flask and cultivate, the temperature of cultivating is 33 DEG C, under 200rpm/min, cultivate 35h, make thalline or spore concentration reach 1 × 10
14individual/liter (CFU/L), obtained starter, then in starter, nutrient substance is added, the weight controlling nutrient substance is 1/5 of described starter weight, described nutrient substance is wort, namely described prozyme is obtained, add described prozyme according to the weight ratio of eel oligopeptides bone soup and prozyme 1000: 1 and carry out biological enzymolysis process, in described biological enzymolysis process, keep pH in hydrolytic decomposition pot to be 7.0, described hydrolysis temperature is 60 DEG C, described enzymolysis time is 21 DEG C, obtains elementary eel small active peptides; (5) aftertreatment: go out described elementary eel small active peptides enzyme, and be rapidly heated to 185 DEG C and keep 25s, then 56 DEG C/s is cooled to 60 DEG C and makes enzyme deactivation; Then supercentrifuge is adopted by enzymolysis solution with the centrifugation 3min of 60000r/min, upper liquid and middle level liquid are merged in input dilution tank body, i.e. enzymolysis parting liquid, subnatant is separated, then adopt hot barrier film cryoconcentration technology to concentrate, thickening temperature is 45 DEG C, and vacuum tightness is 2.0MPa, centrifugal spray drying tower inner drying concentrated solution being transported to 200 DEG C becomes powder, namely obtains described eel small active peptides.
Embodiment 2
A kind of method extracting eel small active peptides from eel bone, comprise the steps: (1) eel bone process: cleaned up by fresh eel bone, fragmentation, in digester, employing temperature is the temperature of 120 DEG C, the pressure boiling 10h of 0.13Mpa, bone slag is discharged, and obtains bone soup, after quick-frozen, form frozen bone soup oil and freeze block, for subsequent use in the freezer being stored in-18 DEG C; (2) pulverize at low temperature process: adopt pulverizer to freeze block at low temperature to described frozen bone soup oil and carry out pulverize at low temperature, pulverizing temperature controls at 0 DEG C, described frozen bone soup oil is frozen block and pulverizes as particle diameter freezes block particle at the frozen bone soup oil of 0.05 millimeter; (3) cell stripping process: adopt the lower 4 DEG C of oxygen-free waters of normal pressure to freeze block particle to the frozen bone soup oil pulverized and carry out infiltration water treatment, the water saturation time is 8 hours, and described oxygen-free water is as the criterion to flood raw material completely; Then fast deep is freezing with thawing process, wherein the Deep-Frozen treatment time is 1h, and freezing temp is subzero 25 DEG C, and then thawing, namely freeze block particle with the lower 4 DEG C of water-baths of normal pressure to the frozen bone soup oil being in Deep-Frozen and carry out quick ice-melt, the ice-melt treatment time is 5h; Freezing with thawing treating processes 3 times of fast deep described in repetition, cell wall structure is destroyed, active substance stripping, forms eel oligopeptides bone soup; (4) biological enzymolysis process: described eel oligopeptides bone soup is imported in hydrolytic decomposition pot, add prozyme, described prozyme preparation method is as follows, first careless genus bacillus, bifid Helicobacter pylori, lactobacillus rhamnosus, photosynthetic bacteria, Bacillus licheniformis are placed in triangular flask and cultivate, the temperature of cultivating is 33 DEG C, under 250rpm/min, cultivate 25h, make thalline or spore concentration reach 1 × 10
14individual/liter (CFU/L), obtained starter, then in starter, nutrient substance is added, the weight controlling nutrient substance is 1/7 of described starter weight, described nutrient substance is wort, namely described prozyme is obtained, add described prozyme according to the weight ratio of eel oligopeptides bone soup and prozyme 1200: 1 and carry out biological enzymolysis process, in described biological enzymolysis process, keep pH in hydrolytic decomposition pot to be 6.8, described hydrolysis temperature is 60 DEG C, described enzymolysis time is 13 DEG C, obtains elementary eel small active peptides; (5) aftertreatment: go out described elementary eel small active peptides enzyme, and be rapidly heated to 185 DEG C and keep 20s, then 50 DEG C/s is cooled to 60 DEG C and makes enzyme deactivation; Then supercentrifuge is adopted by enzymolysis solution with the centrifugation 5min of 80000r/min, upper liquid and middle level liquid are merged in input dilution tank body, i.e. enzymolysis parting liquid, subnatant is separated, then adopt hot barrier film cryoconcentration technology to concentrate, thickening temperature is 50 DEG C, and vacuum tightness is 0.4MPa, centrifugal spray drying tower inner drying concentrated solution being transported to 200 DEG C becomes powder, namely obtains described eel small active peptides.
