Flavobacterium lcy-1 and its use

Abstract

The present invention relates to a Flavobacterium lcy-1 and a use thereof, wherein the Flavobacterium lcy-1 is a Flavobacterium ( Xanthobacter sp. )lcy‑1, deposited in China Center for Type Culture Collection, the deposit date is September 30, 2024, the deposit number is CCTCC NO: M 20242130, the address is: Wuhan University, Wuhan, China, zip code 430072. The Flavobacterium lcy‑1 can grow and reproduce with cyclohexane as the only carbon source and energy source and efficiently degrade the substrate, and has a certain degradation ability for a variety of volatile organic compounds, and performs well in a mixed substrate system. Therefore, the present invention has important significance in the engineering application of biological purification of volatile organic waste gases such as cyclohexane.

Description

Technical Field

[0001] The invention relates to the technical field of microorganisms, and in particular to Flavobacterium lcy-1 and uses thereof. Background Art

[0002] Cyclohexane is a volatile organic compound whose saturated ring structure makes it more stubborn than alkanes or monoaromatic compounds. Currently, there are three methods for treating this volatile organic compound: combustion, absorption, adsorption and biological methods. Among them, the biological method has received extensive attention in the removal of gaseous pollutants because of its mild reaction conditions, low secondary pollution, simple equipment and low operating costs. However, cyclohexane is not easily attacked by microorganisms due to the difficulty of its molecules to degrade; in addition, the low solubility and potential toxicity of cyclohexane also hinder the use of microorganisms. Therefore, there are currently very few microorganisms that can use cyclohexane as the sole carbon source and energy source. A small number of strains in the prior art, such as Nocardia sp. Although the strain can use cyclohexane as a carbon source, it needs to add biotin in the culture medium, and there are also disadvantages of slow degradation rate and slow microbial growth. The present invention screens a high-efficiency degradation strain that uses cyclohexane as the only carbon source and energy from the environment, providing strong support for the biological purification project containing such pollutants. Summary of the invention

[0003] The purpose of the present invention is to overcome the shortcomings of the prior art and to provide a Flavobacterium lcy-1 which uses cyclohexane as the only carbon source and energy source and can efficiently degrade cyclohexane.

[0004] A Flavobacterium lcy-1 of the present invention is a Flavobacterium ( Xanthobacter sp. )lcy-1, deposited in China Center for Type Culture Collection, the deposit date is September 30, 2024, the deposit number is CCTCC NO: M 20242130, address: Wuhan University, Wuhan, China, Postal Code 430072.

[0005] Furthermore, the Flavobacterium lcy-1 described in the present invention is rod-shaped, has a size of (0.4-0.8) μm×(1.3-1.9) μm, is spore-free, has flagella, is aerobic, and is Gram-negative.

[0006] Furthermore, the colonies of the Flavobacterium lcy-1 described in the present invention are dot-shaped, yellow, have neat edges, are translucent, smooth and moist.

[0007] The present invention also provides a use of any of the above-mentioned Flavobacterium lcy-1, wherein the Flavobacterium lcy-1 is used for degrading volatile organic matter.

[0008] Furthermore, in the use described in the present invention, the Flavobacterium lcy-1 is used to prepare a bacterial agent for degrading volatile organic compounds, and the bacterial agent includes Flavobacterium lcy-1.

[0009] Furthermore, in the use of the present invention, the volatile organic compounds include one or more of alkane compounds, cycloalkane compounds, ether compounds, ester compounds and aromatic compounds. Alkane compounds such as hexane, heptane and nonane; cycloalkane compounds such as methylcyclohexane and 1,4-dimethylcyclohexane; ether compounds such as tetrahydrofuran; ester compounds such as ethyl acetate, aromatic compounds such as chlorobenzene and o-xylene.

[0010] Furthermore, in the use described in the present invention, the volatile organic compound includes cyclohexane.

[0011] Furthermore, in the use described in the present invention, the use specifically includes using the bacterial liquid obtained by expanding the culture of Flavobacterium lcy-1 as the enzyme source, cyclohexane as the substrate, and an inorganic salt culture medium as the medium, and degrading cyclohexane under the conditions of pH = 5 to 9 and temperature of 20°C to 40°C.

