Curdlan producing strain and application thereof
A thermo-cured polysaccharide and strain technology, applied in bacteria, microorganisms, microorganisms, etc., can solve the problems of low gel strength, low yield, poor gel properties, etc., and achieve the effects of high gel strength, high yield, and large molecular weight
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Embodiment 1
[0030] Screening of strains
[0031] (1) Weigh 10 g of the soil sample, add 90 mL of sterile water, put a small amount of glass beads into the Erlenmeyer flask, and vibrate on a shaker for 10 min to make a soil suspension.
[0032] (2) After standing for 10 minutes, take 0.1mL and add it to 0.9mL sterile saline, and serially dilute to 10 -4 、10 -5 、10 -6 For the three gradients, take 0.1mL to apply to the screening plate.
[0033] (3) Place the plate in an incubator at 30°C and incubate it upside down for 2-3 days, then observe.
[0034] (4) Select the colonies that turn blue and darker in color, streak on the plate, and pick a single colony for preservation. The strains screened on the plate were further inoculated into the fermentation medium for re-screening by shake flask fermentation. The culture conditions were 30°C, 240rpm, 4d.
[0035] (5) Take 5ml of fermentation broth and heat it in a water bath at 95°C for 10 minutes to observe whether a gel can be formed. The...
Embodiment 2
[0038] Identification of isolated strain DH-6
[0039] ⑴morphological identification
[0040] After cultured on LB plates for 1 day, the colonies were round, with a smooth and moist surface, neat edges, slightly raised colonies, and off-white. Microscopically, the cells were rod-shaped and Gram staining was negative.
[0041] ⑵ Physiological and biochemical identification
[0042] The physiological and biochemical characteristics of strain DH-6 are shown in Table 1.
[0043] Table 1: Physiological and biochemical characteristics of strain DH-6
[0044]
[0045] Among them, "+" indicates a positive reaction, and "-" indicates a negative reaction.
[0046] CD16SrDNA sequence identification
[0047] Extract the total genomic DNA of bacterial strain DH-6, and use this as a template to amplify the 16SrDNA sequence of the bacterial strain, its primer sequence is SEQIDNo.1, F:5'-CAGAGTTTGATCCTGGCT-3', SEQIDNo.2, R:5' - AGGAGGTGATCCAGCCGCA-3', SEQ ID No. 3. The comparison of...
Embodiment 3
[0050] Scrape a ring of strains (Rhizobium sp. DH-6) from the inclined surface, inoculate in a 250ml Erlenmeyer flask with 30ml seed culture medium, culture at 30°C, 180r / min, on a shaker for 20h, to obtain a seed solution . Take the seed solution and transfer it into a 500ml Erlenmeyer flask with 100ml of fermentation medium at an inoculum size of 8%, and culture it on a shaker at 30°C and 240r / min for 4 days.
[0051] Seed medium (g / L): sucrose 20, (NH) 2 HPO 4 3. KH 2 PO 4 1. MgSO 4 .7H 2 O0.5, yeast extract 1, CaCO 3 3. pH=7.0.
[0052] Fermentation medium (g / L): sucrose 60, (NH) 2 HPO 4 2.3、KH 2 PO 4 1. MgSO 4 .7H 2 O0.5, corn steep liquor 1, CaCO 3 3. pH=7.0.
[0053] Take the fermentation broth and centrifuge at 5000rpm for 5min, remove the supernatant, and add water to make up to the original volume. Then add 0.6M NaOH solution at a volume ratio of 1:1, stir evenly, and stand still for 1 hour. Then centrifuge at 10000rpm for 10min, take the supernatant...
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