CN113528599A - Production method of efficient chelating enzyme peptide - Google Patents
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Abstract
The invention belongs to the field of biological feed, and relates to a production method of high-efficiency chelating enzyme peptide. The method comprises the following steps: activating the expanded culture strains to respectively obtain bacillus subtilis, saccharomyces cerevisiae, lactobacillus plantarum and lactobacillus rhamnosus fermentation seed liquid; weighing bean dregs and bean pulp, and uniformly stirring to prepare a solid fermentation culture medium; inoculating bacillus subtilis fermentation seed liquid, adding wheat middling, adjusting water content and carrying out primary aerobic fermentation; and inoculating saccharomyces cerevisiae, lactobacillus plantarum, lactobacillus rhamnosus fermentation seed liquid, protease and inorganic mineral salt for secondary anaerobic fermentation, and obtaining the chelate type enzyme peptide after the fermentation is finished. The invention adopts the mode of aerobic and anaerobic step-by-step fermentation, bacterial enzyme synergistic fermentation and synchronous fermentation and chelation, the production process is simple, the chelation rate of various mineral elements in the obtained chelated enzyme peptide is high, and the chelated Fe is Fe2+Mainly contains low phytic acid content, high small peptide content, organic acid content and probiotics, and has obvious economic benefit.
Description
Technical Field
The invention belongs to the field of biological feed, and particularly relates to a production method of mineral element efficient chelating type enzyme peptide suitable for poultry.
Background
Ca. The mineral elements such as Fe, Cu, Zn, Mn and the like relate to almost all life activity processes such as growth metabolism, signal transduction, immunoregulation, enzyme activation and the like of poultry such as laying hens and the like, and have very important significance for maintaining normal physiological functions, health states and high-efficiency production performance of organisms. When the feed is lack of corresponding mineral elements, the poultry can have the phenomena of slow growth, reduced disease resistance, reduced laying rate, reduced quality of laid eggs and the like.
Poultry mineral element supplementation underwent four generations of development. First generation, inorganic salts, such as: ferrous sulfate, copper sulfate, and the like. The inorganic salt type mineral elements have poor stability and low absorption utilization rate, and different mineral elements have antagonism and are easy to combine with phytic acid and the like to form insoluble salt to influence absorption. Second generation, organic acid type, such as: calcium lactate, zinc gluconate, and the like. The absorption utilization rate of the organic acid mineral elements is improved compared with the first generation, but the problems of high price, poor stability, absorption competition among different mineral elements and the like still exist. Third generation, amino acid chelating, such as: ferrous glycine, zinc methionine, and the like. The amino acid chelated mineral elements have the advantages of high absorption and utilization rate, strong stability, safety, no toxic or side effect, extremely high price and great increase of the breeding cost when being used in large-scale breeding. Fourth generation, small peptide chelation type, such as: soybean peptide chelated calcium, casein chelated iron and the like. The mineral elements are transported and absorbed in a chelate form by virtue of a transport and absorption mechanism of the small peptide. The small peptide chelating mineral element has the advantages of low price, high absorption and utilization rate, strong stability, no toxic or side effect and the like. The chelating type of small peptide also has the biological functions of small peptide ligands, such as: enhancing immunity, resisting bacteria, resisting oxidation, and reducing cholesterol. However, the research on the small peptide chelating mineral elements is mostly in a theoretical stage, the actual industrialized production is less, and the purchase on the market is difficult.
In actual cultivation, in order to control cultivation cost and meet the requirements of poultry on mineral elements such as Ca, Fe, Cu, Zn, Mn and the like, a large amount of inorganic salt type mineral elements are often added. The poultry has low absorption and utilization rate of inorganic mineral salt, and most of the inorganic mineral salt is directly discharged out of the body, thereby causing serious environmental pollution and resource waste. The poultry takes too much inorganic mineral salt, and has huge toxic and side effects on the organism. In addition, Fe in the form of an inorganic salt2+Is very easily oxidized into Fe3+Obviously reduce the Fe of poultry2+The absorption and utilization of the blood lead to the reduction of hemoglobin in the body, the reduction of oxygen transportation capability and the generation of anemia.
