Serratia nematodiphila and acquisition method and application thereof

A technology of Serratia and its acquisition method, which is applied in the field of bioengineering and can solve the problems of less research on environmental hormone microbial degradation characteristics and degradation mechanism, and a late start of research.

Active Publication Date: 2019-10-11
NORTHEAST NORMAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] Regarding the topic of removing estrogen in the environment by biological methods, 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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  • Serratia nematodiphila and acquisition method and application thereof
  • Serratia nematodiphila and acquisition method and application thereof
  • Serratia nematodiphila and acquisition method and application thereof

Examples

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Embodiment 1

[0029] This embodiment provides a method for obtaining Serratia nematophila DH-S01, specifically, comprising the following steps:

[0030] (1) Collect activated sludge and water samples from pharmaceutical factories and sewage treatment plants as the microbial source of Serratia nematophila DH-S01, and refrigerate them and transport them back to the laboratory.

[0031] (2) Take 15mg of estradiol and dissolve it in 30mL of methanol to make a concentration of 500mg / L of estradiol mother liquor, and take 1mL of estradiol mother liquor and add it to a Erlenmeyer flask filled with 99mL of liquid medium , so that the final concentration of estradiol in the liquid medium is 5mg / L; then, put the Erlenmeyer flask into a constant temperature water bath, and bathe in water at a temperature of 60°C for 30min; completely volatilize the methanol in the liquid medium; then, put Seal the mouth of the triangular flask with gauze and kraft paper, put it in an autoclave, and sterilize it at 121...

Embodiment 2

[0038] This example is to identify the Serratia nematophila DH-S01 strain obtained above. Specifically, DNA is amplified by PCR amplification technique using bacterial universal primers. The sequencing results were searched for homology comparison with the 16S rDNA gene sequence in the GenBank database using the BLAST tool in NCBI. The comparison results showed that the 16SrDNA gene sequence of Serratia nematodiphila DH-S01 had high homology with the gene sequence of Serratia nematodiphila. The 16S rDNA sequences of the 20 strains with higher scores in the BLAST results were selected to analyze the phylogenetic tree using MEGA software and Neighbor-Joining, and the unrooted phylogenetic tree (see attached image 3 ), combined with the morphological characteristics and physiological and biochemical indicators of Serratia nematophila strain DH-S01, DH-S01 was preliminarily identified as Serratia nematophila strain, and named Serratia nematophila DH-S01.

[0039] In addition, t...

Embodiment 3

[0041] This example is to verify the ability of Serratia nematophila DH-S01 to degrade estradiol. Specifically, first inoculate the strain into a liquid medium containing 50 mg / L estradiol and cultivate it for 24 hours; then, Pour the bacterial solution into a sterilized centrifuge tube, centrifuge at 4000rpm for 10min, discard the supernatant, and add a phosphate buffer solution with a pH of 7 to wash; then, place it in a centrifuge tube at 4000rpm for 15min , pour off the supernatant. After repeated washing in this way for 3 times, the bacterium was broken up with a vortex mixer to obtain a bacterium liquid; then, the bacterium liquid was diluted with a phosphate buffer solution to an OD600 value of 1.0, and the bacterium suspension was prepared for use. Then, take 200 μL of the bacterial liquid from the bacterial suspension and inoculate it into a liquid medium with estradiol as the sole carbon source, and the concentration of estradiol is 50 mg / L, and continuously in a con...

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Abstract

The invention discloses a serratia nematodiphila and an acquisition method and application thereof, and belongs to the technical field of bioengineering. The serratia nematodiphila is named as serratia nematophila DH-S01 and deposited in the China center for type culture collection on January 14, 2019, and the preservation number is CCTCCM2019038. The screened serratia nematophila DH-S01 has a strong degradation ability to estradiol. When the serratia nematici DH-S01 is cultured in a medium with estradiol as the sole carbon source for 7 days, the estradiol with the concentration of 100 mg/L can be rapidly degraded, and the degradation rate is 99% or above. Therefore, the serratia nematici DH-S01 can degrade estradiol stably and efficiently, and can be applied to the biodegradation and environmental remediation of estradiol in an environment.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to Serratia nematophila and its acquisition method and 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, and have become an urgent problem of environmental pollution worldwide, which 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 disrupt the normal synthesis and metabolism of endocrine hormones, affect growth and sex differentiation, but also cause developmental abnormalities and reproductive disorders in multiple systems such as nerves, endocrine, and immunity, and even cause teratogenicity and carcinogenesis. [0003] Environmental hormones can transfer from low...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N1/02C02F3/34C12R1/425C02F101/34
CPCC12N1/20C12N1/02C02F3/34C02F2101/34C02F2101/345C12R2001/425C12N1/205
Inventor 霍洪亮周东文赵雪莹田克俭孟琪李雪王乐
Owner NORTHEAST NORMAL UNIVERSITY
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