Leclercia adecarboxylata ZJB-17008 and application thereof
A technology of ZJB-17008, non-decarboxylated leukemia, applied in bacteria, microorganisms, biochemical equipment and methods, etc., can solve the problems of high price of trimethylsilicon, waste of raw materials, low product yield and e.e. value, etc.
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Embodiment 1
[0028] Example 1: Screening of non-decarboxylated Lecleria (Leclercia adecarboxylata) ZJB-17008
[0029] 1. Primary screening
[0030] The present invention takes soil samples from all over the country, and takes 80 parts of soil samples altogether. The specific method of screening: Weigh 1g of soil sample and place it in 10mL of 0.85% normal saline, shake it and let it stand still, take the supernatant into the enrichment medium, and culture it at 30°C and 150 r / min for 2-3 days with shaking . Take 1mL of the enrichment solution and add it to 50mL of fresh enrichment medium, and repeat this process 3 times before separation and purification.
[0031] Bromothymol blue filter paper was selected as the indicator filter paper to detect the colony capable of degrading the substrate. The principle is: after the colony containing the target enzyme degrades the substrate, acidic by-products (phenylacetic acid, acetic acid, formic acid, benzoic acid) will be produced, thereby chang...
Embodiment 2
[0047] Embodiment 2: identification of bacterial strain ZJB-17008
[0048] 1. Morphological identification:
[0049] The bacterial strain ZJB-17008 screened in Example 1 was inoculated on a solid medium and cultured at 37°C for 24 hours to form round or nearly round, soft texture, smooth surface, flat, neat edges, and glossy milky white colonies. 2-4mm in diameter. Observation by Gram staining: short pink rods without spores. Composition of solid medium: sodium chloride 10 g / L, peptone 10 g / L, yeast powder 5 g / L, agar 20 g / L, solvent is deionized water.
[0050] 2. Physiological and biochemical identification:
[0051] Using the Biolog (GENⅢ) automatic microbial identification system, 94 kinds of phenotypic tests were carried out on the strain ZJB-17008, including 71 kinds of carbon source utilization detection and 23 kinds of chemical sensitivity detection: the strains were inoculated on BUG plate medium (BIOLOG UNIVERSAL GROWTH AGAR), cultured at a constant temperature o...
Embodiment 3
[0061] Embodiment 3: the preparation of wet thalline
[0062] (1) Incline cultivation:
[0063] Inoculate non-decarboxylated Leclercia (Leclercia adecarboxylata) ZJB-17008 into the slant culture medium and culture at 30°C for 48 hours to obtain slant bacteria;
[0064] The final concentration of the slant culture medium is: lactose 8g / L, peptone 6g / L, Na 2 HPO 4 3.2g / L, K 2 HPO 4 1.5g / L, Na 2 EDTA 0.06g / L, MnSO 4 0.002g / L, MgSO 4 0.001g / L, ZnSO 4 0.002g / L, FeSO 4 0.01g / L, the agar is 20g / L, the solvent is deionized water, and the pH value is 6.8.
[0065] (2) Seed cultivation
[0066] Pick an inoculation loop of bacteria from the slant and inoculate it into the seed medium, and cultivate it at 30°C for 24 hours to obtain the seed liquid;
[0067] The final concentration of the seed medium consists of: lactose 8g / L, peptone 6g / L, Na 2 HPO 4 3.2g / L, K 2 HPO 4 1.5g / L, Na 2 EDTA 0.06g / L, MnSO 4 0.002g / L, MgSO 4 0.001g / L, ZnSO4 0.002 g / L, FeSO 4 0.01g / L,...
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