Biochar-based immobilized microbial agent and preparation method and application thereof

A technology for immobilizing microorganisms and biochar, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problems of easily hindering the contact between bacterial strains and pollutants, high cost, cumbersome operation of modified biochar carriers, etc. , to achieve the effect of less loss, high degradability and favorable preservation

Active Publication Date: 2019-04-16
YONKER ENVIRONMENTAL PROTECTION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, the operation of the modified biochar carrier is cumbersome and the cost is high; while the biochar bacterial agent is made into bacterial balls, which only has certain advantages in the preservation of the bacterial agent. In the actual application process, the external packaging material of the bacterial agent is easy to Hinder the contact between strains and pollutants, and improve the degradation ability of bacterial agents is very limited

Method used

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  • Biochar-based immobilized microbial agent and preparation method and application thereof
  • Biochar-based immobilized microbial agent and preparation method and application thereof
  • Biochar-based immobilized microbial agent and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Fermentation of Mycobacterium sp.CSC-6

[0047] Use an inoculation loop to pick a ring of strains from the LB solid plate of Mycobacterium sp.CSC-6 and inoculate it in 5mL of 1# liquid medium (peptone 10.0g, yeast powder 5.0g, sodium chloride 10.0g, deionized water 1000mL, pH 7.2-7.4), activate for 48h at 30°C in a shaking incubator with a rotation speed of 120r / min.

[0048] Take 8 mL of the activated CSC-6 bacterial solution and add it to 400 mL of 2# liquid medium (glucose 20.0 g, peptone 20.0 g, yeast powder 10.0 g, crude oil 2.0 g, deionized water 1000 mL, pH 7.2-7.4), Expand the culture in a shaking incubator at 30°C with a rotational speed of 120r / min for 48h to obtain seed liquid.

[0049] Take 400mL of seed liquid and add it to a fermenter with 3.6L of 2# liquid medium. The culture conditions are: ventilation ratio 0.5vvm, stirring speed 200r / min, temperature 30°C, pH 7.2-7.4, fermentation time 48h. After the fermentation finishes, centrifuge on a high-speed ...

Embodiment 2

[0051] Weigh 1.0g of 1# bacterial agent and add it into 1000mL of 3# liquid medium (5.0g of sucrose, 3.0g of beef extract, 1.0g of yeast powder, 1000mL of deionized water, 2.0g of petroleum, pH 7.2-7.4), at 30 ℃, in a shaking incubator with a rotational speed of 120r / min for 4 hours, then add 100g of biochar carrier, the microstructure of the biochar carrier is as follows: figure 1 As shown, after continuing to shake for 24 hours, use a disc centrifuge to remove excess 3# liquid medium to obtain 2# bacterial agent.

[0052] Add 50g of nutrient solution (oleic acid 0.4mL, glycerol 1.0g, sodium citrate 0.2g, disodium hydrogen phosphate 1.0g, potassium dihydrogen phosphate 1.0g, ammonium sulfate 0.5g, zinc sulfate 0.5mg, calcium chloride 0.5 mg, copper sulfate 0.5mg, magnesium sulfate 15.0mg) in 2# microbial agent, after 2h of adsorption, put in freeze dryer to remove excess water, get 3# bacterial agent. Microscopic picture of 3# bacterial agent figure 2 shown.

Embodiment 3

[0054] Degradation performance of biochar-based composite bacterial agent

[0055] The 1# bacterial agent, 2# bacterial agent, and 3# bacterial agent prepared in Examples 1 and 2 were used to carry out experiments on degrading petroleum pollutants in water bodies respectively. Prepare the inorganic salt medium (1.00g / L NH 4 Cl, 0.05g / L CaCl 2 , 0.10g / LMgSO 4 .7H 2 O, 1.00g / L K 2 HPO 4 , 1.00g / L KH 2 PO 4 ) 100mL, add 1# bacterial agent 0.01g to be experimental group 1, add 2# bacterial agent 1.0g to be experimental group 2, add 3# bacterial agent 1.0g to be experimental group 3, add 1.0g biochar + nutrient (oleic acid 0.4mL, glycerin 1.0g, sodium citrate 0.2g, disodium hydrogen phosphate 1.0g, potassium dihydrogen phosphate 1.0g, ammonium sulfate 0.5g, zinc sulfate 0.5mg, calcium chloride 0.5mg, copper sulfate 0.5mg, magnesium sulfate 15.0 mg) was the control group 1, adding 1.0 g of biochar was the control group 2, and no bacterial agent was added as the control group...

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Abstract

The invention relates to a biochar-based immobilized microbial agent, a preparation method and application thereof. The biochar-based immobilized microbial agent includes a biochar-based material, a bacterial body adsorbed on the biochar-based materialand a nutrient, the bacterial bodyis mycobacterium sp. CSC-6, and is deposited at the China Center for Type Culture Collection on 2017 November 27,and a deposit number isCCTCC No. M 2017726. The biochar-based immobilized microbial agent is used for degrading a petroleum. Compared with the prior art, the microbial agent prepared by a method of immobilizing an adsorbent andthe nutrient has a higher degradation ability to petroleum hydrocarbons, a degradation rate canreach 83.5%after degradingsewage containing 1.0 wt% of the petroleum for 7 days, the agent has large increase compared with the degradation rate of 52.0% of simple bacteria, and the biochar-based immobilized microbial agentis more conducive to the preservation of active bacteria.

Description

technical field [0001] The invention relates to the technical field of microbial bacterial agents, in particular to a biochar-based immobilized microbial bacterial agent and its preparation method and application. Background technique [0002] As petroleum and its derivatives are more and more widely used in daily life, the environmental pollution problems caused by petroleum extraction, refining and transportation are becoming more and more prominent. Therefore, the task of remediating petroleum pollutants is urgent. At present, bioremediation technology has become the most promising technology in the field of oil pollution remediation due to its advantages of simple operation, low cost, no damage to the soil environment, no secondary pollution and good treatment effect. The traditional bioremediation technology treats pollutants such as oil by directly adding oil-degrading bacteria strains, so as to achieve the purpose of purifying the repaired objects. However, there are...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N11/14C02F3/34C12R1/32
CPCC02F3/343C12N1/20C12N11/14C12N1/205C12R2001/32
Inventor 罗启仕闫淑兰赵秀红崔长征李梓冉雨灵吴晴雯任静
Owner YONKER ENVIRONMENTAL PROTECTION
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