Preparation method of sodium alginate immobilized bacterial combination for polycyclic aromatic hydrocarbon pollution remediation

A technology of sodium alginate and combined bacteria, which is applied in the field of biological treatment of environmental pollutants, and can solve problems such as poor adaptability of single strains and secondary pollution of bacterial groups

Inactive Publication Date: 2019-02-22
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] The purpose of the present invention is to overcome the problem of poor adaptability of single strains existing in the existing immobilized microorganisms to degrade polycyclic aromatic hydrocarbons, and

Method used

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  • Preparation method of sodium alginate immobilized bacterial combination for polycyclic aromatic hydrocarbon pollution remediation
  • Preparation method of sodium alginate immobilized bacterial combination for polycyclic aromatic hydrocarbon pollution remediation
  • Preparation method of sodium alginate immobilized bacterial combination for polycyclic aromatic hydrocarbon pollution remediation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Example 1: Comparison of the degradation effects of different proportions of free combined flora on benzo[a]pyrene

[0054] According to the differences in the degradation ability and growth rate of the 8 strains to benzo[a]pyrene, the 8 strains were divided into two groups, P6 and P7 were the main degrading bacteria, and the other 6 strains were secondary degrading bacteria.

[0055] Take the culture fluid of the 8 strains that have been subcultured for 48 hours and adjust the OD600 to 0.120. The equal volumes of the culture fluids of P6 and P7 were mixed as the inoculum of the main degrading bacteria; The two kinds of inoculums were mixed according to main degrading bacteria: secondary degrading bacteria = 1:0, 9:1, 4:1 and 3:2 respectively as the inoculation solutions of combinations 1-4. Add 0.2mL benzo[a]pyrene-acetone stock solution to a 15mL brown sample bottle, blow nitrogen to dryness, add 1.8mL inorganic salt liquid medium and 0.2mL bacterial inoculum, cultiv...

Embodiment 2

[0057] Example 2: Comparison of the degradation effects of the free combined flora and the sodium alginate immobilized combined flora on the degradation of benzo[a]pyrene

[0058] The strain P4 was inoculated in the R2A liquid medium, and the remaining 7 strains were inoculated in the glucose-inorganic salt liquid medium, and immobilized after culturing in a shaker at 28°C and 160 rpm for about 48 hours. In the ultra-clean workbench, add an equal volume of 4% sodium alginate solution to the bacterial solution of the 8 strains, stir slowly until the bacterial solution is uniform, use a sterile syringe to draw and inject 4% CaCl drop by drop 2 In the solution, let it stand for 5-10 minutes for hardening treatment, remove the pellet, wash it with sterile saline for 3 times, put it into a ground vial, and place it in a refrigerator at 4°C for later use. During this process, except for the sterilizing solution and sterile syringes, other supplies must be autoclaved at 121°C for 30 ...

Embodiment 3

[0061] Example 3: Comparison of the degradation effects of immobilized original flora and combined flora on benzo[a]pyrene

[0062] Weigh 5 g of fresh sediment into 50 mL of inorganic salt liquid medium containing benzo[a]pyrene (concentration: 10 mg / L), and enrich culture at 25 degrees Celsius and 180 rpm on a shaking table for 7 days. After several subcultures, the concentration of benzo[a]pyrene was increased to 50mg / L, so that the polycyclic aromatic hydrocarbon degrading functional bacteria were enriched, and the original flora was obtained.

[0063] In the ultra-clean workbench, add an equal volume of 4% sodium alginate solution to the original flora bacterial solution, stir slowly until the bacterial solution is uniform, use a sterile syringe to absorb and inject 4% CaCl drop by drop 2 In the solution, let it stand for 5-10 minutes for hardening treatment, remove the pellet, wash it with sterile saline for 3 times, put it into a ground vial, and place it in a refrigerat...

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Abstract

The invention discloses a preparation method of sodium alginate immobilized bacterial combination for polycyclic aromatic hydrocarbon pollution remediation. According to the method, benzo[a]pyrene isselected as a representative model of polycyclic aromatic hydrocarbon, sodium alginate pellets are adopted as carriers for immobilizing eight bacteria including rhodobacter P1, flavobacterium P2, rhizobium P3, Mucilaginibacter P4, flavobacterium P5, cupriavidus P6, cupriavidus P7 and cupriavidus P8, the sodium alginate pellets containing the eight bacteria are mixed in a mass ratio of P1, P2, P3,P4, P5, P6, P7 and P8 being 1:1:1:1:1:12:12:1, and a bacterial combination is obtained. The bacterial combination is obtained with a method of first immobilization and later mixing, and a ratio of strains in the bacteria combination is guaranteed while polycyclic aromatic hydrocarbon is prevented from being introduced in a culture process; bacteria are immobilized with a sodium alginate embeddingmethod, an antagonistic effect among the strains in the bacterial combination is reduced, and the efficiency of degrading polycyclic aromatic hydrocarbon by the bacteria combination is improved.

Description

【Technical field】 [0001] The invention relates to a preparation method of sodium alginate immobilized combined flora used for polycyclic aromatic hydrocarbon pollution restoration, and belongs to the technical field of environmental pollutant biological treatment. 【Background technique】 [0002] Polycyclic aromatic hydrocarbons are a class of persistent organic pollutants widely present in the environment, formed by two or more benzene rings in the form of shared ring edges, including naphthalene, phenanthrene, pyrene, benzo[a]pyrene, etc. 500 Various compounds. In my country, benzo[a]pyrene is also included as an environmental indicator in the "Environmental Quality Standards for Surface Water" (GB 3838-2002), "Ambient Air Quality Standards" (GB3095-2012) and "Soil Environmental Quality Agricultural Land Soil Pollution Risk". Control Standards (Trial)” (GB 15618-2018), 8 kinds of PAHs including benzo[a]anthracene and benzo[a]pyrene were included in the “Soil Environmental Q...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N11/10C02F3/34C12R1/01C02F101/32C12R1/20C12R1/41
CPCC12N1/20C02F3/34C02F2101/327C12N11/10
Inventor 黄家明吴群河麦顺之
Owner SUN YAT SEN UNIV
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