[0012] Further, in the use described in the present invention, when the initial bacterial count is OD 600 =0.02, pH=5~8, temperature of 20℃~35℃, cyclohexane concentration of 20~80 mg / L, and degradation time of 45 hours, the degradation rate of cyclohexane is 90%~95%.

[0013] Further, in the use described in the present invention, when the initial bacterial count is OD 600 =0.02, pH=5-8, temperature of 20°C-35°C, and degradation time of 48 hours, the degradation rate of mixed system 1 is greater than or equal to 96%; the degradation rate of mixed system 2 is 65%-75%; wherein mixed system 1 is a mixed system of n-hexane and cyclohexane, and the concentrations of n-hexane and cyclohexane are both 20-80 mg / L; the mixed system 2 is a mixed system of cyclohexane and methylcyclohexane, and the concentrations of cyclohexane and methylcyclohexane are both 20-80 mg / L.

[0014] Beneficial effects of the present invention:

[0015] The present invention provides a highly efficient degradation bacterium of organic matter such as cyclohexane, namely, Flavobacterium lcy-1, which can grow and reproduce with cyclohexane as the only carbon source and energy source and efficiently degrade the substrate. 600 =0.02, it can completely degrade 50 mg / L cyclohexane. The growth temperature and pH range of this strain is wide. 600 =0.02, the pH range of 6.0 to 7.0 was better for the degradation of cyclohexane.

[0016] When the initial bacterial count is OD 600 =0.02, pH=5~8, temperature of 20℃~35℃, cyclohexane concentration of 20~80mg / L, and degradation time of 45 hours, the degradation rate of cyclohexane is 90%~95%.

[0017] It is also very effective in degrading a variety of volatile organic compounds, with the best performance for nonane, methylcyclohexane and ethyl acetate, with a degradation rate of over 89%.

[0018] Moreover, it can degrade systems including multiple mixed substrates. For example, when the initial bacterial count is OD 600 =0.02, pH=5-8, temperature of 20°C-35°C, and degradation time of 48 hours, the degradation rate of mixed system 1 is greater than or equal to 96%; the degradation rate of mixed system 2 is 65%-75%; wherein mixed system 1 is a mixed system of n-hexane and cyclohexane, and the concentrations of n-hexane and cyclohexane are both 20-80 mg / L; the mixed system 2 is a mixed system of cyclohexane and methylcyclohexane, and the concentrations of cyclohexane and methylcyclohexane are both 20-80 mg / L.

[0019] In view of the fact that the strain has a certain degradation ability for a variety of volatile organic compounds in addition to cyclohexane, and performs well in a mixed substrate system, the present invention is of great significance in the engineering application of biological purification of volatile organic waste gases such as cyclohexane. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 This is a morphological diagram of the Flavobacterium lcy-1 of the present invention;

[0021] Figure 2 is a phylogenetic tree of Flavobacterium lcy-1 of the present invention;

[0022] Figure 3 It is a bacterial growth and cyclohexane degradation curve of Flavobacterium lcy-1 of the present invention;

[0023] Figure 4 The effect of different pH values ​​of culture medium on cyclohexane degradation (A), growth (B), and mineralization (C) of Flavobacterium lcy-1;

[0024] Figure 5 The effect of different temperatures on cyclohexane degradation (A), growth (B), and mineralization (C) of Flavobacterium lcy-1;

[0025] Figure 6 The degradation (A), growth (B), and mineralization (C) of Flavobacterium lcy-1 at different initial cyclohexane concentrations;

[0026] Figure 7 This is a diagram showing the degradation effect of Flavobacterium lcy-1 on different substrates;

[0027] Figure 8 Figure 2 shows the degradation (A), mineralization and growth (B) of different substrates of Flavobacterium lcy-1 including mixed system 1 and mixed system 2. DETAILED DESCRIPTION

[0028] In order to have a clearer understanding of the technical features, purposes and effects of the present invention, the specific implementation methods of the present invention are now described in detail with reference to the accompanying drawings. The described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention. If the specific conditions are not specified in the specific implementation method, they shall be carried out according to the conventional conditions or the conditions provided by the manufacturer.

[0029] When the following description refers to the drawings, unless otherwise indicated, the same numbers in different drawings represent the same or similar elements. The embodiments described in the following exemplary embodiments do not represent all embodiments consistent with the present disclosure. Instead, they are merely examples of devices and methods consistent with some aspects of the present disclosure as detailed in the appended claims. The terms used in the present disclosure are only for the purpose of describing specific embodiments and are not intended to limit the present disclosure.

