Epidermis brevibacterium ZJB-07021 and use for preparation of (S)-2,2-dimethyl cyclopropane formamide thereof
A technology of dimethylcyclopropanecarboxamide, ZJB-07021, which is applied to Brevibacterium epidermidis ZJB-07021 and its application in the preparation of (S)-2,2-dimethylcyclopropanecarboxamide, which can solve the problem of Environmental hazards and other issues
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Embodiment 1
[0036] Example 1: Isolation of strains
[0037] Isolation of R-amidase-producing bacteria: Add 1 g of soil sample and appropriate amount of glass beads to 9 mL of 0.85% saline, shake well to make a uniform soil suspension; pipette 0.5 mL of soil suspension to inoculate In a 250-mL Erlenmeyer flask with 30 mL of enriched medium, place it in a shaker at 30°C and 150 rpm for 4 days. After the enriched liquid appears turbid, transfer 0.5 mL to fresh medium and continue to culture for 4 days ; After repeating 4 to 5 cycles, the enrichment solution is diluted with multiple gradients and spread on the separation plate to obtain a single colony;
[0038] The enrichment medium uses 2,2-dimethylcyclopropanamide as the only nitrogen source, and the composition is as follows (final concentration): K 2 HPO 4 ·12H 2 O: 2.5g / L, KH 2 PO 4 :2g / L, MgSO 4 ·7H 2 O: 0.5g / L, FeSO 4 ·7H 2 O: 0.01g / L, CaCl 2 :0.06g / L, 2,2-Dimethylcyclopropanecarboxamide: 1.5g / L, prepared with tap water, pH adjusted to 7....
Embodiment 2
[0040] Example 2: Cultivation of strains
[0041] Slant medium composition (final concentration): beef extract 5g / L, peptone 10g / L, NaCl5g / L, agar 18g / L, prepared with tap water;
[0042] Pick a single colony of CCTCC M.207076 on the plate, inoculate it on the slope, cultivate it in a constant temperature incubator at 30°C for 48 hours, and store it in a refrigerator at 4°C for later use.
Embodiment 3
[0043] Example 3: Obtaining wet bacterial cells
[0044] Medium preparation: glucose 10.0g, yeast powder 13.0g, KH 2 PO 4 0.5g, K 2 HPO 4 0.5g, NaCl1g, acetamide 5g, tap water to make up to 1L, pH6.5;
[0045] 250ml shake flask with 30% liquid volume, inoculate a loop of Brevibacterium epidermidis CCTCC M.207076, shake culture at 20°C, 300rpm for 72h; after the culture, the fermentation broth is centrifuged and washed twice with saline to collect the wet bacterial cells and suspended in 50mM In the phosphate buffer solution of pH 7.8, a bacterial suspension with a wet bacterial cell content of 0.5 g / mL is obtained for use.
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