At present, the supplementation of chelating mineral elements is the most ideal mineral element supplementation mode recognized. According to the traditional production mode of the chelate mineral elements, firstly, proteins are hydrolyzed by methods such as chemical hydrolysis and enzyme hydrolysis, and then, micromolecular substances such as high-purity amino acid and small peptide are obtained through protein purification, or pure amino acid is obtained through an amino acid fermentation process or a chemical synthesis process, and then, free mineral elements and the purified micromolecular substances are chelated through a corresponding chelation process. Finally, the chelating mineral elements with high purity are obtained by methods such as absolute ethyl alcohol washing, liquid chromatography separation and the like. The obtained chelate of the mineral elements has high purity and strong stability. However, the production process of these high-purity chelates is complicated, the equipment requirement is high, the production cost is high, and difficulties are caused in the actual production and wide use of the mineral element chelates. Further, a single complex having a high purity is sought after, and is not necessarily an optimal mineral element supplement. The single mineral element chelate or single amino acid and small peptide chelate can not satisfy the comprehensive mineral element and amino acid nutrition supplement. In addition, the high-price amino acid and small peptide chelate greatly increases the cost of the breeding feed, and is a great resistance to the development of the breeding industry. All types of nutrients have their absorption pathways, which are saturated. Although single amino acids or small peptide chelates are much stronger than inorganic mineral salts, their addition beyond their absorptive capacity also reduces the absorption of mineral elements.
Disclosure of Invention
Aiming at the problems of mineral element supplement of the existing poultry, the invention provides a method for producing mineral element efficient chelating type enzyme peptide by taking bean dregs and bean pulp as raw materials and performing bacterial enzyme synergistic fermentation and chelation. The method has the advantages of simple production process, low cost and remarkable economic benefit. The chelating type enzyme peptide has high chelating rate of various mineral elements, and the chelated Fe is Fe2+Mainly, the content of phytic acid is low, and the content of small peptide, organic acid, probiotics and the like is high.
In order to realize the technical purpose, the invention provides a method for producing mineral element efficient chelating type enzyme peptide by fermenting and chelating bean dregs and bean pulp serving as raw materials, which comprises the following steps:
(1) respectively carrying out activation and propagation on bacillus subtilis, saccharomyces cerevisiae, lactobacillus plantarum and lactobacillus rhamnosus to obtain corresponding fermentation seed liquid;
(2) weighing bean dregs and bean pulp, and uniformly stirring to prepare a solid fermentation culture medium;
(3) inoculating the bacillus subtilis fermentation seed liquid obtained in the step (1) into the solid fermentation culture medium prepared in the step (2), adding wheat middling, adding water according to the water content, uniformly stirring, and performing primary aerobic fermentation to obtain an aerobic fermentation culture medium;
(4) and (3) inoculating the saccharomyces cerevisiae fermentation seed liquid, the lactobacillus plantarum fermentation seed liquid, the lactobacillus rhamnosus fermentation seed liquid, the protease and the inorganic mineral salt obtained in the step (1) into the aerobic fermentation culture medium obtained in the step (3), uniformly stirring the mixture, sealing the mixture for secondary anaerobic fermentation, and obtaining the mineral element efficient chelating type enzyme peptide after the fermentation is finished.
Preferably, the bean dregs in the solid fermentation medium in the step (2) account for 25-70% of the total mass, and the bean dregs account for 30-75% of the total mass.
Preferably, the inoculation amount of the bacillus subtilis in the step (3) is 1-15% of the mass of the solid fermentation culture medium in the step (2); the addition amount of the wheat middling is 5-30% of the mass of the solid fermentation medium in the step (2).
Preferably, the water content in the step (3) is 25-65%.
Preferably, the first-stage aerobic fermentation temperature in the step (3) is 20-40 ℃, and the aerobic fermentation time is 3-48 h.
Preferably, the inoculation amounts of the saccharomyces cerevisiae, lactobacillus plantarum and lactobacillus rhamnosus seed liquid in the step (4) are all 1-10% of the mass of the solid fermentation culture medium in the step (2).
Preferably, the protease in the step (4) is acid protease and papain; the total adding amount of the acid protease and the papain is 0.01-2% of the mass of the solid fermentation medium in the step (2), the adding ratio of the two enzymes is 1:1, and the enzyme activities are both 6 ten thousand u/g.
