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 increased difficulty and incomplete degradation, and achieve the effect of avoiding serious threats, great social significance and economic value, and increasing the degradation rate

Inactive Publication Date: 2013-05-01
NANJING UNIV OF TECH
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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 (S

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

Example Embodiment

[0050] Example 1

[0051] This example illustrates the mutagenesis screening method of 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%, 1.2% agar, 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 , 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%, 1.2% agar, the rest is distilled water, pH 7.4.

[0056] Microcystis indoor culture solution: Inoculate Microcystis aeruginosa seeds on BG-11 medium with an inoculum of 5-15% (v / v), culture in a light incubator for ten days, and a culture temperature of 28-33℃. The light intensity is 4000lx, the light to dark ratio is 12h:12h,...

Example Embodiment

[0069] Example 2

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

[0071] The main morphology and biological characteristics of Bacillus sp. PN-S435 of the present invention are as follows: the cells are rod-shaped, short and thick, facultative anaerobic Gram-positive bacteria, without flagella, and round spores in the cysts. The color of the colony on the beef extract peptone medium is light yellow, the colony size is 1-2.5mm, round colony, the colony is thick, moist, the edges are neat, the surface is smooth, the texture is uniform, easy to pick up, the colony size is 1-2.5mm, Optimal growth temperature: 25℃~28℃, growth temperature range 15-50℃, optimum growth pH: 7.2~7.5, growth pH range 5.6~10.0.

[0072] This example illustrates the mutant bacillus obtained by screening ( Paenibacillus sp. ) The genetic stability of PN-S435. The results of the passage degradation test are shown in Table 2:

[0073] Table 2 Bacillus type ( Paenibacillus s...

Example Embodiment

[0076] Example 3

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

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

[0079] 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%, 1.2% agar, 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%, 1.2% agar, the rest is distilled water, pH 7.4.

[0081] The cultivation method includes the following steps:

[0082] 1) Plate / slant culture: Streak Bacillus PN-S435 on a plate or slant medium for culture, the culture temperature is 25-28℃, and the culture time is 3~4d;

[0083] 2) Expansion culture: Inoculate the Bacillus spp. PN-S435 cultivated on the plate in step 1) into the fermentation medium for expansion culture, the culture temperature is 25~28℃, and the culture time is 3~4 days;

[0084] 3) Degradation application: The Ba...

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