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Klebsiella sp. strain, pyridine biodegradation agent as well as preparation method and application of pyridine biodegradation agent

A technology of Klebsiella and Lebsiella strains, applied in the biological field, can solve the problems of high cost of pyridine, insignificant effect, and salt-tolerant pyridine-degrading bacteria, and achieve good application prospects

Active Publication Date: 2019-03-08
HUNAN CHEM RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In recent years, there are a lot of literatures on pyridine synthesis, but there are few reports on pyridine degradation process. Yu Fengwen et al. used biodiesel as extraction agent to extract and recover pyridine in wastewater, but this technology is expensive and incomplete. , easy to cause secondary pollution
Zhang Liangbo et al. used ultrasonic / Fenton reagents to degrade pyridine in wastewater, but it was difficult to treat the wastewater to the concentration of pyridine in the common biochemical system, and it needed to be diluted in a large proportion before entering the biochemical treatment device.
Qiaolin et al. introduced pyridine-degrading bacteria Paracoccus sp.KT-5 into the sequencing batch reactor of pyridine wastewater to accelerate the start-up of the reactor, but the late effect was not obvious, and the activity of the bacteria was not strong.
In summary, although the research on pyridine removal has been carried out at home and abroad, the physical and chemical methods involved in the removal of pyridine are costly and complicated, and the screened pyridine-degrading bacteria are not resistant to salt and have poor environmental tolerance, making it difficult for practical application

Method used

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  • Klebsiella sp. strain, pyridine biodegradation agent as well as preparation method and application of pyridine biodegradation agent
  • Klebsiella sp. strain, pyridine biodegradation agent as well as preparation method and application of pyridine biodegradation agent
  • Klebsiella sp. strain, pyridine biodegradation agent as well as preparation method and application of pyridine biodegradation agent

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] A high-efficiency pyridine-degrading strain, which is classified as Klebsiella sp. BD2, was deposited in the China Center for Type Culture Collection on October 17, 2018, and the preservation number is CCTCC NO: M 2018685 ; The depository address is Wuhan University, Hubei Province, China.

[0043] The bacterial strain BD2 of the present invention is obtained by screening and separating water samples taken from a wastewater treatment pool of a pesticide and chemical enterprise in Changsha.

[0044] The specific screening methods are:

[0045] (1) Collect 3 samples of water from the wastewater pool of a pesticide chemical factory in Changsha, each sample is about 500ml, and bring it back to the laboratory after marking.

[0046] (2) Get 1ml of the supernatant of the water sample in the wastewater treatment pond, dilute it with concentration gradient dilution method, get 10 -5 、10 -6 、10 -7 1ml of the dilution solution of the three gradients is evenly spread on the py...

Embodiment 2

[0051] A kind of pyridine biodegradation bacterial agent, its preparation method is:

[0052] (1) Take out the Klebsiella BD2 strain stored at -20°C LB solid slant or -80°C ultra-low temperature 70% glycerol cryopreservation, place it at room temperature, then inoculate it into 50 mL of LB liquid medium and culture it on a shaking table for 24 hours, shake The bed speed is 180r / min to complete the activation of the strain.

[0053] (2) Take 2 ml of the activated bacterial liquid into 100 ml of inorganic salt culture medium with a pyridine concentration of 100 mg / L and a pH of 7.5, and culture it for 24 hours at a shaker speed of 180 r / min at 30°C to obtain a bacterial liquid; Figure 5 In the process of cultivating for 32h, the growth curve of the strain is obtained from Figure 5 It can be seen that the growth of the strain reached the maximum at 24h.

[0054] (3) Take 2ml of the bacterial solution in step (2) and put it in 100ml of inorganic salt culture medium with a pyri...

Embodiment 3

[0056] To investigate the salt tolerance of pyridine biodegradation bacteria agent. Get the pyridine biodegradation bacterial agent of 1ml embodiment 2 and add to salinity and be 0% (W / V), 0.5% (W / V), 1% (W / V), 1.5% (W / V), 2% (W / V), 2.5% (W / V) MS medium, shaker speed 180r / min, pH7.0, pyridine concentration 100mg / L, culture 24h under the condition of 30 ℃, ultraviolet spectrophotometer (600nm) The growth of the strain was measured, and the content of pyridine was determined by high performance liquid chromatography.

[0057] See results Image 6 ,from Image 6 As can be seen in the figure, when the salinity is between 0% and 1.5%, the growth amount of the bacterial strain BD2 changes little, and when the salinity is greater than 1.5%, the growth amount drops significantly, and the growth of the bacterial strain is inhibited. Has a certain salt tolerance.

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Abstract

The invention discloses a Klebsiella sp. strain, a pyridine biodegradation agent as well as a preparation method and an application of the pyridine biodegradation agent. The Klebsiella sp. strain is Klebsiella sp. BD2, is preserved in China Center for Type Culture Collection with preservation number of CCTCC NO: M 2018685. The pyridine biodegradation agent is prepared from the Klebsiella sp. strain by activation and culture. The Klebsiella sp. BD2 pyridine has high degradation efficiency, good salt tolerance and good environmental tolerance, and the pyridine biodegradation agent prepared fromKlebsiella sp. BD2 can be applied to degradation of pyridine in wastewater and soil, and has high degradation efficiency.

Description

technical field [0001] The invention relates to the field of biological technology, in particular to a Klebsiella strain, a pyridine biodegradation bacterial agent and a preparation method and application of the pyridine biodegradable bacterial agent. Background technique [0002] Pyridine is a six-membered heterocyclic compound containing a nitrogen heteroatom, molecular formula C 5 h 5 N, colorless or yellowish liquid with foul odor, soluble in water, alcohol, ether and most other organic solvents, highly irritating, its vapor can form explosive mixture with air, it is easy to burn and explode in case of open flame or high heat. Pyridine and its homologues exist in bone tar, coal tar, gas, shale oil, and petroleum. In addition to being used as solvents in industry, they can also be used as denaturants, dyeing aids, and synthetic drugs, disinfectants, dyes, The starting material of food seasonings, adhesives, explosives, etc., as a broad-spectrum, high-efficiency insectic...

Claims

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Application Information

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IPC IPC(8): C12N1/20C02F3/34B09C1/10C12R1/22C02F101/38
CPCB09C1/10C02F3/34C02F2101/38C12N1/20C12N1/205C12R2001/22
Inventor 张海涛邱德跃杨海君欧晓明杜升华陈灿秦岳军刘欢钟若楠张燕干兴利马林
Owner HUNAN CHEM RES INST
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