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Preparation and application of bacterium-enzyme composite preparation for petroleum hydrocarbon degradation

A compound preparation and technology of petroleum hydrocarbons, applied in the field of bioremediation of soil environment, can solve the problems such as short-efficiency and low-efficiency of microbial remediation technology, easy inactivation of biological enzymes, short-term action, etc. Harmony, the effect of improving the degradation efficiency

Active Publication Date: 2021-01-22
SHENYANG INST OF APPL ECOLOGY CHINESE ACAD OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, although the petroleum hydrocarbon degradation process of enzyme preparations is rapid, the effect is short-lived, especially in multi-tropic heterogeneous soil environments, the disadvantages of biological enzymes being easily inactivated by environmental factors such as soil pH, temperature, and electrolyte strength have emerged. undoubtedly
Although the immobilization technology of biological enzymes has effectively improved the side effects of environmental factors on biological enzymes to a certain extent, the application of enzyme preparations is still difficult to make up for the short-term and low-efficiency problems of microbial restoration technology.

Method used

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  • Preparation and application of bacterium-enzyme composite preparation for petroleum hydrocarbon degradation
  • Preparation and application of bacterium-enzyme composite preparation for petroleum hydrocarbon degradation
  • Preparation and application of bacterium-enzyme composite preparation for petroleum hydrocarbon degradation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 2

[0036] The preparation of embodiment 2 compound preparations

[0037] 1) Enzyme preparation of Variovorax sp.JWLC1 (Variovorax sp.JWLC1)

[0038] The screened Variovorax sp. JWLC1 (Variovorax sp. JWLC1) was inoculated in 1L of beef extract peptone medium with a pH of 7.0, and cultured with constant temperature shaking at 30°C and 190rpm for 4 days. When the OD value of the fermentation culture medium reaches 2.1, the fermentation culture is stopped, and the fermentation culture is subjected to ultrasonic crushing treatment under ice bath conditions. The treatment condition is 330W, the total treatment time is 35min, and the broken solid-liquid mixture is processed at 9500rpm Centrifuge for 10 minutes to separate the solid and liquid, and collect the supernatant liquid as the whole enzyme solution; then collect the solid components such as cell fragments, and perform anaerobic high-temperature carbonization treatment at 300°C for 2 hours to obtain the solid components in the en...

Embodiment 3

[0045] The shake flask test function verification of embodiment 3 bacterium-enzyme composite preparation

[0046] The above-mentioned embodiment obtains composite preparation and carries out petroleum hydrocarbon degradation test in shake flask, and the bacteria agent and enzyme agent in the composite preparation add respectively:

[0047] Adopt screening medium formula (that is, the embodiment records inorganic salt screening liquid medium), the petroleum hydrocarbon initial content is 2.5g / L, at first the inoculum in the compound preparation in the embodiment 2 is added (its addition is 3% (according to Bacteria dosing ratio meter, mL / g)), 30 ℃ under 190rpm shaking culture treatment for 7 days, the petroleum hydrocarbon degradation rate reached 51.3%; after 7 days of treatment, add enzyme agent in the compound preparation (the addition of enzyme agent is the same as the embodiment 2 in the compound preparation added in the calculation of the mixing ratio of the two), and the...

Embodiment 4

[0050] Application Verification of Example 4 Bacteria-Enzyme Composite Preparation in Petroleum Hydrocarbon Contaminated Soil Remediation

[0051]Seven treatments were set in the embodiment, which were control group (CK), single microbial treatment group (Bio), single enzyme treatment group (En), secondary microbial replenishment treatment group (Bio+Bio), bacterial agent application combined with killing Live enzyme supplement treatment group (Bio+dEn), bacterial agent and enzyme supplement treatment group (Bio-En), and bacterial agent application combined with enzyme supplement treatment group (Bio+En), the specific parameters of the test settings are shown in Table 1 shown.

[0052] Prepare the bacterial agent according to the operation process of Example 3, cultivate 500mL of the bacterium of the genus Vultiva JWLC1 at 30°C, apply it to 5kg of contaminated soil containing 3.2% of petroleum hydrocarbon content, wherein the OD value of the bacterial solution reaches 2.2, app...

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Abstract

The invention belongs to the field of soil environment bioremediation, and further develops a bacteria-enzyme composite preparation for biodegradation of petroleum hydrocarbon pollutants in petroleumhydrocarbon contaminated soil, so as to realize efficient and rapid petroleum hydrocarbon contaminated soil bioremediation. The composite preparation is a microbial inoculum with petroleum hydrocarbondegradation capability and an enzyme agent obtained by crushing the microbial inoculum; the microbial inoculum is a culture solution which is obtained by carrying out fermentation culture on Variovorax sp. JWLC1 at the temperature of 30 + / -1 DEG C until the OD value is 1.5-2.5, wherein the volume ratio of the fermentation culture solution in the microbial inoculum to the fermentation culture solution in the enzyme agent is 1: 1 to 1: 3. The bacteria-enzyme composite preparation disclosed by the invention remarkably prolongs the microbial efficient degradation process of the petroleum hydrocarbon and improves the bioremediation efficiency of the petroleum hydrocarbon contaminated soil.

Description

technical field [0001] The invention belongs to the field of bioremediation of soil environment, and furthermore, develops a bacteria-enzyme compound preparation for biodegradation of petroleum hydrocarbon pollutants in petroleum hydrocarbon polluted soil, so as to realize efficient and rapid bioremediation of petroleum hydrocarbon polluted soil. Background technique [0002] The continuous progress of the oil industry has contributed to the vigorous development of the social economy and brought considerable economic benefits. However, there have been environmental pollution problems to varying degrees. Among them, in the process of crude oil extraction, transportation, and processing and production of petrochemical products, oil pollution of the soil has occurred, resulting in damage to the ecological environment of the soil and loss of function, which urgently needs to be repaired and treated. In this regard, a variety of remediation technologies for oil-contaminated soil...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20B09C1/10C12R1/01
CPCC12N1/20B09C1/10Y02W10/10
Inventor 王卅郭书海程凤莲荆佳维
Owner SHENYANG INST OF APPL ECOLOGY CHINESE ACAD OF SCI
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