Application of aspergillus in flammable and explosive gas metabolism

Abstract

The invention belongs to the technical field of microorganisms, and particularly relates to application of aspergillus in the aspect of flammable and explosive gas metabolism. The aspergillus S8 has good absorption and metabolism effects on flammable and explosive gases such as hydrogen and methane and mixed gases containing the flammable and explosive gases, and can be metabolized by taking the flammable and explosive gases or the mixed gases containing the flammable and explosive gases as a carbon source and / or an energy source; furthermore, the effect of efficiently treating flammable and explosive gas and mixed gas thereof is achieved, the environmental safety is high, the removal effect is good, use is convenient, no toxic by-product exists, and application and conversion are convenient.

Description

Technical Field

[0001] The invention belongs to the technical field of microorganisms, and particularly relates to the application of Aspergillus in the metabolism of flammable and explosive gases. Background Art

[0002] Flammable and explosive gases refer to gases that can form a combustible mixture with air, with an explosion range of 20% to 80%. Common flammable and explosive gases include hydrogen (H2), methane (CH4), and carbon monoxide (CO). Because flammable and explosive gases are easy to leak and flammable, with low explosion limits, they are extremely dangerous and can cause industrial accidents or fires, posing a serious threat to local ecosystems and human safety. Therefore, these gases need to be distinguished and classified during production, transportation, storage, and use.

[0003] Research has found that microorganisms play an important role in the decomposition and metabolism of flammable and explosive gases, which can further regulate the ecosystem. Furthermore, the process of using microorganisms to decompose flammable and explosive gases is convenient and easy to operate, without any toxic side effects.

[0004] Aspergillus ( Aspergillus ) is a filamentous fungus that is widely distributed in grains, air, soil and various organic substances. It is an important species in the fermentation industry and food processing industry. Nearly 60 species have been used, mainly for winemaking and vinegar making. However, its application in gas metabolism, especially in the metabolism of flammable and explosive gases, is relatively rare. Summary of the Invention

[0005] The purpose of the present invention is to provide an application of Aspergillus in the metabolism of flammable and explosive gases. The Aspergillus S8 described in the present invention has a good absorption and metabolism effect on flammable and explosive gases and mixed gases containing flammable and explosive gases, thereby increasing the biological uses of Aspergillus S8.

[0006] The present invention provides use of Aspergillus S8 in gas metabolism, wherein the gas comprises hydrogen or a mixed gas containing flammable and explosive gases; the flammable and explosive gases comprise hydrogen and / or methane; the Aspergillus S8 has a preservation number of CGMCC NO.40828.

[0007] The present invention also provides the use of a bacterial agent containing Aspergillus S8 in gas metabolism, wherein the gas includes hydrogen or a mixed gas containing flammable and explosive gases; the flammable and explosive gases include hydrogen and / or methane; the preservation number of Aspergillus S8 is CGMCC NO.40828.

[0008] Preferably, the bacterial agent includes a bacterial suspension of Aspergillus S8.

[0009] Preferably, the mixed gas includes any one or more of the following four mixed gases: 1) Hydrogen and carbon dioxide; 2) Hydrogen and methane; 3) methane and carbon dioxide; 4) Hydrogen, methane and carbon dioxide.

[0010] Preferably, in the mixed gas described in 1), the volume ratio of hydrogen to carbon dioxide is (1-2):(1-2).

[0011] Preferably, in the mixed gas described in 2), the volume ratio of hydrogen to methane is (1~2):(1~2).

[0012] Preferably, in the mixed gas described in 3), the volume ratio of methane to carbon dioxide is (1-2):(1-2).

[0013] Preferably, in the mixed gas described in 4), the volume ratio of hydrogen, methane and carbon dioxide is 1:1:1.

[0014] The present invention also provides a method for gas metabolism, which uses Aspergillus S8 or a bacterial agent containing Aspergillus S8 to treat gas; the gas is hydrogen or a mixed gas containing flammable and explosive gases; the flammable and explosive gases include hydrogen and / or methane; the preservation number of Aspergillus S8 is CGMCC NO.40828.

[0015] Preferably, the bacterial agent is a bacterial suspension of Aspergillus S8.