Preferably, the inorganic mineral salt in the step (4) is CaCO3、FeSO4·7H2O、CuSO4·5H2O、MnSO4·H2O and ZnSO4·7H2O。
Preferably, the CaCO in step (4)3The adding amount is 1-12% of the mass of the solid fermentation medium in the step (2); the FeSO4·7H2The adding amount of O is 0.01-1.6% of the mass of the solid fermentation medium in the step (2); the CuSO4·5H2The addition amount of O is 0.001-0.064% of the mass of the solid fermentation medium in the step (2); the MnSO4·H2The addition amount of O is 0.01-1% of the mass of the solid fermentation medium in the step (2); the ZnSO4·7H2The addition amount of O is 0.01-1.2% of the mass of the solid fermentation medium in the step (2).
Preferably, the temperature of the second-stage anaerobic fermentation in the step (4) is 20-40 ℃, and the fermentation time is 2-30 days.
The strain source is as follows: lactobacillus plantarum (CGMCC 1.557) and Lactobacillus rhamnosus (CGMCC 1.2467) are purchased from China general microbiological culture Collection center; bacillus subtilis and Saccharomyces cerevisiae were purchased from Biotech Ltd.
Enzyme sources: both acid protease and papain are available from Xiushang enzyme Biotechnology Ltd.
The invention has the beneficial effects that:
(1) the invention simultaneously performs fermentation and mineral element chelation, has simple and easy operation of fermentation process and low production cost, obviously reduces the feeding cost of the mineral elements of the poultry, and meets the requirement of large-scale production and application.
(2) The chelating type ferment peptide obtained by the invention has high chelating rate of various mineral elements, and the absorption and utilization rate of the mineral elements by poultry is obviously improved.
(3) The Fe in the chelate type ferment peptide obtained by the invention is mainly Fe2+And a chelating enzyme peptide pair Fe2+Has good antioxidant effect and prolongs Fe2+The storage time of the fertilizer is obviously prolonged, and the Fe content of poultry is obviously improved2+The absorption and utilization rate of the feed additive reduces the occurrence of poultry anemia.
(4) The invention obviously reduces the phytic acid content in the chelating type enzyme peptide through the bacterial enzyme synergistic fermentation, avoids the combination of the phytic acid and mineral elements and reduces the absorption of the mineral elements.
(5) According to the invention, the content of small peptides, the content of organic acids and the content of probiotics in the chelating enzyme peptide are obviously increased through the synergistic fermentation of the bacterial enzymes, the nutritional quality of the chelating enzyme peptide is improved, the protein absorption of poultry is facilitated, the health of intestinal flora is maintained, the immunity of poultry is enhanced, and the use of antibiotics is reduced.
Drawings
FIG. 1 is a flow diagram of a fermentation process of the present invention.
The specific implementation mode is as follows:
the invention is further described with reference to specific examples.
Example 1:
(1) activating and culturing bacillus subtilis, saccharomyces cerevisiae, lactobacillus plantarum and lactobacillus rhamnosus and preparing a fermentation seed solution:
A. preparing a strain culture medium:
slant and plate media: 5g of yeast extract, 10g of peptone, 10g of sodium chloride, 10g of glucose, 20g of agar and 1000mL of distilled water, and sterilizing at 121 ℃ for 30 min;
seed liquid nutrient medium: preparing soybean milk from 20g of soybeans, 20g of brown sugar and 1000mL of distilled water, and sterilizing at 121 ℃ for 30 min;
B. activation of each strain and preparation of fermentation seed liquid:
activating bacillus subtilis and preparing a fermentation seed solution: inoculating Bacillus subtilis strain stored at 4 deg.C into slant culture medium, and culturing at 37 deg.C for 36 h. Then, single colonies were streaked on the plate, inverted cultured at 37 ℃ for 36 hours, picked and inoculated into a 250mL conical flask containing 50mL seed liquid culture medium, and cultured at 37 ℃ for 18 hours at 160 r/min. Performing amplification culture at 3% (v/v) to obtain fermentation seed solution, and culturing at 37 deg.C and 160r/min for 24 hr.