Embodiment 3
A kind of method extracting eel small active peptides from eel bone, comprise the steps: (1) eel bone process: cleaned up by fresh eel bone, fragmentation, in digester, employing temperature is the temperature of 125 DEG C, the pressure boiling 8h of 0.14Mpa, bone slag is discharged, and obtains bone soup, after quick-frozen, form frozen bone soup oil and freeze block, for subsequent use in the freezer being stored in-18 DEG C; (2) pulverize at low temperature process: adopt pulverizer to freeze block at low temperature to described frozen bone soup oil and carry out pulverize at low temperature, pulverizing temperature controls at 2 DEG C, described frozen bone soup oil is frozen block and pulverizes as particle diameter freezes block particle at the frozen bone soup oil of 0.04 millimeter; (3) cell stripping process: adopt the lower 4 DEG C of oxygen-free waters of normal pressure to freeze block particle to the frozen bone soup oil pulverized and carry out infiltration water treatment, the water saturation time is 7 hours, and described oxygen-free water is as the criterion to flood raw material completely; Then fast deep is freezing with thawing process, wherein the Deep-Frozen treatment time is 1.5h, and freezing temp is subzero 24 DEG C, and then thawing, namely freeze block particle with the lower 4 DEG C of water-baths of normal pressure to the frozen bone soup oil being in Deep-Frozen and carry out quick ice-melt, the ice-melt treatment time is 4h; Freezing with thawing treating processes 2 times of fast deep described in repetition, cell wall structure is destroyed, active substance stripping, forms eel oligopeptides bone soup; (4) biological enzymolysis process: described eel oligopeptides bone soup is imported in hydrolytic decomposition pot, add prozyme, described prozyme preparation method is as follows, first careless genus bacillus, bifid Helicobacter pylori, lactobacillus rhamnosus, photosynthetic bacteria, Bacillus licheniformis are placed in triangular flask and cultivate, the temperature of cultivating is 29 DEG C, under 220rpm/min, cultivate 27h, make thalline or spore concentration reach 1 × 10
14individual/liter (CFU/L), obtained starter, then in starter, nutrient substance is added, the weight controlling nutrient substance is 1/6 of described starter weight, described nutrient substance is wort, namely described prozyme is obtained, add described prozyme according to the weight ratio of eel oligopeptides bone soup and prozyme 1000-1200: 1 and carry out biological enzymolysis process, in described biological enzymolysis process, keep pH in hydrolytic decomposition pot to be 6.8-7.0, described hydrolysis temperature is 60 DEG C, described enzymolysis time is 13-21 DEG C, obtains elementary eel small active peptides; (5) aftertreatment: go out described elementary eel small active peptides enzyme, and be rapidly heated to 1825 DEG C and keep 23s, then 55 DEG C/s is cooled to 60 DEG C and makes enzyme deactivation; Then supercentrifuge is adopted by enzymolysis solution with the centrifugation 3-5min of 70000r/min, upper liquid and middle level liquid are merged in input dilution tank body, i.e. enzymolysis parting liquid, subnatant is separated, then adopt hot barrier film cryoconcentration technology to concentrate, thickening temperature is 46 DEG C, and vacuum tightness is 0.45MPa, centrifugal spray drying tower inner drying concentrated solution being transported to 210 DEG C becomes powder, namely obtains described eel small active peptides.
The eel small molecule active peptide content obtained in above-described embodiment 1-3 is close to 100%, and the molecular weight of described eel small-molecular peptides is the daltonian accounting of 0-480 is 88.64%, 88.66%, 88.78%, and color and luster is faint yellow.
Above-described embodiment, only for technical conceive of the present invention and feature are described, its object is to person skilled in the art can be understood content of the present invention and implement according to this, can not limit the scope of the invention with this.All equivalences done according to spirit of the present invention change or modify, and all should be encompassed within protection scope of the present invention.
