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A high-efficiency oil-degrading Providencia rettgeri l1 and its application

A high-efficiency technology of Providencia rettgeri, applied in the field of petroleum degradation, achieves broad application prospects and significant degradation effect

Active Publication Date: 2020-05-12
BIOTECH CENT OF SHANDONG ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The above-mentioned strains have a good degradation effect on aniline herbicides, but there is no report that the microorganisms of this genus have a degradation effect on the degradation of petroleum hydrocarbons.

Method used

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  • A high-efficiency oil-degrading Providencia rettgeri l1 and its application
  • A high-efficiency oil-degrading Providencia rettgeri l1 and its application
  • A high-efficiency oil-degrading Providencia rettgeri l1 and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Example 1 Isolation and Identification of Petroleum Degrading Bacteria

[0059] 1. Culture medium formula

[0060] Induction medium ( / L): 1.0g NH 4 NO 3 , 0.5g K 2 HPO 4 , 0.02g MgSO 4 , 1.0g NaCl, 0.02mgCaCl 2 , 0.002mg FeCl 3 , 5.0g yeast extract, 10.0g peptone, sterilized at 121°C for 20min. Add oil before use, and the amount of oil added is 5g / L.

[0061] Nutrient broth medium: peptone 10g, beef powder 3g, sodium chloride 5g, distilled water 1000mL, pH value 7.2±0.2, sterilized at 121°C for 20min, set aside. Add 18.0 g of agar to this medium, and keep the other things unchanged, that is, nutrient agar medium.

[0062] 2. Isolation of strains

[0063] The traditional strain screening method was carried out using shake flask culture experiments. Add crude oil (5g / L) to the induction medium, culture at 30°C and 160rpm for 10 days, draw 10ml of culture solution and transfer to the same liquid shake flask, the concentration of crude oil is 5g / L, continue to cu...

Embodiment 2

[0082] Example 2 Degradation Experiment of Petroleum Using the Biological Enzyme Prepared by Bacterial Strain L1

[0083] 1. Preparation of bacterial suspension

[0084] Put the purified Providencia rettgeri (Providencia rettgeri) L1 into the NB liquid medium containing 5ml for overnight activation and culture to the logarithmic phase, then transfer to the 250ml Erlenmeyer flask containing 50ml induction medium and cultivate to the right For several phases, the bacteria were collected by centrifugation at 5000rpm for 10 minutes, washed with PBS buffer three times, added with PBS buffer at a ratio of 1:15 (W:V), and stored at low temperature for later use.

[0085] 2. Cell fragmentation and degradation of petroleum

[0086] The bio-enzyme was prepared by the method described in Example 1, and the degradation of petroleum by the strain was detected by the bio-enzyme method. The collected mixed enzyme solution and FDH were mixed at a ratio of 3:5 (volume ratio), and 0.1 g of so...

Embodiment 3

[0095] The preparation of embodiment 3 bioremediation bacteria agent

[0096] (1) Seed cultivation

[0097] Inoculate Providencia rettii CGMCC No.16526 stored at -80°C on nutrient agar medium, culture at 28°C for 24 hours, transfer the grown single colony to nutrient broth medium, 28°C, 180rpm Cultivate on a shaking table for 24 hours, and finally obtain the bacterial liquid of Providencia rettgeri CGMCC No.16526.

[0098] (2) Liquid fermentation

[0099] The bacterium liquid prepared in the step (1) is transferred to the liquid fermentation medium according to the ratio of volume ratio 5%, 28 ℃, 180rpm culture 16h, finally obtains the fermentation of Providencia rettii CGMCC No.16526 bacteria liquid. After testing, the number of effective viable bacteria in the bacterial liquid of Providencia rettii CGMCC No.16526 was 2.2×10 9 cfu / ml.

[0100] In the above-mentioned steps (2), the composition of the liquid fermentation medium is as follows: MgSO 4 ·7H 2 O 0.5%, NaCl 0....

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Abstract

The present invention relates to a strain of Providencia rettgeri (Providencia rettgeri) L1 that efficiently degrades petroleum and its application. The strain was preserved in the General Microbiology Center (CGMCC) of the China Committee for the Collection of Microbial Cultures on September 26, 2018, address : Institute of Microbiology, Chinese Academy of Sciences, No. 3, Courtyard No. 1, Beichen West Road, Chaoyang District, Beijing, strain collection number CGMCC No.16526. The present invention discovers for the first time that the bacterial strain of Providencia (Providencia) has the function of degrading petroleum hydrocarbons, and has differential selectivity to the degradation of four components in petroleum, especially for normal alkanes, one of the petroleum components. It has a very significant degradation effect, can be applied to the treatment of petroleum pollutants, and has broad application prospects.

Description

technical field [0001] The invention relates to a strain of Providencia rettgeri (Providencia rettgeri) L1 that efficiently degrades petroleum and its application, belonging to the technical field of petroleum degradation. Background technique [0002] Since the end of the 1970s, with the acceleration of industrialization and the development of society, the production and consumption of oil have been increasing, which in turn has promoted the rapid development of the oil extraction industry and the petrochemical industry. In the process of mining, transportation and processing, oil pollution of the soil is caused by man-made or uncontrollable factors such as leakage, stacking and volatilization. According to relevant statistics, the world's current annual oil production reaches 2.2 billion tons, of which more than 8 million tons of crude oil enter the environment due to accidents and other reasons in the process of exploitation, transportation, processing and storage. As fa...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20B09C1/10C12R1/01
CPCB09C1/10C12N1/20C12N1/205C12R2001/01
Inventor 黄玉杰张新建王加宁高永超张闻陈贯虹赵庆庆王磊磊郑立稳
Owner BIOTECH CENT OF SHANDONG ACAD OF SCI
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