Method for producing heat-resistant superoxide dismutase (SOD) by using thermoleophilum minutum

A superoxide and dismutase technology, applied in the field of bioengineering, can solve the problems of cumbersome extraction process, poor thermal stability of natural SOD, easy to be deactivated by heat, etc. Effect

Inactive Publication Date: 2015-06-10
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

According to current reports, SOD can be produced by microbial fermentation, but mainly intracellular enzymes (Diao Zhimin, Yao Yinxia, ​​Zhang Wenjing. Research on yeast SOD fermentation conditions [J]. Journal of Qinghai Normal University (Natural Science Edition), 2002,4 :48-51; Yang Mingya

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0014] Example 1

[0015] The slant strain was inoculated into a 250ml Erlenmeyer flask containing 50ml culture medium. The composition of the culture medium is: glucose 20g / l, beef extract 10g / l, peptone 10g / l, NH 4 NO 3 10g / l, corn steep liquor 12g / l, natural pH. Fermented at 55°C for 72 hours. After the fermentation, the 50ml fermentation broth was centrifuged at 10,000 rpm to remove the bacteria. The supernatant crude enzyme solution is used to determine the pH and temperature parameters of the enzyme. Take 0.5ml of the crude enzyme solution from the supernatant and place them in 3.5ml of citric acid-phosphate buffer at pH 4, 5, 6, 7, 8, and determine the SOD activity at 55°C to be 600, 1500, 1000, 600, respectively. 300U / ml. Therefore, the optimal reaction pH of the SOD is 5. Take 0.5ml of the supernatant crude enzyme solution and mix it with 3.5ml of pH 5 citric acid-phosphate buffer. The enzyme activities measured at 25, 35, 45, 55, and 65°C are 400, 700, 1200, 1500, r...

Example Embodiment

[0016] Example 2

[0017] The slant strain was inoculated into a 250ml Erlenmeyer flask containing 50ml of seed culture medium. The composition of the seed culture medium was: glucose 20g / l, beef extract 10g / l, peptone 10g / l, NH 4 NO 3 10g / l, corn steep liquor 12g / l, natural pH, 121 sterilization for 20 minutes, shaker rotation speed 150 rpm, incubate at 45°C for 48 hours. There are 10 identical Erlenmeyer flasks in this process, and a total of 500ml seed liquid is obtained.

[0018] Inoculate 500ml of seed liquid into a 50L fermentor. The fermentation medium is composed of: glucose 20g / l, beef extract 10g / l, peptone 3g / l, NH 4 NO 3 10g / l, corn steep liquor 6g / l, natural pH, fermentation temperature 45°C, filling liquid volume 70%, ventilation volume 1:1.5v / v / m. The fermentation time is 3 days. Under these conditions, the final SOD activity in the fermentation broth is 1200U / ml.

Example Embodiment

[0019] Example 3

[0020] Inoculate the cultured 1L seed liquid into a 10L fermentor. The composition of seeds and fermentation medium are: fructose 30g / l, beef extract 10g / l, corn steep liquor 20g / l, K 2 HPO 4 4g / l, MgSO 4 7H 2 0.5g / l, natural pH. The fermentation temperature is 20°C, the liquid volume is 70%, the ventilation volume is 1:0.3v / v / m, and the fermentation time is 5 days. Under these conditions, the final SOD activity in the fermentation broth is 1500 U / ml.

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Abstract

The invention relates to a method for producing superoxide dismutase (SOD) by using thermoleophilum minutum and belongs to the technical field of bioengineering. According to the method, SOD is produced from thermoleophilum minutum under the condition of liquid-state fermentation and is an ectoenzyme. The yield of heat-resistant SOD can reach 1500U/ml to the maximum, so that the method has a good industrial application prospect.

Description

technical field [0001] The production of superoxide dismutase (SOD) by using Thermoleophilum minutum belongs to the technical field of bioengineering. The invention relates to obtaining high-temperature-resistant SOD through liquid state fermentation by using bacterial strain Thermoleophilum minutum whose preservation number is ATCC35268. Background technique [0002] Superoxide dismutase (SOD) has a good defense against oxygen toxicity and can enhance the body's ability to resist radiation damage. In recent years, it has become a new anti-inflammatory drug for the treatment of inflammation, tumors and autoimmune diseases; an additive for sunscreen, anti-aging and anti-inflammatory in the cosmetic and food industries, and a biochemical reagent for certain disease probes. SOD widely exists in animals, plants, all aerobic microorganisms and a few anaerobic microorganisms. At present, SOD is mainly extracted from animal blood (such as pig blood or cow blood) (Yuan Ye. Prepara...

Claims

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Application Information

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IPC IPC(8): C12N9/02C12R1/01
CPCC12N9/0089C12Y115/01001
Inventor 丁彦蕊牟兆琳饶榕王雪芹
Owner JIANGNAN UNIV
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