Nicotine-degrading bacterium and application thereof
A nicotine-degrading bacteria and nicotine-degrading technology can be applied to bacteria, organic fertilizers, microorganisms, etc., and can solve the problems of lowering the quality of compost products.
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Embodiment 1
[0030] Embodiment 1: A nicotine-degrading bacterium of the present invention, which is named Arthrobacter histidinolovorans EA-17 taxonomically, and was preserved in the China Type Culture Collection Center on September 15, 2013. The number is CCTCCM2013405. The industrial production of nicotine-degrading bacteria EA-17 is secondary fermentation. The production method is as follows: inoculate the activated strains in LB liquid medium, cultivate them at 30-37°C for 10-18 hours, and prepare first-grade seeds; 0.1%-0.5% of the inoculum was inoculated in 2M liquid LB medium 3 In the fermenter, the liquid volume of the fermenter is 60%, 30-37°C, the stirring speed is 250rpm, the aeration ratio is 0.2VVM-1VVM, the fermentation is 16-24h, and the biomass reaches 3×10 9 CFU / mL.
[0031] An application of nicotine-degrading bacteria in tobacco stalk piles. The use of nicotine-degrading bacteria EA-17 is as follows: after mixing the fermentation liquid of nicotine-degrading bacteria a...
Embodiment 2
[0032] Embodiment 2: Preparation of a kind of nicotine degrading bacteria EA-17 bacterial liquid of the present invention
[0033] 1 medium
[0034] LB medium: peptone 10g, yeast powder 5g, sodium chloride 10g, agar 20g, distilled water 1000ml, pH7.0-pH7.2, sterilized at 121°C for 30min.
[0035] 2 Bacteria preparation
[0036] 2.1 Strain activation: Nicotine-degrading bacteria EA-17 was inoculated on LB medium (2% agar was added on the basis of LB medium), and cultured at 30°C for 18-24h.
[0037] 2.2 Cultivation of first-class strains: Transfer the above-mentioned cultured nicotine-degrading bacteria EA-17 into liquid LB medium, fill in 50ml / 500ml Erlenmeyer flask, culture at 30°C, shaker speed 180r / min, shake Bed culture for 12h.
[0038] 2.3 Fermentation tank culture: the volume of the fermentation tank is 2M 3 , 1200L culture medium, the above-mentioned primary seed inoculum volume is 2.4L, the stirring speed is 250rpm, the aeration ratio is 0.5VVM, the fermentation t...
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