Method for extracting catalase from ox liver

A technology of catalase and bovine liver, which is applied in the biological field, can solve the problems of no industrial production, and achieve the effects of economical and environmentally friendly enzyme activity, increased yield, and shortened extraction time

Inactive Publication Date: 2014-12-03
西藏天虹科技股份有限责任公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The production of catalase by microbial fermentation in China is only in the laboratory research stage, and there is no indust

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  • Method for extracting catalase from ox liver

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Example Embodiment

[0032] In the method for extracting catalase from bovine liver, the preparation method of the crude catalase extract includes the following steps:

[0033] Step 1. Mash the homogenate, wash the fresh beef liver with water and chop it, mix the chopped beef liver with a phosphate buffer with a pH of 7.0-7.8 in a volume ratio of 1:6-10, and keep the temperature at 25 Use a tissue mashing homogenizer to mash for 20-30 minutes at -30°C, and the speed of the homogenizer is 2000-3000rpm to obtain a beef liver homogenate;

[0034] Step 2. Ultrasonic extraction. The obtained bovine liver homogenate is ultrasonically oscillated for 10-30 minutes at an ultrasonic intensity of 30-80W, so that the cells are broken and the catalase in the cells is quickly dissolved to obtain bovine liver catalase homogenate;

[0035] Step 3. Centrifugation. Centrifuge the obtained bovine liver catalase homogenate in a low-temperature centrifuge at 10-30°C for 15-25 minutes. The speed of the centrifugal separation ...

Example Embodiment

[0037] Example 1

[0038] Wash 20 kg of fresh beef liver with tap water, chop it, add 6 times the volume of phosphate buffer with a pH of 7.2, mash it with a tissue mashing homogenizer, and then extract 20 min with an ultrasonic intensity of 20W to obtain a beef liver homogenate , Take the bovine liver homogenate, and then centrifuge it at 6000 rpm at 15°C for 20 minutes in a low-temperature centrifuge. The supernatant is the crude catalase extract. First flush the ultrafiltration equipment, make sure that the pipes and containers are clean, fill the raw material tank with the material liquid, use Beijing Hyde Energy HU10-90, and perform ultrafiltration and concentration at a flow rate of 10L / h to obtain a catalase concentrate. The over-liquid is recovered in the storage tank, and the pH value is adjusted to 7.2 to replace the phosphate buffer used for mashing. The resulting catalase concentrate was subjected to gel filtration chromatography on a Sephacryl S-200 gel column. Afte...

Example Embodiment

[0039] Example 2

[0040] Wash 20 kg of fresh beef liver with tap water, chop it, add 6-fold volume of phosphate buffer with a pH of 7.4, mash it with a tissue mashing homogenizer, and then extract at 20W for 25 minutes with ultrasonic intensity to obtain a beef liver homogenate , Take the bovine liver homogenate, and then centrifuge it at 6000 rpm at 15°C for 20 minutes in a low-temperature centrifuge. The supernatant is the crude catalase extract. First flush the ultrafiltration equipment, make sure that the pipes and containers are clean, fill the raw material tank with the material liquid, use Beijing Hyde Energy HU10-90 at a flow rate of 15L / h to perform ultrafiltration and concentration to obtain a catalase concentrate, and The liquid is recovered in the storage tank, and the pH is adjusted to 7.4 to replace the phosphate buffer used for mashing. The resulting catalase concentrate was subjected to gel filtration chromatography on a Sephacryl S-200 gel column. After the col...

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Abstract

The invention discloses a method for extracting catalase from ox liver. A catalase crude extracting solution is prepared from ox liver by ultrasonic extraction, low-temperature centrifugation and other techniques; and ultrafiltration concentration and gel chromatography techniques are used for catalase extraction and purification instead of the complex extraction technique using abundant organic solvents, inorganic salt solutions or the like. The extraction technique is easier to operate, the enzyme activity and yield are greatly enhanced, the enzyme activity of the catalase is up to higher than 10000 U/mg, the yield is up to higher than 18%, and the production cost is greatly lowered; the filtrate generated by ultrafiltration concentration is reutilized, thereby avoiding discharge of wastewater; and thus, the method is an environment-friendly production technique.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for extracting catalase from bovine liver. Background technique [0002] Catalase (CAT), also known as catalase, is a type of terminal oxidase that widely exists in animals, plants and microorganisms. It uses hydrogen peroxide as a substrate and catalyzes the transfer of a pair of electrons to eventually Decompose it into water and oxygen, its systematic name is H 2 o 2 :H 2 o 2 Oxidoreductase, the number of the International Enzyme Committee is ECl.11.1.6. The molecular structure of catalase contains an iron porphyrin ring, and one molecular enzyme protein contains 4 iron atoms, and its active group is heme. Catalase is mainly composed of four subunits, each of which contains a hematoxylin derivative called ProtohemeIX. Regardless of the source, its molecular weight is 225. Catalase is one of the key enzymes in the biological defense system established in the process...

Claims

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Application Information

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IPC IPC(8): C12N9/08
CPCC12N9/0065C12Y111/01006
Inventor 张春颖杨旭锦
Owner 西藏天虹科技股份有限责任公司
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