Activation of saccharomyces cerevisiae and preparation of fermentation seed liquid: the saccharomyces cerevisiae strain preserved at 4 ℃ is inoculated in a slant culture medium and cultured for 36h at 28 ℃. Then, single colonies were streaked on the plate, inverted and cultured at 28 ℃ for 36 hours, and the single colonies were selected and inoculated into 250mL Erlenmeyer flasks containing 50mL of seed liquid medium, and cultured at 28 ℃ for 18 hours at 160 r/min. Performing amplification culture at 3% (v/v) to obtain fermentation seed solution, and culturing at 28 deg.C and 160r/min for 24 hr.
Activating lactobacillus plantarum and lactobacillus rhamnosus and preparing a fermented seed liquid: the lactobacillus plantarum and lactobacillus rhamnosus strains preserved at 4 ℃ are respectively inoculated in a slant culture medium and cultured for 48 hours at 37 ℃. And then, single colonies are scratched on the plate, the plate is subjected to inverted culture at 37 ℃ for 48h, the single colonies are selected and inoculated into a 250mL anaerobic bottle filled with 230mL seed liquid culture medium, and the plate is cultured at 37 ℃ for 18 h. Performing amplification culture at 3% (v/v) to obtain fermented seed solution, and culturing at 37 deg.C for 24 hr.
(2) Preparing a solid fermentation culture medium: the weight ratio of each component is as follows: 50% of bean dregs and 50% of bean pulp;
(3) inoculating the prepared bacillus subtilis fermentation seed liquid into a fermentation culture medium according to the inoculation amount of 3% of the mass of the solid fermentation culture medium, adding wheat middling 10% of the mass of the solid fermentation culture medium, adding water to ensure that the water content is 45%, uniformly stirring, and carrying out primary aerobic fermentation at the fermentation temperature of 32 ℃ for 15 hours;
(4) respectively inoculating the obtained Saccharomyces cerevisiae, Lactobacillus plantarum and Lactobacillus rhamnosus fermentation seed liquid into a fermentation culture medium according to the inoculation amount of 3% of the solid fermentation culture medium, adding 0.5% protease (the ratio of acid protease to papain is 1:1) according to the solid fermentation culture medium, and finally adding 6% CaCO3、0.8％FeSO4·7H2O、0.032％CuSO4·5H2O、0.5％MnSO4·H2O、0.6％ZnSO4·7H2And O, uniformly stirring, filling into a fermentation bag with a one-way vent hole, sealing for secondary anaerobic fermentation at 31 ℃ for 15 days, and obtaining the mineral element high-efficiency chelating type enzyme peptide after the fermentation is finished.
Example 2:
(1) the specific operation of the activating and fermenting seed liquid of the bacillus subtilis, the saccharomyces cerevisiae, the lactobacillus plantarum and the lactobacillus rhamnosus is the same as that of the embodiment 1;
(2) preparing a solid fermentation culture medium: the weight ratio of each component is as follows: 60% of bean dregs and 40% of bean pulp;
(3) inoculating the prepared bacillus subtilis fermentation seed liquid into a fermentation culture medium according to the inoculation amount of 8% of the mass of the solid fermentation culture medium, adding wheat middling 24% of the mass of the solid fermentation culture medium, adding water to enable the water content to be 55%, uniformly stirring, and carrying out primary aerobic fermentation at the fermentation temperature of 36 ℃ for 24 hours;
(4) respectively inoculating the prepared saccharomyces cerevisiae, lactobacillus plantarum and lactobacillus rhamnosus fermentation seed liquid into a fermentation culture medium according to the inoculation amount of 10 percent of the mass of the solid fermentation culture medium, and then adding 1.5 percent of protease (the ratio of acid protease to papain is 1:1) and 12 percent of CaCO according to the mass of the solid fermentation culture medium3、0.4％FeSO4·7H2O、0.05％CuSO4·5H2O、0.1％MnSO4·H2O、1.2％ZnSO4·7H2And O, uniformly stirring, filling into a fermentation bag with a one-way vent hole, sealing for secondary anaerobic fermentation at the fermentation temperature of 40 ℃ for 8 days, and obtaining the mineral element high-efficiency chelating type enzyme peptide after the fermentation is finished.