[0030] The present invention is further described in detail below in conjunction with specific embodiments.

[0031] Implementation method 1:

[0032] A Flavobacterium lcy-1, which is a Flavobacterium ( Xanthobacter sp. )lcy-1, deposited in China Center for Type Culture Collection, the deposit date is September 30, 2024, the deposit number is CCTCC NO: M 20242130, address: Wuhan University, Wuhan, China, Postal Code 430072.

[0033] In other embodiments, the Flavobacterium lcy-1 is rod-shaped, with a size of (0.4-0.8) μm×(1.3-1.9) μm, no spores, and flagella; it is aerobic and Gram-negative.

[0034] In other embodiments, the colonies of the Flavobacterium lcy-1 are dot-shaped, yellow, have neat edges, are translucent, smooth and moist.

[0035] Implementation method 2:

[0036] A use of the Flavobacterium lcy-1 as described in any of the above embodiments, wherein the Flavobacterium lcy-1 is used to degrade volatile organic compounds.

[0037] In other embodiments, the Flavobacterium lcy-1 is used to prepare a bacterial agent for degrading volatile organic compounds, and the bacterial agent includes Flavobacterium lcy-1.

[0038] In other embodiments, the volatile organic matter includes one or more of alkane compounds, cycloalkane compounds, ether compounds, ester compounds and aromatic compounds. Alkane compounds such as hexane, heptane and nonane; cycloalkane compounds such as methylcyclohexane and 1,4-dimethylcyclohexane; ether compounds such as tetrahydrofuran; ester compounds such as ethyl acetate, aromatic compounds such as chlorobenzene and o-xylene.

[0039] In other embodiments, the volatile organic compound comprises cyclohexane.

[0040] In other embodiments, the use specifically includes using the bacterial liquid obtained by expanding the culture of Flavobacterium lcy-1 as the enzyme source, cyclohexane as the substrate, and an inorganic salt culture medium as the medium to degrade cyclohexane at pH = 5 to 9 and a temperature of 20°C to 40°C.

[0041] In other embodiments, when the initial bacterial count is OD 600 =0.02, pH=5~8, temperature of 20℃~35℃, cyclohexane concentration of 20~80 mg / L, and degradation time of 45 hours, the degradation rate of cyclohexane is 90%~95%.

[0042] In other embodiments, when the initial bacterial count is OD 600 =0.02, pH=5-8, temperature of 20°C-35°C, and degradation time of 48 hours, the degradation rate of mixed system 1 is greater than or equal to 96%; the degradation rate of mixed system 2 is 65%-75%; wherein mixed system 1 is a mixed system of n-hexane and cyclohexane, and the concentrations of n-hexane and cyclohexane are both 20-80 mg / L; the mixed system 2 is a mixed system of cyclohexane and methylcyclohexane, and the concentrations of cyclohexane and methylcyclohexane are both 20-80 mg / L.

[0043] In the following embodiments:

[0044] The inorganic salt culture medium used is composed of: Na2HPO4·12H2O 4.5 g / L, KH2PO4 1.0 g / L, (NH4)2SO42.5 g / L, MgSO4·7H2O 0.2 g / L, anhydrous CaCl2 0.023 g / L, trace element mother solution 1 mL / L, pH=7.0, and the solvent is deionized water; the concentration composition of the trace element mother solution is: FeSO4·7H2O 1.0 g / L, CuSO4·5H2O 0.02 g / L, H3BO3 0.014 g / L, MnSO4·4H2O 0.10 g / L, ZnSO4·7H2O 0.10 g / L, Na2MoO4·2H2 O 0.02 g / L, CoCl2·6H2O 0.02 g / L, and the solvent is deionized water.

[0045] The R2A solid culture medium is composed of 0.5 g / L yeast powder, 0.5 g / L starch, 0.5 g / L glucose, 0.5 g / L peptone, 0.3 g / L sodium pyruvate, 0.3 g / L K2HPO4, 0.05 g / L MgSO4·7H2O and 28 g / L agar, the solvent is deionized water, and the pH value is natural.

