CN108669310A - A kind of nanometer small peptide chelated iron and preparation method thereof for feed addictive - Google Patents
A kind of nanometer small peptide chelated iron and preparation method thereof for feed addictive Download PDFInfo
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- CN108669310A CN108669310A CN201810490307.2A CN201810490307A CN108669310A CN 108669310 A CN108669310 A CN 108669310A CN 201810490307 A CN201810490307 A CN 201810490307A CN 108669310 A CN108669310 A CN 108669310A
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- small peptide
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/142—Amino acids; Derivatives thereof
- A23K20/147—Polymeric derivatives, e.g. peptides or proteins
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/20—Inorganic substances, e.g. oligoelements
- A23K20/30—Oligoelements
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/107—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length by chemical modification of precursor peptides
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/06—Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products
Abstract
The present invention relates to chemosynthesis technical fields, and in particular to a kind of nanometer small peptide chelated iron and preparation method thereof for feed addictive.It is unfavorable for the problem of pig is to the absorption of ferro element and pollution environment for inorganic iron is added directly in feed, the technical scheme is that, include the following steps：[1] bean cake powder dissolves：Bean cake powder is dissolved in the buffer solution of pH=2.2 8.0, obtains bean cake powder solution；[2] prepared by dry Gly-His-Lys：The bean cake powder solution heat treatment that step [1] is obtained, is then added alkali protease and flavor protease, heating enzymolysis, and then boiling water bath enzyme deactivation activity, centrifugal filtration obtains enzymolysis liquid, is dried to obtain dry Gly-His-Lys；[3] preparation of nanometer small peptide chelated iron：The dry Gly-His-Lys that step [2] obtains are dissolved to obtain small peptide solution, under condition of heating and stirring, inorganic ferrous iron solution is added, small peptide is made to react to form chelate with iron, separation, it is dry after up to nanometer small peptide chelated iron.The present invention is suitable for pannage.
Description
Technical field
The present invention relates to chemosynthesis technical fields, and in particular to a kind of nanometer small peptide chelated iron for feed addictive
And preparation method thereof.
Background technology
Iron is widely distributed in animal body, and nearly all tissue all contains iron.In parenchymatous organ, iron content in spleen, kidney
The amplitude of variation of amount is larger, and in most animals, and the iron-holder of liver is especially high.Iron has important not only for internal enzyme system
Effect and ferroheme and epidermal tissue component part.
The most of presence in the form of protein complex of iron in animal body, participates in vital movement in a variety of forms
Metabolism and energetic supersession.Iron is present in the form of hemoglobin in red blood cell, and muscle is present in the form of myoglobins
In, it is present in serum in the form of transferrin and ferritin, is present in placenta in the form of the ferritin of uterus, with newborn iron egg
White form is present in breast, and being present in liver and spleen in the form of ferritin and flavine ferritin has small part with divalent or trivalent
The form of iron exists.The existence form of iron is different, their characteristic and effect are also different.But the iron of quovis modo is all
Necessary to object.
In the prior art, the mode that iron is added in feed is generally the inorganic salts for directly adding iron.Inorganic salts are not only unfavorable
The absorption of ferro element in animal is to feed, and can largely be drained to be easy to pollute environment by animal.
Invention content
It is unfavorable for animal to the absorption of ferro element and asking for pollution environment for the above-mentioned inorganic iron that added directly in feed
Topic, the present invention provide a kind of nanometer small peptide chelated iron and preparation method thereof for feed addictive, its object is to：Passing through will
Nanometer small peptide chelated iron enhances the nutrient functional of ferro element in feed, improves the biological value of iron as feed addictive,
And reduce environmental pollution.
The technical solution adopted by the present invention is as follows：
A kind of preparation method of nanometer small peptide chelated iron for feed addictive, includes the following steps：
[1] bean cake powder dissolves：Bean cake powder is dissolved in the buffer solution of pH=2.2-8.0, bean cake powder solution is obtained；
[2] prepared by dry Gly-His-Lys：The bean cake powder solution heat treatment that step [1] is obtained, is then added alkali protease and wind
Taste protease, heating enzymolysis, then boiling water bath enzyme deactivation activity, centrifugal filtration obtain enzymolysis liquid, are dried to obtain dry Gly-His-Lys；
[3] preparation of nanometer small peptide chelated iron：The dry Gly-His-Lys that step [2] obtains are dissolved to obtain small peptide solution, heating is stirred
Under the conditions of mixing, inorganic ferrous iron solution is added, small peptide is made to react to form chelate with iron, up to nanometer small peptide chela after separation, drying
Close iron.