Example 3:
(1) the specific operation of the activating and fermenting seed liquid of the bacillus subtilis, the saccharomyces cerevisiae, the lactobacillus plantarum and the lactobacillus rhamnosus is the same as that of the embodiment 1;
(2) preparing a solid fermentation culture medium: the weight ratio of each component is as follows: 40% of bean dregs and 60% of bean pulp;
(3) inoculating the prepared bacillus subtilis fermentation seed liquid into a fermentation culture medium according to the inoculation amount of 10% of the mass of the solid fermentation culture medium, adding wheat middling 18% of the mass of the solid fermentation culture medium, adding water to ensure that the water content is 40%, uniformly stirring, and carrying out primary aerobic fermentation at the fermentation temperature of 25 ℃ for 48 hours;
(4) respectively inoculating the prepared saccharomyces cerevisiae, lactobacillus plantarum and lactobacillus rhamnosus fermentation seed liquid into a solid fermentation culture medium according to the inoculation amount of 5 percent of the mass of the solid fermentation culture mediumAdding protease (acidic protease and papain ratio is 1:1) 1 wt% of the solid fermentation medium, and adding CaCO 4%3、1.6％FeSO4·7H2O、0.064％CuSO4·5H2O、0.01％MnSO4·H2O、0.2％ZnSO4·7H2And O, uniformly stirring, filling into a fermentation bag with a one-way vent hole, sealing for secondary anaerobic fermentation at the fermentation temperature of 37 ℃ for 2 days, and obtaining the mineral element high-efficiency chelating type enzyme peptide after the fermentation is finished.
Example 4:
(1) the specific operation of the activating and fermenting seed liquid of the bacillus subtilis, the saccharomyces cerevisiae, the lactobacillus plantarum and the lactobacillus rhamnosus is the same as that of the embodiment 1;
(2) preparing a solid fermentation culture medium: the weight ratio of each component is as follows: 25% of bean dregs and 75% of bean pulp.
(3) Inoculating the prepared bacillus subtilis fermentation seed liquid into a fermentation culture medium according to the inoculation amount of 1% of the mass of the solid fermentation culture medium, adding wheat middling 30% of the mass of the solid fermentation culture medium, adding water to enable the water content to be 25%, uniformly stirring, and carrying out primary aerobic fermentation at the fermentation temperature of 40 ℃ for 9 hours;
(4) inoculating the obtained Saccharomyces cerevisiae, Lactobacillus plantarum and Lactobacillus rhamnosus fermentation seed liquid into a fermentation culture medium according to the inoculation amount of 8% of the mass of the solid fermentation culture medium, adding 2% of protease (the ratio of acid protease to papain is 1:1) according to the mass of the solid fermentation culture medium, and finally adding 8% of CaCO3、0.01％FeSO4·7H2O、0.01％CuSO4·5H2O、0.8％MnSO4·H2O、0.9％ZnSO4·7H2And O, uniformly stirring, sealing for secondary anaerobic fermentation at the fermentation temperature of 20 ℃ for 21 days, and obtaining the mineral element high-efficiency chelating type enzyme peptide after the fermentation is finished.
Example 5:
(1) the specific operation of the activating and fermenting seed liquid of the bacillus subtilis, the saccharomyces cerevisiae, the lactobacillus plantarum and the lactobacillus rhamnosus is the same as that of the embodiment 1;
(2) preparing a solid fermentation culture medium: the weight ratio of each component is as follows: 70% of bean dregs and 30% of bean pulp;
(3) inoculating the prepared bacillus subtilis fermentation seed liquid into a fermentation culture medium according to the inoculation amount of 15% of the mass of the solid fermentation culture medium, adding water to ensure that the water content is 65%, uniformly stirring, and performing primary aerobic fermentation at the fermentation temperature of 20 ℃ for 3 hours;
(4) respectively inoculating the obtained Saccharomyces cerevisiae, Lactobacillus plantarum and Lactobacillus rhamnosus fermentation seed liquid into a fermentation culture medium according to the inoculation amount of 1% of the mass of the solid fermentation culture medium, adding 0.01% of protease (the ratio of acid protease to papain is 1:1) according to the mass of the solid fermentation culture medium, and finally adding 1% of CaCO3、1.2％FeSO4·7H2O、0.001％CuSO4·5H2O、1％MnSO4·H2O、0.01％ZnSO4·7H2And O, uniformly stirring, sealing for secondary anaerobic fermentation at 26 ℃ for 30 days, and obtaining the mineral element high-efficiency chelating type enzyme peptide after the fermentation is finished.