[0046] The R2A liquid culture medium is composed of 0.5 g / L yeast powder, 0.5 g / L starch, 0.5 g / L glucose, 0.5 g / L peptone, 0.3 g / L sodium pyruvate, 0.3 g / L K2HPO4 and 0.05 g / L MgSO4·7H2O, the solvent is deionized water, and the pH value is natural.

[0047] The room temperature mentioned in the present invention refers to 25-30°C.

[0048] Embodiment 1:

[0049] (1) Isolation and purification of Flavobacterium lcy-1:

[0050] Activated sludge was collected from Zhenhai Refining and Chemical Company (Sinopec Group). The lower layer of sludge after standing was mixed with inorganic salt culture medium at a ratio of 1:1 (v / v). 3 L of the mixture was inoculated into a 5 L sludge acclimation tank. Cyclohexane was used as the carbon source and energy source. 1 mL of cyclohexane was added every morning, noon and evening. The acclimation was carried out at room temperature. Part of the inorganic salt was replaced every two days. After 30 days, 10 mL of sludge was taken from the acclimation tank and added into a shake flask containing 40 mL of inorganic salt culture medium. It was found that the acclimated sludge could stably degrade 20 mg / L cyclohexane per day in the shake flask, and the degradation rate could reach 70-80% (shake flask experimental conditions: 30°C, 160 r / min). This sample was transferred to a shake flask for further enrichment to obtain an acclimated sample.

[0051] 5 mL of the sludge sample in the shake flask was transferred to a shake flask containing 45 mL of inorganic salts for further performance testing. It was found that the transferred sludge could still stably degrade 20 mg / L cyclohexane in the shake flask every day. The sludge sample was further transferred, and 2 mL of the sludge sample in the shake flask was transferred to a shake flask containing 50 mL of inorganic salts for further enrichment. After multiple passages and enrichment (1 mL each time), the sludge was diluted and spread, and the concentration was 10 -1 ~10 -6 The plate was spread at multiple times, and a single colony was picked. Then, the degradation activity was determined using cyclohexane as a substrate. After separation and purification, a strain lcy-1 with cyclohexane degradation activity was obtained.

[0052] (2) Identification of Flavobacterium lcy-1:

[0053] The colonies of the Flavobacterium lcy-1 are small dots, yellow, with neat edges, translucent, smooth and moist, and easy to pick up; the Flavobacterium lcy-1 is rod-shaped, with a size of (0.4-0.8) μm×(1.3-1.9) μm, no spores, and flagella. Figure 1 As shown;

[0054] The physiological and biochemical characteristics of the Flavobacterium lcy-1 are: aerobic and Gram-negative.

[0055] The 16S rDNA sequencing of the Flavobacterium lcy-1 was performed by Sangon Biotech (Shanghai) Co., Ltd. The 16S rDNA sequence of the strain (SEQ ID NO. 1) is as follows:

[0056] (Genbank accession number is PQ517068):

[0057]

[0058] The 16S rDNA sequence of strain lcy-1 was uploaded to the Ezbiocloud.net website and compared with the standard strains on the website. The bacterial phylogenetic tree was constructed using the Neighbor-Joining method using MEGA-X software and evaluated using the Bootstrap method (repeated 1000 times). The constructed phylogenetic tree is shown in Figure 2. Figure 2 , thus determining that strain lcy-1 is Xanthobacter , named Flavobacterium ( Xanthobacter sp. )lcy-1, deposited in China Center for Type Culture Collection, the deposit date is September 30, 2024, the deposit number is CCTCC NO: M 20242130, address: Wuhan University, Wuhan, China, Postal Code 430072.

[0059] (3) Expansion culture of Flavobacterium lcy-1:

[0060] 1) Slant culture: Inoculate Flavobacterium lcy-1 onto slant R2A solid culture medium and culture at 30°C for 36-48 h to obtain slant bacteria.

[0061] 2) Expanded culture: Use an inoculation loop to pick the slant bacteria obtained in step 1) and inoculate them into R2A liquid culture medium. Cultivate at 30°C for 36-48 hours to obtain the OD 600 = 0.1~0.2 of bacterial solution.

