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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 industrial production, and the research on extracting catalase from animal raw materials has only attracted attention in recent years

Method used

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  • Method for extracting catalase from ox liver

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preparation example Construction

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

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

[0034] Step 2, ultrasonic extraction, the obtained bovine liver homogenate is ultrasonically oscillated for 10-30min, the ultrasonic intensity is 30-80W, the cells are broken, and the catalase in the cells is rapidly dissolved to obtain the bovine liver catalase homogenate;

[0035] Step 3, centrifugation, centrifuge the bovine liver catalase homogenate at 10-30°C for 15-25min in a low-temperature centrifuge, the speed of centri...

Embodiment 1

[0038] Wash 20kg of fresh beef liver with tap water, chop it, add 6 times the volume of phosphate buffer solution with a pH value of 7.2, mash it with a tissue masher homogenizer, and then extract it for 20 minutes with an ultrasonic intensity of 20W to obtain a homogenate of beef liver , take bovine liver homogenate, and then centrifuge at 6000 rpm for 20 minutes at 15°C with a low-temperature centrifuge, and the supernatant is the crude extract of catalase. Rinse the ultrafiltration equipment first, and after confirming that the pipes and containers are clean, fill the raw material tank with the feed liquid, and use Beijing Haideneng HU10-90 to conduct ultrafiltration concentration at a flow rate of 10L / h to obtain a concentrated solution of catalase. The excess liquid is recovered in the storage tank, and then adjusted to a pH value of 7.2 to replace the phosphate buffer used for crushing. The catalase concentrated solution that obtains adopts Sephacryl S-200 gel column to ...

Embodiment 2

[0040] Wash 20 kg of fresh beef liver with tap water, chop it, add 6 times the volume of phosphate buffer solution with a pH value of 7.4, mash it with a tissue masher homogenizer, and then extract it for 25 minutes with an ultrasonic intensity of 20W to obtain beef liver homogenate , take bovine liver homogenate, and then centrifuge at 6000 rpm for 20 minutes at 15°C with a low-temperature centrifuge, and the supernatant is the crude extract of catalase. Rinse the ultrafiltration equipment first, after confirming that the pipes and containers are clean, fill the raw material tank with feed liquid, and use Beijing Haideneng HU10-90 to conduct ultrafiltration concentration at a flow rate of 15L / h to obtain a concentrated solution of catalase, and through The liquid is recovered in the storage tank, and then adjusted to a pH value of 7.4 to replace the phosphate buffer used for crushing. The catalase concentrated solution that obtains adopts Sephacryl S-200 gel column to carry o...

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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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