CN105012940A - Preparation method of nanometer collagen peptide zinc chelate - Google Patents
Preparation method of nanometer collagen peptide zinc chelate Download PDFInfo
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Abstract
The invention discloses a preparation method of nanometer collagen peptide zinc chelate of a marine organism source. The preparation method comprises the following steps: taking marine organism source collagen peptide (such as fishskin and scale hydrolyzed collagen peptide) as a raw material, preparing micromolecule collagen peptide with a high metal zinc chelating property by adopting an enzyme method, enabling the micromolecule collagen peptide and zinc salt to be subjected to a nometer chelation reaction to prepare the collagen peptide zinc chelate, wherein the collagen peptide zinc chelate is uniformly dispersed nanometer particles with the size being smaller than 1000nm, and has good water solubility. The preparation method has the advantages that the technology is simple and mild, special equipment or complicated treatment process is not needed, the cost is low, and a novel nanometer zinc supplementation preparation easy to absorb by the human body is provided.
Description
Technical field
The present invention relates to biological technical field, be specifically related to a kind of preparation method being used as the nanometre collagen peptide chelated zinc of zinc supplement agent.
Background technology
Zinc is one of teleorganic trace element, is the important composition of a lot of enzyme, participates in nucleic acid and portein metabolism, by collagen and keratic synthesis required, and the growth of bone growth and brain cell can be promoted.Zinc deficiency easily causes the symptoms such as growth retardation, dysthymic disorder, compromised immune, hypogonadism.In China, zinc deficiency is also commonplace, is mainly present in baby, preschooler, anemia of pregnant woman and old people.Take the method safely and efficiently that zinc supplement agent is the nutriture improving zinc deficiency crowd.
Research shows, the bioavailability of zinc depends primarily on soluble zinc in duodenal absorption, the combining form of zinc and albumen or polysaccharide, it is the most effective form of soluble zinc, this physical property comparatively other zinc salt is stablized, and not by the impact of the factors such as phytic acid in intestinal segment, thus improves the utilization rate of zinc.Secondly, little peptide has the effect promoting zinc-iron alloy solution, metallic zinc ion can by the mechanism of absorption of peptide in body, thus the antagonism between other metal ion avoiding and utilize same passage to absorb is competed, improve the bioavailability of trace element zinc, therefore peptide chelated zinc is desirable zinc supplement agent, is more and more subject to people's attention.At present, prepare the focus that peptide chelated zinc is research and development, if publication number is the patent of invention of CN 101225102A, Liu Wei etc. are raw material with Java tilapia skin, have prepared fishskin polypeptide chelate zinc, and its particle size is about 5.3 μm; Publication number is in the patent of invention of CN 101225104A, and Liu Chengmei etc. have prepared Java tilapia skin small molecular peptide chelate zinc, and particle size is about 9.4 μm etc.
In recent years, along with the fast development of science and technology, nanotechnology penetrates into food, medicine and other fields gradually.By nanotechnology, nanorize process is carried out to trace element, make it directly can be absorbed by intestinal, avoid trace element interact in human body thus reduce toxic and side effects.The trace element of nanorize process supplements except human nutrition under having stock size, can also improve human or animal to the absorption and use efficiency of trace element by high degree.At present, the nanometer microelement developed as nano-calcium carbonate, nanometer selenium etc., but for nanometer zinc supplement agent, also not studies have reported that, therefore, research and development nanometre collagen peptide chelated zinc tool is of great significance.
Summary of the invention
The object of the invention is to solve the not enough problem that traditional zinc supplement agent has side effects and absorption efficiency is low, provide a kind of preparation method of novel nano zinc supplement agent-nanometre collagen peptide chelated zinc, the method is without the need to special equipment, simple to operate.
