A kind of water-insoluble exopolysaccharide of leuconostoc enterococcus and preparation method thereof
A Leuconostoc enterococci, water-insoluble technology, applied in the biological field, can solve the problems of cumbersome preparation methods, single type of glycosidic bond in the connection structure, narrow source of water-insoluble polysaccharides, etc., and achieve high food safety and broad application prospects.
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Embodiment 1
[0032]Example 1 Preparation of the water-insoluble exopolysaccharide of Leuconostoc enterococci
[0033] 1. Materials and methods
[0034] Preparation of seeds (fermented strains): dissolve the lyophilized powder of Leuconostoc mesenteroides BD3749 with a small amount of sterile distilled water, and use an inoculation loop to get a ring and streak it on the M17 solid medium containing 5% sucrose ( Purchased from OXOID, UK), 30°C aerobic culture for 48h, took out a single colony with an inoculation loop and put it into 10mL M17 liquid medium containing 5% sucrose (purchased from OXOID, UK), and used a vortex shaker to homogenize the colony Disperse in the liquid medium, 30°C, 180rpm shaking culture for 24h, take out, inoculate with 2% (v / v) inoculum size in M17 liquid medium containing 5% sucrose (purchased from OXOID, UK), 30°C, 180rpm shaking After culturing for 24 hours, the culture was centrifuged at 15,000rpm for 10 minutes, the supernatant was discarded, the bacteria wer...
Embodiment 2
[0040] Example 2 Preparation of the water-insoluble exopolysaccharide of Leuconostoc enteroenteritis
[0041] 1. Materials and methods
[0042] The preparation of seed (fermentation strain): with embodiment 1.
[0043] Synthetic medium (peptone 1%, yeast extract 0.5%, K 2 HPO 4 0.5%, CaCl 2 0.034%, sucrose 30%, described percentage is the preparation of mass percent): with peptone 10g, yeast extract 5g, sucrose 300g, K 2 HPO 4 20g and CaCl 2 0.034g mixed with 1L distilled water, fully dissolved, with NaHCO 3 Adjust the pH to 8 and sterilize to obtain a synthetic medium with the required sucrose concentration.
[0044] 2. Preparation of water-insoluble exopolysaccharides from Leuconostoc enterolis
[0045] Leuconostoc enterococcus seed solution was inoculated in a 5% (v / v) inoculation amount into a synthetic medium with a sucrose mass percentage of 30% and a pH of 8, 15°C, 300rpm shaking fermentation culture for 24 hours to obtain a fermentation broth, After the fermen...
Embodiment 3
[0048] Example 3 Preparation of the water-insoluble exopolysaccharide of Leuconostoc enteroenteritis
[0049] 1. Materials and methods
[0050] The preparation of seed (fermentation strain): with embodiment 1.
[0051] Synthetic medium (peptone 1%, yeast extract 0.5%, K 2 HPO 4 0.5%, CaCl 2 0.034%, sucrose 5%, the percentage is the preparation of mass percent): with peptone 10g, yeast extract 5g, sucrose 50g, K 2 HPO 4 20g and CaCl 2 Mix 0.034g with 1L of distilled water, fully dissolve, adjust the pH to 5 with NaOH, and sterilize to obtain a synthetic medium with the required sucrose concentration.
[0052] 2. Preparation of water-insoluble exopolysaccharides from Leuconostoc enterolis
[0053] Leuconostoc enterococcus seed solution was inoculated in a 1% (v / v) inoculation amount into a synthetic medium with 5% sucrose mass percentage and a pH of 5, 37°C, 100rpm shaking fermentation culture for 96 hours to obtain a fermentation broth, After lyophilization of the ferme...
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