CN107043789A - A kind of method that octopus residue protein zymolyte regulation and control prepare nanometer selenium - Google Patents
A kind of method that octopus residue protein zymolyte regulation and control prepare nanometer selenium Download PDFInfo
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- CN107043789A CN107043789A CN201710187693.3A CN201710187693A CN107043789A CN 107043789 A CN107043789 A CN 107043789A CN 201710187693 A CN201710187693 A CN 201710187693A CN 107043789 A CN107043789 A CN 107043789A
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Abstract
The present invention relates to the method that a kind of regulation and control of octopus residue protein zymolyte prepare nanometer selenium, comprise the following steps：By octopus residue decontamination, clean after, add water progress tissue mashing homogenate, obtain homogenate；Homogenate is placed in enzymatic vessel, animal protease enzymolysis is added, obtains enzymolysis liquid；Enzymolysis liquid is gone out after enzyme, is centrifuged, is taken and octopus residue protein zymolyte is obtained after supernatant vacuum freeze drying；Take octopus residue protein zymolyte to be mixed with water, add sodium selenite and nanometer selenium solution is made in vitamin C reaction.Nanometer selenium prepared by the present invention has that bioactivity is high, Size Distribution is small, stability is strong, it is nontoxic the features such as, there is good development prospect in fields such as medicine, food, agriculturals.Secondly, the present invention can not only create economic benefit, and improve the value of octopus using octopus residue protein zymolyte as the template adjusting control agent for preparing nanometer selenium, it is to avoid environmental pollution, with important practical significance.
Description
Technical field
The invention belongs to changing rejected material to useful resource and nanometer selenium preparing technical field, and in particular to a kind of octopus residue albumen
The method that zymolyte regulation and control prepare nanometer selenium.
Background technology
Selenium (Se) is required a kind of trace element in humans and animals body, with a series of anti-oxidation function, first in human body
The interwoveness such as the metabolism of shape glandular hormone and immune system.Compared with inorganic selenium and Organic Selenium, red nano selenium has toxicity low, raw
The high feature of thing activity, in terms of being mainly used in animal productiong, medicine and health products.
The preparation of nanometer selenium generally uses reducing process, using reducing agent by the oxygen hydrochlorate containing selenium or oxide (such as selenic acid
Salt, selenite, selenium dioxide) reduction obtain elemental selenium.Reduced form Vc, glutathione are the most frequently used reducing agents, in addition, sub-
Sodium sulphate, hydrazine, sodium thiosulfate also are used as reducing agent, but their bio-toxicities are than larger, are not suitable for preparing biological use
Nanometer selenium material.Secondly, the red nano selenium stability that prepared by reducing process is poor, and bioactivity is low, is easily changed into grey black tool
Virose elemental selenium, therefore, biomolecule of the selection with proper function is used as nanometer selenium particle diameter and shape in preparation process
The template adjusting control agent of looks, the bioactivity or the new biological function of imparting nanometer selenium of reinforced nano selenium, with important economy
And social benefit.
The content of the invention
In view of the shortcomings of the prior art, prepared it is an object of the invention to provide a kind of regulation and control of octopus residue protein zymolyte
The method of nanometer selenium, the problem of effectively solving high nanometer selenium production cost in the prior art, bioactivity and low security, and
Effectively utilize discarded octopus residue, it is to avoid environmental pollution, realize the recycling of discarded object.
