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Rhodococcus DSH capable of degrading environmental hormone bisphenol A, and applications thereof

A technology of environmental hormones and Rhodococcus, applied in the field of bioengineering, can solve the problems of late start of research, less research on the microbial degradation characteristics and degradation mechanism of environmental hormones, and achieve the effect of strong degradation ability

Inactive Publication Date: 2017-10-03
NORTHEAST NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

With regard to the topic of using biological methods to remove estrogen in the environment, domestic research started relatively late, and researchers' research on environmental hormones also mainly focused on the toxicology research of estrogen, the exploration of detection methods, and the risk assessment of estrogen pollution. , there are few studies on the microbial degradation characteristics and degradation mechanism of environmental hormones, and extensive and in-depth research is urgently needed

Method used

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  • Rhodococcus DSH capable of degrading environmental hormone bisphenol A, and applications thereof
  • Rhodococcus DSH capable of degrading environmental hormone bisphenol A, and applications thereof
  • Rhodococcus DSH capable of degrading environmental hormone bisphenol A, and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1 Acquisition and preservation of Rhodococcus sp. DSH strain

[0031] Collect mud and water samples from pharmaceutical factories and sewage treatment plants as microbial sources, and transport them back to the laboratory in cold storage. And the sample is cultivated with the only carbon source. Dissolve 15 mg of bisphenol A in 30 mL of methanol to prepare a bisphenol A mother liquor with a concentration of 500 mg / L. Take 1 mL of bisphenol A mother liquor and add it to an Erlenmeyer flask containing 100 mL of inorganic salt medium to make the final concentration of bisphenol A in the inorganic salt medium 5 mg / L. Composition of inorganic salt medium (g / L): Na 2 HPO 4 4.260; KH 2 PO 4 2.650; MgSO 4 ·7H 2 O0.200; (NH 4 ) 2 SO 4 1.500; CaCl 2 0.020. Adjust the pH to 7.0 with 0.1 mol / L NaOH and 0.1 mol / L HCl, and add 1 mL of trace elements. Trace element composition (g / L): NiCl 2 ·6H 2 O 0.024; CoCl 2 ·6H 2 00.190; H 3 BO 3 0.006; ZnCl 2 0.070; CuCl 2 ·2H 2 O 0.002;...

Embodiment 2

[0032] Example 2 Identification of Rhodococcus sp. DSH strain

[0033] The DNA is amplified by PCR amplification technology using bacterial universal primers. The sequencing results were searched for homology comparison between the BLAST tool in NCBI and the 16S rDNA gene sequence in GenBank database. The comparison results showed that the 16S rDNA gene sequence of Rhodococcus DSH has high homology with the 16S rDNA gene sequences of Rhodococcus and Rhodococcus equi. Select the 16S rDNA gene sequence of 18 strains of Rhodococcus, and select the 16S rDNA gene sequence of 2 strains of Mycobacterium as the outgroup, use MEGA software and perform phylogenetic tree analysis according to Neighbor-Joining, The constructed rootless phylogenetic tree, combined with the morphological characteristics and physiological and biochemical indicators of Rhodococcus DSH, preliminarily identified the strain DSH as Rhodococcus and named it Rhodococcus sp. DSH.

Embodiment 3

[0034] Example 3 Rhodococcus sp. DSH's ability to degrade bisphenol A

[0035] The strain was inoculated into an inorganic salt medium containing 50 mg / L bisphenol A and cultured for 24 hours. Pour the bacterial solution into a sterile centrifuge tube, centrifuge at 4000 rpm for 10 min, discard the supernatant, add a pH 7 phosphate buffer solution to wash, and centrifuge at 4000 rpm for 15 min, and pour the supernatant. Repeat the washing 3 times, use a vortex mixer to disperse the bacteria, and finally dilute the bacteria to OD with phosphate buffer solution 600 The value is 1.0 and is prepared as a bacterial suspension for later use. Take out 200μL of bacterial suspension from each strain of bacteria and inoculate it in an inorganic salt medium with bisphenol A as the sole carbon source and a concentration of 50mg / L. In a constant temperature shaking incubator at 30°C and 120rpm, Cultivation was continued for 7 days in a constant temperature shaking incubator at 120 rpm and a ...

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Abstract

The invention discloses Rhodococcus DSH capable of degrading environmental hormone bisphenol A, and applications thereof, wherein the Rhodococcus DSH belongs to Rhodococcus sp, is named as Rhodococcus DSH, has the preservation number of CGMCC NO. 12392, and is preserved in the China General Microbiological Culture Collection Center, wherein the address is Institute of Microbiology, Chinese Academy of Sciences, 3# Court No.1, Beichen West Road, Chaoyang District, Beijing, and the sequence of the 16S rDNA fragment of the Rhodococcus DSH is represented by SEQ ID NO:1 in the sequence table. According to the present invention, the active sludge of the bisphenol A production drug factory is subjected to enrichment acclimation, purification separation and re-screening to obtain the Rhodococcus DSH, wherein the Rhodococcus DSH can stably and efficiently degrade bisphenol A, and can be used for the bisphenol A biodegradation in the environment, and the environmental remediation.

Description

Technical field [0001] The invention relates to the technical field of biological engineering, in particular to a Rhodococcus DSH capable of degrading environmental hormone bisphenol A and its application. Background technique [0002] In the past half century, environmental hormones have caused serious harm to the natural environment on which humans and wild animals depend for survival, and have become a common environmental pollution problem that needs to be solved urgently on a global scale, and has attracted widespread attention. As a common endocrine disruptor, environmental hormones are similar in structure and function to estrogen secreted by the body under normal conditions. Once it enters the body, it can not only disturb the body's normal synthesis and metabolism of endocrine hormones, affect growth and gender differentiation, but also cause neurological, endocrine, immune and other multi-system development abnormalities and reproductive disorders, and even cause terato...

Claims

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Application Information

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IPC IPC(8): C12N1/20C02F3/34A62D3/02A62D101/28C02F101/34C12R1/01
CPCA62D3/02C02F3/34C02F2101/345C12N1/20C12N1/205C12R2001/01
Inventor 霍洪亮于清淼王平朱遂一杨武邵缓缓马喆
Owner NORTHEAST NORMAL UNIVERSITY
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