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Strain for degrading phenol compounds and application of strain

A technology for phenolic compounds and strains, applied in the field of strains that efficiently degrade phenolic compounds, can solve the problems of high toxicity and less degradation of phenol derivatives, and achieve the effect of enriching diversity

Active Publication Date: 2016-08-31
NANJING UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to provide a kind of efficient and broad-spectrum degrading microorganisms for phenol and its derivatives in view of the problem that industrial phenol-containing wastewater has high toxicity to organisms, but the amount of degradation of phenol derivatives by existing microorganisms is small

Method used

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  • Strain for degrading phenol compounds and application of strain
  • Strain for degrading phenol compounds and application of strain
  • Strain for degrading phenol compounds and application of strain

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Isolation and screening of phenol-degrading strain Sphingomonas paucimobilis DL-10:

[0046] Take 5mL sample of industrial wastewater containing phenol and place it in 100mL inorganic salt culture medium containing 50mg / L phenol, at 30℃, 180r·min -1 Cultivate for 2-3d. Use an ultraviolet scanner to measure the degradation of phenol in the enriched solution. After confirming that the phenol is degraded, insert 10% of the inoculum into the medium containing 75mg / L phenol inorganic salt, continue to enrich and determine the degradation, click here Method until the phenol concentration is increased to 200mg / L, and passage 3 times.

[0047] The phenol enrichment solution was gradually diluted and spread on an inorganic salt medium plate with 100mg / L phenol as the sole carbon source, and cultured in an incubator at 30°C for 2-3 days. The single colonies with different colonies grown on the plate were streaked on a 1 / 3LB plate for purification, and inoculated in an inorganic salt ...

Embodiment 2

[0052] Identification of phenol-degrading strain Sphingomonas paucimobilis DL-10 and its growth characteristics:

[0053] Identification of DL-10:

[0054] Perform 16S rDNA identification on DL-10: use primer 27F: 5`-AGAGTTTGATCCTGGCTCAG-3` and 1492R: 5`-TACCTTGTTACGACTT-3` to amplify the 16S rDNA of strain DL-10, and connect to the clone by T / A cloning Vector pMD19T, construct recombinant cloning vector pMD19T-16S, transform it into the cloning host Escherichcoli DH5α to obtain recombinant microorganism Escherich coli DH5α (pMD19T-16S), sequence the obtained recombinant microorganism foreign fragments, NCBI database compares the 16S rDNA sequence, the strain DL-10 was identified to Sphingomonas at the molecular level, and the nucleotide sequence of its 16S rDNA is shown in SEQ ID NO:1 in the sequence table.

[0055] Growth characteristics of DL-10:

[0056] Sphingomonas paucimobilis DL-10 grows slowly on LB plates. It can form yellow colonies with a diameter of 2mm at 30°C for 96 ho...

Embodiment 3

[0059] Degradation characteristics of phenol degrading bacteria Sphingomonas paucimobilis DL-10:

[0060] Pick single colonies of strain DL-10 from the 1 / 3LB plate and inoculate them in 3mL 1 / 3LB liquid medium at 30℃, shaker 180r·min -1 Cultivate for 48h. Then the culture solution of the strain was transferred to 20 mL of fresh liquid 1 / 3 LB medium, and the culture was continued for 18 h. 6000r·min -1 Centrifuge for 10 min, collect the bacteria, wash once with sterilized inorganic salt medium, and make OD 600nm = 1.0 bacterial suspension, namely seed liquid.

[0061] In an inorganic salt medium with a final concentration of phenol of 100 mg / L, insert OD at 5% inoculum 600nm =1.0 strain DL-10 seed liquid, at 30℃, 180r·min -1 Shake culture, take samples every 2h, measure OD 600nm And the concentration of phenol.

[0062] by Figure 4 It can be seen that the strain DL-10 was inoculated into the culture medium for 2 hours, and then it began to degrade phenol without obvious delay; after...

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Abstract

The invention relates to a strain for degrading phenol compounds and application of the strain. The strain is classified and named as Sphingomonas paucimobilis DL-10 and has been preserved in China Center for Type Culture Collection on March 3, 2014, with a preservation number of CCTCC NO: M 2014058. A 16S rDNA nucleotide sequence of the strain is as shown in SEQ ID NO: 1 in a sequence list. The strain DL-10 is an aerobic microorganism, wherein the optimal growth temperature is 30 DEG C, the optimal growth pH is 6.0, and the growth is obviously limited when the NaCl concentration is higher than 3%. By virtue of the strain DL-10, 100mg / L phenol can be completely degraded within 12h, and by taking the strain DL-10 as a unique carbon source for growth, the phenol compounds such as catechol, hydroquinone, o-methylphenol, m-methylphenol, 2,6-dimethylphenol, 3,5-dimethylphenol, 2,6-dimethyl hydroquinone can be completely degraded. The strain has important application value in the biological treatment of the phenol compounds in industrial wastewater.

Description

Technical field [0001] The invention belongs to the technical field of biological treatment, and specifically relates to a strain capable of efficiently degrading phenol compounds and its application. Background technique [0002] Phenol-containing wastewater has a wide range of sources and complex components, such as pharmaceutical wastewater, coking wastewater, chemical industry, printing and dyeing wastewater, etc. The high concentration of phenolic compounds contained in wastewater can inhibit the growth of microorganisms, thereby increasing the difficulty of biological treatment and becoming a reality. A difficult problem to be solved urgently in the field of phenol-containing wastewater treatment technology at home and abroad. [0003] Biodegradation is an important transformation process of pesticides in the environment. Microorganisms exist in nature in large numbers, and have the characteristics of a wide range of species, rapid reproduction and strong adaptability to the...

Claims

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Application Information

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IPC IPC(8): C12N1/20C02F3/34C12R1/01C02F101/34
CPCC12N1/20C02F3/34C02F2101/345C12R2001/01C12N1/205
Inventor 董维亮马江锋章文明信丰学姜岷
Owner NANJING UNIV OF TECH
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