CN111073931A - Preparation method of protein peptide iron chelate suitable for ruminants - Google Patents
Preparation method of protein peptide iron chelate suitable for ruminants Download PDFInfo
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- CN111073931A CN111073931A CN201811225654.9A CN201811225654A CN111073931A CN 111073931 A CN111073931 A CN 111073931A CN 201811225654 A CN201811225654 A CN 201811225654A CN 111073931 A CN111073931 A CN 111073931A
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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
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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
- C07K1/14—Extraction; Separation; Purification
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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
- C07K1/14—Extraction; Separation; Purification
- C07K1/34—Extraction; Separation; Purification by filtration, ultrafiltration or reverse osmosis
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Abstract
The invention relates to a preparation method of a protein peptide iron chelate suitable for ruminants, which mainly comprises the steps of protein liquid preparation, enzymolysis, ultrafiltration, chelation and spray drying to obtain the protein peptide iron chelate. The method takes soybean protein with wide sources as a protein peptide source and takes inorganic iron salt as an iron source to chelate iron element and protein peptide, has simple process flow and low cost, and is suitable for large-scale industrial production. The product prepared by the invention has high content of small molecular peptides and high content of chelated iron ions, and is a novel high-efficiency iron supplement for feed.
Description
Technical Field
The invention relates to a preparation method of organic trace elements, in particular to a preparation method of a protein peptide iron chelate suitable for ruminants.
Background
Iron is a trace element which is needed by the animal body in the largest quantity, exists in red blood cells in the form of hemoglobin mostly, and is one of the components of muscle and ferrase of the animal body. Iron is also a cofactor for superoxide dismutase and is involved in cellular antioxidant action. In the production process of livestock and poultry, iron-deficiency anemia is the most common nutritional deficiency of animals, and the deficiency of iron trace elements directly causes slow growth and reduction of feed conversion efficiency. At present, the common iron supplement agents in the market comprise inorganic iron and organic iron, inorganic iron ions are easy to generate antagonistic action with partial nutrients, and are easily influenced by Ph value, lipids, proteins, cellulose, oxalic acid, vitamins, phosphate, phytate and the like in the digestion and absorption process, so that the absorption and utilization rate is low. Many researches prove that the organic iron has higher bioavailability than the inorganic iron and has obvious promotion effects on growth, reproduction, health, feed conversion rate and the like of animals. The organic iron mainly comprises organic acid state iron, amino acid chelated iron and protein peptide chelated iron, the protein peptide chelated iron is a third-generation novel iron source supplement which is developed rapidly at home and abroad in recent years, and is a compound with a cyclic structure generated by the coordination reaction of iron ions and small molecular peptides or short peptide substances. Because the protein peptide iron chelate is in a form of iron element which is close to the natural form in the animal body, the protein peptide iron chelate has good chemical stability, higher biological value, easy digestion and absorption, interference resistance, no stimulation and no toxic action, is more superior than the corresponding organic acid state iron and amino acid chelate iron, and is considered as an ideal novel high-efficiency iron supplement for feed at present. Therefore, the development of the protein peptide chelated iron with high absorption and utilization rate, good biochemical stability, high nutritional value and small side effect has very important market significance. However, at present, the research and development of the protein peptide iron chelate are not much in China, and no research report is found about a preparation method of the protein peptide iron chelate suitable for ruminants.
Disclosure of Invention
The invention aims to provide a preparation method of a protein peptide iron chelate suitable for ruminants based on the above situation, and the method takes soy protein as a main raw material, and has the advantages of simple process, low cost and easy implementation.
In order to achieve the purpose, the invention is realized by adopting the following technical scheme: a method for preparing protein peptide iron chelate suitable for ruminant animals comprises the following steps:
(1) preparing protein liquid: adding water into the isolated soy protein powder, stirring and mixing uniformly to prepare a protein solution with the protein concentration of 10-15% (w/w);
(2) enzymolysis: adjusting the pH value of the protein solution to 8.0-8.5, adding protease, performing enzymolysis at 50-55 deg.C for 5-8h, heating the enzymolysis solution to 85 deg.C, and inactivating enzyme for 10 min to obtain enzymolysis solution;
(3) and (3) ultrafiltration: carrying out ultrafiltration on the enzymolysis liquid obtained in the step (2) by an ultrafiltration membrane device with the molecular weight cutoff of 3000Da, and collecting a permeate to obtain a protein peptide solution;
(4) carrying out chelation reaction: adding water-soluble iron salt into the protein peptide solution obtained in the step (3), wherein the addition amount of the water-soluble iron salt is 30-50% of the amount of the protein peptide (based on the content of the protein) in the protein peptide solution, adjusting the pH value of the solution to 5.5-6.0, and carrying out stirring chelation reaction for 2-3 hours at 55-60 ℃ to obtain a protein peptide iron chelation reaction solution;
(5) spray drying: and (4) carrying out spray drying on the protein peptide iron chelate reaction solution obtained in the step (4) to obtain the protein peptide iron chelate.