After the technical program, make ferro element with it is small-peptide chelated so that ferro element be easier absorbed by organism, to
On the one hand the nutrient functional and biological value for improving ferro element, on the other hand reduce micro- in excreta contain
Amount, to reduce environmental pollution.Further, since the absorption rate higher of ferro element, and the raw soybean dregs powder used has
Cheap advantage, thus advantageously reduce cost.Nanometer small peptide chelated iron purity height, the crystalline structure prepared by this method
It is good, to be conducive to accurately control usage amount.
Preferably, buffer solution is formulated by 0.2M disodium phosphate solns and 0.1M citric acid solutions.
Preferably, a concentration of 155-165g/L of bean cake powder solution.
Preferably, the mass ratio of step [2] neutral and alkali protease and flavor protease is 1：1.
Preferably, the gross mass of step [2] neutral and alkali protease and flavor protease is the 10-11% of bean cake powder quality.
Preferably, the detailed process of step [2] is that 10-15min is stirred at 35-45 DEG C；It is heat-treated at 89-92 DEG C again
15-20min；Alkali protease and flavor protease is added, is digested 4-6 hours in 50-60 DEG C, pH=6.5-7.5, oscillation, enzyme
It lives in boiling water bath enzyme deactivation after solution, adjusts pH value to 4-5, centrifugation, filtering obtain enzymolysis liquid, dry, obtain dry Gly-His-Lys.
Preferably, inorganic ferrous iron solution is the ferrous sulfate solution of 15-25%.
Preferably, the detailed process of step [3] is to pour into small peptide solution in reaction vessel, and adjustment pH is 7.5-8.5, temperature
Degree is 50-60 DEG C, is added with stirring ferrous sulfate solution, and the mass ratio of the quality for making dry Gly-His-Lys and sulfuric acid monohydrate iron is 3-5 ︰ 1,
3-8min is reacted after adding again, detaches, be dried to obtain a nanometer small peptide chelated iron.
Preferably, it is 0.1M sodium hydroxide solutions that the reagent used in pH is adjusted in step [3].
The present invention also provides the nanometer small peptide chelated irons prepared using the above method.Using small-peptide chelated iron as feed
Additive is than directly using inorganic iron to have better palatability as additive.
In conclusion by adopting the above-described technical solution, the beneficial effects of the invention are as follows：
1. make ferro element with it is small-peptide chelated so that ferro element be easier absorbed by organism, on the one hand improve iron
The nutrient functional and biological value of element, have the effect of better strengthen immunity, on the other hand reduce excreta
The content of middle trace element, to reduce environmental pollution.
2. due to the absorption rate higher of ferro element, thus advantageously reducing cost.
3. price is relatively high for existing small-peptide chelated object (the especially small-peptide chelated salt of individual event), and the raw material that the present invention uses
Bean cake powder has the advantages that cheap.
4. it is good by nanometer small peptide chelated iron purity height, crystalline structure prepared by this method, to be conducive to accurately control
Usage amount processed.
5. preferably being fitted as feed addictive than directly using inorganic iron to have as additive using small-peptide chelated iron
Mouth property.
6. digesting small peptide using dregs of beans, using Organometallic compounds reaction and crystal polymorph control, and spray-dried it is prepared for
The high nanometer small peptide chelated iron of chelating degree, and provide new research for other nanometers of small peptide microelement chelate research promotions and think
Road.
Specific implementation mode
All features disclosed in this specification or disclosed all methods or in the process the step of, in addition to mutually exclusive
Feature and/or step other than, can combine in any way.
A kind of preparation method of nanometer small peptide chelated iron for feed addictive, includes the following steps：
[1] bean cake powder dissolves：Bean cake powder is dissolved in the buffer solution of pH=2.2-8.0, bean cake powder solution is obtained；
[2] prepared by dry Gly-His-Lys：The bean cake powder solution heat treatment that step [1] is obtained, is then added alkali protease and wind
Taste protease, heating enzymolysis, then boiling water bath enzyme deactivation activity, centrifugal filtration obtain enzymolysis liquid, are dried to obtain dry Gly-His-Lys；
[3] preparation of nanometer small peptide chelated iron：The dry Gly-His-Lys that step [2] obtains are dissolved to obtain small peptide solution, heating is stirred
Under the conditions of mixing, inorganic ferrous iron solution is added, small peptide is made to react to form chelate with iron, up to nanometer small peptide chela after separation, drying
Close iron.