The present invention is achieved through the following technical solutions：
A kind of method that octopus residue protein zymolyte regulation and control prepare nanometer selenium, comprises the following steps：
(1) preparation of octopus residue protein zymolyte
A, octopus residue gone to the removal of impurity, cleaned after, add water, carry out tissue mashing homogenate, obtain homogenate；
B, homogenate is placed in enzymatic vessel, adds animal protease and digested, obtain enzymolysis liquid；
C, enzymatic vessel temperature rises to 90 DEG C of enzyme 30min that go out, and in 4 DEG C, 4500r/min centrifugation 15min are filtered, collected
Supernatant, as octopus residue protein zymolyte, vacuum freeze drying are preserved；
(2) preparation of nanometer selenium solution
A, octopus residue protein zymolyte freeze-dried powder obtained by step (1) is taken, add deionized water, stirring is to completely molten
Solution, adds 1mmol/L sodium selenite solutions, after whirlpool is mixed, 4mmol/L vitamin Cs (Vc) solution is added dropwise, 45 DEG C of constant temperature are placed in
Concussion is incubated 24h in shaking table；
B, reaction solution obtained by step A is transferred in the bag filter that molecular cut off is 500u, 4 DEG C of lucifuges of distilled water are saturating
Analysis, first water is changed every 2h；The outer liquid of bag filter is detected through ICP-AES dialyses without stopping after plasma selenium, institute in bag filter
It is a nanometer selenium solution to obtain sample.
Preferably, octopus residue and water quality volume ratio (quality kg/ volume L) are octopus residue in step (1)：Water
=1：2~10.
Preferably, it is 1 by enzyme amount and homogenate weight ratio in step (1)：200~500 add animal protease.
Preferably, enzymatic hydrolysis condition is in step (1)：Hydrolysis temperature is 40~65 DEG C, and enzymolysis pH is 5~8, and enzymolysis time is
2~7 hours.
Preferably, octopus residue protein zymolyte freeze-dried powder and deionized water solid-liquid ratio are 1~6 in step (2)：1.
Preferably, sodium selenite solution, vitamin C (Vc) solution, octopus residue protein zymolyte solution in step (2)
It is Vc by volume：Na2SeO3：Octopus residue protein zymolyte solution=4：4：1 ratio is mixed.
Nanometer selenium：Nanometer selenium is to realize to obtain selenate or selenite reduction under certain condition by strong reductant
Arrive, or using the microorganism with stronger reducing power, solvable selenium is reduced to biological nano selenium and prepared.With inorganic selenium
Compared with Organic Selenium, nanometer selenium has bioresorbable high, energy high-efficient cleaning removes free radical, and toxicity is low, the features such as active high, in doctor
The fields such as medicine, food, agricultural have huge application potential.Research shows that nanometer selenium is to tumour, Kaschin-Beck disease, diabetes, Chinese mugwort
Growing disease, cataract, breathing problem, low fecundity, Keshan disease, heavy metal poisoning etc. has certain preventive and therapeutic action；
Nanometer selenium health products can increase antioxidant ability of organism, immunocompetence and antitumaous effect；Nanometer selenium Feed Energy enhancing animal resists
Stress ability and immunity, improve meat, improve production performance；Nanometer selenium fertilizer can significantly improve crop anti-adversity, regulation
Plant growth, improves crop quality, improves crop Se content and yield.
Octopus residue：Octopus Mollusca, Cephalopoda, Octopodida, octopus section, popular name " octopus " is a kind of high
The precious marine resources of economic value.Octopus residue is a large amount of discarded objects produced in octopus process, such as internal organ, neck
Son, tail point and tooth etc., account for more than the 13% of the total processing capacity of octopus.Detection shows have in octopus residue containing a large amount of poles
The natural active matter of value, such as amino acids, nitrogenous non-protein class, protein-based, unsaturated fatty acids, research hair
Existing, 18 kinds of amino acid contents are up to 52.7% (butt) in octopus internal organ.Therefore, high added value is extracted from octopus residue
Natural active matter, is the effective way for realizing octopus aquatic products comprehensive utilization of resources, with important practical significance.The present invention
Obtained octopus residue protein zymolyte is digested with animal protease, with the spy such as safe, adhesion is strong, degradable
Property, as the template adjusting control agent for preparing nanometer selenium, tied by regulating and controlling the particle diameter of formed nano-particle, changing its surface
Structure or charging property etc., can greatly improve water solubility, bio-compatibility and the interbiotic recognition capability of nano-particle, prevent from receiving
Rice Se particle is reunited or precipitated in preparation, preservation and application process.