To achieve these goals, solution of the present invention is:
A preparation method for nanometre collagen peptide chelated zinc, comprises the following steps:
(1) by soluble in water for marine organisms derived collagen polypeptide, the collagen polypeptide aqueous solution that concentration is 1 ~ 10 mg/ml is made;
(2) in above-mentioned collagen polypeptide aqueous solution, the protease adding 1 ~ 10 mg/ml carries out enzymolysis, and hydrolysis temperature is 40 ~ 60 DEG C, and enzyme digestion reaction pH value controls 6 ~ 8, enzymolysis 3 ~ 6 hours; Be warming up to 80 ~ 90 DEG C after enzymolysis terminates, constant temperature 20 ~ 30 min enzyme denaturing, after centrifugal, obtain the micromolecule collagen peptide aqueous solution with high metallic zinc sequestering activity;
(3) prepare zinc salt solution, the mol ratio of zinc ion and collagen peptide is 0.5 ~ 1;
(4) the aqueous solution pH 4.0 ~ 7.0 of micromolecule collagen peptide is regulated, slowly joined by zinc salt solution in micromolecule collagen peptide aqueous solution, the continuous agitating heating of whole process, after zinc solution adds, continue stirring 60 ~ 120 min again, obtain nanometre collagen peptide chelated zinc particle;
(5) through membrane filtration remove impurity and obtain after drying white or flaxen nanometre collagen peptide chelated zinc powder.
Marine organisms derived collagen polypeptide in described step (1) is the collagen peptide be hydrolyzed by fish skin, fish scale marine organisms source, molecular weight≤3000 Da.
The protease adopted in described step (2) is trypsin, marine product hydrolysis specific enzyme, food flavor enzyme or alkaline protease, and the micromolecule collagen peptide after these four kinds of protease hydrolyzeds has good Nano-Zinc sequestering power.
Zinc salt in described step (3) is zinc sulfate, zinc chloride or zinc acetate.
In described step (4), zinc salt solution is dropwise added drop-wise in micromolecule collagen peptide aqueous solution, and nanometer chelatropic reaction temperature is 60 ~ 80 DEG C.
The filter membrane that the nanometre collagen peptide chelated zinc remove impurity adopted in described step (5) adopts is the NF membrane of molecular cut off 200 ~ 1000 Da.
Nanometre collagen peptide chelated zinc particle diameter < 1000 nm prepared in described step (5).
The nanometre collagen peptide chelated zinc particle dispersion adopting said method to prepare is even, particle diameter < 1000 nm, water solublity is good, and its preparation technology is simply gentle, without the need to special installation and numerous and diverse treatment process, with low cost, provide a kind of novel nanometer zinc supplement preparation be easily absorbed by the body.
Accompanying drawing explanation
Fig. 1 is the transmission electron microscope picture of marine organisms source nanometre collagen peptide chelated zinc particle.
Detailed description of the invention
Present invention is disclosed a kind of preparation method of nanometre collagen peptide chelated zinc, comprise the following steps:
(1) by soluble in water for marine organisms derived collagen polypeptide, the collagen polypeptide aqueous solution that concentration is 1 ~ 10 mg/ml is made; Marine organisms derived collagen polypeptide is wherein the collagen peptide be hydrolyzed by fish skin, fish scale marine organisms source, molecular weight≤3000 Da.
(2) in above-mentioned collagen polypeptide aqueous solution, the protease adding 1 ~ 10 mg/ml carries out enzymolysis, and hydrolysis temperature is 40 ~ 60 DEG C, and enzyme digestion reaction pH value controls 6 ~ 8, enzymolysis 3 ~ 6 hours; Be warming up to 80 ~ 90 DEG C after enzymolysis terminates, constant temperature 20 ~ 30 min enzyme denaturing, after centrifugal, obtain the micromolecule collagen peptide aqueous solution with high metallic zinc sequestering activity; The protease wherein adopted is trypsin, marine product hydrolysis specific enzyme, food flavor enzyme or alkaline protease, and the micromolecule collagen peptide after these four kinds of protease hydrolyzeds has good Nano-Zinc sequestering power.
(3) prepare zinc salt solution, the mol ratio of zinc ion and collagen peptide is 0.5 ~ 1; Zinc salt is wherein zinc sulfate, zinc chloride or zinc acetate.
(4) the aqueous solution pH 4.0 ~ 7.0 of micromolecule collagen peptide is regulated, slowly joined by zinc salt solution in micromolecule collagen peptide aqueous solution, the continuous agitating heating of whole process, after zinc solution adds, continue stirring 60 ~ 120 min again, obtain nanometre collagen peptide chelated zinc particle; Zinc salt solution is dropwise added drop-wise in micromolecule collagen peptide aqueous solution herein, and nanometer chelatropic reaction temperature is 60 ~ 80 DEG C.
