CN105012940B - A kind of preparation method of nanometre collagen peptide chelated zinc - Google Patents
A kind of preparation method of nanometre collagen peptide chelated zinc Download PDFInfo
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Abstract
The invention discloses a kind of preparation method of marine organisms source nanometre collagen peptide chelated zinc.With marine organisms source collagen polypeptide(Such as fish-skin, fish scale hydrolytic collagen peptide)For raw material, small molecule collagen peptide with high metal zinc sequestering activity is prepared using enzyme process, this small molecule collagen peptide and zinc salt are subjected to nanometer chelatropic reaction, the collagen peptide chelated zinc for preparing gained is finely dispersed nano-particle, 1000 nm of particle diameter <, have good water solubility.Present invention process is simply gentle, of low cost without special installation and numerous and diverse treatment process, there is provided a kind of new nanometer zinc supplementation preparation being easily absorbed by the body.
Description
Technical field
The present invention relates to biological technical field, and in particular to a kind of preparation of the nanometre collagen peptide chelated zinc as zinc supplementation agent
Method.
Background technology
Zinc is one of teleorganic trace element, is the important composition of many enzymes, participates in the metabolism of nucleic acid and protein
Effect, the synthesis for collagen and keratin is required, and can promote the development of bone growth and brain cell.Zinc deficiency is easy
Cause the symptoms such as growth retardation, emotional handicap, compromised immune, hypogonadism.In China, zinc deficiency also compares
Generally, it is primarily present in baby, preschool child, pregnant woman and the elderly.It is to improve the battalion of zinc deficiency crowd to take zinc supplementation agent
The safe and efficient method for the situation of supporting.
Research shows that the bioavailability of zinc depends primarily upon soluble zinc in duodenal absorption, zinc and albumen or more
The combining form of sugar, is the most effective form of soluble zinc, the more other zinc salts of the physical property are stablized, from phytic acid etc. in intestinal segment
The influence of factor, so as to improve the utilization rate of zinc.Secondly, small peptide has the function that to promote zinc-iron alloy solution, and metal zinc ion can be borrowed
Help mechanism of absorption of the peptide in body, thus avoid and using same passage absorb other metal ions between antagonism it is competing
Strive, improve the bioavailability of trace element zinc, therefore peptide chelated zinc is preferable zinc supplementation agent, is increasingly subject to the weight of people
Depending on.At present, the hot spot that peptide chelated zinc is research and development is prepared, such as the patent of invention of Publication No. CN 101225102A, Liu Wei etc.
It is raw material with Java tilapia skin, is prepared for fishskin polypeptide chelate zinc, its particle size is about 5.3 μm;Publication No. CN
In the patent of invention of 101225104A, Liu Chengmei etc. is prepared for Java tilapia skin small molecular peptide chelate zinc, and particle size is about 9.4 μ
M etc..
In recent years, as the fast development of science and technology, nanometer technology gradually penetrate into food, medicine and other fields.By receiving
Rice technology, carries out nanosizing processing to trace element, it is directly absorbed by enteron aisle, avoids trace element in human body
Interaction is so as to reduce toxic side effect.Nanosizing is handled micro- in addition to human nutrition is supplemented under with stock size,
Can also high degree improve human or animal to trace element absorption and use efficiency.At present, the nanometer microelement developed is such as
Nano-calcium carbonate, nanometer selenium etc., but be directed to nanometer zinc supplementation agent, also not studies have reported that, therefore, research and develop nanometre glue former peptide
Chelated zinc tool is of great significance.
The content of the invention
Present invention aim to address the low insufficient problem of traditional zinc supplementation agent generation side effect and absorption efficiency, there is provided one
The preparation method of kind novel nano zinc supplementation agent-nanometre collagen peptide chelated zinc, this method are easy to operate without special equipment.
To achieve these goals, solution of the invention is:
A kind of preparation method of nanometre collagen peptide chelated zinc, comprises the following steps:
(1)Marine organisms source collagen polypeptide is soluble in water, it is water-soluble that the collagen polypeptide that concentration is 1~10 mg/ml is made
Liquid;
(2)Into above-mentioned collagen polypeptide aqueous solution, the protease for adding 1~10 mg/ml is digested, and hydrolysis temperature is
40~60 DEG C, the control of enzyme digestion reaction pH value is 6~8, when enzymolysis 3~6 is small;80~90 DEG C are warming up to after enzymolysis, constant temperature
20~30 min enzyme deactivations, obtain the small molecule collagen peptide aqueous solution with high metal zinc sequestering activity after centrifugation;
(3)The molar ratio of preparation zinc salt solution, zinc ion and collagen peptide is 0.5~1;
(4)The aqueous solution pH 4.0~7.0 of small molecule collagen peptide is adjusted, zinc salt solution is slowly added into small molecule glue
In former peptide aqueous solution, whole process is stirred continuously heating, after zinc solution adds, is further continued for 60~120 min of stirring,
Obtain nanometre collagen peptide chelated zinc particle;
(5)White or flaxen nanometre collagen peptide chelated zinc powder is obtained after membrane filtration cleans and dries.