Beneficial effects of the present invention：
1st, the present invention closes protein zymolyte and selenizing, by regulating and controlling the particle diameter of formed nano-particle, changing its table
Face structure or charging property etc., can greatly improve water solubility, bio-compatibility and the interbiotic recognition capability of nano-particle, prevent
Only nanometer Se particle is reunited or precipitated in preparation, preservation and application process.
2nd, the present invention can not only create warp using octopus residue protein zymolyte as the template adjusting control agent for preparing nanometer selenium
Help benefit, and improve the value of octopus, it is to avoid environmental pollution, with important practical significance.
3rd, the nanometer selenium for preparing of the present invention have that bioactivity is high, Size Distribution is small, stability is strong, it is nontoxic the features such as,
The fields such as medicine, food, agricultural have good development prospect.
Embodiment
In order to make the purpose , technical scheme and advantage of the present invention be clearer, with reference to embodiments, to the present invention
It is further elaborated.It should be appreciated that the specific embodiments described herein are merely illustrative of the present invention, it is not used to
Limit the present invention.
Embodiment 1
A kind of method that octopus residue protein zymolyte regulation and control prepare nanometer selenium, comprises the following steps：
(1) preparation of octopus residue protein zymolyte
A, octopus residue gone to the removal of impurity, cleaned after, be 1 by solid-liquid ratio：10 add water, carry out tissue mashing homogenate,
Obtain homogenate；
B, homogenate is placed in enzymatic vessel, is 1 by enzyme amount and homogenate weight ratio：500 add animal protease, in temperature
Spend for 40 DEG C, digested 2 hours under the conditions of pH 8.0, obtain enzymolysis liquid；
C, enzymatic vessel temperature rises to 90 DEG C of enzyme 30min that go out, and in 4 DEG C, 4500r/min centrifugation 15min are filtered, collected
Supernatant, as octopus residue protein zymolyte, vacuum freeze drying are preserved；
(2) preparation of nanometer selenium solution
A, octopus residue protein zymolyte freeze-dried powder obtained by step (1) is taken, by solid-liquid ratio 1：1 adds deionized water,
Stirring adds 1mmol/L sodium selenite solutions to being completely dissolved, and after whirlpool is mixed, 4mmol/L vitamin Cs (Vc) solution is added dropwise,
Wherein, sodium selenite solution, vitamin C (Vc) solution, walnut protein zymolyte liquor capacity ratio are 4：4：1, it is placed in 45 DEG C of perseverances
Concussion is incubated 24h in warm shaking table；
B, reaction solution obtained by step A is transferred in the bag filter that molecular cut off is 500u, 4 DEG C of lucifuges of distilled water are saturating
Analysis, first water is changed every 2h；The outer liquid of bag filter is detected through ICP-AES dialyses without stopping after plasma selenium, institute in bag filter
It is a nanometer selenium solution to obtain sample.
Embodiment 2
A kind of method that octopus residue protein zymolyte regulation and control prepare nanometer selenium, comprises the following steps：
(1) preparation of octopus residue protein zymolyte
A, octopus residue gone to the removal of impurity, cleaned after, be 1 by solid-liquid ratio：3 add water, carry out tissue mashing homogenate, obtain
Homogenate；
B, homogenate is placed in enzymatic vessel, is 1 by enzyme amount and homogenate weight ratio：300 add animal protease, in temperature
Spend for 55 DEG C, digested 5 hours under the conditions of pH 6.0, obtain enzymolysis liquid；
C, enzymatic vessel temperature rises to 90 DEG C of enzyme 30min that go out, and in 4 DEG C, 4500r/min centrifugation 15min are filtered, collected
Supernatant, as octopus residue protein zymolyte, vacuum freeze drying are preserved；
(2) preparation of nanometer selenium solution
A, octopus residue protein zymolyte freeze-dried powder obtained by step (1) is taken, by solid-liquid ratio 4：1 adds deionized water,
Stirring adds 1mmol/L sodium selenite solutions to being completely dissolved, and after whirlpool is mixed, 4mmol/L vitamin Cs (Vc) solution is added dropwise,
Wherein, sodium selenite solution, vitamin C (Vc) solution, walnut protein zymolyte liquor capacity ratio are 4：4：1, it is placed in 45 DEG C of perseverances
Concussion is incubated 24h in warm shaking table；
B, reaction solution obtained by step A is transferred in the bag filter that molecular cut off is 500u, 4 DEG C of lucifuges of distilled water are saturating
Analysis, first water is changed every 2h；The outer liquid of bag filter is detected through ICP-AES dialyses without stopping after plasma selenium, institute in bag filter
It is a nanometer selenium solution to obtain sample.
