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

Active Publication Date: 2009-09-09
CHINA TOBACCO ZHEJIANG IND +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] The strains involved in the above three patents have effective degradation properties for nicotine and are suitable for the treatment of waste nicotine. The only disadvantage is that they all involve the degradation of nicotine in a liquid state, and whether they have an effect on solid tobacco leaves has not yet been experimentally confirmed.

Method used

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  • Pseudomonad ZCJ bacterial strain applied to nicotine degradation of tobacco and screening method and application thereof
  • Pseudomonad ZCJ bacterial strain applied to nicotine degradation of tobacco and screening method and application thereof
  • Pseudomonad ZCJ bacterial strain applied to nicotine degradation of tobacco and screening method and application thereof

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Experimental program
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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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Abstract

The invention relates to a technology of biological degradation of nicotine in tobacco, in particular to a pseudomonad ZCJ bacterial strain applied to nicotine degradation of tobacco and a screening method and an application thereof. The pseudomonad ZCJ bacterial strain (Pseudomonas sp. ZCJ) was preserved in the China Center for Type Culture Collection on March, 30th, 2009 with a preservation number of CCTCC NO. M209064. The invention also discloses screening and application of the bacterial strain. In the invention, nicotine degradation bacterial strain screened from tobacco materials has relatively high biological activity and is applicable to degradation of nicotine in tobacco in a solid state, thus achieving the purpose of the degradation of the nicotine in tobacco.

Description

technical field [0001] The invention relates to a technology for biodegrading tobacco nicotine, in particular to a Pseudomonas ZCJ strain used for degradation of tobacco nicotine and its screening method and application. Background technique [0002] Affected by factors such as weather, climate, and cultivation, the nicotine (nicotine) content of tobacco leaves fluctuates to a certain extent, which easily leads to instability of the inherent quality of cigarettes. At the same time, compared with reducing tar and harmful components of smoke, the reduction of nicotine (nicotine) in tobacco has not yet achieved breakthrough progress. [0003] Compared with the method of reducing nicotine by agricultural field management, microorganisms are easy to cultivate, and the degradation of nicotine is quick, selective and controllable, and will have other beneficial effects on the quality of cigarettes; The nicotine degradation ability of bacteria is not very strong, and the tolerance ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14A24B3/18C12R1/38
Inventor 舒明杨君钟卫鸿朱晨静朱申康朱雪娜
Owner CHINA TOBACCO ZHEJIANG IND
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