Pseudomonas putida and method for producing nicotinic acid or isonicotinic acid through converting Pseudomonas putida
A technology of Pseudomonas putida and isonicotinic acid, applied in the directions of microorganism-based methods, biochemical equipment and methods, bacteria, etc., can solve problems such as no reports of Pseudomonas putida, and achieve high yield and reaction conditions. Gentle, environmentally friendly effect
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[0035] Example 1: Isolation and identification of Pseudomonas putida CGMCC3830 (Pseudomonas putida)
[0036] (1) Enrichment screening process of Pseudomonas putida CGMCC3830 (Pseudomonas putida).
[0037] Collect soil samples from 5-15 cm around the nitrile compound production plant. Take 1g of soil sample and put it into a triangular flask, add 15ml of normal saline, add glass beads, and shake on a shaker for 30min. Take 0.2 mL of soil suspension and spread it on a solid screening medium with 3-cyanopyridine as the sole nitrogen source. The composition of the solid screening medium is: glucose 0.5%, potassium dihydrogen phosphate 0.1%, magnesium sulfate 0.01%, sulfuric acid Ferrous iron 0.002%, calcium chloride 0.002%, sodium chloride 0.1%, 3-cyanopyridine 0.1%, agar 2%, pH 7.0. Incubate at 30°C for 3 days, pick colonies of different bacteria and continue streaking to a single colony .
[0038] Pick 20 purified strains and transfer them to liquid medium for re-screening. ...
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[0041] Example 2: Shake flask fermentation culture of Pseudomonas putida CGMCC3830 (Pseudomonas putida)
[0042] Prepare shake flask fermentation medium separately
[0043] A: 1% of glucose, 0.5% of soybean peptone, 0.3% of yeast powder, 0.3% of malt extract, 5% of NaCl, pH 7.2.
[0044] B: glycerol 1%, tryptone 1%, yeast powder 0.5%, NaCl 0.5%, pH 7.2.
[0045] C: glycerol 1%, tryptone 1%, yeast powder 0.5%, NaCl 0.1%, KH 2 PO 4 10.1%, pH 7.2.
[0046] D: glycerol 1%, tryptone 1%, yeast powder 0.5, NaCl 0.1%, KH 2 PO 4 10.1%, 1% urea, pH 6.0.
[0047] Divide the above-mentioned culture media into separate packages, three of each type in parallel, and sterilize at 121°C for 20 min. Pseudomonas putida CGMCC3830 (Pseudomonas putida) was inoculated into each fermentation medium, cultured at 30° C. and 120 rpm, and the fermentation was completed for 36 h. After the fermentation, the cells were collected by centrifugation at 10,000 rpm for 10 min, and the cells were washed ...
Example Embodiment
[0050] Example 3: Fermentation culture of Pseudomonas putida CGMCC3830 (Pseudomonas putida) in a fermenter
[0051] Pseudomonas putida CGMCC3830 (Pseudomonas putida) was inoculated into the seed medium, cultivated at 30° C. and 120 rpm for 24 hours to obtain seed liquid; the seed medium was composed as follows: peptone 0.5%, yeast powder 0.5%, NaCl 0.5%, pH7.0.
[0052] The fermentation medium was inoculated into the fermentation medium with an inoculation amount of 1% by volume, and cultured at 30° C. for 36 hours to obtain a fermentation broth. The composition of the fermentation medium is as follows: glycerol 1%, tryptone 1%, yeast extract 0.5%, potassium dihydrogen phosphate 0.1%, sodium chloride 0.1%, urea 0.1%. 30°C, stirring speed 150rpm-600rpm, ventilation rate 0.5-3mL / min, pH was controlled at 7.0 with acid-base, after culturing for 24h, the cells in the fermentation broth were collected for reaction.
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