Kineococcus cavernae sp. and application thereof
A technology of dynamic coccus and cave, applied in the field of microorganisms, can solve the problem of no public report of dynamic coccus cave, and achieve the effects of expanding the richness, rounding the smoke, and enriching the aroma of the smoke
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Embodiment 1
[0021] ——Isolation of Kinetococcus burrowing bacteria SYSUZL-1
[0022] Soil samples were collected from karst caves adjacent to the Yinyanzhuang tobacco field in Shilin County, Yunnan Province, sealed in plastic bags, and stored at 4°C until use. Accurately weigh 10g of soil sample, add 90mL of sterile water, shake at 30°C and 180rpm for 30min. Dilute to 10 with sterile water -1 、10 -2 、10 -3 、10 -4 、10 -5 Concentration, respectively take 0.2mL spread on LB plate culture medium, set 3 replicates for each. After culturing at 30°C for 48 hours, different single colonies were picked from the plates of each soil sample, purified on the corresponding solid medium, added glycerol at 20% of the volume of the bacterial solution, and stored in a -80°C refrigerator for later use.
[0023] The isolated microorganisms were respectively inoculated into LB liquid medium, shaken and cultivated at 30°C and 160 rpm on a shaker, and the bacteria were cultured overnight until the concentr...
Embodiment 2
[0025] ——Identification of Kinetococcus burrowing bacteria SYSUZL-1
[0026] The isolated and purified SYSUZL-1 was identified by morphology, physiology, biochemistry and molecular biology. The classification of SYSUZL-1 was named Kineococcus cavernae sp.nov.
[0027] The micrographs and electron micrographs of SYSUZL-1 on R2A medium are attached figure 1 .
[0028] SYSUZL-1 grows well on most media, and the hyphae in the substrate are reddish brown to brown. Gelatin liquefaction, milk coagulation and peptonization are positive, starch hydrolysis and nitric acid reduction are negative, hydrogen sulfide is not produced, and melanin is formed. Can utilize glucose, galactose, fructose xylose, raffinose, rhamnose, melibiose, sweet alcohol, glycerin, mannitol, sorbitol, sodium acetate. The cell wall contains L-diaminopimelic acid and glycine. (Table 1-3)
[0029] Table 1 Culture characteristics of SYSUZL-1
[0030]
[0031] Table 2 Physiological and biochemical characteri...
Embodiment 3
[0040] ——Application of Kinetococcus burrowing bacteria SYSUZL-1 in the preparation of tobacco flavor by fermentation
[0041] The application of the above-screened Kinetococcus burrowing bacteria SYSUZL-1 in the fermentative production of tobacco spices, the specific steps are as follows:
[0042] (1) Slant culture: medium: glucose 0.5g; yeast extract 0.5g; peptone 0.5g; acid hydrolyzed casein 0.5g; soluble starch 0.5g; sodium pyruvate 0.3g; g; agar 15g; distilled water 1000ml, pH7.2; culture temperature is 28°C, culture time is 7d.
[0043] (2) Seed culture: pick part of the mycelium from the slope and put it into the seed liquid, and shake the flask for 36 hours. Seed medium: dextrin 120g; soybean powder 40g; yeast extract 2g; tryptophan 0.5g; β-alanine 5g; magnesium sulfate 0.5g; ammonium phosphate 0.2g; ℃.
[0044] (3) Fermentation culture: Inoculate 10% of the inoculum into the fermentation medium, and culture in shake flasks for 6 days. Medium: soybean powder 10g; g...
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