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A bacterial strain for degrading phenolic compounds and its application

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: 2019-08-30
NANJING TECH UNIV
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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

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  • A bacterial strain for degrading phenolic compounds and its application
  • A bacterial strain for degrading phenolic compounds and its application
  • A bacterial strain for degrading phenolic compounds and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

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

[0046] Take 5mL of phenol-containing industrial wastewater samples and place them in 100mL of inorganic salt medium containing 50mg / L phenol, at 30℃, 180r min -1 Culture 2-3d. Use an ultraviolet scanner to measure the degradation of phenol in the enrichment solution. After confirming that the phenol is degraded, insert it into the medium containing 75mg / L phenol inorganic salt with a 10% inoculum size, continue to enrich and measure the degradation, press here Methods until the phenol concentration increased to 200mg / L, and passaged for 3 times.

[0047] The phenol-enriched solution was serially diluted and spread on an inorganic salt medium plate with 100mg / L phenol as the sole carbon source, and cultured in a 30°C incubator for 2-3 days. Streak the single colonies with different colonies grown on the plate on 1 / 3LB plates for purification, and inoculate them in the inorganic salt m...

Embodiment 2

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

[0053] Identification of DL-10:

[0054] 16S rDNA identification of DL-10: Use primers 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 the recombinant cloning vector pMD19T-16S, transform it into the cloning host strain Escherich coli DH5α to obtain the recombinant microorganism Escherich coli DH5α (pMD19T-16S), sequence the exogenous fragment of the recombinant microorganism obtained, and compare the 16S with the NCBI database 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, and can form yellow colonies ...

Embodiment 3

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

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

[0061] In the inorganic salt medium with a final concentration of phenol of 100mg / L, insert OD at 5% inoculum 600nm =1.0 Strain DL-10 seed liquid, at 30°C, 180r·min -1 Shake the culture, take a sample every 2 hours, and measure the OD 600nm and phenol concentrations.

[0062] Depend on Figure 4 It can be seen that the strain DL-10 began to degrade phenol within 2 hours after being inoculated in the medi...

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Abstract

The invention relates to a bacterial strain for degrading phenolic compounds and its application. The bacterial strain is classified as Sphingomonas paucimobilis DL‑10 was deposited in China Center for Type Culture Collection CCTCC on March 3, 2014, and the deposit number is: CCTCC NO: M 2014058. The nucleotide sequence of its 16S rDNA is shown in SEQ ID NO: 1 in the sequence listing. Strain DL‑10 is an aerobic microorganism with an optimum growth temperature of 30°C and an optimum growth pH of 6.0. When the NaCl concentration is higher than 3%, its growth is significantly inhibited. Strain DL‑10 can completely degrade 100 mg / L phenol within 12 h, and use it as the sole carbon source for growth, and can completely degrade catechol, hydroquinone, o-cresol, m-cresol, 2,6-dimethylphenol, 3,5-dimethylphenol, 2,6-dimethylhydroquinone and other phenolic compounds are degraded. The invention has important application value for biological treatment of phenolic compounds in industrial waste water.

Description

technical field [0001] The invention belongs to the technical field of biological treatment, and in particular relates to a bacterial strain for efficiently degrading phenolic compounds and an application thereof. Background technique [0002] Phenol-containing wastewater has a wide range of sources and complex components, such as pharmaceutical wastewater, coking wastewater, chemical industry and printing and dyeing wastewater, etc. The high concentration of phenolic compounds contained in wastewater will inhibit the growth of microorganisms, thus increasing the difficulty of biological treatment At this stage, it is a difficult problem to be solved urgently in the field of phenolic wastewater treatment technology at home and abroad. [0003] Biodegradation is an important transformation process of pesticides in the environment. Microorganisms play an important role in the biodegradation of pesticides because they exist in large numbers in nature and have the characteristi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C02F3/34C12R1/01C02F101/34
CPCC02F3/34C02F2101/345C12N1/20C12N1/205C12R2001/01
Inventor 董维亮马江锋章文明信丰学姜岷
Owner NANJING TECH UNIV
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