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Pseudomonas sp.HZN6 and application thereof to nicotine degradation

A Pseudomonas, nicotine technology, applied in bacteria, microorganism-based methods, water/sludge/sewage treatment, etc., can solve the problems of strong irritation, toxicity, environmental hazards, etc., and achieves a simple preparation process, Good application prospect and low cost effect

Active Publication Date: 2012-11-28
菏泽建数智能科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, nicotine is an important precursor of tobacco-specific nitrosamines (TSNA), the main carcinogenic component in tobacco leaves and smoke. At the same time, nicotine is also an environmentally toxic substance. Pure nicotine is a colorless, bitter, and strongly irritating oily liquid at room temperature, and it is easily oxidized into dark gray in the air.
The flue gas environment contains a large amount of nicotine. At present, the content of nicotine in some tobacco leaves in my country is too high, especially in the upper tobacco leaves, which brings great challenges to tobacco leaf production.
At the same time, tobacco will produce high-concentration nicotine waste during the processing process. This waste is considered to be "toxic and hazardous waste" and causes great harm to the environment.

Method used

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  • Pseudomonas sp.HZN6 and application thereof to nicotine degradation
  • Pseudomonas sp.HZN6 and application thereof to nicotine degradation
  • Pseudomonas sp.HZN6 and application thereof to nicotine degradation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Embodiment 1: Screening and identification of strains

[0043] 1) culture medium

[0044] Preparation of inorganic salt medium: NaCl 1g, K 2 HPO 4 1.5g, KH 2 PO 4 0.5g, (NH 4 ) 2 SO 4 1.5g, MgSO 4 0.1g, 1ml trace element solution, made up to 1000ml with distilled water, mixed and stirred evenly, natural pH value, prepared after high-pressure steam sterilization (121°C, 20min), and each liter of trace element solution contains MnSO 4 ·H 2 O 0.13g, ZnCl 2 0.23g, CuSO 4 ·H 2 O0.03g, CoCl 2 ·6H 2 O 0.42g, Na 2 MoO 4 2H 2 O 0.15g, AlCl 3 ·6H 2 O 0.05g, make up to 1000ml with distilled water.

[0045] Enriched culture solution: Add nicotine to the inorganic salt culture solution so that the final concentration of nicotine is 200mg / L.

[0046] Preparation of LB liquid medium: Yeast powder 10g, peptone 5.0g, sodium chloride 10.0g, distilled water to make up to 1000ml, mix and stir evenly, natural pH value, high pressure steam sterilization (121°C, 20min...

Embodiment 2

[0053] Embodiment 2: the preparation that contains bacterial cell suspension

[0054] (1) Slant culture: Inoculate Pseudomonas HZN6 on the slant medium and culture at 30°C for 6 days to obtain the slant; the final concentration of the slant medium consists of: yeast powder 10g, peptone 5.0g, chlorine Sodium chloride 10.0g, agarose 2.0g, water 1000ml;

[0055] (2) Seed culture: Pick an inoculation loop from the slant of the bacteria in step (1) and inoculate it into the inorganic salt medium, and cultivate it at 30°C for 6 days to obtain the seed liquid; the final concentration of the inorganic salt medium is the same as Embodiment 1;

[0056] (3) Expansion culture: Inoculate the seed solution obtained in step (2) into LB liquid medium (100mL) with an inoculum volume concentration of 15%, culture at 30°C and 150rpm until the logarithmic growth phase, and obtain the bacterial solution. Centrifuge the bacterial solution (8000rpm, 5min), discard the supernatant, and suspend the ...

Embodiment 3

[0057] Embodiment 3: Nicotine degradation experiment

[0058] 1) Detection of bacterial concentration and nicotine content in inorganic salt culture solution:

[0059] The growth of bacteria is detected by ultraviolet spectrophotometer, which is expressed by measuring the absorbance value of bacteria in the culture solution at 600nm.

[0060] In this experiment, reversed-phase high-performance liquid chromatography was used to detect the residual amount of nicotine in the inorganic salt culture solution. Reversed-phase high-performance liquid chromatography detection conditions: mobile phase is methanol: 1mM H 2 SO 4 =10:90 (volume ratio), the analytical column is a Grace Alltima C18 column (4.6×250mm, 5μm), the flow rate is 0.6ml / min, the injection volume is 20μl, and the column temperature is 30°C.

[0061] 2) Nicotine degradation experiment:

[0062] The nicotine standard substance is dissolved in sterile water and prepared as a 100mg / L standard solution, and the standa...

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Abstract

The invention discloses Pseudomonas sp.HZN6 and application thereof to nicotine degradation. In application, with nicotine as a substrate, an inorganic salt nutrient solution as a reaction medium and Pseudomonas sp.HZN6 as an enzyme, the reaction is carried out for 8-15h under the conditions that temperature is 25-45 DEG C and a Ph value is 5.5-9.0 to ensure that the content of nicotine in the reaction solution is less than 0.1 percent by mass, thus the purpose of degrading nicotine is reached. The Pseudomonas sp.HZN6 can be directly added to a water body and soil to degrade nicotine in the water body and soil, and can be used for safely, efficiently and rapidly degrading residual nicotine on objects such as water and soil. A bactericide containing the Pseudomonas sp.HZN6 is prepared by a simple process with low cost, is convenient to use, and has a good application prospect.

Description

(1) Technical field [0001] The invention relates to a novel high-efficiency nicotine-degrading bacterium, in particular to a novel high-efficiency Pseudomonas nicotine-degrading bacterium, that is, Pseudomonas HZN6 and application thereof. (2) Background technology [0002] Nicotine, commonly known as nicotine, is the unique and most important alkaloid in various tobaccos, accounting for about 1%-2% of the dry weight of tobacco, and is one of the important factors affecting the quality of tobacco leaves. One of the main harmful ingredients of cigarettes. The molecular formula for nicotine is C 10 h 14 N 2 , the structural formula is shown in formula (I). [0003] [0004] Because nicotine is a psychotropic drug, human beings have maintained the habit of smoking tobacco for a long time. However, nicotine is an important precursor of tobacco-specific nitrosamines (TSNA), the main carcinogenic component in tobacco leaves and smoke. At the same time, nicotine is also an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C02F3/34A62D3/02C12R1/38C02F101/38A62D101/26
Inventor 马云邱吉国刘学虎魏银
Owner 菏泽建数智能科技有限公司
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