CN111073933A - Preparation method of protein peptide selenium chelate suitable for ruminants - Google Patents
Preparation method of protein peptide selenium chelate suitable for ruminants Download PDFInfo
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- 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 selenium chelate suitable for ruminants, which mainly comprises the steps of crushing soybean meal, carrying out enzymolysis, carrying out filter pressing, carrying out ultrafiltration, chelating, and carrying out spray drying to obtain the protein peptide selenium chelate. The method of the invention uses the bean pulp with low price and wide source as the protein peptide source, uses the low-toxicity and high-efficiency nano selenium as the selenium source, chelates the nano selenium and the protein peptide, has simple process flow and low cost, and is suitable for large-scale industrial production. The product prepared by the invention has the characteristics of safety, stability, direct absorption, strong biological activity, lasting selenium supplement and the like, can be used as an animal feed additive, improves the feed utilization rate, enhances the immunity and disease resistance of animals, and promotes the growth of the animals.
Description
Technical Field
The invention relates to a preparation method of organic trace elements, in particular to a preparation method of a protein peptide selenium chelate suitable for ruminants.
Background
Selenium is used as a composition component of glutathione peroxidase (GSH-Px), can remove lipid peroxidation free radical intermediate products in organisms, prevent lipid peroxidation of biological membranes and maintain normal structure and function of the biological membranes; selenium is also involved in the synthesis of coenzyme A and coenzyme Q, and plays an important role in the tricarboxylic acid cycle and the electron transfer process in vivo. Selenium exists in selenoprotein in two forms of selenocysteine and selenomethionine, and exerts biological functions through the influence of the selenoprotein on the free radical metabolism, the antioxidant function, the immune function, the reproductive function, the apoptosis, the endocrine system and the like of animal organisms. Nutritional muscular atrophy can occur when selenium of animals is deficient, dyspnea and skeletal muscle stiffness can occur in the early stage, white muscle disease occurs in young animals, nutritional liver necrosis also occurs in pigs, susceptibility to diseases is increased when selenium of adult animals is deficient, reproductive dysfunction easily occurs in female animals and the like. Inorganic selenium has high toxicity and low absorption and utilization rate, and organic selenium and nano selenium have the advantages of low toxicity, easy absorption, high efficiency, high activity and the like, and are good selenium supplement sources.
The protein peptide microelement chelate is a new third-generation microelement additive, is easy to be absorbed by animals, has high biological value, is safe and efficient, supplements microelements while supplementing bioactive peptides and meeting the nutritional requirements of proteins, plays a very important role in the growth of animals, particularly the healthy growth of young animals, and is the development direction of the feed additive in the post-antibiotic age. However, at present, research and development on protein peptide trace element chelate at home and abroad are few, no mature technology exists, and particularly, no research report is found on a preparation method of a protein peptide selenium chelate suitable for ruminants.
Disclosure of Invention
The invention aims to provide a preparation method of a protein peptide selenium chelate compound suitable for ruminants based on the above situation, the method takes soybean meal as a main raw material, the process is simple, the cost is low, the implementation is easy, and the prepared protein peptide selenium chelate compound is used as a trace element selenium additive, so that the feed utilization rate can be obviously improved, the immunity and disease resistance of animals are enhanced, and the growth of the animals is promoted.
