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Shinella sp. HZN1 capable of effectively degrading nicotine and application thereof

A technology of S. brucei and nicotine, which is applied to S. bacillus HZN1 capable of efficiently degrading nicotine and its application field, can solve the problems of strong irritation, toxicity, inhibition of human central nervous system, etc., and achieves a simple preparation process and good application prospects. , the effect of rapid degradation

Inactive Publication Date: 2010-12-29
ZHEJIANG UNIV OF TECH
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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. Severe cases can be fatal; nicotine is also an environmentally toxic substance. Pure nicotine is a colorless, bitter, and strongly irritating oily liquid at room temperature. 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 production.
At the same time, tobacco will produce high-concentration nicotine waste during the processing process. This waste is considered to be "toxic and dangerous waste" and causes great harm to the environment.

Method used

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  • Shinella sp. HZN1 capable of effectively degrading nicotine and application thereof
  • Shinella sp. HZN1 capable of effectively degrading nicotine and application thereof
  • Shinella sp. HZN1 capable of effectively degrading nicotine and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Embodiment 1: Screening and identification of strains

[0018] Medium:

[0019] Inorganic salt medium: NaCl 1g, K 2 HPO 4 1.5g, KH 2 PO 4 0.5g, (NH4) 2 SO 4 1.5g, MgSO 4 0.1g, 1ml trace element solution, made up to 1000ml with distilled water, mixed and stirred evenly, the natural pH value, made after high-pressure steam sterilization (121°C, 20min), and the trace element solution was prepared as follows: 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.

[0020] Enriched culture solution: Add nicotine to the inorganic salt culture solution so that the concentration of nicotine is 200mg / L.

[0021] LB culture medium: yeast powder 10g, peptone 5.0g, sodium chloride 10.0g, distilled water to make up to 1000ml, mixed and stirred evenly, natural pH value, prepared after high-pressure steam sterilization (121°C, 20min).

[0022] LB solid ...

Embodiment 2

[0028] Embodiment 2: bacterial agent preparation

[0029] 1. Inoculate the strains stored in the liquid test tube in 40ml of inorganic salt culture solution for activation and culture for 4 days;

[0030] 2. Inoculate the activated bacteria into 100ml of LB liquid medium containing 50-500mg / L nicotine, culture at 30°C and 150rpm until logarithmic growth phase;

[0031] 3. Centrifuge (6000rpm) the above-mentioned bacterial solution in logarithmic growth phase for 3 minutes, discard the supernatant, and suspend the bacterial cells with an appropriate amount of phosphate buffer solution with a pH value of 7.0, which is the microbial agent.

[0032] The formula of 0.2mol / L phosphate buffer solution with a pH of 7.0 is: take 39ml of 0.2mol / L sodium dihydrogen phosphate and 61ml of 0.2mol / L disodium hydrogen phosphate, dilute to 1000ml with ultrapure water, and steam under high pressure It can be obtained after sterilization (121°C, 20min).

Embodiment 3

[0033] Embodiment 3: Nicotine degradation experiment

[0034] Detection of bacterial concentration and nicotine content in inorganic salt culture solution:

[0035] In this experiment, the growth of bacteria was detected by ultraviolet spectrophotometer, which was expressed by measuring the absorbance value of bacteria at 600nm.

[0036] In this experiment, reversed-phase high-performance liquid chromatography was used to detect the residual amount of nicotine in the inorganic salt culture solution. High performance liquid chromatography (HPLC) detection condition: mobile phase is methanol: 1mM H 2 SO 4 =10:90 (volume ratio), the analytical column is 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.

[0037] Use different concentrations of nicotine solutions for high-performance liquid chromatography detection, and use the nicotine concentration as the abscissa and the absorption peak are...

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Abstract

The invention provides a novel Shinella sp. HZN1 capable of effectively degrading nicotine and application thereof. The Shinella sp. HZN1 is preserved in China Center for Type Culture Collection; the address is 430072, Wuhan University, Wuhan, China; the preservation number is CCTCC No:M 2010154; and the preservation date is June 23rd, 2010. The nicotine-degrading bacterium can be directly thrown in a water body to degrade nicotine, and can safely, efficiently and quickly degrade the residual nicotine in the water body, soil and the like. Besides, the microbial inoculum containing the strain has the advantages of simple preparation process, low cost and convenient use, thereby having favorable application prospects.

Description

(1) Technical field [0001] The invention relates to a strain of Shennella HZN1 capable of efficiently degrading nicotine and its application. (2) Background technology [0002] Nicotine, commonly known as nicotine, is the unique and most important alkaloid in various tobaccos, accounting for 1% to 2% of the weight of tobacco, and is one of the important factors affecting the quality of tobacco leaves. One of the main harmful ingredients. The molecular formula for nicotine is C 10 h 14 N 2 , the structure is as follows: [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. Severe cases can be fatal; nicotine is also an environmentally toxic substance. Pure nicotine is a colorless, bitter, and strongly irritating oily liquid at room temperature, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C02F3/34B09C1/10A62D3/02C12R1/01C02F101/38A62D101/26
Inventor 马云江吉红邱吉国吴立飞陈连升刘维屏
Owner ZHEJIANG UNIV OF TECH
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