[0062] Embodiment 2:

[0063] Cyclohexane degradation using Flavobacterium lcy-1 described in Example 1:

[0064] Cyclohexane was used as the sole carbon source for Flavobacterium lcy-1, and the OD 600 =0.1 bacterial solution, inoculated into 50 mL of fresh inorganic salt medium containing 50 mg / L cyclohexane, so that the initial bacterial concentration is OD 600 The pH value was 0.02 and 7. The culture was placed in a shaker at 30°C and 160 r / min. Samples were taken every 6 hours to determine the cyclohexane degradation rate. A portion of the bacterial solution was drawn out with a 5 mL syringe to determine the bacterial OD value. The results are shown in the table. Figure 3 It can be seen that with the extension of time, the bacterial concentration gradually increased. When cyclohexane was used as the substrate, the bacterial concentration reached a maximum of about OD 600 =0.1395, and the degradation rate of cyclohexane reached more than 95%. This example shows that Flavobacterium lcy-1 can use cyclohexane as the only carbon source and energy source for growth and reproduction, and has the ability to degrade cyclohexane stably and efficiently.

[0065] Embodiment 3:

[0066] The difference between Example 3 and Example 2 is that the inorganic salt medium is adjusted with 1 mol / L NaOH aqueous solution or 1 mol / L H2SO4 aqueous solution, and the bacterial solution prepared by the method of Example 1 is inoculated under the condition that the initial cyclohexane concentration is 50 mg / L, so that the initial bacterial cell concentration in each parallel sample is OD 600 The calculated value is 0.02, pH = 6. The results are as follows Figure 4 As shown in the figure, the degradation rate of cyclohexane by Flavobacterium lcy-1 was over 95%. At the same time, the growth amount and CO2 value produced by Flavobacterium lcy-1 also showed a consistent trend with the cyclohexane degradation rate.

[0067] Embodiment 4:

[0068] The difference between Example 4 and Example 3 is that pH = 5. Figure 4 shown.

[0069] Embodiment 5:

[0070] The difference between Example 5 and Example 3 is that pH = 8. Figure 4 shown.

[0071] Embodiment 6:

[0072] The only difference between Example 6 and Example 3 is that the sample is placed in a shaker at 20°C for constant temperature shaking culture. Figure 5 shown.

[0073] Embodiment 7:

[0074] The difference between Example 7 and Example 6 is that the sample is placed in a shaker at 25°C for constant temperature shaking culture. Figure 5 shown.

[0075] Embodiment 8:

[0076] The difference between Example 8 and Example 6 is that the sample is placed in a shaker at 35°C for constant temperature shaking culture. Figure 5 shown.

[0077] Embodiment 9:

[0078] The difference between Example 9 and Example 3 is that different concentrations of cyclohexane were added to the fresh inorganic salt medium to make the initial substrate concentration 20 mg / L. Figure 6 shown.

[0079] Embodiment 10:

[0080] The difference between Example 10 and Example 3 is that different concentrations of cyclohexane were added to the fresh inorganic salt medium to make the initial substrate concentration 80 mg / L. Figure 6 shown.

[0081] Embodiment 11:

[0082] At the initial bacterial count OD 600 = 0.02, hexane, heptane, nonane, methylcyclohexane, 1,4-dimethylcyclohexane, tetrahydrofuran, ethyl acetate, chlorobenzene and o-xylene were added to the inorganic salt medium respectively, and the final concentration was controlled to be 50 mg / L. After constant temperature shaking culture for 48 h, the substrate degradation and strain growth were observed. Figure 7 As shown in the figure, the substrate utilization range of Flavobacterium lcy-1 is relatively wide. Except for tetrahydrofuran, different degrees of turbidity can be observed in inorganic salts containing other substrates, indicating that Flavobacterium lcy-1 can use the above substrates to grow and reproduce. Although Flavobacterium lcy-1 is separated and purified using cyclohexane as the only carbon source, it has a certain ability to degrade a variety of volatile organic compounds. Among them, it has the best degradation performance for nonane, methylcyclohexane and ethyl acetate, with a degradation rate of more than 89%; it has a certain degradation ability for hexane and 1,4-dimethylcyclohexane; and it has a certain tolerance to heptane, chlorobenzene and o-xylene.

