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Nornicotine degrading bacteria and microbial agent produced by same

A technology of methyl nicotine and degrading bacteria, which is applied in the high biological field, can solve problems affecting drug efficacy and toxicity, achieve low production and use costs, protect the ecological environment, and have good removal effects

Active Publication Date: 2019-08-16
镇江蔚绿生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Nornicotine can directly induce abnormal glycosylation of proteins in the plasma of smokers, and can covalently react with commonly used steroids, affecting drug efficacy and toxicity

Method used

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  • Nornicotine degrading bacteria and microbial agent produced by same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] 1.1 Isolation of demethylnicotine degrading bacteria W1-1 (CCTCC NO.M 2018758)

[0028] The soil sample used to enrich the demethylated nicotine-degrading strain was taken from the soil of a certain tobacco plantation site. Take 3.0g of soil and add 150ml to 500mg·L -1 In the basal salt medium with demethylnicotine as the sole carbon source, 30°C, 170r·min -1 Cultivate for 3 days, transfer to the same medium with 4% inoculum size, and serially dilute the enrichment solution after transferring 4 times, take 10 -5 ~10 -8 Each 0.1mL of the diluted enrichment solution was applied to add 500mg·L -1 On the solid medium plate of demethylnicotine, after culturing at 30°C for 2 days, pick the grown single colony and inoculate it in a medium containing 500mg·L -1 Demethylnicotine inorganic salt medium, 30°C, 170r·min -1 Incubate on a shaking table for 2 days to detect its degradation effect.

[0029] The basic salt medium formula is (1L): 1.0g NH 4 NO 3 , 1.0g NaCl, 1.5g K...

Embodiment 2

[0036] Activate the original seed of the microorganism W1-1 of the present invention on a petri dish, measure its degradation performance, and inoculate it on the inclined surface of a test tube for later use. The test tube seeds were inoculated into 1000ml shake flasks containing 200ml broth medium, cultured with constant temperature shaking until logarithmic phase, and prepared to inoculate seed tanks. 1 ton of seed tank, 700 liters of feeding capacity, medium formula is: glucose 1.5%, (NH4 2 ) SO 4 1.5%, K 2 HPO 4 0.4%, MgSO 4 0.06%, NaCl 1.5%, CaCO 3 0.1%, yeast extract 0.04%, pH 7.2-7.5. After feeding, sterilize under high pressure at 115°C, cool to 33°C, inoculate 5% of the cultured shake flask strains into a 1 ton liter seed tank, and cultivate to the logarithmic growth phase with a stirring speed of 200°C. RPM, sterile air intake rate is 1:0.8. The seed solution that has reached the logarithmic phase is inserted into the production tank according to the inoc...

Embodiment 3

[0038] Inoculate the tobacco processing enterprise waste water (the concentration of demethylnicotine in the waste water has fluctuations, and the average amount is at 400mg L with the inoculation amount prepared by Example 2) -1 ) biochemical treatment pool, carry out aeration, and take samples to measure the removal rate of demethylnicotine in the wastewater at different times after the addition. As can be seen from Table 1, the use of demethylnicotine degradation bacteria can make formazan in the wastewater The removal rate of nicotine is maintained above 95%, and the invention has great significance for protecting the ecological environment and people's health.

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Abstract

The invention discloses nornicotine degrading bacteria and a microbial agent produced by the same. The nornicotine degrading bacteria is a gram-positive Pseudarthrobacter sp. strain W1-1 collected ina China Center for Type Culture Collection, and the collection number of the strain is CCTCC NO. M 2018758. The strain W1-1 can degrade nornicotine, the nornicotine degrading microbial agent producedby the strain has the advantages of low production and use cost, convenience in use and good removing effects and is suitable for being used in a biochemical treatment tank for tobacco processing enterprise wastewater. The starting time of a biochemical treatment system can be shortened, nornicotine degrading effects are improved, nornicotine removing rate is kept above 95%, and the nornicotine degrading bacteria have an excellent application prospect and great significance for protecting ecological environments and physical health of people.

Description

technical field [0001] The invention provides a demethyl nicotine degrading bacterium and a bacterial agent produced therefrom, which belong to the high-tech field of biology and use microorganisms to degrade demethyl nicotine, and are suitable for wastewater treatment of tobacco processing enterprises. Background technique [0002] Nicotine and demethylnicotine are the main alkaloids in plants of the Nicotiana genus. Nicotine accounts for 95% of the alkaloid content of most commercial tobacco varieties, while its structural analogue, nornicotine, accounts for only about 3%. But once the tobacco leaves are aged, the content of demethylated nicotine will increase significantly. Studies have shown that during tobacco aging and curing processing, a large amount of nicotine in tobacco leaves will be converted to demethyl nicotine. [0003] Nornicotine can directly induce abnormal glycosylation of proteins in the plasma of smokers, and can covalently react with commonly used st...

Claims

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Application Information

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IPC IPC(8): C12N1/20C02F3/34C02F101/38C12R1/01
CPCC12N1/20C02F3/34C02F2101/38C12R2001/01C12N1/205
Inventor 张磊洪青
Owner 镇江蔚绿生物科技有限公司
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