Enterobacteriaceae and their uses
A technology for Enterobacteriaceae and its uses, which is applied in the field of Enterobacteriaceae and its uses, and can solve problems such as the conversion of stevioside into Enterobacteriaceae that have not yet been known.
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Embodiment 1
[0044] Example 1. Fermentation of Enterobacter
[0045] 1) Transfer an inoculated loop of Enterobacter S618-1 to 100ml LB liquid medium for enrichment culture at 200r / min and 30°C for 24h; to obtain enriched culture solution (seed solution).
[0046] 2). Connect the enriched culture solution obtained in step 1) to the fermentation medium at 200r / min, 30℃ for fermentation culture, the culture time is 72h, and the volume ratio of enriched culture solution: fermentation medium=1:9 ;
[0047] The resulting fermentation broth was centrifuged at 8000 rpm at 4°C for 15 min, and the supernatant was collected.
[0048] 3) Filter the supernatant with a 0.22μm filter membrane; use a micro-injector to take 20μl for HPLC detection.
[0049] Experiment 1. Detecting active substances in fermentation broth by HPLC
[0050] Chromatographic conditions: Yilite 3200 high performance liquid chromatograph, Agilent Agilent reversed-phase chromatography column, EclipseXDB-C18, 4.6x150mm, UV detection at 210nm,...
Embodiment 2
[0060] Example 2. The method for converting stevioside into sweet tea glycosides (that is, the method for producing sweet tea glycosides using Enterobacter bacteria), followed by the following steps:
[0061] 1). Activate the strains stored in the glycerol tube with LB liquid medium:
[0062] An inoculated loop of Enterobacter S618-1 stored on a modified inorganic salt medium was inoculated into 100ml LB liquid medium at a rotation speed of 200r / min and cultivated at 30°C for 24h to obtain a seed solution.
[0063] 2) Centrifuge the seed liquid at 5000 rpm for 10 minutes to remove the supernatant (about 80% of the seed liquid);
[0064] Add 50ml of fermentation medium I to the obtained precipitate for fermentation; the fermentation conditions are: fermentation culture is carried out at 200r / min, 30°C, and the culture time is 72h; the fermentation broth is obtained.
[0065] Fermentation medium I is: stevioside 10g / L, bran 10g / L, soybean meal 5g / L, potassium dihydrogen phosphate 5g / L, di...
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