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Microbial agent containing halophilic denitrifying bacteria YL5-2 and application of microbial agent

A technology of microbial agent and denitrifying bacteria, which is applied in the field of microbial agent containing halophilic denitrifying bacteria YL5-2, can solve the problem of less halophilic denitrifying bacteria, achieve nitrogen nutrient consumption and inhibit algal overgrowth , cost reduction effect

Active Publication Date: 2019-03-12
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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  • Microbial agent containing halophilic denitrifying bacteria YL5-2 and application of microbial agent
  • Microbial agent containing halophilic denitrifying bacteria YL5-2 and application of microbial agent
  • Microbial agent containing halophilic denitrifying bacteria YL5-2 and application of microbial agent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

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

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

[0033]LB liquid medium with NaCl concentration of 20% was prepared, and 250 mg / L of glycerol, 250 mg / L of glucose and 50 mg / L of methanol were added, and the sedimentary soil of Chaerhan Salt Lake was enriched and cultured at 30℃ for 72 hours. The strains in the enriched culture medium were isolated 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; t...

Embodiment 2

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

[0036] The genomic DNA of strain YL5-2 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 1518 bp, as shown in SEQ ID NO. 1, and the GenBank accession number is MF782425.

[0038] Whole-genome sequencing of strain YL5-2 was performed using the IlluminaMiSeq 2000 high-throughput sequencing platform of Shanghai Paisenuo Biotechnology Co., Ltd. Raw sequencing data were filtered and corrected using PRINSEQ (version number v 0.20.4) software, followed by base pairing of genomes using SOAP denovo software (version number v1.05) software with default parame...

Embodiment 3

[0041] Example 3 Identification of phenotypic characteristics 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 the exponentially growing medium, stained with 0.5% uranyl acetate, and photographed under a microscope (Tecnai Spirit, FEI, Hillsboro, OR, USA).

[0045] Oxidase activity was measured using an oxidase kit (bioMérieux) by adding 3.0% H 2 O 2 The solution was poured into bacterial colonies and observed for bubble generation to determine catalase activity.

[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. The strain...

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Abstract

The invention discloses a microbial agent containing halophilic denitrifying bacteria YL5-2 and application of the microbial agent. The microbial agent contains the halophilic denitrifying bacteria YL5-2, and the YL5-2 is a novel species of Halovibrio genus, and is preserved in the China General Microbiological Culture Collection Centre with a preservation number of CGMCC NO.16315 on August 20, 2018. The invention further discloses application of the microbial agent in high-salt wastewater treatment. The YL5-2 can be used for biological denitrification treatment of high-salt wastewater with salt content greater than 10%, and denitrification treatment cost of the wastewater is reduced. According to the invention, the microbial agent prepared from the halophilic denitrifying bacteria YL5-2 can be used for degradation, conversion and biological denitrification processes of pollutants under a high-salt condition, and the processes comprise high-salt wastewater treatment, polluted seawatergovernance, saline-alkali land repair, nitrogen nutrition consumption, inhibition on excessive multiplication of alga, water body purification, substrate improvement, and the like.

Description

technical field [0001] The invention relates to the field of environmental protection microorganisms, in particular to a microbial inoculum 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 water environment quality and preventing water eutrophication. Nitrogen-containing wastewater discharged from petroleum, chemical, food processing, fertilizer and other industries is characterized by high salt. Some waste water from chemical industry and food processing has more salt content than sea water. The traditional biological method has advantages in the treatment of low-salinity nitrogen-containing wastewater, but when the wastewater concentration is too high, the metabolism of denitrifying microorganisms will be inhibited. The existence of salt-tolerant and halophilic de...

Claims

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Application Information

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Patent Type & Authority Applications(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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