After the technical program, make ferro element with it is small-peptide chelated, on the one hand so that
Preferably, buffer solution is formulated by 0.2M disodium phosphate solns and 0.1M citric acid solutions.
Preferably, a concentration of 155-165g/L of bean cake powder solution.
Preferably, the mass ratio of step [2] neutral and alkali protease and flavor protease is 1：1.
Preferably, the gross mass of step [2] neutral and alkali protease and flavor protease is the 10-11% of bean cake powder quality.
Preferably, the detailed process of step [2] is that 10-15min is stirred at 35-45 DEG C；It is heat-treated at 89-92 DEG C again
15-20min；Alkali protease and flavor protease is added, is digested 4-6 hours in 50-60 DEG C, pH=6.5-7.5, oscillation, enzyme
It lives in boiling water bath enzyme deactivation after solution, adjusts pH value to 4-5, centrifugation, filtering obtain enzymolysis liquid, dry, obtain dry Gly-His-Lys.
Preferably, inorganic ferrous iron solution is the ferrous sulfate solution of 15-25%.
Preferably, the detailed process of step [3] is to pour into small peptide solution in reaction vessel, and adjustment pH is 7.5-8.5, temperature
Degree is 50-60 DEG C, is added with stirring ferrous sulfate solution, and the mass ratio of the quality for making dry Gly-His-Lys and sulfuric acid monohydrate iron is 3-5 ︰ 1,
3-8min is reacted after adding again, detaches, be dried to obtain a nanometer small peptide chelated iron.
Preferably, it is 0.1M sodium hydroxide solutions that the reagent used in pH is adjusted in step [3].
The present invention also provides the nanometer small peptide chelated irons prepared using the above method.
Technical scheme of the present invention is further described below by specific embodiment and test example.
Embodiment 1
A kind of preparation method of feed addictive nanometer small peptide chelated iron, includes the following steps：
(1) preparation of buffer solution：The buffer solution of pH2.2~8.0 is prepared with 0.2M disodium hydrogen phosphates and 0.1M citric acids
1000mL。
(2) prepared by dry Gly-His-Lys：It takes 155g bean cake powders (granularity≤0.6mm) to be dissolved in the buffer solution of 1000ml, adjusts pH value extremely
4.3, the acid processing 10min at 35-40 DEG C；Again 15min is heat-treated at 89-90 DEG C；Then in alkali protease and flavor egg
White enzyme mass ratio is 1：1, temperature is maintained at 50-55 DEG C, pH 7.0, and enzyme concentration (M/S) is placed in constant temperature under conditions of being 10.5%
Oscillation enzymolysis 4 hours on oscillator, then live 10min in boiling water bath enzyme deactivation, adjust pH value to 4-4.5, then in 3000r/min,
10min is centrifuged, and filters to obtain enzymolysis liquid, is spray-dried, is obtained dry Gly-His-Lys.
(3) preparation of small peptide solution：It is small peptide solution dry Gly-His-Lys to be dissolved in tap water, saturated solution is made.
(4) preparation of ferrous solution：The accurate ferrous sulfate heptahydrate powder for weighing 25g is moved into originally water dissolution on a small quantity
In 100mL volumetric flasks, with tap water constant volume, shake up.
(5) preparation of 100kg nanometers of small peptide chelated irons：Small peptide solution is at the uniform velocity poured into the container with agitating device,
It is 8.50 to adjust pH, and temperature is 50-55 DEG C, mixing speed 120r/min, rate of charge (enzymolysis small peptide and ferrous sulfate heptahydrate it
Than) it is 3：1, ferrous solution is first at the uniform velocity added with the speed of 100mL/min in small peptide solution, waits for that the 1/20 of total amount is added in ferrous solution
Afterwards, then with the speed of 1000mL/min it at the uniform velocity adds, to the end of reacting 3min again after material adds.
(6) wait for that reaction finishes separation and is drying to obtain a nanometer small peptide chelated iron, output capacity 99.88% measures average particle size
For 219nm.
