Pseudomonad ZCJ bacterial strain applied to nicotine degradation of tobacco and screening method and application thereof
A pseudomonas, screening method technology, applied in the direction of microorganism-based methods, applications, biochemical equipment and methods, etc., can solve the problem of unproven solid tobacco leaves, achieve the maintenance of brand image, improve usability, improve quality effect
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Embodiment 1
[0025] Example 1 Isolation, Purification and Identification of Pseudomonas ZCJ Strain (Pseudomonas sp.ZCJ)
[0026] 1. Separation and purification of tobacco leaf nicotine (nicotine) degrading bacteria
[0027] Add 10 g of aged tobacco leaves (provided by Zhejiang China Tobacco Industry Co., Ltd.) into 100 mL of physiological saline, shake at 30° C. and 180 rpm for 30 min. Take 1mL shaking liquid and add to 100mL enrichment medium (enrichment medium: Na 2 HPO 4 5.57g·L -1 , KH 2 PO 4 2.44g·L -1 , (NH 4 ) 2 SO 4 1g·L -1 , MgCl 2 ·6H 2 O0.2g L -1 , MnCl 2 4H 2 O 0.0004g·L -1 , FeCl 3 ·6H 2 O 0.001g·L -1 , CaCl 2 0.001g·L -1 , Nicotine 10g·L -1 , pH=7.0), 30°C, shaking at 180rpm for 48h. An appropriate amount of enriched culture solution was taken, spread on BSM agar medium (nicotine content 2g / L), and cultured at 28°C for 48h. Pick a single colony and streak it on fresh BSM agar medium to obtain a purified strain.
[0028] 2. Morphological structure o...
Embodiment 2
[0037] Nicotine degradation characteristics of embodiment 2 Pseudomonas ZCJ bacterial strain (Pseudomonas sp.ZCJ)
[0038] 1. Growth of Pseudomonas sp.ZCJ in liquid medium and its degradation effect on nicotine
[0039] Inoculate 1 mL of bacterial suspension (dry cell weight about 0.3125 mg) into 100 mL of BSM medium containing nicotine 2 g / L, culture at 180 rpm at different temperatures (25, 28, 37°C). After 4 days, the dry weight of the bacteria and the degradation of nicotine were measured.
[0040] Take 30mL of bacterial suspension, centrifuge at 4000rpm for 15min, remove the supernatant, and dry in an oven at low temperature until constant weight. It was found that as the culture temperature increased, the growth of the bacteria increased (such as image 3 shown). The nicotine content was detected by HPLC, and it was found that when the incubation temperature was 37°C, the nicotine was degraded the most (such as Figure 4 shown).
[0041] 2. The effect of Pseudomonas...
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