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A strain bw02 with low-temperature salt tolerance and high-efficiency pyrene degradation function and its application

A high-efficiency, strain-based technology, applied in the field of microorganisms, can solve the problems of utilization and difficult general microorganisms, and achieve good degradation effect, good degradation performance, and high pyrene degradation rate.

Active Publication Date: 2020-05-08
DALIAN NATIONALITIES UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In general, high molecular weight PAHs have a stable chemical structure and are difficult to be utilized by general microorganisms

Method used

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  • A strain bw02 with low-temperature salt tolerance and high-efficiency pyrene degradation function and its application
  • A strain bw02 with low-temperature salt tolerance and high-efficiency pyrene degradation function and its application
  • A strain bw02 with low-temperature salt tolerance and high-efficiency pyrene degradation function and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Strain BW02 culture

[0034] LB medium (g / L): tryptone (Tryptone) 10g; agar (Agar) 15g; yeast extract powder (YeastExtract) 5g, NaCl 10g, deionized water 1L, adjust the pH to 7.0, liquid medium does not add agar .

[0035] ASM inorganic salt culture (g / L): NaCl 30g; MgSO 4 , 0.35g; NH 4 NO 3 1g; KCI 0.7g, Na 2 HPO 4 .12H 2 O5g; KH 2 PO 4 0.2245g; pH7.5, finally add 100ul trace element solution.

[0036] Trace element solution (mg / L): CaCl 2 2 mg; FeCl 3 .6H 2 O5mg; GuSO 4 0.5 mg; MnCl 2 .4H 2 00.5mg; ZnSO 4 .7H 2 O 10mg; distilled water 1000ml.

[0037] The purified strains were inoculated on LB solid medium and cultured for 48 hours to observe the colony morphology. At the same time, drop the bacterial suspension of the strain to be tested at an appropriate concentration on a 5mm×4mm silicon wafer, solidify with 2.5% glutaraldehyde for 2-4h, then wash with phosphate buffer solution with pH=7.3, dehydrate with ethanol gradient, and finally use acetic a...

Embodiment 2

[0044] Determination of Growth Curve of Efficient Pyrene Degrading Strain BW02

[0045] The experiment uses the visible spectrophotometry value method to measure the growth curve of the high-efficiency degradation strain of pyrene. The detection wavelength is 600nm, and the sterile liquid medium is used as the blank control. , OD corresponding to 72, 78, 84h 600 . Each sample was measured three times, and the average value was taken (if the concentration of the bacterial suspension is too thick, it should be properly diluted to make the OD 600 value less than 1).

[0046] Under suitable conditions, the strain BW02 will have four stages of bacterial growth, lag phase, logarithmic phase, stable phase, and decay phase. The experimental results showed that: 0-20h was the lag phase, and the bacteria grew slowly; 20-60h was the logarithmic phase, and the bacteria grew rapidly; 60-90h was the stable period, and the bacteria growth gradually became stable. like Figure 4 .

Embodiment 3

[0048] Effect of Bacteria Age on Pyrene Degradation Performance of Strain BW02

[0049] According to the growth curve, the bacterial strains with bacterial ages of 24h, 48h, 72h and 96h were selected as research objects, and the bacterial suspension of bacterial strain BW02 was prepared, and the bacterial concentration was adjusted to cfu / ml=10 9 . Inoculate 4% of the inoculum into 50ml of ASM liquid medium with a pyrene content of 1mg / ml, adjust the pH to 7.5, and culture at 15°C for 15 days. Each sample is replicated three times, and no inoculation is used as a blank control. The effects of different bacterial ages on the pyrene degradation performance of the strain BW02 were determined by gas chromatography.

[0050] In the experiment, the strains of bacterial strain BW02 (Rhodococcus) in different periods were selected for analysis, which were respectively 24h, 48h, 72h, and 96h, as Figure 5 The degradation performance is the best when the bacteria age is 48h, which can...

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Abstract

The invention belongs to the technical field of microbes, and specifically relates to a strain BW02 with low-temperature, salt-tolerant, and high-efficiency pyrene-degrading functions and its application. The preservation number of the strain is CGMCC No.10354, and its 16SrRNA sequence is shown in SEQ ID No:1. The strain BW02 screened by the present invention has a high-efficiency pyrene degradation function. The strain has good degradation performance under neutral and slightly alkaline conditions, and has a relatively low demand for oxygen. It degrades pyrene at a low temperature of 15°C and a salt concentration of 3%. It has a high efficiency, is suitable for the seabed environment, and has a good degradation effect on polycyclic aromatic hydrocarbons in marine sediment pollutants.

Description

technical field [0001] The invention belongs to the technical field of microbes, and specifically relates to a strain BW02 with low-temperature, salt-tolerant, and high-efficiency pyrene-degrading functions and its application. Background technique [0002] Polycyclic aromatic hydrocarbons (Polycyclic Aromatic Hydrocarbons referred to as PAHs): Refers to condensed ring compounds in which two or more benzene rings are combined with each other and arranged in a linear, angular or clustered form. The basic unit is a benzene ring, and the structure is stable and difficult Degradation, and can cause serious health hazards to human production and life through bioaccumulation and biomagnification. Containing 2-3 benzene rings, they are called low-molecular-weight PAHs (Low-molecular-weight, LMW-PAHs for short), such as naphthalene, anthracene and phenanthrene. High-molecular-weight PAHs (High-molecular-weight, HMW-PAHs for short) with 4-6 benzene rings, such as pyrene, benzo-R and...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C02F3/34C12R1/01C02F101/32
CPCC02F3/34C02F2101/327C12N1/205C12R2001/01
Inventor 刘秋李佳怡肖升月袁峰皓于基成
Owner DALIAN NATIONALITIES UNIVERSITY
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