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A kind of extraction method of hemoglobin

An extraction method and heme technology, applied in fermentation and other directions, can solve problems such as complex process, and achieve the effect of small equipment investment and simple and controllable production process

Active Publication Date: 2018-01-16
成都天屹生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The present invention aims to solve the technical problems of the existing hemoglobin extraction method with complex process and the use of a large amount of organic solvents. The invention provides a novel and environmentally friendly heme extraction method

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Add 150kg of isolated fresh pig blood cells to the jacketed enzymolysis tank, add 150kg of water to mix, stir rapidly (frequency 30 Hz) to rupture the membrane for 1 hour, then raise the temperature of the enzymolysis tank to 50°C and adjust with dilute hydrochloric acid solution When the pH reached 7.0, 4.5 kg of trypsin was added. After dissolving, the stirring speed was reduced (frequency 14 Hz). After 10 hours of enzymolysis, the pH of the enzymolysis solution was adjusted to 5.0, and heated to 80°C for 30 minutes. The treatment ended. After cooling, it was separated with a decanter centrifuge at a speed of 4000 rpm and centrifuged for 20 minutes to collect the precipitated heme crude product with a purity of 30%.

[0027] Put 10kg of crude heme into an enzymolysis tank, add 200kg of 0.5% sodium hydroxide solution and heat to 90°C, stir rapidly to dissolve it. After the solution is cooled, add 6 kg of sodium carboxymethylcellulose and stir. Disc centrifuge centrifu...

Embodiment 2

[0029] Add 150kg of isolated fresh pig blood cells to the jacketed enzymolysis tank, add 300kg of water to mix, stir rapidly (frequency 30 Hz) to rupture the membrane for 1 hour, then raise the temperature of the enzymolysis tank to 40°C and adjust with dilute hydrochloric acid solution When the pH reached 5.0, 4.5 kg of trypsin was added, and after dissolving, the stirring speed was reduced (frequency 14 Hz). After 15 hours of enzymolysis, the pH of the enzymolysis solution was adjusted to 8.0, and heated to 80°C for 30 minutes, and the treatment ended. After cooling, it was separated with a decanter centrifuge at a speed of 4000 rpm and centrifuged for 20 minutes to collect the precipitated heme crude product with a purity of 30%.

[0030] Put 10kg of crude heme into an enzymolysis tank, add 150kg of 2% sodium hydroxide solution and heat to 70°C, stir rapidly to dissolve it. After the solution is cooled, add 10 kg of sodium carboxymethylcellulose and stir. Disc centrifuge c...

Embodiment 3

[0033] Add 150kg of isolated fresh pig blood cells to the jacketed enzymolysis tank, add 200kg of water to mix, stir rapidly (frequency 30 Hz) to rupture the membrane for 1 hour, then raise the temperature of the enzymolysis tank to 70°C and adjust with dilute hydrochloric acid solution When the pH reached 9.0, 4.5 kg of trypsin was added, and after dissolving, the stirring speed was reduced (frequency 14 Hz). After 15 hours of enzymolysis, the pH of the enzymolysis solution was adjusted to 8.0, and heated to 90°C for 30 minutes, and the treatment ended. After cooling, it was separated with a decanter centrifuge at a rotation speed of 4000 rpm for 20 min, and the precipitate was collected to obtain crude heme with a purity of 29%.

[0034] Put 10kg of crude heme into an enzymolysis tank, add 300kg of 0.1% sodium hydroxide solution and heat to 70°C, stir rapidly to dissolve it. After the solution is cooled, add 20 kg of sodium carboxymethylcellulose and stir. Disc centrifuge c...

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Abstract

The invention discloses a method for extracting heme, which comprises the following steps: centrifuging fresh pig blood, collecting pig blood cells; mixing pig blood cells with water in a certain proportion, stirring at high speed to rupture the membrane of pig blood cells, and then adjusting the temperature to 30~70℃, pH4.0~9.0, then add trypsin for enzymolysis for 6~24 hours, then adjust the pH of the enzymolysis solution after enzymolysis to 4.0~9.0, and heat to a temperature of 50~90℃; then enzymolysis Liquid centrifugation to collect the precipitate, which is crude heme; add sodium hydroxide solution to the crude heme, heat to 50-90°C, and stir to dissolve the crude heme; after the solution is cooled, add CMC and stir evenly; centrifuge , collect and obtain CMC-heme; use ultrafiltration membrane to separate CMC and heme and dry it, which is the finished product of heme. The method does not need any organic solvent, is safe and environment-friendly, has a simple and controllable production process, and has a purity of over 85%.

Description

technical field [0001] The invention relates to a method for extracting heme. Background technique [0002] Pig blood contains 0.5% heme. It can not only be used as a coloring agent in meat products, but also as a semi-synthetic bilirubin raw material. It is also a special anti-cancer drug and can also be used as an iron supplement in clinical practice. , to treat anemia caused by iron deficiency. At present, domestic research on heme is relatively popular, but the separation and extraction of production has not yet broken through the traditional method. To obtain high-purity heme, it still relies on the acidic acetone precipitation method. In this method, chloroform is used to precipitate the hemoglobin in porcine blood cells first, then the precipitate is collected by filtration, 5 times the volume of 2.5% concentrated hydrochloric acid acetone solution is added, stirred and extracted, the acetone heme solution is collected, and finally the acetone is recovered by vacuum ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P17/18
Inventor 郝刚
Owner 成都天屹生物科技有限公司