Para-chloro nitrobenzene degrading testosterone coma monad and its use

A technology of Comamonas, chlorinated nitrobenzene, applied in the field of microbial biotechnology and environmental biology

Active Publication Date: 2005-05-18
INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, apart from individual reports from abroad, little is known about the degradation ability and degradation properties of single or mixed strains of chloronitrobenzene

Method used

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  • Para-chloro nitrobenzene degrading testosterone coma monad and its use
  • Para-chloro nitrobenzene degrading testosterone coma monad and its use
  • Para-chloro nitrobenzene degrading testosterone coma monad and its use

Examples

Experimental program
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Effect test

Embodiment 1

[0016] Example 1: Culture and biological characteristics of Comamonas testosteroni CNB1 CGMCC No.1028.

[0017] Comamonas testosteroni CNB1 CGMCC No.1028 was inoculated in inorganic salt medium, the medium composition: disodium hydrogen phosphate 1g, potassium dihydrogen phosphate 0.5g, MgSO 4 ·7H 2 O 0.03g, trace element solution 5mL, p-chloronitrobenzene 200mg, distilled water 1000mL, pH7.0, add 15g / L agar to the solid medium. Sterilize at 121°C for 30 minutes. Shake the culture on a shaker at 28-30°C and 130rpm. The bacterium has the following characteristics: (1) The size of the colony cultivated on the LB plate for 2 days is 1-2 mm in diameter, and the colony is round, with a smooth surface, neat edges, raised, and colorless. (2) Cell morphology characteristics of Comamonas testosteroni (Comamonastestosteroni) CNB1 CGMCC No.1028: suitable for normal temperature culture in the range of pH 6-11, the cells are short rod-shaped, with 2 extreme hairs, Gram staining negative...

Embodiment 2

[0024] Embodiment 2: PCR amplification and sequence determination of the 16S rRNA gene of Comamonas testosteroni CNB1 CGMCC No.1028

[0025] Comamonas testosteroni CNB1 CGMCC No.1028 was inoculated on the LB plate, directly picked a ring of bacteria from the plate, added 100 μL sterile redistilled water, vortex mixed, boiled water bath for 2 minutes, centrifuged at 12000r / min for 5 minutes, and the supernatant was directly for PCR. The primers used for the PCR reaction of 16SrDNA amplification are a pair of universal primers. Forward primer Pf: 5'-AGAGTTTGATCCTGGCTCAG-3'; reverse primer Pr: 5'-ACGGCTACCTTGTTACGACT-3', corresponding to 8-27 and 1495-1514 bases of the 16S rRNA gene of Escherichia coli, respectively. The PCR reaction system (50μL) is: 10×buffer 5μL, 25mmol / L MgCl 2 4 μL, 10 mmol / L dNTPs 1 μL, 30 pmol / L primers 1 μL, BSA 0.5 μL, ddH 2 0.37 μL, TaqDNase 0.4 μL. The PCR reaction conditions were: 95°C for 4min, 95°C for 1min, 48°C for 1min, 72°C for 1min, 30 cyc...

Embodiment 3

[0026] Embodiment 3: Comamonas testosteroni CNB1 CGMCC No.1028 provided by the present invention is to the degradation of p-chloronitrobenzene

[0027] Comamonas testosteroni CNB1CGMCCNo.1028 strain culture fluid (OD 600 =0.2) Inoculate in the culture medium that contains p-chloronitrobenzene (20mg / L) by 1% of the inoculation amount, at 30 DEG C, shake culture on the shaker of 130r / min, regularly take samples to measure thalline growth, p-chlor Nitrobenzene content and chloride ion concentration, the results are as follows figure 1 shown. figure 1 It shows that the growth of CNB1 bacteria and the degradation of p-chloronitrobenzene have a short lag period, and then enter the logarithmic phase, and reach the highest growth amount in the 20th hour, and the degradation rate reaches 87% at this time, and the p-chloronitrobenzene The decrease in moles of benzene is comparable to the increase in moles of chloride ions.

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Abstract

The present invention relates to one Comamonas testosteroni CNB1CGMCC No. 1028 strain, whose growing cell, cell suspension and immobile cell can degrade para-chloronitrobenzene. The strain can grow with para-chloronitrobenzene as unique carbon source, nitrogen source and energy source, so as to degrade and utilize para-chloronitrobenzene completely. During its growth, the strain first reduces nitro group into hydroxylamine, and produces 2-aminophenol 1, 6-bioxygenase to open the ring of 2-amino-5-chlorophenol into hydrocarbon through meta ring opening. The strain is suitable for the biological treatment of industrial effluent from para-chloronitrobenzene production and the biological repair of para-chloronitrobenzene polluted soil.

Description

technical field [0001] The invention belongs to the field of microbial biotechnology and environmental biotechnology, in particular, it relates to a Comamonas testosteroni CNB1 CGMCC No.1028 bacterial strain, growth cells, cell suspensions, immobilized cells of the bacterial strain The bacteria can degrade p-chloronitrobenzene; the strain can grow with p-chloronitrobenzene as the only carbon source, nitrogen source and energy source, and completely degrade and utilize p-chloronitrobenzene. When the strain grows with p-chloronitrobenzene as the only carbon source, nitrogen source and energy source, it first reduces the nitro group to hydroxylamine, and then under the action of 2-aminophenol 1,6-dioxygenase produced by the strain , through meta ring opening, 2-amino-5-chlorophenol is ring-opened into hydrocarbons, which can be further degraded and utilized. The bacterial strain is suitable for biological treatment of industrial wastewater producing p-chloronitrobenzene and bior...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20
Inventor 刘双江吴建峰刘志培王保军姜成英沈锡辉
Owner INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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