Method for producing heat-resistant superoxide dismutase (SOD) through Marinamoeba thermophila
A superoxide and dismutase technology, applied in the field of bioengineering, can solve the problems of easy thermal inactivation, cumbersome extraction process, poor thermal stability of natural SOD, etc. Effect
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[0014] Example 1
[0015] Inoculate the slant strains into a 250ml Erlenmeyer flask containing 50ml of culture medium. The culture medium consists of: glucose 20g / l, beef extract 10g / l, peptone 10g / l, NH 4 NO 3 10g / l, corn syrup powder 12g / l, pH natural. 45°C, ferment for 72 hours. After the fermentation is over, 50 ml of the fermentation broth is centrifuged at 10,000 rpm to remove bacteria. The supernatant crude enzyme solution was used to determine the pH and temperature parameters of the enzyme. Take 0.5ml of the supernatant crude enzyme solution, place them in 3.5ml of citric acid-phosphate buffer solution with pH 4, 5, 6, 7, and 8 respectively, and measure the SOD activity at 55°C as 1600, 1700, 2000, 1700, 1600U / ml. Therefore, the optimal reaction pH of the SOD is 6. Take 0.5ml of the supernatant crude enzyme solution and mix it with 3.5ml of pH 6 citric acid-phosphate buffer solution. The enzyme activities measured at 25, 35, 45, 55, and 65°C are 1600, 1800, 2000...
Example Embodiment
[0016] Example 2
[0017] Inoculate the slant strain into a 250ml Erlenmeyer flask with 50ml of seed medium, the composition of the seed medium is: glucose 20g / l, beef extract 10g / l, peptone 10g / l, NH 4 NO 3 10g / l, corn syrup powder 12g / l, natural pH, sterilized at 121 for 20min, shaker at 150rpm, cultured at 45°C for 48 hours. This process has 10 identical Erlenmeyer flasks, obtains 500ml seed liquid altogether.
[0018] Inoculate 500ml of seed solution into a 50L fermenter, and the composition of the fermentation medium is: glucose 20g / l, beef extract 10g / l, peptone 3g / l, NH 4 NO 3 10g / l, corn syrup powder 6g / l, natural pH, fermentation temperature 45°C, liquid volume 70%, ventilation volume 1:1.5v / v / m. The fermentation time is 3 days. Under this condition, the final SOD activity in the fermentation broth was 2000U / ml.
Example Embodiment
[0019] Example 3
[0020] Inoculate 1L of the cultivated seed solution into a 10L fermenter. The composition of seeds and fermentation medium are: fructose 30g / l, beef extract 10g / l, corn syrup powder 20g / l, K 2 HPO 4 4g / l, MgSO 4 7H 2 0.5g / l, pH natural. 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 1300U / ml.
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