Novel bacterial strain for nicotine degradation-Pseudomonas ZUTSKD and uses thereof

A technology of Pseudomonas and Pseudomonas, applied in the field of environmental biology, can solve problems such as paralysis of the heart, fatality, and danger

Inactive Publication Date: 2009-09-09
ZHEJIANG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Long-term smoking will lead to dependence on nicotine, and excessive inhalation of nicotine will inhibit the central nervous system, paralyze the heart, and even be fatal

Method used

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  • Novel bacterial strain for nicotine degradation-Pseudomonas ZUTSKD and uses thereof
  • Novel bacterial strain for nicotine degradation-Pseudomonas ZUTSKD and uses thereof
  • Novel bacterial strain for nicotine degradation-Pseudomonas ZUTSKD and uses thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1: Isolation, Identification and Nicotine Degradation Properties of Pseudomonas sp. ZUTSKD

[0023] 1. Isolation of strains

[0024] (1) Collect samples

[0025] Sampling location: The sample tobacco powder was taken from the warehouse of Hangzhou Liqun Environmental Protection Paper Co., Ltd.

[0026] (2) Strain isolation

[0027] Add 10g of tobacco powder to 100mL of distilled water, shake at 200r / min for 10min, take 10mL of leaching solution, add it to 100mL of enrichment medium, and culture at 200r / min at 30°C for 7 days. Dilute the enriched medium by 10 -5 ~10 -8 , spread on BSM agar medium, and cultured at 30°C for 48h. Pick a single colony and streak it on fresh BSM agar medium until purified bacteria are obtained.

[0028] 2. Identification of strains

[0029] (1) The morphological characteristics of the nicotine-degrading bacteria ZUTSKD and its colonies

[0030] The nicotine-degrading bacteria ZUTSKD has an ellipsoidal shape, with a size of (0....

Embodiment 2

[0039] Embodiment 2: the application of Pseudomonas (Pseudomonas sp.) ZUTSKD in waste tobacco dust treatment

[0040] Soak 4 g of dried waste tobacco powder in 60 mL of sterile water (or 0.02 mol / L phosphate buffer (pH=7.0) is also acceptable), inoculate with 2 mL of bacterial suspension (dry weight 7.1 mg / mL), and add Sterile water 2mL, 200r / min, 30 ℃ culture, continuous measurement of nicotine degradation. The result is as Figure 6 , it was found that nicotine continued to dissolve into the aqueous phase. Since the phosphate buffer can maintain the pH value of the solution, it is beneficial for strain ZUTSKD to degrade nicotine.

Embodiment 3

[0041] Example 3: Analysis of intermediate products of nicotine degradation by Pseudomonas sp. ZUTSKD

[0042] Inoculate 0.1mL of bacterial suspension into 100mL of BSM (nicotine 2g / L), culture at 200r / min, 30°C. At 6h and 12h, 10mL of the culture was taken respectively, and the supernatant after centrifugation (12000r / min, 10min, 4°C) was extracted twice with 1 / 2 volume of benzene, the benzene liquid was combined, and the benzene liquid was subjected to GC- MS analysis. After the culture was centrifuged (12000r / min, 10min, 4°C), the supernatant was extracted with benzene, and the benzene liquid was analyzed by GC-MS, and 3-(3,4-dihydro-2H-pyrrol-5 Accumulation of -yl)-Pyridine (I), 2,3'-Bipyridine (II), cotinine (III) and 3-Pyridinecarboxylic acid (IV).

[0043]

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Abstract

The invention provides a new strain of nicotine degrading bacteria—Pseudomonas ZUTSKD and its application. Pseudomonas sp. ZUTSKD (Pseudomonas sp. ZUTSKD), preserved in China Center for Type Culture Collection, date of preservation is June 18, 2007, preservation number CCTCCNO: M 207083. The bacteria can degrade nicotine to produce intermediate products such as 2,3'-Bipyridine, cotinine, 3-(3,4-dihydro-2H-pyrrol-5-yl)-Pyridine and 3-Pyridinecarboxylic acid, and can tolerate the highest nicotine With a concentration of 5.5g / L, it can degrade nicotine in waste tobacco powder, and has a certain application prospect in industry.

Description

(1) Technical field [0001] The invention relates to a new nicotine-degrading bacterium—pseudomonas ZUTSKD and its application, belonging to the field of environmental biotechnology. (2) Background technology [0002] Nicotine (nicotine, [1-methyl-2-(3-pyridyl-pyrrolidine), C 10 h 14 N 2 ]) are the main alkaloids in tobacco. Long-term smoking can lead to dependence on nicotine, and excessive inhalation of nicotine can inhibit the central nervous system, paralyze the heart, and even be fatal. When smoking, nicotine is also nitrosated to generate strong carcinogen N-nitrosamine (TSNA). At the same time, nicotine is also an environmentally toxic substance. Many wastes with high nicotine concentration will be produced in the tobacco processing process. -1 The dry weight of waste is classified as "toxic hazard". Therefore, controlling the nicotine content in tobacco and tobacco waste is of great significance to human health. [0003] The use of microorganisms to control the...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A62D3/02C12R1/38
Inventor 钟卫鸿孙柯丹朱晨静柳华贵李艺潇邱乐泉吴石金陈建孟汪新张艾晓
Owner ZHEJIANG UNIV OF TECH
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