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A kind of microbial agent containing halophilic denitrifying bacteria yl5-2 and its application

A technology of microbial agent and microbial strain, applied in the field of environmental protection microorganisms, can solve the problem of less halophilic denitrifying bacteria, achieve the effects of consuming nitrogen nutrients, inhibiting excessive algae reproduction, and reducing costs

Active Publication Date: 2021-10-26
BLUESTAR LEHIGH ENG INST CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] There are few reports on halophilic denitrifying bacteria in the prior art, especially halophilic denitrifying bacteria under the condition of salinity>10%

Method used

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  • A kind of microbial agent containing halophilic denitrifying bacteria yl5-2 and its application
  • A kind of microbial agent containing halophilic denitrifying bacteria yl5-2 and its application
  • A kind of microbial agent containing halophilic denitrifying bacteria yl5-2 and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 Isolation and preservation of the halophilic denitrifying bacteria

[0032] The halophilic denitrifying bacteria YL5-2 was isolated from the sedimentary soil of Golmucharhan Salt Lake (36°51′N, 94°95′E) in Qinghai Province, China. The water of Chaerhan Salt Lake is saturated or close to saturated salt concentration all year round.

[0033]Prepare LB liquid medium with NaCl concentration of 20%, add glycerol 250mg / L, glucose 250mg / L, methanol 50mg / L, and enrich and cultivate the depositional soil of Chaerhan Salt Lake at 30°C for 72h. The strains in the enriched culture medium were isolated by using YL solid medium. 1L medium contains the following components: glucose: 0.6g, trisodium citrate 0.5g, glycerol 2mL, yeast extract 0.8g, peptone 1.6g, dipotassium hydrogen phosphate 0.35g, potassium dihydrogen phosphate 0.1g, ammonium sulfate 0.25g, ammonium chloride 0.25g, MgSO 4 0.5g, CaCl 2 0.1g, NaCl 180g; trace element SL-4 10mL, pH 7.0-7.2; agar 2.5%.

[...

Embodiment 2

[0035] Example 2 Analysis of 16S rRNA sequence and whole gene sequence of halophilic denitrifying bacteria YL5-2

[0036] Strain YL5-2 genomic DNA was extracted using TaKaRa kit (TaKaRa MiniBEST Bacteria Genomic DNA Extraction 68Kit Ver.3.0).

[0037] 16S rRNA was amplified using universal bacterial primers 27F (5’-AGAGTTTGATCMTGGCTCA G-3’) and 1492R (5’-TACGGYTACCTTGTTACGACTT-3’). PCR sequencing was entrusted to Shanghai Sangon Biotechnology Co., Ltd. The complete 16S rRNA sequence of strain YL5-2 is 1431bp, as shown in SEQ ID NO.1.

[0038] The whole genome of strain YL5-2 was sequenced using the IlluminaMiSeq 2000 high-throughput sequencing platform of Shanghai Passino Biotechnology Co., Ltd. The original sequencing data was filtered and corrected using PRINSEQ (version number v 0.20.4) software, and then SOAP denovo software (version number v1.05) software with default parameters was used for genome base pairing, and then CheckM software (version number 1.03) Assess the...

Embodiment 3

[0041] Example 3 Identification of phenotypic and physiological and biochemical characteristics of halophilic denitrifying bacteria YL5-2

[0042] Gram staining properties were tested using the BD Gram staining kit.

[0043] Cell motility was determined using half MA medium (0.5% agar, w / v).

[0044] Cell morphology was detected by transmission electron microscopy (TEM) analysis. That is, cells were picked from exponentially growing culture medium, stained with 0.5% uranyl acetate, and photographed under a microscope (Tecnai Spirit, FEI, Hillsboro, OR, USA).

[0045] Oxidase activity using the oxidase kit (bioMérieux), by adding 3.0% H 2 o 2 Catalase activity was determined by pouring the solution onto bacterial colonies and observing the generation of bubbles.

[0046] The temperature growth conditions were carried out on YL liquid agar medium, the temperatures were 4, 10, 15, 20, 25, 30, 33, 37, 40, 45 and 50 ° C, and the pH was constant at 7.5, and the strain YL5- 2 T...

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Abstract

The invention discloses a microbial agent containing halophilic denitrifying bacteria YL5‑2 and an application thereof. A microbial agent containing halophilic denitrifying bacteria YL5‑2, YL5‑2 is a new species of the genus Halovibrio, preserved in the China General Microorganism Culture Collection and Management Center, the preservation number is CGMCC NO.16315, and the preservation date is 2018 August 20. The application of the microbial agent of the invention in the treatment of high-salt wastewater. YL5‑2 can be used for biological denitrification treatment of high-salt wastewater with a salt content greater than 10%, reducing the cost of denitrification treatment of such wastewater. The microbial bacterial agent prepared by the halophilic denitrifying bacteria YL5-2 disclosed by the present invention can be used in the degradation, transformation and biological denitrification process of pollutants under high-salt conditions, including high-salt wastewater treatment, polluted seawater treatment, saline-alkali land restoration, Consume nitrogen nutrients, inhibit algae overgrowth, purify water, improve substrate, etc.

Description

technical field [0001] The invention relates to the field of environment-friendly microorganisms, in particular to a microbial agent containing halophilic denitrifying bacteria YL5-2 and its application. Background technique [0002] Nitrogen pollution is one of the important causes of water eutrophication. The denitrification treatment of wastewater plays an important role in maintaining the quality of water environment and preventing eutrophication of water body. Nitrogen-containing wastewater discharged from petroleum, chemical, food processing, chemical fertilizer and other industries has the characteristics of high salt. The salt content of some waste water produced by chemical industry and food processing even exceeds that of sea water. Traditional biological methods have advantages in treating low-salinity nitrogen-containing wastewater, but when the concentration of wastewater is too high, it will inhibit the metabolism of nitrogen-removing microorganisms. The exi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C02F3/34C12R1/01
CPCC02F3/34C12N1/20
Inventor 徐军王开春李坤王强田凤蓉洪磊张璐璐孙文妮
Owner BLUESTAR LEHIGH ENG INST CO LTD
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