In order to achieve the purpose, the invention is realized by adopting the following technical scheme: a preparation method of a protein peptide selenium chelate suitable for ruminants comprises the following process steps:
(1) crushing the soybean meal: crushing the defatted soybean meal and sieving the crushed defatted soybean meal with a 100-mesh sieve to obtain soybean meal powder;
(2) enzymolysis: adding deionized water into the soybean meal powder obtained in the step (1), stirring and uniformly mixing to prepare a soybean meal liquid with a protein concentration of 8-10% (w/w), adjusting the pH value of the soybean meal liquid to 8.0-9.0, adding protease, performing enzymolysis at the temperature of 50-55 ℃ for 4-6 hours, heating the enzymolysis liquid to 85 ℃, and inactivating the enzyme for 10 minutes to obtain the soybean meal enzymolysis liquid;
(3) and (3) filter pressing: performing pressure filtration on the soybean meal enzymatic hydrolysate obtained in the step (2) by using a plate-and-frame filter press, wherein the mesh number of filter cloth is 200 meshes to obtain filter press liquid and filter cake, and drying and crushing the filter cake to prepare soybean residue feed;
(4) and (3) ultrafiltration: ultrafiltering the pressure filtrate obtained in the step (3) by an ultrafiltration membrane device with the molecular weight cutoff of 5000Da, and collecting the permeate to obtain a protein peptide solution;
(5) carrying out chelation reaction: adding nano selenium into the protein peptide solution obtained in the step (4), wherein the addition amount of the nano selenium is 1.5-3% of the amount of the protein peptide (based on the content of protein) in the protein peptide solution, adjusting the pH value of the solution to 8.0-8.5, and carrying out stirring chelation reaction for 1-1.5 hours at 45-50 ℃ to obtain a protein peptide selenium chelation reaction solution;
(6) spray drying: and (4) carrying out spray drying on the protein peptide selenium chelate reaction solution obtained in the step (5) to obtain the protein peptide selenium chelate.
The defatted soybean meal in the step (1) is defatted soybean meal with protein content more than or equal to 43 percent.
The protease in the step (2) is subtilisin, the enzyme activity is 200000-220000U/ml, and the addition amount of the protease is 1.0-1.5% of the mass of the protein in the soybean meal liquid.
And (5) mixing and grinding sodium selenite and citric acid according to the mass ratio of 1:2 to prepare the nano-selenium.
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 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 the bean pulp with low price and wide source as the protein peptide source, takes the low-toxicity and high-efficiency nano selenium as the selenium source, chelates the nano selenium and the protein peptide, mutually promotes the physiological functions of the protein peptide and the nano selenium, synergizes, and the obtained product has the characteristics of direct absorption, strong biological activity, safety, stability, lasting selenium supplement and the like, can obviously improve the growth performance of animals, and enhances the immunity and disease resistance of the animals.
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) Crushing the soybean meal: weighing 100Kg of defatted soybean meal (with the crude protein content of 45 percent), crushing and sieving by a 100-mesh sieve to obtain 100Kg of soybean meal;
(2) enzymolysis: adding 460L of deionized water into the soybean meal powder obtained in the step (1), uniformly stirring to prepare a soybean meal liquid with a protein concentration of 8% (w/w), adjusting the pH value of the soybean meal liquid to 8.0, adding protease with a protein mass of 1.0% in the soybean meal liquid, performing enzymolysis at 52 ℃ for 6 hours, heating the enzymolysis liquid to 85 ℃, and inactivating the enzyme for 10 minutes to obtain the soybean meal enzymolysis liquid;
(3) and (3) filter pressing: performing pressure filtration on the soybean meal enzymatic hydrolysate obtained in the step (2) by using a plate-and-frame filter press, wherein the mesh number of filter cloth is 200 meshes to obtain filter press liquid and filter cake, and drying and crushing the filter cake to prepare soybean residue feed;
(4) and (3) ultrafiltration: ultrafiltering the pressure filtrate obtained in the step (3) by an ultrafiltration membrane device with the molecular weight cutoff of 5000Da, and collecting the permeate to obtain a protein peptide solution 292L;
(5) carrying out chelation reaction: adding nano selenium with the protein peptide amount (based on the protein content) of 1.5% into the protein peptide solution obtained in the step (4), wherein the nano selenium is prepared by mixing sodium selenite and citric acid according to the mass ratio of 1:2 and then grinding, adjusting the pH value of the solution to 8.0, and stirring and reacting at 45 ℃ for 1.5 hours to obtain protein peptide selenium chelate reaction liquid 285L;
(6) spray drying: and (4) carrying out spray drying on the protein peptide selenium chelate reaction solution obtained in the step (5) to obtain 37.2KG of large protein peptide selenium chelate, wherein the detected selenium content is 0.177%.
