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Achromobacter strain JD417 and application thereof

A colorless bacillus and bacterial strain technology, applied in the field of microorganisms, can solve the problems of limited application, and achieve the effects of low energy consumption, easy operation, and mild reaction conditions

Pending Publication Date: 2022-04-08
厦门众仁合美生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there is only one patent (CN201710474535.6) in China that discloses the effect of sulfa-tolerant Achromobacter sp. on sewage denitrification, but this strain can only be used under the condition of 0-20 μg / L sulfa antibiotic concentration. Maintain the integrity of the cell structure and the cell membrane is not damaged, so the application is very limited

Method used

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  • Achromobacter strain JD417 and application thereof
  • Achromobacter strain JD417 and application thereof
  • Achromobacter strain JD417 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Example 1: Enrichment, isolation and purification of bacterial strains

[0049] The strains were isolated from the sludge in the aeration tank of the wastewater treatment aeration tank of Baiyunshan Chemical Pharmaceutical Factory in Guangzhou City, Guangdong Province. Take 50g of sludge and put it into a Erlenmeyer flask filled with 50mL of sterile water (with several glass beads in it), shake it on a shaker for 30 minutes to fully disperse the sludge sample. Take 5mL of the suspended mud sample after breaking up, inoculate it into 100mL of the liquid primary screening medium, and incubate with shaking at 30-37°C and 120-150rpm for 24-48h. The pH of the liquid primary screening medium is 7.2 and consists of: peptone 10g / L, yeast extract 5g / L, NaCl 5g / L, sulfamethazine 10-50mg / L.

[0050] Then transfer to the liquid re-screening medium with sulfamethazine as the only carbon source, and acclimatize and cultivate for 3 to 5 times. Shake culture in a shaker at 30-37°C an...

Embodiment 2

[0051] Embodiment 2: the screening of bacterial strain

[0052] Use sulfamethazine degradation efficiency as an index to screen strains, take each single colony and culture it in liquid screening medium, measure the concentration of sulfamethazine in the culture medium every 12 to 48 hours, and screen the degradation effect of sulfamethazine The best strain.

[0053] Described re-screening medium consists of: Na 2 HPO 4 12H 2 O 15.58g / L, KH 2 PO 4 675mg / L, MgSO 4 ·H 2 O 112mg / L, NH 4 Cl 20.4mg / L, FeSO 4 ·7H 2 O 1mg / L, MnSO 4 ·H 2 O 1 mg / L, CuCl 2 2H 2 O0.25mg / L, Na 2 MoO 4 0.25mg / L, CaCl 2 0.015mg / L, SM2 10~100mg / L, pH 7.2. The component of described solid re-screening medium is to add 15g / L agar on the basis of liquid re-screening medium.

[0054] The strain with the best degradation effect on sulfamethazine was named JD417.

Embodiment 3

[0055] Example 3 strain identification

[0056] 1. Morphological identification

[0057] Strain JD417 is a gram-negative bacterium ( figure 1 A). In plate culture, the colony is round, smooth and flat, with neat edges, light yellow, opaque, and the diameter of the colony is about 2-3mm ( figure 1 B), observed under a scanning electron microscope (7000×), the bacteria are in the shape of a small cylinder with a length of about 1-1.5 μm ( figure 1 C).

[0058] 2. Molecular identification

[0059] According to the sequencing results, the strain JD417 is Achromobacter sp., and its 16S rDNA sequence is homologous to Achromobacter spanius MYb73 (accession number CP023270.1), with 99% similarity. The phylogenetic tree of this strain is shown in figure 2 . The Achromobacter spanius strain JD417 was deposited in the China Center for Type Culture Collection on July 2, 2021, with the preservation number CCTCC NO: M 2021813, and the preservation address is Wuhan University, Wuhan,...

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Abstract

The invention relates to an Achromobacter sanius strain JD417 and application thereof, the strain is preserved in the China Center for Type Culture Collection on July 2, 2021, and the preservation number is CCTCC NO: M 2021813. The invention also relates to an application of the Achromobacter sanius strain JD417. The strain JD417 can tolerate sulfonamide compounds (antibiotics) with high concentration (50mg / L), and not only can degrade the sulfonamide compounds, but also can degrade nitrogen substances. The strain can maintain survival by taking nitrate nitrogen as a sole nitrogen source and sulfamethazine as a sole carbon source, effectively reduces total nitrogen, nitrate nitrogen and sulfonamide compounds in a water body, and has good ecological efficiency and application prospects in the aspect of treatment of environmental pollution such as sewage and the like.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, and in particular relates to Achromobacter strain JD417 and its cultivation method and application. Background technique [0002] Sulfonamide compounds (sulfonamide antibiotics) are widely used in the prophylactic treatment of human and animal diseases, as well as in the growth promotion of animals in aquaculture and animal husbandry. In recent years, with the detection of different levels of sulfa antibiotic residues in groundwater, sediment, soil and even food, including sulfamethazine (SM2), sulfadiazine (SDZ), sulfamethazine Sulfamethoxazole (SMX) and other sulfonamides are gradually attracting people's attention for their harm to the environment and human health. [0003] Sulfa antibiotics have environmental toxicity and environmental enrichment effects. After ingestion by the body, most of them will enter the environment in the form of parent or metabolites. The long-term existence...

Claims

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Application Information

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IPC IPC(8): C12N1/20C02F3/34C12R1/025C02F101/38C02F101/30C02F101/16
Inventor 陶然曾露苹杨扬张毅欣王晓燕李婉旋邰义萍
Owner 厦门众仁合美生物科技有限公司
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