Table 1 shows the chelating ratio of each mineral element in the chelating type enzyme peptides in each example
| Mineral elements | Example 1 | Example 2 | Example 3 | Example 4 | Example 5 |
| Ca(％) | 89.87 | 82.35 | 85.74 | 87.86 | 93.14 |
| Fe(％) | 97.73 | 98.16 | 97.13 | 98.79 | 97.38 |
| Cu(％) | 76.42 | 74.24 | 73.85 | 77.43 | 78.29 |
| Mn(％) | 96.84 | 96.92 | 97.46 | 96.25 | 96.18 |
| Zn(％) | 96.28 | 95.63 | 96.37 | 95.82 | 97.93 |
Table 2 shows the contents of the relevant substances in the chelating type enzyme peptides in the respective examples
Example 6: experiment of feeding
The experimental site was a layer farm of Zhejiang Kangxing biotechnology limited, and the highly effective chelating type enzyme peptide used was prepared in example 1. Taking 72 local three-yellow layer chickens, randomly combining into two groups of 36 chickens, feeding the experimental layer chickens with the feed formula shown in table 3 for 60 days, and feeding the experimental results shown in tables 4, 5 and 6.
As is clear from the results in tables 4 to 6, the amount of the highly effective chelating enzyme peptide of the present invention added was 5%. The chelating enzyme peptide fed to the laying hens can obviously improve the average daily feed intake, the laying rate, the average egg weight and the eggshell thickness of the laying hens and reduce the soft egg breaking rate, which indicates that the chelating enzyme peptide obtained by the invention improves the production performance of the laying hens. Meanwhile, the content of Ca, Fe, Cu, Mn and Zn in the excrement of the laying hens is obviously reduced by feeding the chelating type enzyme peptide, and the chelating type enzyme peptide has the effects of improving the absorption of the laying hens on mineral elements and reducing the discharge of the mineral elements along with the excrement. The erythrocyte, hemoglobin and hematocrit of the laying hens fed with the chelate type enzyme peptide are all obviously higher than those of the control group, which indicates that the feeding of the chelate type enzyme peptide improves the Fe content of the laying hens2+、Cu2+The absorption of the feed additive and the like enhances the transportation of oxygen in the laying hens and avoids the anemia of the laying hens.
Table 3 shows the feed formula of experimental laying hens
| Composition of | Experimental group (%) | Control group (%) |
| Corn (corn) | 62 | 62 |
| Bean pulp | 26.28 | 26.28 |
| Multi-element vitamin | 0.05 | 0.05 |
| DL methionine | 0.12 | 0.12 |
| Stone | 5 | 5 |
| Calcium hydrogen phosphate | 1.4 | 1.4 |
| Vc | 0.1 | 0.1 |
| Phytase | 0.05 | 0.05 |
| Mineral elements | 5 (chelating enzyme peptide) | 5 (unfermented feed) |
| Total up to | 100 | 100 |
Note: the unfermented feed correspondingly added in the mineral elements of the control group is different from the chelate type enzyme peptides added in the experimental group in that no strain and protease are added in the control group, the fermentation operation is not carried out, and the rest added substances and the use amount are the same as those in the experimental group.
Table 4 shows the results of the experimental laying hen feeding performance test
Table 5 shows the content of mineral ions in the feces of experimental laying hens
| Mineral elements | Experimental group | Control group |
| Ca | 18.99 | 25.8 |
| Fe | 0.44 | 0.70 |
| Cu | 0.059 | 0.078 |
| Mn | 0.18 | 0.26 |
| Zn | 0.15 | 0.21 |
Table 6 shows the results of conventional tests of blood in experimental layer chickens fed with the test
| Mineral elements | Experimental group | Control group |
| White blood cell | 2.48 | 2.19 |
| Red blood cell | 3.52 | 2.24 |
| Hemoglobin | 179 | 137 |
| Hematocrit of red blood cells | 43.8 | 30.6 |
Description of the drawings: the above embodiments are only used to illustrate the present invention and do not limit the technical solutions described in the present invention; thus, while the present invention has been described in detail with reference to the foregoing embodiments, it will be understood by those skilled in the art that various changes may be made and equivalents may be substituted; all such modifications and variations are intended to be included herein within the scope of this disclosure and the present invention and protected by the following claims.