[0083] Embodiment 12:

[0084] At the initial bacterial count OD 600 = 0.02, design substrate mixed system 1: n-hexane and cyclohexane mixed system, and set single substrate system (n-hexane, cyclohexane and methylcyclohexane) as the control group, control the concentration of single substrate in each system to 50 mg / L. During the experiment, design 2 parallel samples, and culture them in a shaker at 30℃ and 160 r / min, and measure the concentration of each substrate and the final OD of the system. 600 , CO2 value. Figure 8 As shown in the figure, in the mixed system 1, the degradation rate of cyclohexane in 48 h reached 96.7%, which was slightly higher than the degradation rate of cyclohexane in the control group (96.1%), while the degradation rate of n-hexane was lower, indicating that Flavobacterium lcy-1 preferentially degraded cyclohexane and had a stronger degradation ability for cyclohexane. In addition, n-hexane did not compete with cyclohexane and had little effect on the degradation of cyclohexane by Flavobacterium lcy-1. Figure 8 Medium OD 600 It can also be proved that the decrease in mineralization rate may be due to the inhibition of the mineralization process by degradation products.

[0085] Embodiment 13:

[0086] The difference between Example 13 and Example 12 is that the substrate mixed system 2 is designed as a mixed system of cyclohexane and methylcyclohexane. The results are the same as Figure 8 As shown. It can be seen that compared with the single substrate system, the degradation rates of methylcyclohexane and cyclohexane in mixed system 2 are reduced to varying degrees. This may be due to the similarity of the two structures. During the degradation process, they compete for the active site of the same enzyme, resulting in a decrease in enzyme activity, which in turn affects the degradation efficiency. Figure 8 It can be seen that the competitive inhibition between the two pollutants also affected the mineralization process. Although the degradation rate of each pollutant was reduced, Flavobacterium lcy-1 could degrade methylcyclohexane and cyclohexane at the same time, and the degradation rate could be stabilized at 65%~75% in 48 h.

[0087] Comparative Example 1

[0088] The difference between this comparative example 1 and example 3 is that pH = 9. Figure 4 As shown, the degradation rate was low, only 16%.

[0089] Comparative Example 2

[0090] The difference between this comparative example 2 and example 6 is that the sample is placed in a shaker at 40°C for constant temperature shaking culture. Figure 5 As shown, the degradation rate is also low, about 70%.

[0091] The present invention is described by the above specific embodiments, and those skilled in the art should understand that various changes and equivalent substitutions can be made to the present invention without departing from the scope of the present invention. The parts not described in detail in the present specification are well-known technologies to those skilled in the art. In addition, various modifications can be made to the present invention for specific situations or specific circumstances without departing from the scope of the present use. Therefore, the present invention is not limited to the specific embodiments disclosed, but should include all implementation methods falling within the scope of the claims of the present invention.

Claims

1. A Flavobacterium lcy-1, characterized in that The Flavobacterium lcy-1 is a Flavobacterium ( Xanthobacter sp. )lcy-1, deposited in China Center for Type Culture Collection, the deposit date is September 30, 2024, the deposit number is CCTCC NO: M 20242130, address: Wuhan University, Wuhan, China, Postal Code 430072.

2. A use of Flavobacterium lcy-1 as claimed in claim 1, characterized in that: The Flavobacterium lcy-1 is used for degrading one or more of hexane, heptane, nonane, methylcyclohexane, 1,4-dimethylcyclohexane, cyclohexane, ethyl acetate, chlorobenzene, n-hexane and o-xylene.

3. The use according to claim 2, characterized in that: The Flavobacterium lcy-1 is used to manufacture a bacterial agent for degrading one or more of hexane, heptane, nonane, methylcyclohexane, 1,4-dimethylcyclohexane, cyclohexane, ethyl acetate, chlorobenzene, n-hexane and o-xylene, and the bacterial agent includes the Flavobacterium lcy-1.

4. The use according to claim 3, characterized in that The use specifically includes using the bacterial liquid obtained by expanding the culture of Flavobacterium lcy-1 as the enzyme source, cyclohexane as the substrate, and an inorganic salt culture medium as the medium to degrade cyclohexane under the conditions of pH=5-9 and temperature of 20°C-40°C.

5. The use according to claim 4, characterized in that The initial bacterial count was OD 600 =0.02, pH=5~8, temperature of 20℃~35℃, cyclohexane concentration of 20~80 mg / L, and degradation time of 45 hours, the degradation rate of cyclohexane is 90%~95%.

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