Method of extracting crude enzyme preparation for degrading polycyclic aromatic hydrocarbons

An extraction method and polycyclic aromatic hydrocarbon technology, applied in the field of environmental restoration, can solve the problem of high cost, and achieve the effect of low cost, quick effect and great application value

Inactive Publication Date: 2010-09-29
INST OF SOIL SCI CHINESE ACAD OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Purpose of the invention: Aiming at the problem of high cost of laccase in environmental restoration, this patent proposes a method for extracting oxidase such as laccase from mushroom dregs. This crude enzyme preparation can be used for the degradation of polycyclic aromatic hydrocarbons efficiently

Method used

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  • Method of extracting crude enzyme preparation for degrading polycyclic aromatic hydrocarbons
  • Method of extracting crude enzyme preparation for degrading polycyclic aromatic hydrocarbons
  • Method of extracting crude enzyme preparation for degrading polycyclic aromatic hydrocarbons

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0014] Embodiment 1: the extraction of crude enzyme liquid

[0015] 1.1 Mushroom dregs tested

[0016] Mushroom residues of Agaricus bisporus and Pleurotus eryngii were cultivated in our laboratory, and samples of mushroom residues of Pleurotus ostreatus and Coprinus comatus were purchased from Henan Academy of Agricultural Sciences. The samples were stored at 4°C after acquisition until use.

[0017] 1.2 Extraction of crude enzyme solution

[0018] The extraction step is: adding an acidic buffer solution with a pH of 4 to 6 in Agaricus bisporus, Pleurotus eryngii, Pleurotus eryngii or Coprinus comatus slag, the acidic buffer is acetic acid, phosphoric acid or citric acid (preferably pH 5, 50mM acetic acid buffer solution), shake at 20-40°C for 2 hours at a speed of 100-200 rpm, centrifuge at 11,000 g for 10 minutes at a high speed, and the obtained supernatant is the crude enzyme solution.

[0019] Or, take 50g of fresh mushroom dregs into a 500mL conical flask, add 250mL ...

Embodiment 2

[0020] Embodiment 2: Crude enzyme liquid is to the degradation kind of 15 kinds of PAHs

[0021] 2.1 Test method

[0022] Take 4.5 mL of the crude enzyme solution obtained in step 1.2 and place it in a 15 mL brown reagent bottle with a stopper, add 0.5 mL of acetonitrile solution containing 15 kinds of PAHs (see Table 1 for the initial concentration of PAHs in the reaction system), tighten the stopper, shake well, and place Incubate in a dark room for 48 hours (25° C.), and add 5 mL of acetonitrile to terminate the reaction. Shake well, let stand for 1h (25°C, 150rpm), centrifuge at high speed (11,000g), pass through a 0.22μm filter membrane, and perform HPLC determination. As a control, the crude enzyme solution was added in the form of inactivation (boiling for 30 min), and each treatment was repeated three times.

[0023] 2.2 Sample analysis

[0024]The samples were analyzed by Shimadzu UFLC-20 ultra-high pressure liquid chromatography. The analytical column is a macrop...

Embodiment 3

[0027] Example 3: Degradation of high concentration anthracene and benzo[a]pyrene by crude enzyme solution

[0028] 3.1 Test method

[0029] Put 4.5 mL of the crude enzyme solution obtained in step 1.2 into a 15 mL brown reagent bottle with a stopper, add 0.5 mL of acetonitrile solution containing anthracene and benzo[a]pyrene, and make the anthracene and benzo[a]pyrene in the reaction system reach 1000 μgl -1 , other steps are the same as 2.1.

[0030] 3.2 Sample analysis

[0031] Same as 2.2.

[0032] 3.3 Statistics

[0033] Same as 2.3.

[0034] Table 1 is the Chinese name, ring number, solubility and initial concentration in the reaction system of the 15 polycyclic aromatic hydrocarbons to be tested.

[0035] Table 1 Initial concentrations of 15 PAHs

[0036]

[0037] Table 2 shows the degradation rate of 15 kinds of PAHs by the extracted crude enzyme solution. The values ​​in the table are: mean ± standard deviation, and those marked with the same l...

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Abstract

The invention discloses a method of extracting a crude enzyme preparation for degrading polycyclic aromatic hydrocarbons, which comprises the following steps of: adding acidic buffer with pH of between 4 and 6 into fresh spent mushroom substrates; oscillating the mixed solution for 2 hours at the temperature of between 20 and 40 DEG C and at the rotating speed of between 100 and 200 rpm; and centrifuging the oscillated product at a high speed for 10 minutes with a centrifugal force of 11, 000 gravity to obtain supernatant which is the crude enzyme liquid. The invention provides a polycyclic aromatic hydrocarbon removing method which has good and quick effects, low cost and a great application value in the field of environmental remediation. Meanwhile, laccase contained in the crude enzyme preparation has various types of degradant substrates, so the method can be applied in the remediation of pollution caused by polycyclic aromatic hydrocarbon pollutants and the remediation of the pollution caused by other organic pollutants such as polychlorinated biphenyl and chlorophenol.

Description

1. Technical field [0001] The invention belongs to the technical field of environmental restoration, and in particular relates to an extraction method for removing polycyclic aromatic hydrocarbon polluted enzyme preparations. 2. Background technology [0002] Persistent organic pollutants (POPs) are a class of natural or synthetic organic pollutants that are toxic, bioaccumulative, and refractory and can persist in the environment (Lohmann R, Breivik K, Dachs J and Muir D. Global fate of POPs: Current and future research directions. Environmental Pollution. 2007(150): 150-165). With the increase of human activities, POPs are inevitably dispersed into the atmosphere, soil, water and sediments, posing risks to the ecological environment (El-Shahawi MS, Hamza A, Bashammakh AS and Al-Saggaf WT.An overview on the accumulation, distribution, transformations, toxicity and analytical methods for the monitoring of persistent organic pollutants. Talanta. 2010(80): 1587-1597). Polycy...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/02A62D3/02A62D101/20
Inventor 李烜桢林先贵张晶
Owner INST OF SOIL SCI CHINESE ACAD OF SCI
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