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Production process for modifying SOD

A production method and enzyme solution technology, applied in the production process of modifying superoxide dismutase, in the field of biochemistry, can solve the problems of large loss of SOD activity, decrease of lauroyl chloride, increase of production cost, etc., and achieve less loss of SOD activity and lower pH The value fluctuates little and the effect of saving production cost

Inactive Publication Date: 2012-04-25
江忠丽
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Professor Yan Jiaqi’s modification method needs to activate lauric acid, and activation is a complex chemical reaction process. Incomplete reaction may easily lead to residues of chemical reagents such as thionyl chloride and pyridine, which will affect the safety of the product.
Professor Zou Guolin’s modification method is to use SOD dry powder as raw material, which increases the production cost and loses the enzyme activity; and in the modification process, lauroyl chloride is added to the SOD enzyme solution at one time, which will cause the pH of the SOD enzyme solution to increase. The value drops sharply and the local concentration of lauroyl chloride is too high, causing the activity loss of SOD to be large, at about 20%, the reaction process is not easy to control and is not suitable for large-scale production
[0008] In addition, in the current modification method, toxic acetone is used to precipitate and collect SOD after modification, which affects the safety of the product and reduces the activity of SOD

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] 1. 0.5 tons of pig blood clots were collected through emulsification, hemolysis, continuous centrifugation, ultrafiltration concentration, heat denaturation, column chromatography and other production processes, and the activity was 103,300 U / ml, A 280 For the blue-green SOD enzyme solution with a value of 14.6 and a volume of 2.65L, it is estimated that the amount of lauroyl chloride added is 46ml.

[0025] 2. Pour the SOD enzyme solution into a jacketed 10L glass reaction kettle, start the stirring device, and add 2M NaOH solution dropwise to adjust the pH value to 9.0. Then the hot water in the jacket was circulated and heated to 40°C, and kept at a constant temperature; then, 46ml of lauroyl chloride was continuously and slowly added dropwise, and 2M NaOH solution was added dropwise at the same time, so that the pH value of the SOD enzyme solution was always maintained at about 9.0, and the reaction was carried out for 1 hour. The addition of acid chloride was compl...

Embodiment 2

[0028] 1. 2.0 tons of pig blood clots were collected through emulsification, hemolysis, continuous centrifugation, ultrafiltration concentration, heat denaturation, column chromatography and other production processes, and the activity was 152,700 U / ml, A 280 For the blue-green SOD enzyme solution with a value of 30.8 and a volume of 6.1L, the amount of lauroyl chloride added is calculated to be 112ml.

[0029] 2. Pour the SOD enzyme solution into a jacketed 10L glass reaction kettle, start the stirring device, and add 2M KOH solution dropwise to adjust the pH value to 9.0. Then the hot water in the jacket is circulated and heated to 42°C, and kept at a constant temperature; then 112ml of lauroyl chloride is continuously and slowly added dropwise, and 2M KOH solution is added dropwise at the same time, so that the pH value of the SOD enzyme solution is always maintained at about 9.0, and the reaction is carried out for 1 hour. The addition of acid chloride was completed in the...

Embodiment 3

[0032] 1. 2.5 tons of bovine blood clots were collected through emulsification, hemolysis, continuous centrifugation, ultrafiltration concentration, heat denaturation, column chromatography and other production processes, and the activity was 164,000 U / ml, A 280 For the blue-green SOD enzyme solution with a value of 42.3 and a volume of 7.8L, it is estimated that the amount of lauroyl chloride added is 147ml.

[0033] 2. Pour the SOD enzyme solution into a jacketed 10L glass reaction kettle, start the stirring device, and add 2M NaOH solution dropwise to adjust the pH value to 9.0. Then the hot water in the jacket was circulated and heated to 45°C, and kept at a constant temperature; then 147ml of lauroyl chloride was continuously and slowly added dropwise, and 2M NaOH solution was added dropwise at the same time, so that the pH value of the SOD enzyme solution was always maintained at about 9.0, and the reaction was carried out for 1 hour. The addition of acid chloride was co...

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Abstract

The invention discloses a production process for modifying SOD. The production process comprises the following three steps: preparing SOD enzyme solution by using animal blood; performing modification reaction of the SOD enzyme solution; and preparing SOD freeze-dry powder by using the SOD enzyme solution. In the production process, the SOD modification process and the SOD extraction-purificationprocess are realized on the same production line, so that the production ring is simplified, the production cost is saved by about 20 percent, and the SOD activity is better protected by the process than by the method for preparing raw solution by dissolving the SOD dry powder; the addition of lauroyl chloride and alkali solution changes the environment and the process of the modification chemical reaction, but hardly causes the fluctuation of the pH value of the SOD enzyme solution, the reaction process is easily controlled, the production process is suitable for mass production, and the activity loss of the SOD is less; in the whole production process, the acetone addition process in the conventional process is eliminated, so that the product safety is guaranteed, and the activity loss of the SOD is less.

Description

technical field [0001] The invention belongs to the technical field of biochemistry, in particular to the technical field of a production process for modifying superoxide dismutase (SOD). Background technique [0002] The theory of free radicals is a theory about the aging of living organisms that is more and more widely accepted in the field of life sciences. This theory believes that free radicals are an important cause of aging of living organisms and many diseases. It is estimated that 80-90% of aging and degenerative diseases are related to free radicals, including cancer, Alzheimer's disease, Parkinson's disease, skin pigmentation, cataracts, heart disease, etc., so removing free radicals is very important for maintaining a healthy body and maintaining a youthful body is essential. [0003] SOD, superoxide dismutase, is a specific free radical scavenger. SOD is a biological enzyme that widely exists in animals, plants and microorganisms, and can catalyze the dispropo...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/96C12N9/08
Inventor 江忠丽莫宏春
Owner 江忠丽