Example 2
(1) Crushing the soybean meal: weighing 100Kg of defatted soybean meal (with the crude protein content of 45 percent), crushing and sieving by a 100-mesh sieve to obtain 100Kg of soybean meal;
(2) enzymolysis: adding 400L of deionized water into the soybean meal powder obtained in the step (1), uniformly stirring to prepare a soybean meal liquid with a protein concentration of 9% (w/w), adjusting the pH value of the soybean meal liquid to 8.5, adding protease with a protein mass of 1.2% of that of the soybean meal liquid, carrying out enzymolysis for 4h at a temperature of 50 ℃, heating the enzymolysis liquid to 85 ℃, and inactivating the enzyme for 10 min to obtain the soybean meal enzymolysis liquid;
(3) and (3) filter pressing: performing pressure filtration on the soybean meal enzymatic hydrolysate obtained in the step (2) by using a plate-and-frame filter press, wherein the mesh number of filter cloth is 200 meshes to obtain filter press liquid and filter cake, and drying and crushing the filter cake to prepare soybean residue feed;
(4) and (3) ultrafiltration: ultrafiltering the pressure filtrate obtained in the step (3) by an ultrafiltration membrane device with the molecular weight cutoff of 5000Da, and collecting the permeate to obtain 233L of protein peptide solution;
(5) carrying out chelation reaction: adding nano selenium with the protein peptide amount (based on the protein content) of 2.0% into the protein peptide solution obtained in the step (4), wherein the nano selenium is prepared by mixing sodium selenite and citric acid according to the mass ratio of 1:2 and then grinding, adjusting the pH value of the solution to 8.5, and stirring and reacting at 50 ℃ for 1.0 hour to obtain 226L protein peptide selenium chelate reaction liquid;
(6) spray drying: and (4) carrying out spray drying on the protein peptide selenium chelate reaction solution obtained in the step (5) to obtain 35.6KG of protein peptide selenium chelate, and detecting that the selenium content is 0.244%.
Example 3
(1) Crushing the soybean meal: weighing 100Kg of defatted soybean meal (with the crude protein content of 45 percent), crushing and sieving by a 100-mesh sieve to obtain 100Kg of soybean meal;
(2) enzymolysis: adding 350L of deionized water into the soybean meal powder obtained in the step (1), uniformly stirring to prepare a soybean meal liquid with a protein concentration of 10% (w/w), adjusting the pH value of the soybean meal liquid to 8.5, adding protease with a protein mass of 1.5% in the soybean meal liquid, carrying out enzymolysis at 53 ℃ for 5 hours, heating the enzymolysis liquid to 85 ℃, and inactivating the enzyme for 10 minutes to obtain the soybean meal enzymolysis liquid;
(3) and (3) filter pressing: performing pressure filtration on the soybean meal enzymatic hydrolysate obtained in the step (2) by using a plate-and-frame filter press, wherein the mesh number of filter cloth is 200 meshes to obtain filter press liquid and filter cake, and drying and crushing the filter cake to prepare soybean residue feed;
(4) and (3) ultrafiltration: ultrafiltering the pressure filtrate obtained in the step (3) by an ultrafiltration membrane device with the molecular weight cutoff of 1000Da, and collecting the permeate to obtain 210L of protein peptide solution;
(5) carrying out chelation reaction: adding nano selenium with the protein peptide amount (based on the protein content) of 2.5% into the protein peptide solution obtained in the step (4), wherein the nano selenium is prepared by mixing sodium selenite and citric acid according to the mass ratio of 1:2 and grinding, adjusting the pH value of the solution to 8.3, and stirring and reacting at 45 ℃ for 1.0 hour to obtain 198L protein peptide selenium chelate reaction solution;
(6) spray drying: and (4) carrying out spray drying on the protein peptide selenium chelate reaction solution obtained in the step (5) to obtain 36.1KG of protein peptide selenium chelate, and detecting that the selenium content is 0.302%.
The selenium content, the crude protein content and the peptide molecular weight of the finished product obtained in the above embodiment are respectively detected.
The selenium content is determined by atomic absorption spectrophotometry, the crude protein content is determined by GB/T6432-94 method, the peptide molecular weight distribution is determined by high performance gel filtration chromatography (HPLC) method, and known molecular weight protein is used as reference.