Embodiment 2
A kind of preparation method of feed addictive nanometer small peptide chelated iron, is as follows：
(1) preparation of buffer solution：The buffer solution of pH2.2~8.0 is prepared with 0.2M disodium hydrogen phosphates and 0.1M citric acids.
(2) prepared by dry Gly-His-Lys：Bean cake powder is dissolved in the buffer solution of step (1), makes a concentration of 165g/L of bean cake powder,
40-45 DEG C of stirring 15min；Again 20min is heat-treated at 90-92 DEG C；Alkali protease and flavor protease is added, in 55-60
DEG C, pH=7.0, oscillation enzymolysis 6 hours, live in boiling water bath enzyme deactivation after enzymolysis, adjust pH value to 4.5, centrifugation, filtering obtain enzyme
Liquid is solved, it is dry, obtain dry Gly-His-Lys；
(3) preparation of small peptide solution：It is small peptide solution dry Gly-His-Lys to be dissolved in water, saturated solution is made；
(4) preparation of ferrous solution：By ferrous sulfate heptahydrate be dissolved in water be configured to mass fraction be 21% ferrous sulfate it is molten
Liquid；
(5) preparation of the small-peptide chelated ferrous solution of nanometer：Small peptide solution is poured into reaction vessel, adjustment pH is 7.9, temperature
It is 57-60 DEG C, is added with stirring ferrous sulfate solution, the mass ratio of the quality and ferrous sulfate heptahydrate that make dry Gly-His-Lys is 5 ︰ 1, is added
8min is reacted after complete again；
(6) it detaches, be drying to obtain a nanometer small peptide chelated iron.
Test example 1
1. Basic drawing is with reference to NRC (2012) pig nutritional need and China《Swine rearing standard》(NY/T 65-2004) matches
System, basal diet use corn-soybean meal, control group to be used as nutritional supplementation using inorganic microelement (ferrous sulfate heptahydrate)
Agent；Nanometer small peptide chelated iron prepared by test group addition embodiment 1 substitutes the ferrous sulfate heptahydrate of same price, Basic drawing group
At and every nutritive index be shown in Table 1.
1 diet of table forms and trophic level (air bells dry basis)
1)Premix provides for every kilogram of diet：FeSO4·7H2O 500mg/ nanometers of small peptide chelated irons 80mg, ZnSO4·
7H2O 60mg, CuSO4·5H2O 19.8mg, MgSO310mg, KI 0.20mg, MnSO4·H25 400IU of O 10.2mg, VA,
VD3110IU, VB12mg, VB215mg, VB120.03mg, VE 18IU, citric acid 12mg, carnitine 0.5mg, antioxidant
5mg, mould inhibitor 12.5mg, chromium picolinate 5mg, calcium dihydrogen phosphate 285mg, mountain flour 300mg, carrier are zeolite powder.
2)Trophic level is calculated value.
2. being designed using single factor experiment, sodium selenite 80 similar in ± 1 day 40 days weight is selected, is randomly divided into 2
Group, every group of 8 repetitions each repeat 10 piglets.Above-mentioned 2 kinds of diets are fed respectively, and experimental period is piglet 40 days to 75 days, examination
It tests piglet grouping situation and is shown in Table 2.
Table 2 tests piglet and is grouped situation
3. testing index and method：Testing index and method
(1) production performance：Initial counterpoise, last counterpoise, ADFI, ADG and F/G；Growth performance measure and calculation method is as follows：
ADFI=(feeding doses-remains doses)/experiment number of days；ADG=(last weight-starting weight)/experiment number of days；F/G=, which is averaged, searches for food day
Amount/average daily gain.(2) serum immunoglobulin G (IgG) content is analyzed.The result is shown in table 3, tables 4.
3 weaned piglets of table
Note：Colleague indicates that difference is not notable (P ＞ 0.05) without letter, different lowercase letter indication differences significantly (P ＜
0.05)。
Table 4 is same.
4 Serum Biochemical Index in Piglets of table
4. as shown in Table 3 and Table 4, compared with the control group, the ADG and ADFI of piglet are to reflect that body growth is developed important
Index, and F/G is closely related with pig production benefit, and there is also stronger correlations between ADG, ADFI and F/G three
Property.Notebook data shows the nanometer small peptide chelated iron replacement inorganic iron with low dosage, and son is not influenced while reducing environmental pollution
The growth performance of pig, while the IgG content in piglet serum can be improved, it is beneficial to improve the immunity of piglet.
