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CN-122004359-A - Preparation method of small peptide manganese feed additive for enhancing development of white-feather broilers

CN122004359ACN 122004359 ACN122004359 ACN 122004359ACN-122004359-A

Abstract

The invention discloses a preparation method of a small peptide manganese feed additive for enhancing development of white feather broilers, and relates to the technical field of feed additives, the method comprises the steps of taking zein as a raw material, and obtaining a small peptide solution with molecular weight concentrated at 500-1200 daltons through synergistic enzymolysis of complex enzyme; under the protection of nitrogen, carrying out directional chelation according to the mol ratio of small peptide carboxyl to manganese ion of 3:1, removing free manganese through ultrafiltration purification, spray drying to obtain a powder product, and verifying the controllable release performance of the powder product through simulating the digestion environment of poultry. The invention aims to solve the technical problems of low absorption efficiency, limited growth promotion effect, poor batch stability and inadaptation to the physiological characteristics of the digestive tract of poultry of the existing small peptide chelated manganese feed additive in application of white feather broilers, realize high chelation rate, excellent fluidity and storage stability of feed, obviously improve daily gain of white feather broilers, ash content of tibia and feed conversion rate, and has green process and suitability for industrial continuous production.

Inventors

  • HE XI
  • SONG ZEHE
  • ZUO GANG

Assignees

  • 北京德元顺生物科技有限公司

Dates

Publication Date
20260512
Application Date
20260128

Claims (10)

  1. 1. The preparation method of the small peptide manganese feed additive for enhancing the development of white feather broilers is characterized by comprising the following specific steps of: Step1, zein is selected as a small peptide precursor raw material, the zein is dispersed in deionized water to form a suspension with the mass concentration of 8%, the pH value of the system is regulated to 5.6, a compound enzyme preparation consisting of alkaline protease and flavourzyme is added at the temperature of 45 ℃ for carrying out synergistic enzymolysis, the enzymolysis duration is 3 hours, the stirring rotation speed is maintained at 180 revolutions per minute, the temperature is raised to 95 ℃ after the reaction is finished, the temperature is kept for 10 minutes, and enzyme inactivation is realized, so that a small peptide solution with the molecular weight distribution concentrated at 500-1200 daltons is obtained; Dissolving manganese sulfate in deionized water to prepare a manganese ion solution with the molar concentration of 0.4mol/L, slowly adding the manganese ion solution into the small peptide solution under the protection of nitrogen, controlling the molar ratio of small peptide carboxyl to manganese ion to be 3:1, adjusting the pH value of a mixed system to 6.2, and carrying out constant-temperature concussion reaction for 4 hours at 50 ℃ under the condition of avoiding light to enable the manganese ion to be directionally chelated with small peptide side chain carboxyl and amino to generate a small peptide manganese chelate precursor; Step 3, performing ultrafiltration treatment on the precursor solution, adopting a hollow fiber membrane module with a molecular weight cut-off of 1000 daltons, setting the operating pressure to 0.25MPa, continuously replacing the volume of the dialysate by 5 times of the volume of the stock solution, removing free manganese ions and low-molecular impurities which do not participate in chelation, and collecting the cut-off solution; Step 4, concentrating the trapped fluid to 1/3 of the original volume through rotary evaporation, and then performing spray drying treatment to obtain a light yellow powdery small peptide manganese feed additive end product with good fluidity; And 5, carrying out mass characterization on the final product, measuring the chelating rate by an ultraviolet-visible spectrophotometry, analyzing molecular weight distribution by utilizing a high-efficiency gel permeation chromatography, measuring the total manganese content and a soluble manganese release curve by combining an inductively coupled plasma mass spectrometry, and confirming that the product meets the set standard.
  2. 2. The method for preparing the small peptide manganese feed additive for enhancing development of white feather broilers according to claim 1, wherein the initial particle size of zein in step 1 is controlled below 80 microns after ball milling pretreatment, the mass ratio of alkaline protease to flavourzyme is 2:1, and the total enzyme addition amount accounts for 1.2% of the mass of a substrate.
  3. 3. The method for preparing the small peptide manganese feed additive for enhancing development of white feather broilers according to claim 1, wherein the small peptide molecular weight distribution of the enzymolysis product in the step 1 is that the component with the molecular weight of less than 700 daltons accounts for not less than 45%, the component with the molecular weight of 700-1200 daltons accounts for not more than 55%, and the overall polydispersity is lower than 1.15.
  4. 4. The method for preparing the small peptide manganese feed additive for enhancing development of white feather broilers according to claim 1, wherein the dropping speed of the manganese ion solution in the step 2 is 2 milliliters per minute, the dropping process lasts for 40 minutes, and the concentration of dissolved oxygen in a reaction system is maintained below 2 mg/L.
  5. 5. The method for preparing small peptide manganese feed additive for enhancing development of white feather broiler chicken according to claim 1, wherein the solution obtained after the chelation reaction in step 2 is centrifuged at 4000 rpm for 10 minutes to remove trace insoluble substances, and the supernatant is used for the subsequent purification step.
  6. 6. The method for preparing the small peptide manganese feed additive for enhancing development of white feather broilers according to claim 1, wherein manganese ion concentration of permeate is detected every 30 minutes in the ultrafiltration process in the step 3, and when the detected values are lower than 0.01mg/L in two consecutive times, the dialysis end point is judged.
  7. 7. The method for preparing the small peptide manganese feed additive for enhancing development of white feather broilers, as claimed in claim 1, is characterized in that the moisture content of powder obtained by spray drying in the step 4 is controlled within 5%, the apparent density is 0.45g/mL to 0.55g/mL, the repose angle is less than 40 ℃, the air inlet temperature of the drying treatment is controlled to 160 ℃, the air outlet temperature is 85 ℃, and the rotating speed of an atomizer is 25000 r/min.
  8. 8. The method for preparing the small peptide manganese feed additive for enhancing development of white feather broilers according to claim 1, wherein the chelation rate in the step 5 is calculated according to a formula of chelation rate= (total manganese content-extractable free manganese content)/total manganese content×100%, wherein the chelation rate of the final product is required to be 92% or more as measured after the extractable free manganese content is separated by a trichloroacetic acid precipitation method.
  9. 9. The method for preparing the small peptide manganese feed additive for enhancing development of white feather broilers according to claim 1, wherein the determination of the release behavior of soluble manganese in the step 5 is carried out by adopting a method for simulating the foregut digestion environment of white feather broilers, namely, placing 0.1g of additive sample into 10mL of simulated digestive fluid, wherein the digestive fluid consists of pepsin 0.2%, trypsin 0.1% and bile salt 0.3%, the pH gradient is set to gradually rise from 5.8 to 6.8, sampling is carried out every 30 minutes, manganese concentration in filtrate is measured after filtration through a 0.22 micrometer filter membrane, and an accumulated release curve is drawn, wherein the release proportion is required to reach 60-75% in the first 2 hours, and complete release is required in 4 hours.
  10. 10. The method for preparing the small peptide manganese feed additive for enhancing development of white feather broilers according to claim 1, further comprising the steps of establishing an on-line quality monitoring system based on near infrared spectrum, collecting spectrum data of different batches of products and constructing partial least squares regression models, realizing rapid prediction of key quality attributes of chelation rate, moisture content and particle size distribution, and determining coefficients by model cross validation Greater than 0.95, the prediction error is less than 3%.