The results are shown in Table 1:
table 1: protein content and peptide molecular weight distribution of protein peptide selenium chelate
As can be seen from Table 1, the selenium content and crude protein content of the final product obtained in the examples are both above 0.1% and above 76%, wherein the proportion of small-molecule peptides with relative molecular weight between 200-1500Da to the total protein is above 60%.
Claims (4)
1. A method for preparing protein peptide selenium chelate suitable for ruminants comprises the following steps:
(1) crushing the soybean meal: crushing the defatted soybean meal and sieving the crushed defatted soybean meal with a 100-mesh sieve to obtain soybean meal powder;
(2) enzymolysis: adding deionized water into the soybean meal powder obtained in the step (1), stirring and uniformly mixing to prepare a soybean meal liquid with a protein concentration of 8-10% (w/w), adjusting the pH value of the soybean meal liquid to 8.0-9.0, adding protease, performing enzymolysis at the temperature of 50-55 ℃ for 4-6 hours, heating the enzymolysis liquid to 85 ℃, and inactivating the enzyme for 10 minutes to obtain the soybean meal enzymolysis liquid;
(3) and (3) filter pressing: performing pressure filtration on the soybean meal enzymatic hydrolysate obtained in the step (2) by using a plate-and-frame filter press, wherein the mesh number of filter cloth is 200 meshes to obtain filter press liquid and filter cake, and drying and crushing the filter cake to prepare soybean residue feed;
(4) and (3) ultrafiltration: ultrafiltering the pressure filtrate obtained in the step (3) by an ultrafiltration membrane device with the molecular weight cutoff of 5000Da, and collecting the permeate to obtain a protein peptide solution;
(5) carrying out chelation reaction: adding nano selenium into the protein peptide solution obtained in the step (4), wherein the addition amount of the nano selenium is 1.5-3% of the amount of the protein peptide (based on the content of protein) in the protein peptide solution, adjusting the pH value of the solution to 8.0-8.5, and carrying out stirring chelation reaction for 1-1.5 hours at 45-50 ℃ to obtain a protein peptide selenium chelation reaction solution;
(6) spray drying: and (4) carrying out spray drying on the protein peptide selenium chelate reaction solution obtained in the step (5) to obtain the protein peptide selenium chelate.
2. The method for preparing protein peptide selenium chelate for ruminant animals as claimed in claim 1, wherein said method comprises the steps of: the defatted soybean meal in the step (1) is defatted soybean meal with protein content more than or equal to 43 percent.
3. The method for preparing protein peptide selenium chelate for ruminant animals as claimed in claim 1, wherein said method comprises the steps of: the protease in the step (2) is subtilisin, the enzyme activity is 200000-220000U/ml, and the addition amount of the protease is 1.0-1.5% of the mass of the protein in the soybean meal liquid.
4. The method for preparing protein peptide selenium chelate for ruminant animals as claimed in claim 1, wherein said method comprises the steps of: and (5) mixing and grinding sodium selenite and citric acid according to the mass ratio of 1:2 to prepare the nano-selenium.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111302858A (en) * | 2020-03-09 | 2020-06-19 | 沃博特生物科技有限公司 | Chelation method of medium trace elements in optical carbon ribozyme |
| CN113383960A (en) * | 2021-06-10 | 2021-09-14 | 广州大学 | High-stability protein polypeptide-nano selenium and preparation method and application thereof |
| CN116035219A (en) * | 2023-02-03 | 2023-05-02 | 贵州国科锌硒科技有限公司 | Zinc-selenium tea collagen peptide and preparation process and application thereof |
-
2018
- 2018-10-21 CN CN201811225714.7A patent/CN111073933A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111302858A (en) * | 2020-03-09 | 2020-06-19 | 沃博特生物科技有限公司 | Chelation method of medium trace elements in optical carbon ribozyme |
| CN113383960A (en) * | 2021-06-10 | 2021-09-14 | 广州大学 | High-stability protein polypeptide-nano selenium and preparation method and application thereof |
| CN116035219A (en) * | 2023-02-03 | 2023-05-02 | 贵州国科锌硒科技有限公司 | Zinc-selenium tea collagen peptide and preparation process and application thereof |
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Application publication date: 20200428 |