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Bacillus capable of degrading naphthenic acids and application of bacillus capable of degrading naphthenic acids

A bacillus, naphthenic acid technology, applied in the direction of bacteria, microorganism-based methods, biochemical equipment and methods, etc.

Inactive Publication Date: 2019-03-12
NORTHEAST NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, the research on the microbial degradation of naphthenic acid is mainly concentrated in foreign countries, and there are few reports on domestic related research, which is still in its infancy.

Method used

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  • Bacillus capable of degrading naphthenic acids and application of bacillus capable of degrading naphthenic acids
  • Bacillus capable of degrading naphthenic acids and application of bacillus capable of degrading naphthenic acids
  • Bacillus capable of degrading naphthenic acids and application of bacillus capable of degrading naphthenic acids

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1. Isolation, purification and identification of strain CR1.

[0027] Strain CR1 was isolated from oil sands, and the specific enrichment, separation, and purification processes were as follows:

[0028] Basic inorganic salt medium (MSM medium) consists of: KH 2 PO 4 1.36g, (NH 4 ) 2 SO 4 0.66g, Na 2 HPO 4 0.43g, MgSO 4 ·7H 2 O 0.25g, trace element solution 10mL, distilled water 990mL.

[0029] The composition of the trace element solution is: Ca(NO 3 ) 2 4H 2 0800mg / L, FeSO 4 ·7H 2 O 400mg / L, CuSO 4 ·5H 2 O 80mg / L, ZnSO 4 ·7H 2 O 40mg / L, MnSO 4 4H 2 O 40mg / L, NaMO 4 2H 2 O 8mg / L, H 3 BO 3 6mg / L, K 2 HPO 4 500mg / L, the rest is distilled water.

[0030] Add agarose with a mass percent content of 1.5-1.8% to the MSM medium, and sterilize under the condition of 0.06MPa for 20min. After the sterilized MSM culture medium was cooled to 50-60°C, trans-4-methyl-cyclohexane acid was added to make the concentration 50 mg / L. At the same time, th...

Embodiment 2

[0039] Example 2. The degradation profile of bacterial strain CR1 to naphthenic acid

[0040] In order to study the degradation spectrum of naphthenic acid by strain CR1, MSM was used as the basic medium, and the growth of the strain on five kinds of naphthenic acid with different chemical structures as the only external carbon source was investigated respectively. The experiment adopts the method of solid culture, at 28 ℃, static culture for one week, observe the situation of colony formation. Table 1 shows the results of strain CR1 degrading five different naphthenic acids.

[0041] Table 1 The degradation and utilization of naphthenic acid by strain CR1

[0042]

[0043] Note: "+" in the table indicates the growth of bacteria on the corresponding naphthenic acid plate.

[0044] The more "+" means the better the growth of the bacteria and the better the biodegradation of naphthenic acid

[0045] It can be seen from Table 1 that: (1) bacterial strain CR1 has a wide degr...

Embodiment 3

[0046] Example 3. Biodegradation of trans-4-methyl-cyclohexanecarboxylic acid by strain CR1

[0047] Inoculate the preserved bacteria in the LB enriched medium containing 100mg / L trans-4-methyl-cyclohexanecarboxylic acid, grow to the optimum bacterial age, then centrifuge the bacteria at 4000rpm for 10min to collect the bacteria, wash Afterwards, the bacteria were inoculated into the MSM medium containing 100mg / L trans-4-methyl-cyclohexanecarboxylic acid, samples were taken at regular intervals, and the residual concentration of trans-4-methyl-cyclohexanecarboxylic acid was measured to draw the Degradation curve, see the results image 3 . After 144 hours of reaction, the final concentration of trans-4-methyl-cyclohexanecarboxylic acid was 6.84mg / L, and the degradation rate could reach 93.2%. It can be seen that the screened bacterial strain has a high degradation efficiency to trans-4-methyl-cyclohexanecarboxylic acid.

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Abstract

The invention relates to a bacillus capable of degrading naphthenic acids and application of the bacillus capable of degrading the naphthenic acids and particularly relates, separates and identifies abacillus CR1 (in a preservation number of CGMCC NO.7368) capable of biologically degrading five types of naphthenic acids in different chemical structures. The bacillus grows by taking trans-4-methyl-cyclohexanecarboxylic acid, cis-4-methyl-cyclohexanecarboxylic acid, trans-4-tert-butylcyclohexanecarboxylic acid, trans-4-pentyl-cyclohexanecarboxylic acid and dicyclohexyl acetic acid as a unique carbon source, and trans-4-methyl-cyclohexanecarboxylic acid degrading rate and trans-4-pentyl-cyclohexanecarboxylic acid degrading rate are 93.2% and 42.2% respectively, and the bacillus can be applied to biodegradation of naphthenic acids.

Description

technical field [0001] The invention relates to a bacillus strain capable of degrading naphthenic acid and its application. Background technique [0002] Acidic substances in petroleum include naphthenic acid, aromatic acid, fatty acid and phenols, etc. These acidic substances are collectively called petroleum acid. Among them, naphthenic acid is the most important, accounting for more than 90% of the total mass of petroleum acid. Therefore, petroleum acids are often referred to as naphthenic acids without distinction in the petroleum industry. The chemical composition of naphthenic acid is complex, usually composed of a variety of aliphatic carboxylic acids and branched monocyclic or polycyclic alicyclic carboxylic acids. The empirical formula of naphthenic acid is C n h 2n +ZO 2 , where n is the number of carbons (generally 11 to 30); Z is the degree of hydrogen unsaturation, which is 0 or a negative even number. [0003] The pollution of naphthenic acid wastewater i...

Claims

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Application Information

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IPC IPC(8): C12N1/20A62D3/02A62D101/26C12R1/07
CPCA62D3/02A62D2101/26C12N1/205C12R2001/07
Inventor 王小雨
Owner NORTHEAST NORMAL UNIVERSITY
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