Nicotine-degrading bacterium and application thereof
A technology of nicotine-degrading bacteria and tobacco stalks, which 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] Example 1: A nicotine-degrading bacterium of the present invention, which is taxonomically named Arthrobacter histidinolovorans (Arthrobacter histidinolovorans) EA-17, and was deposited in the China Type Culture Collection 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 bacteria in LB liquid medium, cultivate at 30-37°C for 10-18 hours, and prepare them into first-level seeds; 0.1%-0.5% of the inoculum is inoculated into 2M with liquid LB medium 3 In the fermentation tank, the liquid volume in the fermentation tank is 60%, 30-37℃, stirring speed 250rpm, aeration ratio 0.2VVM-1VVM, fermentation 16-24h, biomass up to 3×10 9 CFU / mL.
[0031] A kind of application of nicotine degrading bacteria in tobacco stalk piles. The use of nicotine degrading bacteria EA-17 is as follows: nicotine degrading bacteria fermentation broth a...
Embodiment 2
[0032] Example 2: Preparation of a nicotine-degrading bacterium EA-17 bacterial solution 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, 121℃, sterilized for 30min.
[0035] 2 Bacteria preparation
[0036] 2.1 Strain activation: Nicotine degrading bacteria EA-17 was inoculated on LB medium (2% agar added to LB medium), and cultured at 30°C for 18-24h.
[0037] 2.2 Primary strain culture: transfer the above-mentioned nicotine-degrading bacteria EA-17 to liquid LB medium, fill 50ml / 500ml Erlenmeyer flask, culture temperature 30℃, shaker speed 180r / min, shake Bed culture for 12h.
[0038] 2.3 Fermentation tank culture: Fermentation tank volume 2M 3 , 1200L medium, the above-mentioned primary seed inoculum is 2.4L, the stirring speed is 250rpm, the aeration ratio is 0.5VVM, the fermentation temperature is 30℃, and the fermentation is 20h.
[0039] 3 fermentation broth biomass
[00...
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