(5) through membrane filtration remove impurity and obtain after drying white or flaxen nanometre collagen peptide chelated zinc powder; The filter membrane that the nanometre collagen peptide chelated zinc remove impurity wherein adopted adopts is the NF membrane of molecular cut off 200 ~ 1000 Da, prepared nanometre collagen peptide chelated zinc particle dispersion is even, particle diameter < 1000 nm, the good (see figure 1) of water solublity.
Embodiment 1:
Getting 20 ml concentration is 10 mg/ml marine organisms derived collagen polypeptid solutions, and the marine product hydrolysis specific enzyme adding 10 mg/ml carries out enzymolysis, and hydrolysis temperature is 50 DEG C, and regulate pH 7.5, enzymolysis 6 hours, is warming up to 85 DEG C after enzymolysis terminates, constant temperature 20 min enzyme denaturing.The micromolecule collagen peptide aqueous solution with high metallic zinc sequestering activity is obtained after 10000 rpm are centrifugal.Prepare 4 ml solder(ing)acids, the mol ratio of zinc ion and collagen peptide is 0.5, regulate the aqueous solution pH 6.0 of micromolecule collagen peptide, constant temperature 80 DEG C, be dropwise added drop-wise to by solder(ing)acid in micromolecule collagen peptide aqueous solution, whole process constantly stirs, after solder(ing)acid adds, continue agitating heating 60 min again, obtain nanometre collagen peptide chelated zinc particle, particle size range is at 100 ~ 300 nm.Be the nanofiltration membrane remove impurity of 200 Da through molecular cut off and after drying, obtain flaxen nanometre collagen peptide chelated zinc powder.
Embodiment 2:
Getting 100 ml concentration is 10 mg/ml marine organisms derived collagen polypeptid solutions, and the food flavor enzyme adding 1 mg/ml carries out enzymolysis, and hydrolysis temperature is 50 DEG C, and regulate pH 7.0, enzymolysis 6 hours, is warming up to 85 DEG C after enzymolysis terminates, constant temperature 20 min enzyme denaturing.The micromolecule collagen peptide aqueous solution with high metallic zinc sequestering activity is obtained after 10000 rpm are centrifugal.Prepare 100 ml zinc sulfate solutions, the mol ratio of zinc ion and collagen peptide is 0.5, regulate the aqueous solution pH 6.0 of micromolecule collagen peptide, constant temperature 70 DEG C, be dropwise added drop-wise to by zinc sulfate solution in micromolecule collagen peptide aqueous solution, whole process constantly stirs, after zinc sulfate solution adds, continue agitating heating 120 min again, obtain nanometre collagen peptide chelated zinc particle, particle size range is at 500 ~ 1000 nm.Be the nanofiltration membrane remove impurity of 700 Da through molecular cut off and after drying, obtain flaxen nanometre collagen peptide chelated zinc powder.
Embodiment 3:
Getting 100 ml concentration is 10 mg/ml marine organisms derived collagen polypeptid solutions, and the trypsin adding 10 mg/ml carries out enzymolysis, and hydrolysis temperature is 40 DEG C, and regulate pH 7.0, enzymolysis 6 hours, is warming up to 85 DEG C after enzymolysis terminates, constant temperature 20 min enzyme denaturing.The micromolecule collagen peptide aqueous solution with high metallic zinc sequestering activity is obtained after 10000 rpm are centrifugal.Prepare 20 ml solder(ing)acids, the mol ratio of zinc ion and collagen peptide is 0.5, regulate the aqueous solution pH 7.0 of micromolecule collagen peptide, constant temperature 75 DEG C, be dropwise added drop-wise to by solder(ing)acid in micromolecule collagen peptide aqueous solution, whole process constantly stirs, after solder(ing)acid adds, continue agitating heating 120 min again, obtain nanometre collagen peptide chelated zinc particle, particle size range is at 20 ~ 100 nm.Be the nanofiltration membrane remove impurity of 200 Da through molecular cut off and after drying, obtain flaxen nanometre collagen peptide chelated zinc powder.