The step(1)In marine organisms source collagen polypeptide be by fish-skin, fish scale marine organisms source hydrolyze collagen peptide,
The Da of molecular weight≤3000.
The step(2)The middle protease used hydrolyzes specific enzyme, food flavor enzyme or basic protein for trypsase, marine product
Enzyme, the small molecule collagen peptide after this four kinds of protease hydrolyzeds have good Nano-Zinc sequestering power.
The step(3)In zinc salt be zinc sulfate, zinc chloride or zinc acetate.
The step(4)Middle zinc salt solution is added drop-wise to dropwise in small molecule collagen peptide aqueous solution, nanometer chelatropic reaction
Temperature is 60~80 DEG C.
The step(5)It is middle use nanometre collagen peptide chelated zinc removal of impurities used by filter membrane for molecular cut off 200~
The NF membrane of 1000 Da.
The step(5)In prepared 1000 nm of nanometre collagen peptide chelated zinc particle diameter <.
The nanometre collagen peptide chelated zinc particle prepared using the above method is uniformly dispersed, 1000 nm of particle diameter <, water-soluble good
It is good, and its preparation process is simply gentle, it is of low cost without special installation and numerous and diverse treatment process, there is provided Yi Zhongxin
The nanometer zinc supplementation preparation being easily absorbed by the body of type.
Brief description of the drawings
Fig. 1 is the transmission electron microscope picture of marine organisms source nanometre collagen peptide chelated zinc particle.
Embodiment
Present invention is disclosed a kind of preparation method of nanometre collagen peptide chelated zinc, comprise the following steps:
(1)Marine organisms source collagen polypeptide is soluble in water, it is water-soluble that the collagen polypeptide that concentration is 1~10 mg/ml is made
Liquid;Marine organisms source therein collagen polypeptide is by the collagen peptide of fish-skin, the hydrolysis of fish scale marine organisms source, molecular weight≤3000
Da。
(2)Into above-mentioned collagen polypeptide aqueous solution, the protease for adding 1~10 mg/ml is digested, and hydrolysis temperature is
40~60 DEG C, the control of enzyme digestion reaction pH value is 6~8, when enzymolysis 3~6 is small;80~90 DEG C are warming up to after enzymolysis, constant temperature
20~30 min enzyme deactivations, obtain the small molecule collagen peptide aqueous solution with high metal zinc sequestering activity after centrifugation;Wherein use
Protease be trypsase, marine product hydrolyze specific enzyme, food flavor enzyme or alkali protease, after this four kinds of protease hydrolyzeds
Small molecule collagen peptide has good Nano-Zinc sequestering power.
(3)The molar ratio of preparation zinc salt solution, zinc ion and collagen peptide is 0.5~1;Zinc salt therein for zinc sulfate,
Zinc chloride or zinc acetate.
(4)The aqueous solution pH 4.0~7.0 of small molecule collagen peptide is adjusted, zinc salt solution is slowly added into small molecule glue
In former peptide aqueous solution, whole process is stirred continuously heating, after zinc solution adds, is further continued for 60~120 min of stirring,
Obtain nanometre collagen peptide chelated zinc particle;Zinc salt solution is added drop-wise to dropwise in small molecule collagen peptide aqueous solution herein, nanometer
Chelatropic reaction temperature is 60~80 DEG C.
(5)White or flaxen nanometre collagen peptide chelated zinc powder is obtained after membrane filtration cleans and dries;Wherein adopt
The NF membrane that filter membrane is 200~1000 Da of molecular cut off used by nanometre collagen peptide chelated zinc cleans, it is prepared
Nanometre collagen peptide chelated zinc particle is uniformly dispersed, 1000 nm of particle diameter <, water-soluble good(As shown in Figure 1).
