Bacillus subtilis SWB8 for producing diosgenin
A technology of Bacillus subtilis and diosgenin, which is applied in the field of applied microorganisms, can solve problems such as environmental pollution and backward technology, and achieve the effects of avoiding environmental pollution and stabilizing ability.
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Embodiment 1
[0025] Embodiment one .Bacillus subtilis SWB8 ( Bacillus subtilis SWB8) Isolation and identification of CCTCC M 2010271
[0026] 1. Isolation and purification of strains
[0027] (1) Separation. Medium: 0.3% beef extract; 1% peptone; 0.5% NaCl; 2% agar; 1% turmeric dry powder (the turmeric is dried and crushed into powder after removing the brown skin); distilled water; pH7.0. Select the underground stems of the fresh two-year-old turmeric plant, wash them, burn them with 95% alcohol, peel off the brown skin under aseptic conditions, soak them in 70% alcohol for 10 minutes, wash them with sterile distilled water, and then peel off the outer layer tissue with a thickness of 2-3mm. The internal tissues were cut into 2-3 mm thick slices and pasted on the culture medium, and cultured at a constant temperature at 32°C.
[0028](2) Purification. Beef extract peptone medium: 0.3% beef extract; 1% peptone; 0.5% NaCl; 2% agar; 1% turmeric dry powder; distilled water; pH7.0. Cz...
Embodiment 2
[0039] Embodiment two . Bacillus subtilis SWB8 ( Bacillus subtilis SWB8) CCTCC M 2010271 Diosgenin produced by liquid fermentation
[0040] The standard strains of Bacillus megaterium SWB15 (an endophyte isolated from turmeric) and Bacillus subtilis (ATCC6633) were selected as the control strains of Bacillus subtilis SWB8, and were fermented under the same conditions, and the fermentation products were extracted for the detection of diosgenin.
[0041] 1. Strain activation
[0042] Pick and preserve the strains, dilute and mix them with normal saline, spread them on the seed solid medium (0.3% beef extract; 1% peptone; 0.5% NaCl; 2% agar; distilled water; pH7.0), 30~35℃, Cultivate at constant temperature for 48-72 hours. Under the same conditions, replant once to rejuvenate the strain.
[0043] 2. Liquid fermentation
[0044] Pick the bacterial strains on the seed medium and dilute and mix with normal saline, draw 10 5 Inoculate the bacterial liquid of each cell int...
Embodiment 3
[0051] Embodiment Three . Bacillus subtilis SWB8 ( Bacillus subtilis SWB8) Antibacterial activity of diosgenin produced by CCTCC M 2010271
[0052] Pick the strains to be tested (Staphylococcus aureus, Enterococcus faecalis, Escherichia coli, Salmonella typhi, Salmonella paratyphi A, Shigella dysenteriae) diluted with normal saline and coated on corresponding common peptone solids On the culture medium, culture at 37°C for 48h. Using dimethyl sulfoxide (DMSO) as a solvent, a certain concentration of Bacillus subtilis SWB8 ( Bacillus subtilis The detection solution of diosgenin produced by SWB8) absorbs the The paper sheet of the detection solution is pasted on the coated with 10 7 ~10 8 Incubate at 37°C for 24 hours on the solid medium of each test bacterium with the number of bacterium, and measure the diameter of the inhibition zone. DMSO solution was set as the control in the experiment. The results showed that Bacillus subtilis SWB8 ( Bacillus subtilis SWB8) f...
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