The soybean protein isolate in the step (1) is a soybean protein isolate with the protein content of more than or equal to 90 percent.
The protease in the step (2) is alkaline protease, the enzyme activity is 250000-300000U/ml, and the addition amount of the protease is 1.5-2.0% of the protein mass in the protein solution.
The water-soluble iron salt in the step (4) is any one of ferrous fumarate, ferrous sulfate, ferrous citrate, ferrous lactate, ferrous chloride and ferrous carbonate.
Through the implementation of the technical scheme, the invention has the following beneficial effects:
(1) the method comprehensively applies modern bioengineering technologies and modern processing technologies such as biological enzymolysis, ultrafiltration, molecular chelation, spray drying and the like, and has the characteristics of simple process flow, high production efficiency, low cost and suitability for industrial mass production.
(2) The method disclosed by the invention has the advantages that soybean protein with wide and easily-obtained sources is taken as a peptide source, inorganic iron salt is taken as an iron source, iron ions and protein peptide are chelated to prepare the protein peptide iron chelate, the content of small molecular peptide (the relative molecular weight is between 180 and 1000 Da) in the obtained product is up to more than 70%, the content of chelated iron ions is more than 7%, and the chelation rate is more than 91%.
Detailed Description
The present invention will be described in further detail with reference to specific examples, but the embodiments of the present invention are not limited to the scope of the examples.
Example 1
(1) Preparing protein liquid: weighing 100 kg of soybean protein isolate powder (the content of crude protein is 92%), adding 800 kg of water, and uniformly stirring to obtain a protein solution;
(2) enzymolysis: adjusting the pH value of the protein solution to 8.0, adding 1.5 kg of protease, carrying out enzymolysis at 55 ℃ for 8 hours, heating the enzymolysis solution to 85 ℃ to inactivate the enzyme for 10 minutes to obtain enzymolysis solution;
(3) and (3) ultrafiltration: performing ultrafiltration on the enzymolysis liquid by an ultrafiltration membrane device with the molecular weight cutoff of 3000Da, and collecting the permeate to obtain a protein peptide solution;
(4) carrying out chelation reaction: adding 25 kg of ferrous sulfate into the protein peptide solution, adjusting the pH value of the solution to 5.5 under continuous stirring, heating to 55 ℃, and carrying out heat preservation reaction for 2 hours to obtain protein peptide iron chelate reaction solution;
(5) spray drying: and (4) carrying out spray drying on the protein peptide iron chelate reaction solution obtained in the step (4) to obtain the protein peptide iron chelate.
Example 2
(1) Preparing protein liquid: weighing 100 kg of soybean protein isolate powder (the content of crude protein is 90%), adding 550 kg of water, and uniformly stirring to obtain a protein solution;
(2) enzymolysis: adjusting the pH value of the protein solution to 8.5, adding 1.8 kg of protease, carrying out enzymolysis at the temperature of 52 ℃ for 5 hours, and heating the enzymolysis solution to 85 ℃ to inactivate the enzyme for 10 minutes to obtain enzymolysis solution;
(3) and (3) ultrafiltration: performing ultrafiltration on the enzymolysis liquid by an ultrafiltration membrane device with the molecular weight cutoff of 3000Da, and collecting the permeate to obtain a protein peptide solution;
(4) carrying out chelation reaction: adding 30 kg of ferrous sulfate into the protein peptide solution, adjusting the pH value of the solution to 5.8 under continuous stirring, heating to 60 ℃, and carrying out heat preservation reaction for 3 hours to obtain protein peptide iron chelate reaction solution;
(6) spray drying: and (3) carrying out spray drying on the obtained protein peptide iron chelate reaction solution to obtain the protein peptide iron chelate.
Example 3
(1) Preparing protein liquid: weighing 100 kg of soybean protein isolate powder (the crude protein content is 91%), adding 600 kg of water, and uniformly stirring to obtain a protein solution;
(2) enzymolysis: adjusting the pH value of the protein solution to 8.2, adding 1.62 kg of protease, carrying out enzymolysis at 50 ℃ for 6 hours, heating the enzymolysis solution to 85 ℃ to inactivate the enzyme for 10 minutes to obtain enzymolysis solution;
(3) and (3) ultrafiltration: performing ultrafiltration on the enzymolysis liquid by an ultrafiltration membrane device with the molecular weight cutoff of 3000Da, and collecting the permeate to obtain ultrafiltration permeate;
(4) carrying out chelation reaction: adding 38 kg of ferrous sulfate into the protein peptide solution, continuously stirring to adjust the pH value of the solution to 6.0, heating to 58 ℃, and carrying out heat preservation reaction for 2.5 hours to obtain protein peptide iron chelate reaction solution;
(6) spray drying: and (3) carrying out spray drying on the obtained protein peptide iron chelate reaction solution to obtain the protein peptide iron chelate.