Embodiment 3
A kind of method that octopus residue protein zymolyte regulation and control prepare nanometer selenium, comprises the following steps：
(1) preparation of octopus residue protein zymolyte
A, octopus residue gone to the removal of impurity, cleaned after, be 1 by solid-liquid ratio：2 add water, carry out tissue mashing homogenate, obtain
Homogenate；
B, homogenate is placed in enzymatic vessel, is 1 by enzyme amount and homogenate weight ratio：200 add animal protease, in temperature
Spend for 65 DEG C, digested 7 hours under the conditions of pH 5.0, obtain enzymolysis liquid；
C, enzymatic vessel temperature rises to 90 DEG C of enzyme 30min that go out, and in 4 DEG C, 4500r/min centrifugation 15min are filtered, collected
Supernatant, as octopus residue protein zymolyte, vacuum freeze drying are preserved；
(2) preparation of nanometer selenium solution
A, octopus residue protein zymolyte freeze-dried powder obtained by step (1) is taken, by solid-liquid ratio 6：1 adds deionized water,
Stirring adds 1mmol/L sodium selenite solutions to being completely dissolved, and after whirlpool is mixed, 4mmol/L vitamin Cs (Vc) solution is added dropwise,
Wherein, sodium selenite solution, vitamin C (Vc) solution, walnut protein zymolyte liquor capacity ratio are 4：4：1, it is placed in 45 DEG C of perseverances
Concussion is incubated 24h in warm shaking table；
B, reaction solution obtained by step A is transferred in the bag filter that molecular cut off is 500u, 4 DEG C of lucifuges of distilled water are saturating
Analysis, first water is changed every 2h；The outer liquid of bag filter is detected through ICP-AES dialyses without stopping after plasma selenium, institute in bag filter
It is a nanometer selenium solution to obtain sample.
Comparative example 1
Without octopus residue protein zymolyte in nanometer selenium preparation process, its preparation process is as follows：
(1) 1mmol/L sodium selenite solutions, 4mmol/L vitamin Cs (Vc) solution and deionized water are taken, according to volume ratio
For 4：4：1 mixes, and is placed in concussion in 45 DEG C of constant-temperature tables and is incubated 24h；
(2) reaction solution obtained by step (1) is transferred in the bag filter that molecular cut off is 500u, 4 DEG C of lucifuges of distilled water
Dialysis, first water is changed every 2h；The outer liquid of bag filter is detected through ICP-AES dialyses without stopping after plasma selenium, in bag filter
Gained sample is a nanometer selenium solution.
Experimental example 1
The measure of nanometer selenium average grain diameter
1st, test objective：Detection prepares the side of nanometer selenium by a kind of octopus residue protein zymolyte regulation and control of the present invention
Nanometer selenium average grain diameter obtained by method.
2nd, sample is detected：The nanometer selenium solution example prepared by above-described embodiment 1~3 and comparative example 1.
3rd, detection method：Detection sample is placed in nano particle size instrument and analyzes its particle diameter distribution.
4th, testing result：Testing result is as shown in table 1 below.
