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A high-efficiency petroleum hydrocarbon degrading bacterium pa16_9 and its screening method and application

The technology of Halomonas and microbial strains is applied in the field of high-efficiency petroleum hydrocarbon degrading bacteria PA16-9 and its screening. And other issues

Active Publication Date: 2021-11-23
THIRD INST OF OCEANOGRAPHY MINIST OF NATURAL RESOURCES +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Limited by the imperfection of the current technical process and management concepts, petroleum products containing a large amount of hydrocarbon components flow directly or indirectly (natural leakage, accidental spillage, and widespread use of refined petroleum products) into the soil, ocean and other ecological circles (El-Sheshtawy , Khalil, Ahmed, & Abdallah, 2014), causing a series of serious and persistent pollution problems
Saturated hydrocarbons (alkanes) are the main components of petroleum (Tissot & Welte, 1984). The leaked hydrocarbons are poured into the pores of the soil, affecting the permeability of the soil, destroying the soil water, gas, and solid phase structures of crude oil, and affecting soil The growth of microorganisms in the soil hinders the respiration of plant roots in the soil and the absorption of water and nutrients, and even causes the plant roots to rot and necrosis, seriously endangering the growth of plants and reducing the value of land cultivation in this area

Method used

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  • A high-efficiency petroleum hydrocarbon degrading bacterium pa16_9 and its screening method and application
  • A high-efficiency petroleum hydrocarbon degrading bacterium pa16_9 and its screening method and application
  • A high-efficiency petroleum hydrocarbon degrading bacterium pa16_9 and its screening method and application

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Embodiment 1: strain morphological characteristics

[0030] Streak inoculate a single colony onto an HLB solid plate, place the plate upside down in a constant temperature incubator, and incubate at 32°C for 48 hours. The colony is round, beige translucent, smooth and moist, with regular edges, no halos, and slightly convex in the center from 2 to 4 mm in diameter, see image 3 . Take primary flakes and observe with transmission electron microscope, see Figure 4 , the strain is a gram-negative bacterium, facultatively aerobic, rod-shaped, 1.6-2 μm in length, with pili on the periphery, and mainly relies on flagella for movement.

Embodiment 2

[0031] Embodiment 2: Screening and identification of bacterial strains

[0032] (1) Pacific deep-sea sediment samples were taken from the KW1 sea area of ​​the second voyage of Dayang 45 (sample number 45II-KW1-S37-BC29, 22-26cm away from the sediment surface, W 154°14.9909′, N 9°29.9927′ ), the sediment is yellow-brown, odorless, weakly viscous, and the surface layer is semi-fluid, with a slight silty feeling when rubbed by hand, and the consistency increases downward, and yellow-white lumps below 10cm are mixed in it. After the sediment samples were aseptically collected, they were cryopreserved and then transported to the laboratory for the next stage of research.

[0033] (2) Take a 156mL anaerobic bottle, wash it, add 100mL basic salt medium, pass N 2 Oxygen in the residual space was removed, and cysteine ​​(-HCl) was added to remove residual oxygen in the saline solution. Seal the rubber stopper with an aluminum cover, and sterilize at 121°C for 20 minutes. After the s...

Embodiment 3

[0035] Example 3: Determination of partial hydrocarbon degradation ability of bacterial strains

[0036] (1) Prepare anaerobic liquid medium, the carbon source concentration is 0.33% (n-hexadecane, about 0.07734g), the culture system is 30mL, scrape an appropriate amount of bacteria from the HLB solid plate and dissolve it in the liquid medium, vortex Mix well, inoculate into the anaerobic medium, the inoculum amount is 1%, and the three experimental groups are protected from light and cultured at 32°C.

[0037] (2) After cultivating for 3 months, open the bottle mouth, use n-hexane to repeatedly extract the n-hexadecane in the medium (extract four times in total), combine the extracts, N 2 After blowing, ensure that there is 0.5ml (about 10 drops) of n-hexadecane to prevent oxidation. Obtain the mass spectrogram of sample by GC-MS (gas chromatography-mass spectrometry instrument), record n-hexadecane residual amount, obtain degradation rate 76.7%~86.5%, the anaerobic degrada...

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Abstract

A high-efficiency petroleum hydrocarbon degrading bacterium PA16_9 and its screening method and application relate to offshore petroleum hydrocarbon degrading bacteria. The colony of Halomonas titanicae HT PA16_9, a high-efficiency petroleum hydrocarbon degrading bacterium, is round, beige translucent, smooth and wet, with regular edges, no halos, slightly raised in the center, and a diameter of 2 ~4mm; a strain of Halomonas titanic can be used in anaerobic degradation of organic compounds. It has been identified that a high-efficiency petroleum hydrocarbon degrading bacterium (Halomonas titanicae) PA16-9 can degrade n-hexadecane with an initial concentration of about 3.867g / L at 32°C, under anaerobic and light-proof static culture, and it was tested after 3 months The obtained degradation rate is 76.7%-86.5%, the degradation rate of pyrene can reach 58% under anaerobic condition, and can reach 74.5% under aerobic condition.

Description

technical field [0001] The invention relates to marine petroleum hydrocarbon degrading bacteria, in particular to a high-efficiency petroleum hydrocarbon degrading bacteria PA16-9 and its screening method and application. Background technique [0002] With the continuous development of our country's social economy, the demand and dependence on oil are increasing, which leads to the continuous prosperity of oil-related industries, but also causes the increasingly serious environmental pollution problem. Limited by the imperfection of the current technical process and management concepts, petroleum products containing a large amount of hydrocarbon components flow directly or indirectly (natural leakage, accidental spillage, and widespread use of refined petroleum products) into the soil, ocean and other ecological circles (El-Sheshtawy , Khalil, Ahmed, & Abdallah, 2014), causing a series of serious and persistent pollution problems. Saturated hydrocarbons (alkanes) are the ma...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N1/02C02F3/34B09C1/10C12R1/01C02F101/32C02F101/36
CPCB09C1/10C02F3/34C02F2101/32C02F2101/327C02F2101/36C12N1/02C12N1/20C12N1/205C12R2001/01
Inventor 邵宗泽王万鹏李振宇
Owner THIRD INST OF OCEANOGRAPHY MINIST OF NATURAL RESOURCES
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