And (3) detecting the molecular weight of the protein peptide, the total content of iron elements, the content of chelated iron and the chelation rate of the finished product obtained in the embodiment.
Detecting the molecular weight of the peptide: the protein is determined by high performance gel filtration chromatography (HPLC) with known molecular weight as reference.
And (3) detecting the content of iron element: and respectively measuring the total content of the iron element in the protein peptide iron chelate and the content of chelated iron by using an atomic absorption spectrophotometer, and calculating the chelation rate of the protein peptide iron chelate according to the total content of the iron element in the protein peptide iron chelate and the content of chelated iron.
The results are shown in tables 1 and 2:
table 1: molecular weight distribution of protein peptide in protein peptide iron chelate
Table 2: iron element content and chelated iron content in protein peptide iron chelate
| Group of | Total content of iron element | Iron content in chelated form | Rate of chelation |
| Example 1 | 8.27% | 7.64% | 92.5% |
| Example 2 | 11.43% | 10.70% | 93.6% |
| Example 3 | 13.62% | 12.42% | 91.2% |
As can be seen from Table 1, the proportion of small-molecule peptides with relative molecular weight between 180-1000Da in the final product obtained in the example is more than 70% of the total protein. As can be seen from Table 2, the iron chelation rate of the finished product obtained in the example was more than 91%.
Claims (4)
1. A method for preparing protein peptide iron chelate suitable for ruminant animals comprises the following steps:
(1) preparing protein liquid: adding water into the isolated soy protein powder, stirring and mixing uniformly to prepare a soy protein solution with the protein concentration of 10-15% (w/w);
(2) enzymolysis: adjusting the pH value of the protein solution to 8.0-8.5, adding protease, performing enzymolysis at 50-55 deg.C for 5-8h, heating the enzymolysis solution to 85 deg.C, and inactivating enzyme for 10 min to obtain enzymolysis solution;
(3) and (3) ultrafiltration: carrying out ultrafiltration on the enzymolysis liquid obtained in the step (2) by an ultrafiltration membrane device with the molecular weight cutoff of 3000Da, and collecting a permeate to obtain a protein peptide solution;
(4) carrying out chelation reaction: adding water-soluble iron salt into the protein peptide solution obtained in the step (3), wherein the addition amount of the water-soluble iron salt is 30-50% of the amount of the protein peptide (based on the content of the protein) in the protein peptide solution, adjusting the pH value of the solution to 5.5-6.0, and carrying out stirring chelation reaction for 2-3 hours at 55-60 ℃ to obtain a protein peptide iron chelation reaction solution;
(5) spray drying: and (4) carrying out spray drying on the protein peptide iron chelate reaction solution obtained in the step (4) to obtain the protein peptide iron chelate.
2. The method for preparing a protein peptide iron chelate compound suitable for ruminants according to claim 1, wherein the method comprises the following steps: the soybean protein isolate powder in the step (1) is soybean protein isolate with the protein content of more than or equal to 90 percent.
3. The method for preparing a protein peptide iron chelate compound suitable for ruminants according to claim 1, wherein the method comprises the following steps: the protease in the step (2) is alkaline protease, the enzyme activity is 250000-300000U/ml, and the addition amount of the protease is 1.5-2.0% of the protein mass in the protein solution.
4. The method for preparing a protein peptide iron chelate compound suitable for ruminants according to claim 1, wherein the method comprises the following steps: the water-soluble iron salt in the step (4) is any one of ferrous fumarate, ferrous sulfate, ferrous citrate, ferrous lactate, ferrous chloride and ferrous carbonate.
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| CN201811225654.9A CN111073931A (en) | 2018-10-21 | 2018-10-21 | Preparation method of protein peptide iron chelate suitable for ruminants |
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| CN201811225654.9A CN111073931A (en) | 2018-10-21 | 2018-10-21 | Preparation method of protein peptide iron chelate suitable for ruminants |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2022006649A1 (en) * | 2020-07-08 | 2022-01-13 | Yessinergy Holding S/A | Method for producing chelated mineral concentrates with soy amino acids and/or proteins, and said product |
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- 2018-10-21 CN CN201811225654.9A patent/CN111073931A/en active Pending
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2022006649A1 (en) * | 2020-07-08 | 2022-01-13 | Yessinergy Holding S/A | Method for producing chelated mineral concentrates with soy amino acids and/or proteins, and said product |
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Application publication date: 20200428 |