Claims (9)
1. The production method of the high-efficiency chelating type enzyme peptide is characterized by comprising the following steps:
(1) respectively carrying out activation and propagation on bacillus subtilis, saccharomyces cerevisiae, lactobacillus plantarum and lactobacillus rhamnosus to obtain corresponding fermentation seed liquid;
(2) weighing bean dregs and bean pulp, and uniformly stirring to prepare a solid fermentation culture medium;
(3) inoculating the bacillus subtilis fermentation seed liquid obtained in the step (1) into the solid fermentation culture medium prepared in the step (2), adding wheat middling, adding water according to the water content, uniformly stirring, and performing primary aerobic fermentation to obtain an aerobic fermentation culture medium;
(4) and (3) inoculating the saccharomyces cerevisiae fermentation seed liquid, the lactobacillus plantarum fermentation seed liquid, the lactobacillus rhamnosus fermentation seed liquid, the protease and the inorganic mineral salt obtained in the step (1) into the aerobic fermentation culture medium obtained in the step (3), uniformly stirring, sealing for secondary anaerobic fermentation, and obtaining the efficient chelating enzyme peptide after the fermentation is finished.
2. The method for producing highly effective chelating enzyme peptide according to claim 1, wherein the solid fermentation medium in step (2) contains 25-70% of bean dregs and 30-75% of bean dregs.
3. The method for producing highly effective chelating type enzyme peptide according to claim 1, wherein the amount of Bacillus subtilis inoculation in step (3) is 1-15% of the mass of the solid fermentation medium in step (2); the addition amount of the wheat middling is 5-30% of the mass of the solid fermentation medium in the step (2).
4. The method for producing highly effective chelating enzyme peptide according to claim 1, wherein the water content in step (3) is 25-65%; the first-stage aerobic fermentation temperature is 20-40 ℃, and the aerobic fermentation time is 3-48 h.
5. The method for producing highly effective chelating type enzyme peptide according to claim 1, wherein the inoculation amount of the seed liquid of Saccharomyces cerevisiae, Lactobacillus plantarum and Lactobacillus rhamnosus in step (4) is 1-10% of the mass of the solid fermentation medium in step (2).
6. The method for producing highly potent chelating type enzyme peptide according to claim 1, wherein the protease in the step (4) is acid protease and papain; the total adding amount of the acid protease and the papain is 0.01-2% of the mass of the solid fermentation medium in the step (2), the adding ratio of the two enzymes is 1:1, and the enzyme activities are both 6 ten thousand u/g.
7. The method for producing highly potent chelating enzyme peptide according to claim 1, wherein the inorganic mineral salt in step (4) is CaCO3、FeSO4·7H2O、CuSO4·5H2O、MnSO4·H2O、ZnSO4·7H2O; the CaCO3The addition amount is 1 to 12 percent of the mass of the solid fermentation culture medium in the step (2); the FeSO4·7H2O is added in an amount of0.01-1.6% of the mass of the solid fermentation medium in the step (2); the CuSO4·5H2The addition amount of O is 0.001 to 0.064 percent of the mass of the solid fermentation medium in the step (2); the MnSO4·H2The addition amount of O is 0.01-1% of the mass of the solid fermentation medium in the step (2); the ZnSO4·7H2The addition amount of O is 0.01-1.2% of the mass of the solid fermentation medium in the step (2).
8. Use of the highly effective chelating enzyme peptides produced by the method according to any one of claims 1 to 7 for poultry feeding to improve the absorption of mineral elements and the laying rate of poultry.
9. The use according to claim 8, wherein said highly effective chelating enzyme peptide is used in combination with poultry feed in an amount of 5 to 10% by weight of the poultry feed.
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