Description

Preparation method of small peptide manganese feed additive for enhancing development of white-feather broilers The invention belongs to the technical field of feed additives, and particularly relates to a preparation method of a small peptide manganese feed additive for enhancing development of white feather broilers. Background With the continuous improvement of the requirements of livestock and poultry raising industry on the functionality and bioavailability of the feed additive, the small peptide chelated manganese as an efficient and environment-friendly organic trace element additive has good application prospect in the aspect of promoting the growth and development of animals. Especially in the fast growth period of white feather broilers, manganese element has key effects on skeletal development, enzyme system activation and metabolism regulation. However, the existing small peptide chelated manganese feed additive is designed for monogastric animals such as pigs, and the preparation process, the proportioning composition and the release characteristics of the existing small peptide chelated manganese feed additive cannot be fully matched with the physiological characteristics and the nutritional requirements of white feather broilers, so that the problems of insufficient absorption efficiency, limited growth promotion effect and the like exist in practical application. The preparation of the small peptide chelated manganese needs to take the biological activity of the small peptide source, the chelation stability of manganese ions and the release behavior of the product in the poultry digestive tract environment into consideration. The current mainstream technology path includes chelation of manganese after catalytic synthesis of small peptide with strong acid resin, or metal complexation after obtaining small peptide with vegetable proteolysis. However, the former is easy to introduce chemical residues which are difficult to remove, and affects the feed safety, while the latter adopts natural raw materials, the wide pH control range and the non-specific enzymolysis condition of the former are difficult to ensure the consistency of the small peptide sequence and the directional exposure of the manganese chelating site, so that the performance fluctuation among batches of products is obvious. More importantly, the method does not consider the unique physiological characteristics of high pH value of the foregut of white feather broilers, short intestinal transit time, concentration of manganese absorption window period in brooding to early stage of brooding and the like, so that the prepared additive cannot realize high-efficiency targeted delivery and sustained release of manganese element in vivo. The defects cause that the existing product is difficult to synchronously improve the bone mineralization intensity, daily gain rate and feed conversion rate in the feeding practice of white feather broilers, and a preparation method of a special small peptide manganese feed additive for the high-growth-performance poultry species is needed to realize accurate matching of manganese nutrition supply and broiler development rhythm. Disclosure of Invention The invention aims to provide a preparation method of a small peptide manganese feed additive for enhancing development of white feather broilers, which aims to solve the technical problems that the existing small peptide chelated manganese feed additive is insufficient in absorption efficiency, limited in growth promoting effect, poor in batch stability and incapable of adapting to the physiological characteristics of poultry digestive tracts in white feather broilers. In order to solve the technical problems, the invention provides the following technical scheme: The preparation method of the small peptide manganese feed additive for enhancing the development of white feather broilers comprises the following specific steps: Step1, zein is selected as a small peptide precursor raw material, the zein is dispersed in deionized water to form a suspension with the mass concentration of 8%, the pH value of the system is regulated to 5.6, a compound enzyme preparation consisting of alkaline protease and flavourzyme is added at the temperature of 45 ℃ for carrying out synergistic enzymolysis, the enzymolysis duration is 3 hours, the stirring rotation speed is maintained at 180 revolutions per minute, the temperature is raised to 95 ℃ after the reaction is finished, the temperature is kept for 10 minutes, and enzyme inactivation is realized, so that a small peptide solution with the molecular weight distribution concentrated at 500-1200 daltons is obtained; Dissolving manganese sulfate in deionized water to prepare a manganese ion solution with the molar concentration of 0.4mol/L, slowly adding the manganese ion solution into the small peptide solution under the protection of nitrogen, controlling the molar ratio of small peptide carboxyl to manganese ion to be