Claims (8)
1. from eel bone, extract a method for eel small active peptides, it is characterized in that, comprise the steps: (1) eel bone process; (2) pulverize at low temperature process; (3) cell stripping process; (4) biological enzymolysis process; (5) aftertreatment.
2. the method for claim 1, it is characterized in that, specifically comprise the steps: (1) eel bone process: cleaned up by fresh eel bone, broken, in digester, employing temperature is the temperature of 120-150 DEG C, the pressure boiling 6-10h of 0.13-0.15Mpa, and bone slag is discharged, obtain bone soup, after quick-frozen, form frozen bone soup oil and freeze block, for subsequent use in the freezer being stored in-18 DEG C; (2) pulverize at low temperature process: adopt pulverizer to freeze block at low temperature to described frozen bone soup oil and carry out pulverize at low temperature, pulverizing temperature controls between 0-4 DEG C, described frozen bone soup oil is frozen block pulverizing and freezes block particle for particle diameter at the frozen bone soup oil of 0.02-0.1 millimeter; (3) cell stripping process: adopt the lower 4 DEG C of oxygen-free waters of normal pressure to freeze block particle to the frozen bone soup oil pulverized and carry out infiltration water treatment, the water saturation time is 6-8 hour, and described oxygen-free water is as the criterion to flood raw material completely; Then fast deep is freezing with thawing process, wherein the Deep-Frozen treatment time is 1-2h, and freezing temp is subzero 22-25 DEG C, and then thawing, namely freeze block particle with the lower 4 DEG C of water-baths of normal pressure to the frozen bone soup oil being in Deep-Frozen and carry out quick ice-melt, the ice-melt treatment time is 3-5h; Freezing with thawing treating processes 2-3 time of fast deep described in repetition, cell wall structure is destroyed, active substance stripping, forms eel oligopeptides bone soup; (4) biological enzymolysis process: described eel oligopeptides bone soup is imported in hydrolytic decomposition pot, add prozyme, described prozyme preparation method is as follows, first careless genus bacillus, bifid Helicobacter pylori, lactobacillus rhamnosus, photosynthetic bacteria, Bacillus licheniformis are placed in triangular flask and cultivate, the temperature of cultivating is 28-33 DEG C, under 200-250rpm/min, cultivate 25-35h, make thalline or spore concentration reach 1 × 10
14individual/liter (CFU/L), obtained starter, then in starter, nutrient substance is added, the weight controlling nutrient substance is the 1/5-1/8 of described starter weight, described nutrient substance is wort, namely described prozyme is obtained, add described prozyme according to the weight ratio of eel oligopeptides bone soup and prozyme 1000-1200: 1 and carry out biological enzymolysis process, in described biological enzymolysis process, keep pH in hydrolytic decomposition pot to be 6.8-7.0, described hydrolysis temperature is 60 DEG C, described enzymolysis time is 13-21 DEG C, obtains elementary eel small active peptides; (5) aftertreatment: go out described elementary eel small active peptides enzyme, and be rapidly heated to 180-185 DEG C and keep 20-25s, then 50-60 DEG C/s is cooled to 60 DEG C and makes enzyme deactivation; Then supercentrifuge is adopted by enzymolysis solution with the centrifugation 3-5min of 60000-80000r/min, upper liquid and middle level liquid are merged in input dilution tank body, i.e. enzymolysis parting liquid, subnatant is separated, then adopt hot barrier film cryoconcentration technology to concentrate, thickening temperature is 45-50 DEG C, and vacuum tightness is 0.4-2.0MPa, centrifugal spray drying tower inner drying concentrated solution being transported to 200-220 DEG C becomes powder, namely obtains described eel small active peptides.
3. method as claimed in claim 2, is characterized in that: in step (1), employing temperature is the temperature of 130 DEG C, the pressure boiling 7h of 0.14Mpa, bone slag is discharged, and obtains bone soup, after quick-frozen, form frozen bone soup oil and freeze block, for subsequent use in the freezer being stored in-18 DEG C; Step 2) in, pulverize temperature and control at 2 DEG C, described frozen bone soup oil is frozen block and pulverizes as particle diameter freezes block particle at the frozen bone soup oil of 0.05 millimeter.
