CN111096395A - Preparation method of protein peptide calcium chelate suitable for ruminants - Google Patents
Preparation method of protein peptide calcium chelate suitable for ruminants Download PDFInfo
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- CN111096395A CN111096395A CN201811247974.4A CN201811247974A CN111096395A CN 111096395 A CN111096395 A CN 111096395A CN 201811247974 A CN201811247974 A CN 201811247974A CN 111096395 A CN111096395 A CN 111096395A
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/10—Feeding-stuffs specially adapted for particular animals for ruminants
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/14—Pretreatment of feeding-stuffs with enzymes
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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
- A23K20/24—Compounds of alkaline earth metals, e.g. magnesium
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Abstract
The invention relates to a preparation method of a protein peptide calcium chelate suitable for ruminants, which mainly comprises the steps of protein liquid preparation, enzymolysis, ultrafiltration, chelation and spray drying to obtain the protein peptide calcium chelate. The method takes soybean protein with wide sources as a protein peptide source and inorganic calcium salt as a calcium source, chelates iron element with 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 calcium ions, and is a novel high-efficiency calcium supplement for feed.
Description
Technical Field
The invention relates to a preparation method of an organic macroelement, in particular to a preparation method of a protein peptide calcium chelate suitable for ruminants.
Background
Calcium is a mineral essential for the growth and development of animals and for maintaining physiological functions, and is also the most abundant inorganic element in the animal body. 99% of the calcium in animal bodies exists in bones and teeth, which constitute a body scaffold, which becomes a reservoir of calcium in the body, and 1% of the calcium exists in soft tissues, intercellular spaces and blood, which are collectively called a miscible calcium pool, and is in dynamic balance with bone calcium. Bone calcium exists primarily as amorphous calcium hydrogen phosphate (CaHPO4) and crystalline hydroxyapatite (3Ca3PO4 xca (oh)2), and bone maintains its dynamic balance with blood calcium through constant osteogenesis and osteolysis. Calcium plays an important role in animal metabolism and reproductive activities, and calcium plays an important physiological regulatory role in the exertion of all cellular functions. Calcium is an activator of more than 200 enzymes in a body, so that each organ can normally operate, calcium element participates in metabolism of the body, so that calcium needs to be supplemented every day, and the deficiency of the calcium content in the body of the animal can seriously affect the growth and health of the animal, so that the calcium supplement in daily ration of the livestock is very important.
At present, common calcium supplements in the market are mainly inorganic calcium, and inorganic calcium ions are easily affected by phytic acid, amino acid, cellulose, sugar compounds, vitamins, unsaturated fatty acid and the like in feed in the digestion and absorption process, so that the absorption and utilization rate is low. The research proves that the organic calcium has higher bioavailability than inorganic calcium and has obvious promotion effect on the growth, reproduction, health, feed conversion rate and the like of animals. The protein peptide chelated calcium is a novel organic calcium source supplement which develops rapidly at home and abroad in recent years, and is a compound with a cyclic structure generated by the coordination reaction of calcium ions and protein peptides. Because the protein peptide chelated calcium is a form of calcium element which is close to the natural form in the animal body, the protein peptide chelated calcium has good chemical stability, higher biological value, easy digestion and absorption, interference resistance, no stimulation and no toxic action, is more superior to the corresponding inorganic calcium ion, and is considered as an ideal calcium supplement for feed at present. Therefore, the development of a protein peptide chelated calcium 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 protein peptide calcium chelate are not much in China, and no research report is found about a preparation method of the protein peptide calcium chelate suitable for ruminants.
Disclosure of Invention
The invention aims to provide a preparation method of a protein peptide calcium chelate suitable for ruminants based on the above situation, and the method takes soy protein as a main raw material, has simple process and low cost and is easy to implement.
In order to achieve the purpose, the invention is realized by adopting the following technical scheme: a method for preparing protein peptide calcium 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 calcium salt into the protein peptide solution obtained in the step (3), wherein the addition amount of the water-soluble calcium 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 a chelation reaction for 2-3 hours at 55-60 ℃ under stirring to obtain a protein peptide calcium chelation reaction solution;
(5) spray drying: and (4) carrying out spray drying on the protein peptide calcium chelate reaction solution obtained in the step (4) to obtain the protein peptide calcium 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 calcium salt in the step (4) is any one of calcium chloride, calcium nitrate, calcium chlorate, calcium bicarbonate, calcium dihydrogen phosphate and calcium lactate.
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 of the invention takes soybean protein with wide and easily available sources as a peptide source and inorganic calcium salt as a calcium source to chelate calcium ions and protein peptide to prepare the protein peptide calcium 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 percent, the content of chelated calcium ions is more than 7 percent, and the chelating rate is up to more than 90 percent.
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 calcium chloride 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 calcium chelating reaction solution;
(5) spray drying: and (4) carrying out spray drying on the protein peptide calcium chelate reaction solution obtained in the step (4) to obtain the protein peptide calcium 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 calcium chloride 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 calcium chelating reaction solution;
(6) spray drying: and (3) carrying out spray drying on the obtained protein peptide calcium chelate reaction solution to obtain the protein peptide calcium 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 calcium chloride 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 calcium chelating reaction liquid;
(6) spray drying: and (3) carrying out spray drying on the obtained protein peptide calcium chelate reaction solution to obtain the protein peptide calcium chelate.
And (3) detecting the molecular weight of the protein peptide, the total content of calcium elements, the content of chelated calcium 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 and the content of chelated calcium in the protein peptide calcium chelate by using an atomic absorption spectrophotometer, and calculating the chelation rate of the protein peptide calcium chelate according to the total content of the iron element and the content of chelated calcium in the protein peptide calcium chelate.
The results are shown in tables 1 and 2:
table 1: molecular weight distribution of protein peptide in protein peptide iron chelate
Table 2: calcium element content and chelated calcium content in protein peptide calcium chelate
| Group of | Total content of calcium element | Chelated calcium content | 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 calcium sequestration rate in the final product obtained in the examples reaches more than 90%.
Claims (4)
1. A method for preparing protein peptide calcium 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 calcium salt into the protein peptide solution obtained in the step (3), wherein the addition amount of the water-soluble calcium 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 a chelation reaction for 2-3 hours at 55-60 ℃ under stirring to obtain a protein peptide calcium chelation reaction solution;
(5) spray drying: and (4) carrying out spray drying on the protein peptide calcium chelate reaction solution obtained in the step (4) to obtain the protein peptide calcium chelate.
2. The method for preparing protein peptide calcium chelate suitable for ruminants according to claim 1, wherein the protein peptide calcium chelate is prepared by 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 protein peptide calcium chelate suitable for ruminants according to claim 1, wherein the protein peptide calcium chelate is prepared by 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 protein peptide calcium chelate suitable for ruminants according to claim 1, wherein the protein peptide calcium chelate is prepared by the following steps: the water-soluble calcium in the step (4) is any one of calcium chloride, calcium nitrate, calcium chlorate, calcium bicarbonate, calcium dihydrogen phosphate and calcium lactate.
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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
- 2018-10-25 CN CN201811247974.4A patent/CN111096395A/en active Pending
Cited By (2)
| 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 |
| GB2601858A (en) * | 2020-07-08 | 2022-06-15 | Yessinergy Holding S A | Process for production of concentrates of chelated minerals |
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