Embodiment 1:
It is 10 mg/ml marine organisms source collagen polypeptide aqueous solutions to take 20 ml concentration, adds the marine product water of 10 mg/ml
Solution specific enzyme is digested, and hydrolysis temperature is 50 DEG C, adjusts pH 7.5, when enzymolysis 6 is small, 85 DEG C is warming up to after enzymolysis, perseverance
Warm 20 min enzyme deactivations.The small molecule collagen peptide aqueous solution with high metal zinc sequestering activity is obtained after 10000 rpm centrifugations.Match somebody with somebody
Make 4 ml solder(ing)acids, the molar ratio of zinc ion and collagen peptide is 0.5, adjusts the aqueous solution pH of small molecule collagen peptide
6.0,80 DEG C of constant temperature, solder(ing)acid is added drop-wise to dropwise in small molecule collagen peptide aqueous solution, and whole process is stirred continuously,
After solder(ing)acid adds, 60 min of agitating and heating is further continued for, obtains nanometre collagen peptide chelated zinc particle, particle diameter model
It is trapped among 100~300 nm.Flaxen nanometer is obtained after the nanofiltration membrane that molecular cut off is 200 Da cleans and dries
Collagen peptide chelated zinc powder.
Embodiment 2:
It is 10 mg/ml marine organisms source collagen polypeptide aqueous solutions to take 100 ml concentration, add 1 mg/ml food flavor enzyme into
Row enzymolysis, hydrolysis temperature are 50 DEG C, adjust pH 7.0, when enzymolysis 6 is small, are warming up to 85 DEG C after enzymolysis, 20 min of constant temperature goes out
Enzyme.The small molecule collagen peptide aqueous solution with high metal zinc sequestering activity is obtained after 10000 rpm centrifugations.Prepare 100 ml sulphur
The molar ratio of sour zinc aqueous solution, zinc ion and collagen peptide is 0.5, adjusts the aqueous solution pH 6.0 of small molecule collagen peptide, constant temperature 70
DEG C, zinc sulfate solution is added drop-wise to dropwise in small molecule collagen peptide aqueous solution, whole process is stirred continuously, water-soluble in zinc sulfate
After liquid adds, be further continued for 120 min of agitating and heating, obtain nanometre collagen peptide chelated zinc particle, particle size range 500~
1000 nm.Flaxen nanometre glue former peptide chela is obtained after the nanofiltration membrane that molecular cut off is 700 Da cleans and dries
Close zinc powder.
Embodiment 3:
It is 10 mg/ml marine organisms source collagen polypeptide aqueous solutions to take 100 ml concentration, adds the trypsase of 10 mg/ml
Digested, hydrolysis temperature is 40 DEG C, adjusts pH 7.0, when enzymolysis 6 is small, is warming up to 85 DEG C after enzymolysis, 20 min of constant temperature
Enzyme deactivation.The small molecule collagen peptide aqueous solution with high metal zinc sequestering activity is obtained after 10000 rpm centrifugations.Prepare 20 ml
The molar ratio of solder(ing)acid, zinc ion and collagen peptide is 0.5, adjusts the aqueous solution pH 7.0 of small molecule collagen peptide, constant temperature
75 DEG C, solder(ing)acid is added drop-wise to dropwise in small molecule collagen peptide aqueous solution, whole process is stirred continuously, in zinc chloride
After aqueous solution adds, 120 min of agitating and heating is further continued for, obtains nanometre collagen peptide chelated zinc particle, particle size range is 20
~100 nm.Flaxen nanometre glue former peptide chela is obtained after the nanofiltration membrane that molecular cut off is 200 Da cleans and dries
Close zinc powder.
Embodiment 4:
It is 5 mg/ml fishskin collagen polypeptide aqueous solutions to take 40 ml concentration, and the trypsase for adding 1 mg/ml is digested,
Hydrolysis temperature is 40 DEG C, adjusts pH 7.0, when enzymolysis 6 is small, is warming up to 80 DEG C after enzymolysis, 20 min enzyme deactivations of constant temperature.Through
The small molecule collagen peptide aqueous solution with high metal zinc sequestering activity is obtained after 10000 rpm centrifugations.Prepare 10 ml zinc acetate water
The molar ratio of solution, zinc ion and collagen peptide is 0.5, adjusts the aqueous solution pH 5.0 of small molecule collagen peptide, 75 DEG C of constant temperature will
Zinc acetate aqueous solution is added drop-wise in small molecule collagen peptide aqueous solution dropwise, and whole process is stirred continuously, in zinc acetate aqueous solution plus
After entering, 120 min of agitating and heating is further continued for, obtains nanometre collagen peptide chelated zinc particle, particle size range is 200~500
nm.Flaxen nanometre collagen peptide chelated zinc powder is obtained after the nanofiltration membrane that molecular cut off is 450 Da cleans and dries
End.