4. method as claimed in claim 2, it is characterized in that: cell stripping process in step (3): adopt the lower 4 DEG C of oxygen-free waters of normal pressure to freeze block particle to the frozen bone soup oil pulverized and carry out infiltration water treatment, the water saturation time is 7 hours, and described oxygen-free water is as the criterion to flood raw material completely; Then fast deep is freezing with thawing process, wherein the Deep-Frozen treatment time is 2h, and freezing temp is subzero 25 DEG C, and then thawing, namely freeze block particle with the lower 4 DEG C of water-baths of normal pressure to the frozen bone soup oil being in Deep-Frozen and carry out quick ice-melt, the ice-melt treatment time is 3h; Freezing with thawing treating processes 3 times of fast deep described in repetition, cell wall structure is destroyed, active substance stripping, forms eel oligopeptides bone soup.
5. method as claimed in claim 2, it is characterized in that: step is cultivated in (4) in triangular flask, the temperature of cultivation is 30 DEG C, under 200rpm/min, cultivate 28h, makes thalline or spore concentration reach 1 × 10
14individual/liter (CFU/L), add described prozyme according to the weight ratio of eel oligopeptides bone soup and prozyme 1000: 1 and carry out biological enzymolysis process, in described biological enzymolysis process, keep pH in hydrolytic decomposition pot to be 7.0, described hydrolysis temperature is 60 DEG C, described enzymolysis time is 21 DEG C, obtains elementary eel small active peptides.
6. method as claimed in claim 2, is characterized in that: aftertreatment in step (5): go out described elementary eel small active peptides enzyme, and be rapidly heated to 180 DEG C and keep 25s, then 60 DEG C/s is cooled to 60 DEG C and makes enzyme deactivation; Then supercentrifuge is adopted by enzymolysis solution with the centrifugation 4-5min of 60000-70000r/min, upper liquid and middle level liquid are merged in input dilution tank body, i.e. enzymolysis parting liquid, subnatant is separated, then adopt hot barrier film cryoconcentration technology to concentrate, thickening temperature is 45 DEG C, and vacuum tightness is 2.0MPa, centrifugal spray drying tower inner drying concentrated solution being transported to 200 DEG C becomes powder, namely obtains described eel small active peptides.Preferred supercentrifuge by enzymolysis solution with the centrifugation 4min of 60000r/min.
7. eel small-molecular peptides prepared by the method as described in any one of claim 1-6; It is characterized in that: the molecular weight of described eel small-molecular peptides is that the daltonian accounting of 0-480 is no less than 88.2%, the molecular weight of concrete described eel small-molecular peptides is the daltonian accounting of 0-480 is 88.61%, the daltonian accounting of 480-1000 is 9.43%, being greater than 1000 daltonian accountings is 1.96%, and color and luster is faint yellow.
8. the application of eel small-molecular peptides as claimed in claim 7; It is characterized in that: described eel small-molecular peptides is applicable to diagnosis peptide, peptide vaccine, antibacterial active peptide, cytokine simulating peptide, antiviral peptide or natineoplaston, and peptide health product.
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| CN107385004A (en) * | 2017-09-08 | 2017-11-24 | 长乐聚泉食品有限公司 | The technique that eel accessory substance makes eel Gly-His-Lys |
| CN109806383A (en) * | 2019-03-22 | 2019-05-28 | 深圳大学 | A kind of sea eel peptide promotes the application in immune food, drug or health care product in preparation |
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| CN114146219A (en) * | 2021-12-29 | 2022-03-08 | 上海璞聚生物科技有限公司 | Soft tissue filler and preparation method thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107385004A (en) * | 2017-09-08 | 2017-11-24 | 长乐聚泉食品有限公司 | The technique that eel accessory substance makes eel Gly-His-Lys |
| CN109806383A (en) * | 2019-03-22 | 2019-05-28 | 深圳大学 | A kind of sea eel peptide promotes the application in immune food, drug or health care product in preparation |
| CN109806383B (en) * | 2019-03-22 | 2022-08-16 | 深圳大学 | Application of eel peptide in preparing food, medicine or health product for promoting immunity |
| CN110178964A (en) * | 2019-05-31 | 2019-08-30 | 湖北清江鲟龙渔业有限公司 | The preparation process of sturgeon protein peptides |
| CN114146219A (en) * | 2021-12-29 | 2022-03-08 | 上海璞聚生物科技有限公司 | Soft tissue filler and preparation method thereof |
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