Antarctic sea ice bacterium exopolysaccharide with antioxidative activity, and preparation method thereof
An anti-oxidative activity, Antarctic sea ice bacteria technology, applied in the field of medicine and biology, to achieve wide application prospects, inhibition of auto-oxidation rate, and good antioxidant activity.
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Embodiment 1
[0023] Embodiment 1, the preparation of bacterial suspension:
[0024] Antarctica NJ-B3 was obtained from the sea ice samples collected by China’s 24th Antarctic Scientific Expedition Uruguay Station (S62°11’50.52”, W58°55.’50.4”). It was screened and isolated by Zobel 2216E medium. 16S rDNA identified as Pseudoalteromonas genus. NJ-B3 was stored in a 4°C refrigerator with Zobell2216E solid medium, and NJ-B3 was inserted into a Erlenmeyer flask containing 2216E liquid medium, shaken at 10°C and 150rpm for 48 hours, and used as a bacterial suspension.
[0025] Zobell2216E solid medium: peptone 5g, yeast powder 1g, agar powder 15g, aged seawater 1000ml;
[0026] Zobell2216E liquid medium: peptone 5g, yeast powder 1g, aged sea water 1000ml;
Embodiment 2
[0027] Embodiment 2, the preparation of exopolysaccharide of Antarctica NJ-B3:
[0028] 1. The seed medium is: 25g sucrose, 12.5g soybean powder, K 2 HPO 4 0.2g, Na 2 SO 4 0.1g, FeSO4 ·7H 2 O 0.01g, CaCO 3 2g, 1000mL aged sea water, pH value 7.2, sterilized at 121°C for 20min; add 5% of the bacterial suspension of NJ-B3, culture at 10°C, 120rpm for 48h for later use.
[0029] 2. Preparation of exopolysaccharide fermentation broth by submerged fermentation:
[0030] Medium: 50g sucrose, 25g soybean powder, K 2 HPO 4 0.2g, Na 2 SO 4 0.1g, FeSO 4 ·7H 2 O 0.01g, CaCO 3 3g, aged sea water 1000mL, pH value 7.2, sterilized at 121°C for 20min. The feeding volume of the 100L fermenter is 70L, and the NJ-B3 seed liquid is added at a ratio of 5 to 7%. The fermentation temperature is controlled by a circulating refrigerator, and the temperature in the tank is kept at 10±1°C; Inject compressed air to provide the oxygen needed during the fermentation process; the stirring spee...
Embodiment 3
[0033] Embodiment 3, the antioxidant test of NJ-B3 exopolysaccharide:
[0034] (1) NJ-B3 polysaccharide on O· - The clearing effect of:
[0035] Add pH7.8, 0.05mol / L phosphate buffer, 0.13mol / L methionine, 2×10 -5 mol / L riboflavin, sample polysaccharide solutions with different concentrations or 1.3×10 -3 mol / L ascorbic acid, 7×10 -4 mol / L Nitro blue tetrazolium chloride (NBT) each 0.4mL. (No methionine was added to the blank, no scavenger was added to both the blank and the control), and diluted to 4.0 mL with double distilled water. The mixture was reduced at a constant temperature of 35°C under 4000 lx light for 20 min, and the reaction was stopped in the dark. Use a 721 spectrophotometer to measure the light absorption value of each tube solution at 560 nm. Calculate the average clearance d:
[0036] d=(A control-A sample)×100% / A control
[0037] B3 polysaccharide on O 2 · - The removal efficiency is shown in Table 1.
[0038] Table 1 NJ-B3 polysaccharide to O2 ·...
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