The specific implementation mode of the application above described embodiment only expresses, the description thereof is more specific and detailed, but simultaneously
Cannot the limitation to the application protection domain therefore be interpreted as.It should be pointed out that for those of ordinary skill in the art
For, under the premise of not departing from technical scheme design, various modifications and improvements can be made, these belong to this
The protection domain of application.
Claims (10)
1. a kind of preparation method of nanometer small peptide chelated iron for feed addictive, which is characterized in that include the following steps：
[1] bean cake powder dissolves：Bean cake powder is dissolved in the buffer solution of pH=2.2-8.0, bean cake powder solution is obtained；
[2] prepared by dry Gly-His-Lys：The bean cake powder solution heat treatment that step [1] is obtained, is then added alkali protease and flavor egg
White enzyme, heating enzymolysis, then boiling water bath enzyme deactivation activity, centrifugal filtration obtain enzymolysis liquid, are dried to obtain dry Gly-His-Lys；
[3] preparation of nanometer small peptide chelated iron：The dry Gly-His-Lys that step [2] obtains are dissolved to obtain small peptide solution, heating stirring item
Under part, inorganic ferrous iron solution is added, small peptide is made to react to form chelate with iron, up to nanometer small peptide chelated iron after separation, drying.
2. a kind of preparation method of nanometer small peptide chelated iron for feed addictive described in accordance with the claim 1, feature
It is：The buffer solution is formulated by 0.2M disodium phosphate solns and 0.1M citric acid solutions.
3. a kind of preparation method of nanometer small peptide chelated iron for feed addictive described in accordance with the claim 1, feature
It is：A concentration of 155-165g/L of the bean cake powder solution.
4. a kind of preparation method of nanometer small peptide chelated iron for feed addictive described in accordance with the claim 1, feature
It is：The mass ratio of step [2] neutral and alkali protease and flavor protease is 1：1.
5. a kind of preparation method of nanometer small peptide chelated iron for feed addictive described in accordance with the claim 1, feature
It is：The gross mass of step [2] neutral and alkali protease and flavor protease is the 10-11% of bean cake powder quality.
6. according to a kind of preparation side of nanometer small peptide chelated iron for feed addictive described in any one of claim 1 to 5
Method, it is characterised in that：The detailed process of step [2] is that 10-15min is stirred at 35-45 DEG C；Again 15- is heat-treated at 89-92 DEG C
20min；Alkali protease and flavor protease is added, is digested 4-6 hours in 50-60 DEG C, pH=6.5-7.5, oscillation, enzymolysis knot
It lives in boiling water bath enzyme deactivation after beam, adjusts pH value to 4-5, centrifugation, filtering obtain enzymolysis liquid, dry, obtain dry Gly-His-Lys.
7. a kind of preparation method of nanometer small peptide chelated iron for feed addictive described in accordance with the claim 1, feature
It is：The inorganic ferrous iron solution is the ferrous sulfate solution of 15-25%.
8. special according to a kind of preparation method of nanometer small peptide chelated iron for feed addictive described in claim 1 or 7
Sign is：The detailed process of step [3] is to pour into small peptide solution in reaction vessel, and adjustment pH is 7.5-8.5, temperature 50-
60 DEG C, it is added with stirring ferrous sulfate solution, the quality for making dry Gly-His-Lys and the mass ratio of sulfuric acid monohydrate iron are 3-5 ︰ 1, after adding
3-8min is reacted again, is detached, is dried to obtain a nanometer small peptide chelated iron.
9. the preparation method of a kind of nanometer small peptide chelated iron for feed addictive according to claim 8, feature
It is：It is 0.1M sodium hydroxide solutions that the reagent used in pH is adjusted in step [3].
10. prepared by a kind of preparation method described in accordance with the claim 1 for the nanometer small peptide chelated iron of feed addictive
Nanometer small peptide chelated iron.
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| CN113826764A (en) * | 2021-09-24 | 2021-12-24 | 江苏三仪生物工程有限公司 | Preparation method of soybean small peptide chelated iron |
| CN115316505A (en) * | 2022-08-09 | 2022-11-11 | 山西北正农生物工程有限公司 | Preparation method of livestock and poultry feed additive |
| CN115595276A (en) * | 2022-10-26 | 2023-01-13 | 湖北贯美科技有限公司（Cn） | Moisturizing and whitening cosmetic and preparation method thereof |
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Application publication date: 20181019 |