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Method for comprehensively utilizing porcine hemoglobin

A porcine hemoglobin and hemoglobin technology, applied in organic chemistry, fermentation, etc., can solve problems such as high cost and difficult process, and achieve the effects of high environmental protection value, simplified process, and high product yield

Active Publication Date: 2015-07-01
ANHUI KEBAO BIOLOGICAL ENG CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, after the above treatment, if the denatured hemoglobin monomer needs to continue to be used to produce polypeptide products, acetone and sodium acetate must be removed, which is difficult and costly.
[0008] In addition, in the prior art, there are many literature reports on heme extraction and purification, and hemoglobin enzymatic hydrolysis, but there are few reports on the comprehensive consideration of the above two aspects and the downstream methods to maximize the benefits of hemoglobin.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Add 0.8% trisodium citrate to 1000 mL of fresh pig blood for anticoagulation, centrifuge at 4000 g for 20 min to separate red blood cells, wash twice with 0.9% NaCl saline, and record the volume as 413 mL after the last centrifugation, which is 434 g.

[0032] Add 1 volume of distilled water to the red blood cells, and stir continuously for 30 minutes for hemolysis. After centrifugation at 4000 g for 30 min, the supernatant was taken as the hemoglobin solution. Alkaline protease (200,000 U.g -1 , Guangbo Nanning Pangbo Biological Engineering Co., Ltd.) for enzymatic hydrolysis.

[0033] The enzymatic hydrolysis process is: the amount of enzyme added is 4000U.mL -1 , temperature 50°C, pH 9.0, time 5h, use 2mol.L every half hour during enzymolysis -1 The pH was adjusted once to 9.0 with NaOH. After enzymolysis, heat the enzymolysis solution to 90°C, adjust the pH to 9.0, add a total volume of 5% NaCl, and stir thoroughly for 30 minutes to promote the complete separati...

Embodiment 2

[0040] Add 0.8% trisodium citrate to 1000 mL of fresh pig blood for anticoagulation, centrifuge at 4000 g for 20 min to separate red blood cells, wash twice with 0.9% NaCl saline, and record the volume after the last centrifugation as 396 mL, 415 g in total.

[0041] Add 2 times the volume of distilled water to red blood cells, and stir continuously for 30 minutes to hemolyze. After centrifugation at 4000 g for 30 min, the supernatant was taken as the hemoglobin solution. Add trypsin (Sinopharm Group Chemical Reagent Co., Ltd., casein conversion power >25) to the hemoglobin solution for enzymatic hydrolysis.

[0042] The enzymatic hydrolysis process is as follows: the amount of enzyme added is 15g, the temperature is 40°C, the pH is 8.5, the time is 6h, and 2mol.L is used every half hour -1 NaOH adjusted the pH once to 8.5. After enzymolysis, heat the enzymolysis solution to 80°C, adjust the pH to 9.0, add 10% NaCl in total volume, and stir thoroughly for 30 minutes to promo...

Embodiment 3

[0049] Add 0.8% trisodium citrate to 1000 mL of fresh pig blood for anticoagulation, centrifuge at 4000 g for 20 minutes to separate red blood cells, wash twice with 0.9% NaCl saline, and record the volume as 425 mL after the last centrifugation, which is 446 g.

[0050] Red blood cells were added with 3 times the volume of distilled water, and stirred continuously for 30 minutes for hemolysis. After centrifugation at 4000 g for 30 min, the supernatant was taken as the hemoglobin solution. Add neutral protease (200,000 U.g -1 , Guangbo Nanning Pangbo Biological Engineering Co., Ltd.) for enzymatic hydrolysis.

[0051] The enzymatic hydrolysis process is: the amount of enzyme added is 4000U.mL -1 , temperature 30°C, pH 7.5, time 5h, use 2mol.L every half hour -1 NaOH adjusted the pH once to 7.5. After enzymolysis, heat the enzymolysis solution to 70°C, adjust the pH value to 9 with concentrated ammonia water, add 15% NaCl in total volume, and stir thoroughly for 30 minutes ...

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PUM

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Abstract

The invention discloses a method for comprehensively utilizing porcine hemoglobin. The method comprises the steps that complete hemoglobin is subjected to enzymolysis; an enzymatic hydrolysate is heated and treated with salt at a certain concentration in an alkaline solution to facilitate protoheme to be dissociated from polypeptide; and proteome is separated from polypeptide and small peptide by ultrafiltration and nanofiltration methods, so that the purpose of comprehensively utilizing porcine hemoglobin is finally achieved. The method is significant in economic benefit, and high in social and environmental protection values; the method does not use organic solvent, so that the process is simplified, the cost is lowered, and no waste is generated; protoheme is separated from polypeptide and small peptide thoroughly, so that the yield and the purity of a product are high; and the industrialization application prospect is broad.

Description

Technical field: [0001] The invention belongs to the technical field of food processing, in particular to a method for comprehensive utilization of porcine hemoglobin. Background technique: [0002] my country's pig production ranks first in the world. Except for a small amount of pig blood produced by intensive pig slaughtering, which is made into pig blood tofu for consumption, the rest is directly discharged without being used, which wastes resources and pollutes the environment. Porcine hemoglobin accounts for more than 75% of the total pig blood protein, which is of great significance to its efficient use. Now some enterprises have begun to use this waste resource to dry plasma and red blood cells respectively by spray drying to prepare pig plasma powder and red blood cell powder for animal production. Feed functional protein additive. [0003] Porcine hemoglobin is a globin with a quaternary structure that binds the heme prosthetic group and consists of four subunits....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/06C07D487/22
Inventor 陈群张敏程江华张洁闫晓明
Owner ANHUI KEBAO BIOLOGICAL ENG CO LTD
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