Method for producing low-temperature superoxide dismutase through marine microbial fermentation

A technology of marine microorganisms and microbial fermentation, used in biochemical equipment and methods, enzymes, enzymes, etc.

Inactive Publication Date: 2012-09-05
DALIAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the research on the production of SOD by fermentation of marine microorganisms is still in its infancy, an...

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0014] (1) Culture medium preparation

[0015] ① Strain activation medium: glucose 10.0g, beef extract 14.0g, peptone 7.0g, yeast powder 4.0g, agar 20.0g, distilled water 1.0L, pH value natural, sterilized by high pressure steam at 121℃ for 30min.

[0016] ② Strain-directed acclimation medium: glucose 8.0-16.0g, beef extract 12.0-24.0g, peptone 4.0-12.0g, yeast powder 2.0-8.0g, NaCl 5.0-10.0g, agar 15.0-30.0g, distilled water 1.0L, pH value 7.0, sterilized by high pressure steam at 121°C for 30min.

[0017] ③Liquid seed medium: glucose 8.0-15.0g, beef extract 8.0-16.0g, peptone 5.0-15.0g, yeast powder 2.0-10.0g, NaCl 5.0-10.0g, tap water 1.0L, pH value 7.0, 121℃ high-pressure steam Sterilize for 30min.

[0018] ④ Fermentation enzyme production medium: glucose 15.0-25.0g, (NH 4 ) 2 SO 4 3.0~10.0g, corn steep liquor 5.0~15.0g, yeast powder 2.0~8.0g, NaCl 5.0~10.0g, CaCO 3 1.0~6.0g, MgSO 4 0.1~0.9g, 1.0L tap water, pH value 7.0, sterilize by high pressure steam at 121℃ for...

Embodiment 2

[0027] (1) Culture medium preparation

[0028] ① Strain activation medium: 10.0g glucose, 14.0g beef extract, 7.0g peptone, 4.0g yeast powder, 20.0g agar, 1.0L distilled water, natural pH, sterilized by high pressure steam at 121℃ for 30min.

[0029] ② Strain-directed acclimation medium: glucose 8.0-16.0g, beef extract 12.0-24.0g, peptone 4.0-12.0g, yeast powder 2.0-8.0g, NaCl 5.0-10.0g, agar 15.0-30.0g, distilled water 1.0L, pH value 7.0, sterilized by high pressure steam at 121°C for 30min.

[0030] ③Liquid seed medium: glucose 8.0-15.0g, beef extract 8.0-16.0g, peptone 5.0-15.0g, yeast powder 2.0-10.0g, NaCl 5.0-10.0g, tap water 1.0L, pH value 7.0, 121℃ high-pressure steam Sterilize for 30min.

[0031] ④ Fermentation enzyme production medium: glucose 15.0-25.0g, (NH 4 ) 2 SO 4 3.0~10.0g, corn steep liquor 5.0~15.0g, yeast powder 2.0~8.0g, NaCl 5.0~10.0g, CaCO 3 1.0~6.0g, MgSO 4 0.1~0.9g, 1.0L tap water, pH value 7.0, sterilize by high pressure steam at 121℃ for 30min...

Embodiment 3

[0040] (1) Culture medium preparation

[0041] ① Strain activation medium: glucose 10.0g, beef extract 14.0g, peptone 7.0g, yeast powder 4.0g, agar 20.0g, distilled water 1.0L, pH value natural, sterilized by high pressure steam at 121℃ for 30min.

[0042] ② Strain-directed acclimation medium: glucose 8.0-16.0g, beef extract 12.0-24.0g, peptone 4.0-12.0g, yeast powder 2.0-8.0g, NaCl 5.0-10.0g, agar 15.0-30.0g, distilled water 1.0L, pH value 7.0, sterilized by high pressure steam at 121°C for 30min.

[0043] ③Liquid seed medium: glucose 8.0-15.0g, beef extract 8.0-16.0g, peptone 5.0-15.0g, yeast powder 2.0-10.0g, NaCl 5.0-10.0g, tap water 1.0L, pH value 7.0, 121℃ high-pressure steam Sterilize for 30min.

[0044] ④ Fermentation enzyme production medium: glucose 15.0-25.0g, (NH 4 ) 2 SO 4 3.0~10.0g, corn steep liquor 5.0~15.0g, yeast powder 2.0~8.0g, NaCl 5.0~10.0g, CaCO 3 1.0~6.0g, MgSO 4 0.1~0.9g, 1.0L tap water, pH value 7.0, sterilize by high pressure steam at 121℃ for...

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Abstract

The invention discloses a method for producing low-temperature superoxide dismutase through marine microbial fermentation. The method comprises the following steps of: performing directional acclimation of the marine microorganism for producing superoxide dismutase so that the marine microorganism grows well in low-temperature environment; performing gradual amplification culture of the superoxide dismutase producing bacteria after the directional acclimation at 8-14 DEG C; inoculating the bacteria into a liquid fermentation medium according to an inoculation amount of 4-10% of the fermentation broth volume, and culturing at 8-14 DEG C for 60-96 hours to end the marine microbial fermentation for producing low-temperature superoxide dismutase; centrifuging the fermentation broth at 4,000-8,000rpm and collecting the bacteria; washing the bacteria for multiple times and collecting the precipitate; suspending the precipitate in a buffer solution and adding quartz sand and grinding; centrifuging at 10,000-14,000rpm and collecting the supernate which is the crude enzyme liquid; and performing further concentration, separation and purification of the crude enzyme liquid according to different needs and different objects to obtain the enzymic preparations different in activity, purity and form.

Description

technical field [0001] The invention relates to the fields of microbiology, enzyme engineering, fermentation engineering, biochemistry and the like, and in particular relates to a low-temperature SOD marine microbial fermentation production method. The low-temperature SOD produced by this method is used as a superoxide anion radical cleanser in organisms, and is mainly used in industries such as medicine, food, and cosmetics, especially in radiation protection, anti-aging, anti-inflammatory, tumor suppression, cancer, and autoimmune therapy. unique advantages. Background technique [0002] Superoxide dismutase (EC 1.15.1.1, SOD for short) is a metalloenzyme widely present in organisms, which can catalyze the dismutation reaction of superoxide anion free radicals to generate hydrogen peroxide and molecular oxygen, thereby eliminating biological Oxygen free radicals in the body protect organisms from harm (Zi Junqing, Journal of Dali Teachers College, 1998). In 1969, Mccord ...

Claims

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Application Information

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IPC IPC(8): C12N9/08
Inventor 张庆芳迟乃玉窦少华王晓辉于爽马莉
Owner DALIAN UNIV
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