The nanometer selenium average grain diameter measurement result of table 1
| Embodiment 1 | Embodiment 2 | Embodiment 3 | Comparative example 1 | |
| Nanometer selenium average grain diameter (nm) | 175 | 87 | 92 | 279 |
5th, conclusion
After testing, compared with comparative example 1, the average grain of nanometer selenium prepared by the embodiment of the present invention 1, embodiment 2, embodiment 3
Footpath reduces 104nm, 192nm, 187nm respectively, wherein the best results of embodiment 2, shows octopus residue proteolysis of the present invention
Thing has significant regulating and controlling effect to nanometer selenium particle diameter, has a extensive future.
Claims (6)
1. a kind of method that octopus residue protein zymolyte regulation and control prepare nanometer selenium, it is characterised in that comprise the following steps：
(1) preparation of octopus residue protein zymolyte
A, octopus residue gone to the removal of impurity, cleaned after, add water, carry out tissue mashing homogenate, obtain homogenate；
B, homogenate is placed in enzymatic vessel, adds animal protease and digested, obtain enzymolysis liquid；
C, enzymatic vessel temperature rises to 90 DEG C of enzyme 30min that go out, and in 4 DEG C, supernatant is collected in 4500r/min centrifugation 15min, filtering
Liquid, as octopus residue protein zymolyte, vacuum freeze drying are preserved；
(2) preparation of nanometer selenium solution
A, take octopus residue protein zymolyte freeze-dried powder obtained by step (1), add deionized water, stirring to being completely dissolved,
1mmol/L sodium selenite solutions are added, after whirlpool is mixed, 4mmol/L vitamin Cs (Vc) solution is added dropwise, is placed in 45 DEG C of constant temperature and shakes
Concussion is incubated 24h in bed；
B, reaction solution obtained by step A is transferred in the bag filter that molecular cut off is 500u, 4 DEG C of lucifuges dialysis of distilled water, often
First water is changed every 2h；The outer liquid of bag filter is detected through ICP-AES dialyses without stopping after plasma selenium, gained sample in bag filter
As nanometer selenium solution.
2. the method that a kind of octopus residue protein zymolyte regulation and control according to claim 1 prepare nanometer selenium, it is special
Levy and be, octopus residue and water quality volume ratio (quality kg/ volume L) are octopus residue in the step (1)：Water=1：
2~10.
3. the method that a kind of octopus residue protein zymolyte regulation and control according to claim 1 prepare nanometer selenium, it is special
Levy and be, it is 1 that enzyme amount is pressed in the step (1) with homogenate weight ratio：200~500 add animal protease.
4. the method that a kind of octopus residue protein zymolyte regulation and control according to claim 1 prepare nanometer selenium, it is special
Levy and be, enzymatic hydrolysis condition is in the step (1)：Hydrolysis temperature is 40~65 DEG C, and enzymolysis pH is 5~8, and enzymolysis time is 2~7
Hour.
5. the method that a kind of octopus residue protein zymolyte regulation and control according to claim 1 prepare nanometer selenium, it is special
Levy and be, octopus residue protein zymolyte freeze-dried powder and deionized water solid-liquid ratio are 1~6 in the step (2)：1.
6. the method that a kind of octopus residue protein zymolyte regulation and control according to claim 1 prepare nanometer selenium, it is special
Levy and be, sodium selenite solution, vitamin C (Vc) solution, octopus residue protein zymolyte solution press body in the step (2)
Product is than being Vc：Na2SeO3：Octopus residue protein zymolyte solution=4：4：1 ratio is mixed.
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Cited By (2)
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| CN105876809A (en) * | 2016-04-18 | 2016-08-24 | 厦门三九生物科技有限公司 | Marine polypeptide-selenium chelate preparation method and application |
| CN110423791A (en) * | 2019-08-06 | 2019-11-08 | 广州大学 | A kind of reproducibility polypeptide and its preparation method and application |
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| CN105876809A (en) * | 2016-04-18 | 2016-08-24 | 厦门三九生物科技有限公司 | Marine polypeptide-selenium chelate preparation method and application |
| CN110423791A (en) * | 2019-08-06 | 2019-11-08 | 广州大学 | A kind of reproducibility polypeptide and its preparation method and application |
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Application publication date: 20170815 |