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Method for preparing ABP (antibacterial peptide) from porcine hemoglobin

A porcine hemoglobin and antibacterial peptide technology, applied in the biological field, achieves the effects of scientific and reasonable process, increased added value and simple operation

Inactive Publication Date: 2015-11-18
CHONGQING DOUHAO BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the technical methods and products for extracting antimicrobial peptides from pig blood have not been reported yet.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] (1) Collect 100L of fresh pig blood, anticoagulate with 5g / L sodium citrate, centrifuge at 4°C and 8000r / min for 10min, collect the lower sediment to obtain about 45L of red blood cells (solid content 30%); add 45L of distilled water, ultrasonic cell disruption assisted hemolysis for 20min, and then centrifuged at 4°C and 4000r / min for 10min to obtain 90L of hemoglobin solution.

[0019] (2) Take an appropriate amount of hemoglobin solution, add 410g trypsin to enzymatically hydrolyze (the specific activity of the enzyme is 10×10 4 u / g), stirring, 1mol / L NaoH solution to maintain the pH of the system at 8.0, 50°C, 5h; the enzymolysis reaction product was quickly heated to 85°C and kept at this temperature for 15 minutes, and then rapidly cooled to 0-30°C to terminate the enzyme hydrolysis reaction, adjust the pH value to about 7.0, centrifuge at 8000r / min for 15min, and obtain 90L of supernatant enzymatic hydrolysis solution.

[0020] (3) Add 15kg of activated carbon t...

Embodiment 2

[0023] (1) Collect 50L of fresh pig blood, anticoagulate with 5g / L sodium citrate, centrifuge at 4°C, 8000r / min for 10min, collect the lower sediment to obtain about 23L of red blood cells (solid content 30%); add 23L of distilled water, ultrasonic cell disruption assisted hemolysis for 20min, and then centrifuged at 4°C and 4000r / min for 10min to obtain 46L of hemoglobin solution.

[0024] (2) Take an appropriate amount of hemoglobin solution, add 200g of trypsin for enzymatic hydrolysis (the specific activity of the enzyme is 10×10 4 u / g), stirring, 1mol / L NaoH solution to maintain the pH of the system at 8.0, 50°C, 5h; the enzymolysis reaction product was quickly heated to 85°C and kept at this temperature for 15 minutes, and then rapidly cooled to 0-30°C to terminate the enzyme For the hydrolysis reaction, adjust the pH value to about 7.0, centrifuge at 8000r / min for 15min, and obtain 46L of supernatant enzymatic hydrolysis solution.

[0025] (3) Add 10kg of activated car...

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PUM

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Abstract

The invention discloses a method for preparing an ABP (antibacterial peptide) from porcine hemoglobin. The method comprises steps as follows: porcine blood corpuscles are obtained through separation of porcine whole blood and then subjected to hemolysis, enzymolysis, decoloration, ultrafiltration, vacuum concentration and freeze-drying, and the ABP is obtained; the dosage of trypsin is 3000 U / g, and the enzymolysis reaction time is 4-6 h under conditions that the pH is 8 and the temperature ranges from 45 DEG C to 55 DEG C. The method for preparing the ABP from the porcine hemoglobin is low in cost and good in effect.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for preparing antibacterial peptides from porcine hemoglobin. Background technique [0002] Antibacterial peptides (antibacterialpeptide, ABP) is an important part of the natural immune defense system of organisms. Antimicrobial peptides have many advantages such as broad antibacterial spectrum, high thermal stability, safety and high efficiency. The preparation of food-source protein antimicrobial peptides has become one of the research hotspots in the field of biology. [0003] Pig blood is one of the main by-products in the process of pig slaughtering and processing. It has high nutritional value and is known as "liquid meat". my country's pig blood resources are very rich, but the utilization rate is low, no more than 10%. If it is processed into As a feed additive, blood meal is used in the production of heme and peptone in biochemical pharmaceuticals. These...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/06C07K1/34
Inventor 宁成勇宁发子宁成刚
Owner CHONGQING DOUHAO BIOTECH
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