A luminobacterium fc615 strain and its culture method and application
A technology of photobacteria and liquid culture medium, which is applied in the field of microorganisms, can solve problems such as sources, and achieve the effect of broad application prospects
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Embodiment 1
[0040] Example 1. Acquisition of Photobacterium sp. FC615
[0041] Take sea mud leachate, add 1 mL of supernatant to 9 mL of sterile water, and dilute to 10 -1 , 10 -2 , 10 -3 , 10 -4 , 10 -5 , 10 -6 Then, by conventional dilution method or streaking method, it was inoculated on the sole carbon source solid medium, incubated at 30°C for 1 day, the colonies were counted, and then the colonies with obvious differences in colony morphology were selected, and the streaking was repeated. It was inoculated on the corresponding complete nutrient agar plate until a single colony was obtained after purification, and then transferred to the corresponding agar slant for later use.
[0042] The cultured strains were respectively inoculated onto the only carbon source liquid medium for cultivation. Incubate at 200rpm and 30°C for 72h, observe the turbidity of the bacterial liquid, and take the culture supernatant for carbazole reaction to detect the consumption of carbon source. The...
Embodiment 2
[0047] Embodiment 2, the cultivation of luminescent bacteria FC615 bacterial liquid
[0048] The culturing method of luminescent bacteria FC615 bacterial liquid, the steps are as follows:
[0049] (1) Inoculate the luminobacterium FC615 strain into the liquid medium, and under the condition that the temperature is 28~30°C and the number of revolutions is 200rpm, the seed liquid is obtained by shaking the culture for 10 hours;
[0050] (2) Take the seed solution obtained in step (1), inoculate it in a liquid medium at a volume percentage of 7%, at a temperature of 25 to 30° C. and dissolved oxygen (that is, dissolved oxygen saturation) under the conditions of 30% , expanded and cultured for 2 hours to obtain luminescent bacteria FC615.
[0051] The components per liter of the above liquid medium are as follows:
[0052] Tryptone 10g, yeast extract 5g, seawater 33g, water 1000mL, pH 7.2.
Embodiment 3
[0053] Example 3. Application of Photobacillus FC615 in the preparation of glycosaminoglycan lyase
[0054] The application of luminobacterium FC615 in the preparation of glycosaminoglycan lyase, the steps are as follows:
[0055] (1) Inoculate the luminobacterium FC615 strain into the liquid medium, under the condition that the temperature is 28~30 DEG C and the number of revolutions is 300 rpm, shake the culture for 16 hours to obtain the seed liquid;
[0056] (2) Take the seed solution obtained in step (1), inoculate it in a liquid medium at a volume percentage of 8%, at a temperature of 25 to 30° C. and dissolved oxygen (that is, dissolved oxygen saturation) under the conditions of 25% , expanded and cultured for 5 hours, added chondroitin sulfate to make the mass concentration of chondroitin sulfate 0.01-0.02%, and continued to culture for 72 hours to obtain luminobacterium FC615 fermentation broth;
[0057] (3) Take the photobacillus FC615 fermentation broth obtained in...
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