Embodiment 5:
It is 1 mg/ml scale collagen polypeptide aqueous solutions to take 50 ml concentration, and the alkali protease for adding 1 mg/ml carries out enzyme
Solution, hydrolysis temperature are 45 DEG C, adjust pH 8.0, when enzymolysis 6 is small, are warming up to 85 DEG C after enzymolysis, 20 min enzyme deactivations of constant temperature.Through
The small molecule collagen peptide aqueous solution with high metal zinc sequestering activity is obtained after 10000 rpm centrifugations.Prepare 10 ml zinc chloride water
The molar ratio of solution, zinc ion and collagen peptide is 1.0, adjusts the aqueous solution pH 4.0 of small molecule collagen peptide, 80 DEG C of constant temperature will
Solder(ing)acid is added drop-wise in small molecule collagen peptide aqueous solution dropwise, and whole process is stirred continuously, in solder(ing)acid plus
After entering, 120 min of agitating and heating is further continued for, obtains nanometre collagen peptide chelated zinc particle, particle size range is 200~500
nm.Flaxen nanometre collagen peptide chelated zinc powder is obtained after the nanofiltration membrane that molecular cut off is 300 Da cleans and dries
End.
Claims (4)
1. a kind of preparation method of nanometre collagen peptide chelated zinc, is made of following steps:
(1)Marine organisms source collagen polypeptide is soluble in water, the collagen polypeptide aqueous solution that concentration is 1~10 mg/ml is made;Institute
It is the collagen peptide hydrolyzed by fish-skin, fish scale marine organisms source to state marine organisms source collagen polypeptide, the Da of molecular weight≤3000;
(2)Into above-mentioned collagen polypeptide aqueous solution, the protease for adding 1~10 mg/ml is digested, hydrolysis temperature for 40~
60 DEG C, the control of enzyme digestion reaction pH value is 6~8, when enzymolysis 3~6 is small;It is warming up to 80~90 DEG C after enzymolysis, constant temperature 20~
30 min enzyme deactivations, obtain the small molecule collagen peptide aqueous solution with high metal zinc sequestering activity after centrifugation;
(3)The molar ratio of preparation zinc salt solution, zinc ion and collagen peptide is 0.5~1;
(4)The aqueous solution pH 4.0~7.0 of small molecule collagen peptide is adjusted, zinc salt solution is slowly added into small molecule collagen peptide
In aqueous solution, whole process is stirred continuously heating, after zinc salt solution adds, is further continued for 60~120 min of stirring, obtains
To nanometre collagen peptide chelated zinc particle;The zinc salt solution is added drop-wise to dropwise in small molecule collagen peptide aqueous solution, nanometer chela
It is 60~80 DEG C to close reaction temperature;
(5)White or flaxen nanometre collagen peptide chelated zinc powder is obtained after membrane filtration cleans and dries;The nanometre glue
Filter membrane is the NF membrane of 200~1000 Da of molecular cut off used by former peptide chelated zinc cleans.
A kind of 2. preparation method of nanometre collagen peptide chelated zinc as claimed in claim 1, it is characterised in that:The step(2)
The middle protease used hydrolyzes specific enzyme, food flavor enzyme or alkali protease for trypsase, marine product.
A kind of 3. preparation method of nanometre collagen peptide chelated zinc as claimed in claim 1, it is characterised in that:The step(3)
In zinc salt be zinc sulfate, zinc chloride or zinc acetate.
A kind of 4. preparation method of nanometre collagen peptide chelated zinc as claimed in claim 1, it is characterised in that:The step(5)
In prepared 1000 nm of nanometre collagen peptide chelated zinc particle diameter <.
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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 |
CN107034260B (en) * | 2017-06-27 | 2021-07-02 | 钦州学院 | Preparation method of nano-zinc oyster powder |
CN111925411B (en) * | 2017-06-29 | 2022-04-08 | 安徽省农业科学院农产品加工研究所 | Cys-Ser-containing 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 |
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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CN101225102A (en) * | 2008-01-21 | 2008-07-23 | 南昌大学 | Method for preparing tilapia fishskin polypeptide chelate zinc salt |
CN101481266A (en) * | 2009-02-13 | 2009-07-15 | 杜振宁 | Plant nutrient agent, preparation and use |
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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 |
CN101481266A (en) * | 2009-02-13 | 2009-07-15 | 杜振宁 | Plant nutrient agent, preparation and use |
Non-Patent Citations (1)
Title |
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纳米微量元素的吸收、转运特点及其在动物生产中的应用;宋明明等;《中国饲料》;20150315;第2015年卷(第5期);29-31 * |
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