Embodiment 4:
Getting 40 ml concentration is 5 mg/ml fishskin collagen polypeptide aqueous solutions, and the trypsin adding 1 mg/ml carries out enzymolysis, and hydrolysis temperature is 40 DEG C, and regulate pH 7.0, enzymolysis 6 hours, is warming up to 80 DEG C after enzymolysis terminates, constant temperature 20 min enzyme denaturing.The micromolecule collagen peptide aqueous solution with high metallic zinc sequestering activity is obtained after 10000 rpm are centrifugal.Prepare 10 ml zinc acetate aqueous solutions, the mol ratio of zinc ion and collagen peptide is 0.5, regulate the aqueous solution pH 5.0 of micromolecule collagen peptide, constant temperature 75 DEG C, be dropwise added drop-wise to by zinc acetate aqueous solution in micromolecule collagen peptide aqueous solution, whole process constantly stirs, after zinc acetate aqueous solution adds, continue agitating heating 120 min again, obtain nanometre collagen peptide chelated zinc particle, particle size range is at 200 ~ 500 nm.Be the nanofiltration membrane remove impurity of 450 Da through molecular cut off and after drying, obtain flaxen nanometre collagen peptide chelated zinc powder.
Embodiment 5:
Getting 50 ml concentration is 1 mg/ml scale collagen polypeptide aqueous solution, and the alkaline protease adding 1 mg/ml carries out enzymolysis, and hydrolysis temperature is 45 DEG C, and regulate pH 8.0, enzymolysis 6 hours, is warming up to 85 DEG C after enzymolysis terminates, constant temperature 20 min enzyme denaturing.The micromolecule collagen peptide aqueous solution with high metallic zinc sequestering activity is obtained after 10000 rpm are centrifugal.Prepare 10 ml solder(ing)acids, the mol ratio of zinc ion and collagen peptide is 1.0, regulate the aqueous solution pH 4.0 of micromolecule collagen peptide, constant temperature 80 DEG C, be dropwise added drop-wise to by solder(ing)acid in micromolecule collagen peptide aqueous solution, whole process constantly stirs, after solder(ing)acid adds, continue agitating heating 120 min again, obtain nanometre collagen peptide chelated zinc particle, particle size range is at 200 ~ 500 nm.Be the nanofiltration membrane remove impurity of 300 Da through molecular cut off and after drying, obtain flaxen nanometre collagen peptide chelated zinc powder.
Claims (7)
1. a preparation method for nanometre collagen peptide chelated zinc, comprises the following steps:
(1) by soluble in water for marine organisms derived collagen polypeptide, the collagen polypeptide aqueous solution that concentration is 1 ~ 10 mg/ml is made;
(2) in above-mentioned collagen polypeptide aqueous solution, the protease adding 1 ~ 10 mg/ml carries out enzymolysis, and hydrolysis temperature is 40 ~ 60 DEG C, and enzyme digestion reaction pH value controls 6 ~ 8, enzymolysis 3 ~ 6 hours; Be warming up to 80 ~ 90 DEG C after enzymolysis terminates, constant temperature 20 ~ 30 min enzyme denaturing, after centrifugal, obtain the micromolecule collagen peptide aqueous solution with high metallic zinc sequestering activity;
(3) prepare zinc salt solution, the mol ratio of zinc ion and collagen peptide is 0.5 ~ 1;
(4) the aqueous solution pH 4.0 ~ 7.0 of micromolecule collagen peptide is regulated, slowly joined by zinc salt solution in micromolecule collagen peptide aqueous solution, the continuous agitating heating of whole process, after zinc solution adds, continue stirring 60 ~ 120 min again, obtain nanometre collagen peptide chelated zinc particle;
(5) through membrane filtration remove impurity and obtain after drying white or flaxen nanometre collagen peptide chelated zinc powder.
2. the preparation method of a kind of nanometre collagen peptide chelated zinc as claimed in claim 1, is characterized in that: the marine organisms derived collagen polypeptide in described step (1) is the collagen peptide be hydrolyzed by fish skin, fish scale marine organisms source, molecular weight≤3000 Da.
3. the preparation method of a kind of nanometre collagen peptide chelated zinc as claimed in claim 1, is characterized in that: the protease adopted in described step (2) is trypsin, marine product hydrolysis specific enzyme, food flavor enzyme or alkaline protease.
4. the preparation method of a kind of nanometre collagen peptide chelated zinc as claimed in claim 1, is characterized in that: the zinc salt in described step (3) is zinc sulfate, zinc chloride or zinc acetate.
