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Pseudomonas fluorescens preparation and application thereof

A technology of Pseudomonas fluorescens and preparations, applied in the field of microorganisms, can solve the problem that no Pseudomonas fluorescens can degrade nicotine and the like, and achieve the effects of good nicotine degradation, improved quality and low preparation cost

Active Publication Date: 2012-06-27
YUNNAN ACAD OF TOBACCO AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

According to available information, there is no report that Pseudomonas fluorescens can degrade nicotine

Method used

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  • Pseudomonas fluorescens preparation and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Embodiment 1: (for liquid containing nicotine)

[0023] Prepare Pseudomonas fluorescens preparations:

[0024] The preparation production strain is Pseudomonas fluorescens 1206, CGMCC NO.3149;

[0025] a. Fermentation strain culture: Inoculate Pseudomonas fluorescens 1206 slant strains into a culture solution with a formula of 3% beef extract, 0.5% peptone, 0.5% sodium chloride, and tap water, pH 7.0 or 7.2, shake The rotation speed of the bottle is 150 rpm, and the culture is carried out at a temperature of 28° C., 29° C. or 30° C. for 18 hours to obtain liquid fermented seeds.

[0026] b. Fermentation process: inoculate the liquid fermented seeds into a fermenter equipped with a liquid fermentation system, medium formula (g / L): maltose 35, yeast extract 4.03, ammonium sulfate 4.72, K 2 HPO 4 0.4, MgSO 4 .7H 2 O 0.075, FeSO 4 .7H 2 O 0.0005, CaCl 20.015, NaCl 0.1. The fermentation temperature is 28°C, the inoculum size is 8%, the defoamer is 0.08%, the stirrin...

Embodiment 2

[0028] Embodiment 2: (for cigarette factory waste water)

[0029] The preparation of the Pseudomonas fluorescens preparation was the same as in Example 1.

[0030] Application of the nicotine-degrading preparation of the present invention: Inoculate the nicotine-reducing preparation (1% inoculum size) into tobacco factory wastewater containing nicotine, cultivate in a shaking flask at 30° C. at 150 rpm, and take a sample to detect the nicotine content of the solution after 48 hours. The result of the application is: the degradation rate of nicotine in the waste water of the tobacco factory is 89% after 48 hours at the dosage of 1% of the preparation, that is, without adding any other substances, the preparation of the present invention can make most of the nicotine in the waste water be degraded.

Embodiment 3

[0031] Embodiment 3: (for flue-cured tobacco leaf processing)

[0032] The preparation of the Pseudomonas fluorescens preparation is basically the same as in Example 1.

[0033] Application of the nicotine degradation preparation of the present invention: adopt the half-leaf test method, cut the tobacco leaves from the midrib, half of them are used for the bacterial agent treatment of the present invention, the preparation of the present invention is diluted 10 times with water, sprayed on the tobacco leaves, and the other half is sprayed Shi clear water was used as a control. The excess solution was naturally drained from the treated and control tobacco leaves, then loaded into a curing room, and cured according to a conventional curing method. The cured tobacco leaves are tested for nicotine, and the results show that the nicotine content of the nicotine degradation preparation of the present invention is reduced by 24% compared with that of the control.

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Abstract

The invention relates to a pseudomonas fluorescens preparation and application thereof, and belongs to the field of biotechnology. A preparation method of the pseudomonas fluorescens preparation comprises the following steps of 1, selecting pseudomonas fluorescens 1206 as a production bacterial strain, wherein the pseudomonas fluorescens 1206 has a preservation number of CGMCC NO.3149, 2, carrying out strain culture through inoculating a culture solution with the pseudomonas fluorescens 1206 subjected to inclined plane culture, wherein the culture solution comprises 3% of beef extract, 0.5% of peptone, 0.5% of sodium chloride and the balance distilled water, has a pH value of 7.0 to 7.2 and is put in a shaking bottle, and carrying out culture at a shaking bottle rotate speed of 150rpm at a temperature of 28 to 30 DEG C for 18 hours to obtain a liquid fermented strain, and 3, carrying out fermentation tank fermentation through inoculating a liquid fermentation system in a fermentation tank with the liquid fermented strain, wherein the liquid fermentation system comprises 35g / L of maltose, 4.03g / L of extract yeast, 4.72g / L of ammonium sulfate, 0.4g / L of K2HPO4, 0.075g / L of MgSO4.7H2O, 0.0005g / L of FeSO4.7H2O, 0.015g / L of CaCl2 and 0.1g / L of NaCl, and carrying out fermentation under the conditions of a fermentation temperature of 27 to 29 DEG C, an inoculation amount of 5 to 10%,a fermentation tank stirring speed of 120 to 150rpm, an aeration rate of 5 to 10% and fermentation time of 24 to 48 hours. The pseudomonas fluorescens preparation has a short fermentation period and a high nicotine degradation rate, can improve tobacco quality and can be utilized for degradation of nicotine of tobacco or tobacco waste.

Description

Technical field: [0001] The invention relates to a Pseudomonas fluorescens preparation capable of degrading nicotine and its application, belonging to the technical field of microorganisms. Background technique: [0002] Nicotine (1-methyl-2-(3-pyridyl)pyrrolidine) is an important plant alkaloid in tobacco, the content of which is about 0.17-4.93% of the dry weight of tobacco leaves. China is the largest tobacco producer and consumer in the world, accounting for about 30-35% of the world. The satisfaction of tobacco to the human body is mainly due to the effect of nicotine. An appropriate concentration of nicotine guarantees the smoking properties of tobacco. During the production process of cigarettes, excessive nicotine content will increase the irritation of smoke, affect the taste of cigarettes, and have poor safety. How to control the nicotine content of tobacco to the expected range is a hot topic of research by tobacco scientists and environmentalists. On the othe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A62D3/02A24B15/20C02F3/34C12R1/39A62D101/26
Inventor 夏振远雷丽萍吴玉萍莫笑晗
Owner YUNNAN ACAD OF TOBACCO AGRI SCI
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