Paenibacillus sp. PN-S435 and application thereof in microcystins (MCs) degradation

A technology of microcystin and bacillus, applied in the field of environmental microorganisms, can solve the problems of incomplete degradation and increased difficulty, achieve great social significance and economic value, avoid serious threats, and have good genetic stability.

Inactive Publication Date: 2014-06-25
NANJING TECH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are microorganisms capable of degrading MCs in both natural water bodies and sediments, and some special microbial strains capable of degrading MCs have been reported, such as pure strains of Ralstonia solanacearum (Ralstonia solanacearum), acid-loving Dell Fordella (Delftia acidovorans), Pseudomonas aeruginosa (Pseudomonas aerugiosa) and Sphingomonas (Sphingomonas), although Some of these strains can degrade algal toxins as high as 50mg / L.Day, but these strains do not degrade MCs completely, and will produce other intermediate degradation products, and after laboratory experiments, these strains cannot use MCs as the only one C source and N source increase the difficulty of applying to the actual environment

Method used

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  • Paenibacillus sp. PN-S435 and application thereof in microcystins (MCs) degradation
  • Paenibacillus sp. PN-S435 and application thereof in microcystins (MCs) degradation
  • Paenibacillus sp. PN-S435 and application thereof in microcystins (MCs) degradation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] This example illustrates the mutagenesis screening method for Paenibacillus PN-S435.

[0052] The medium used in the mutagenesis process is as follows:

[0053] Plate / slant medium: glucose 0.5%, beef extract 0.5%, tryptone 0.5%, yeast extract 0.5%, MgSO 4 0.05%, NaCl 0.2%, K 2 HPO 4 0.02%, agar 1.2%, the rest is distilled water, pH 7.4.

[0054] Lithium chloride plate medium: glucose 0.5%, beef extract 0.5%, tryptone 0.5%, yeast extract 0.5%, MgSO 4 0.05%, NaCl 0.2%, K 2 HPO 4 0.02%, agar 1.2%, lithium chloride 1.0moL·L -1 , and the rest is distilled water, pH 7.4.

[0055] MCs medium: MCs 1.8%, MgSO 4 0.05%, NaCl 0.2%, K 2 HPO 4 0.02%, agar 1.2%, the rest is distilled water, pH 7.4.

[0056] Microcystis indoor culture medium: Inoculate Microcystis aeruginosa species on BG-11 medium with an inoculation amount of 5-15% (v / v), and cultivate in a light incubator for ten days at a temperature of 28-33°C. The light intensity is 4000lx, the light-dark ratio ...

Embodiment 2

[0070] This example illustrates the identification and genetic stability of bacterial strains

[0071] The main morphology and biological characteristics of the bacterial strain Paenibacillus PN-S435 of the present invention are as follows: the cells are rod-shaped, shorter and thicker, facultative anaerobic Gram-positive bacteria, without flagella, and round spores are arranged in the cyst. On the beef extract peptone medium, the color of the colony is light yellow, the size of the colony is 1-2.5mm, the colony is round, the colony is thick, moist, the edge is neat, the surface is smooth, the texture is uniform, easy to provoke, the size of the colony is 1-2.5mm, The optimum temperature for growth: 25°C~28°C, the temperature range for growth is 15-50°C, the optimum pH for growth: 7.2~7.5, and the pH range for growth is 5.6~10.0.

[0072]This embodiment illustrates the mutant Paenibacillus obtained by screening ( Paenibacillus sp. ) Genetic stability of PN-S435. Passage deg...

Embodiment 3

[0077] This example illustrates the cultivation method of the mutant Paenibacillus PN-S435.

[0078] The medium formula described in this embodiment (% is mass percent):

[0079] Plate / slant medium: 0.5% glucose, 0.5% beef extract, 0.5% tryptone, 0.5% yeast extract, MgSO 4 0.05%, NaCl 0.2%, K 2 HPO 4 0.02%, agar 1.2%, the rest is distilled water, pH 7.4.

[0080] Fermentation medium: MCs 1.8%, MgSO 4 0.05%, NaCl 0.2%, K 2 HPO 4 0.02%, agar 1.2%, the rest is distilled water, pH 7.4.

[0081] The cultivation method comprises the steps of:

[0082] 1) Plate / slant culture: inoculate Paenibacillus PN-S435 onto a plate or slant culture medium by streaking at 25-28°C for 3-4 days;

[0083] 2) Expansion cultivation: Inoculate the Paenibacillus PN-S435 cultured on the plate in step 1) into the fermentation medium for expansion cultivation, the cultivation temperature is 25~28°C, and the cultivation time is 3~4 days;

[0084] 3) Degradation application: make the Paenibacill...

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Abstract

The invention discloses a Paenibacillus sp. PN-S435, with the preservation number: CCTCC M 2012422, microcystins (MCs) are the only carbon source and nitrogen source, and MCs can be effectively degraded. The Paenibacillus sp. PN-S435 can be obtained through the performing combination of compound mutation of low-energy N+-LiCl with compound mutation of UV-LiCl for original strain PV-3, and then performing preliminary screening and secondary screening. According to the invention, the capability of the strain degrading MCs reaches 30.2 mg / L per day, which is improved by 64.1percent compared with original strain; and the strain greatly improves the capability of degrading MCs in a water body, hereditary stability tests prove that the Paenibacillus sp. PN-S435 can be applied to practical application, controls water body with serious algal bloom, removes even avoids danger of algal toxin to drinking water safety.

Description

technical field [0001] The invention relates to a strain of Paenibacillus and its application in degrading microcystins, belonging to the field of environmental microorganisms. technical background [0002] Due to environmental pollution, the degree of eutrophication in water bodies has intensified in recent years, and frequent outbreaks of cyanobacteria blooms. One of the most important hazards brought about is that some cyanobacteria cells produce and release a variety of different types of algal toxins into the water body. Microcystins (microcystins, MCs) is a type of algae toxins with the highest frequency, the largest production and the most serious harm. It is a kind of hepatotoxin with animal liver as its target organ. It can specifically inhibit the activity of protein phosphatase and induce a series of diseases such as cancer. Microcystis is the most common type of bloom cyanobacteria in freshwater, and the microcystins (MCs) released by it are the most widely dist...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C02F3/34C12R1/01C02F101/38
Inventor 虞龙钱梦宇李玉燕于洋
Owner NANJING TECH UNIV
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