5. the preparation method of a kind of nanometre collagen peptide chelated zinc as claimed in claim 1, is characterized in that: in described step (4), zinc salt solution is dropwise added drop-wise in micromolecule collagen peptide aqueous solution, and nanometer chelatropic reaction temperature is 60 ~ 80 DEG C.
6. the preparation method of a kind of nanometre collagen peptide chelated zinc as claimed in claim 1, is characterized in that: the filter membrane that the nanometre collagen peptide chelated zinc remove impurity adopted in described step (5) adopts is the NF membrane of molecular cut off 200 ~ 1000 Da.
7. the preparation method of a kind of nanometre collagen peptide chelated zinc as claimed in claim 1, is characterized in that: nanometre collagen peptide chelated zinc particle diameter < 1000 nm prepared in described step (5).
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Cited By (8)
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| CN106307540A (en) * | 2016-08-17 | 2017-01-11 | 孙元盛 | Polypeptide capsule for preventing zinc deficiency and preparation method thereof |
| CN107034260A (en) * | 2017-06-27 | 2017-08-11 | 钦州学院 | A kind of preparation method of Nano-Zinc ostreae testa pulverata |
| CN107337709A (en) * | 2017-06-29 | 2017-11-10 | 安徽省农业科学院农产品加工研究所 | A kind of high zinc sequestering activity zinc chelating peptide and its application |
| CN107502642A (en) * | 2017-08-24 | 2017-12-22 | 浦江县美泽生物科技有限公司 | A kind of small-peptide chelated zinc of Fish protein |
| CN107668474A (en) * | 2017-09-07 | 2018-02-09 | 青岛农业大学 | A kind of nisin peptide nanoparticles and its preparation method and application |
| CN105707902B (en) * | 2016-01-14 | 2018-06-29 | 华中农业大学 | A kind of rice polypeptide chelate zinc and preparation method thereof |
| CN109619597A (en) * | 2019-01-10 | 2019-04-16 | 大连工业大学 | Mature vinegar nanoparticle-chelates of zinc preparation method |
| CN111387508A (en) * | 2020-03-12 | 2020-07-10 | 大连工业大学 | Zinc chelating peptide gel and preparation method thereof |
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| CN101225102A (en) * | 2008-01-21 | 2008-07-23 | 南昌大学 | Method for preparing tilapia fishskin polypeptide chelate zinc salt |
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Cited By (11)
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|---|---|---|---|---|
| CN105707902B (en) * | 2016-01-14 | 2018-06-29 | 华中农业大学 | A kind of rice polypeptide chelate zinc and preparation method thereof |
| CN106307540A (en) * | 2016-08-17 | 2017-01-11 | 孙元盛 | Polypeptide capsule for preventing zinc deficiency and preparation method thereof |
| CN107034260A (en) * | 2017-06-27 | 2017-08-11 | 钦州学院 | A kind of preparation method of Nano-Zinc ostreae testa pulverata |
| CN107034260B (en) * | 2017-06-27 | 2021-07-02 | 钦州学院 | Preparation method of nano-zinc oyster powder |
| CN107337709A (en) * | 2017-06-29 | 2017-11-10 | 安徽省农业科学院农产品加工研究所 | A kind of high zinc sequestering activity zinc chelating peptide and its application |
| CN107337709B (en) * | 2017-06-29 | 2021-03-02 | 安徽省农业科学院农产品加工研究所 | High-zinc-chelating-activity zinc chelating peptide and application thereof |
| CN107502642A (en) * | 2017-08-24 | 2017-12-22 | 浦江县美泽生物科技有限公司 | A kind of small-peptide chelated zinc of Fish protein |
| CN107668474A (en) * | 2017-09-07 | 2018-02-09 | 青岛农业大学 | A kind of nisin peptide nanoparticles and its preparation method and application |
| CN107668474B (en) * | 2017-09-07 | 2020-11-03 | 青岛农业大学 | Nisin peptide nanoparticles and preparation method and application thereof |
| CN109619597A (en) * | 2019-01-10 | 2019-04-16 | 大连工业大学 | Mature vinegar nanoparticle-chelates of zinc preparation method |
| CN111387508A (en) * | 2020-03-12 | 2020-07-10 | 大连工业大学 | Zinc chelating peptide gel and preparation method thereof |
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