[0016] Beneficial effects: The present invention provides the use of Aspergillus sp. S8 in gas metabolism, wherein the gas includes hydrogen or a mixed gas containing flammable and explosive gases; the flammable and explosive gases include hydrogen and / or methane; the Aspergillus sp. S8 has a deposit number of CGMCC No. 40828. The present invention finds that Aspergillus sp. S8 has a good absorption and metabolism effect on flammable and explosive gases such as hydrogen and methane, as well as mixed gases containing flammable and explosive gases. It can metabolize flammable and explosive gases or mixed gases containing flammable and explosive gases as a carbon source and / or energy source, thereby achieving the effect of efficiently treating flammable and explosive gases and mixed gases thereof, and has high environmental safety, good removal effect, convenience, no toxic byproducts, and is easy to apply and transform. DETAILED DESCRIPTION

[0017] The present invention provides the use of Aspergillus sp. S8 in gas metabolism, wherein the gas includes hydrogen or a mixed gas containing flammable and explosive gases; the flammable and explosive gases include hydrogen and / or methane; the Aspergillus sp. S8 has a deposit number of CGMCC No. 40828. The Aspergillus sp. S8 described in the present invention is previously disclosed in patent application number CN202311542973.3.

[0018] The present invention also provides the use of a bacterial agent containing Aspergillus S8 in gas metabolism, wherein the gas includes hydrogen or a mixed gas containing flammable and explosive gases; the flammable and explosive gases include hydrogen and / or methane; the preservation number of Aspergillus S8 is CGMCC NO.40828.

[0019] In one embodiment, the bacterial agent is a suspension of Aspergillus sp. S8. In another embodiment, the Aspergillus sp. S8 suspension has an effective viable bacterial concentration of 2% (vol / vol). In another embodiment, the resuspending agent for the Aspergillus sp. S8 suspension is PBS phosphate buffer, wherein the PBS phosphate buffer is 1× PBS phosphate buffer with a pH of 6.8.

[0020] As an embodiment, the preparation method of the bacterial suspension of Aspergillus S8 is: inoculating Aspergillus S8 into an inorganic salt culture medium and shaking and culturing it to obtain an Aspergillus S8 fermentation liquid; separating the solid and liquid of the Aspergillus S8 fermentation liquid, taking the precipitate and resuspending it to obtain the bacterial suspension of Aspergillus S8. In one embodiment, the inorganic salt culture medium includes KH2PO4 0.5 g / L, Na2HPO4 0.5 g / L, NaCl 0.4 g / L, KNO3 1.0 g / L, NH4Cl 0.5 g / L, MgSO4·7H2O 1.0 g / L, CaCl2 0.2 g / L, FeSO4·7H2O 0.004 g / L, CuSO4·5H2O 0.00 4 g / L, MnSO4·H2O 0.004 g / L, ZnSO4·7H2O 0.004 g / L, and NaMoO4·2H2O 0.00024 g / L. In one embodiment, the temperature of the shaking culture is 28°C. In one embodiment, the rotation speed of the shaking culture is 170 rpm. In one embodiment, the shaking culture time is 3 days.

[0021] As an embodiment, the mixed gas includes a mixed gas of any one or more of the following four items: 1) hydrogen and carbon dioxide; 2) hydrogen and methane; 3) methane and carbon dioxide; 4) hydrogen, methane and carbon dioxide.

[0022] As an embodiment, in the mixed gas described in 1), the volume ratio of hydrogen to carbon dioxide is (1~2):(1~2); as another embodiment, the volume ratio of hydrogen to carbon dioxide is 1:1, 1:2 or 2:1. As an embodiment, in the mixed gas described in 2), the volume ratio of hydrogen to methane is (1~2):(1~2); as another embodiment, the volume ratio of hydrogen to methane is 1:1, 1:2 or 2:1. As an embodiment, in the mixed gas described in 3), the volume ratio of methane to carbon dioxide is (1~2):(1~2); as another embodiment, the volume ratio of methane to carbon dioxide is 1:1, 1:2 or 2:1. As an embodiment, in the mixed gas described in 4), the volume ratio of hydrogen, methane and carbon dioxide is 1:1:1.

[0023] The present invention also provides a method for gas metabolism, which uses Aspergillus S8 or a bacterial agent containing Aspergillus S8 to treat gas; the gas is hydrogen or a mixed gas containing flammable and explosive gases; the flammable and explosive gases include hydrogen and / or methane; the preservation number of Aspergillus S8 is CGMCC NO.40828.

[0024] In one embodiment, the bacterial agent is a suspension of Aspergillus S8; the relevant characteristics of the Aspergillus S8 suspension are defined in the above technical solution and will not be repeated here. In one embodiment, the volume of the Aspergillus S8 suspension is 2% of the volume of the gas.

[0025] In order to further illustrate the present invention, the technical solutions provided by the present invention are described in detail below in conjunction with the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0026] The experimental methods and detection methods in the following examples are conventional methods unless otherwise specified; the reagents and materials can be purchased through conventional commercial channels unless otherwise specified; Inorganic salt culture medium: KH2PO4 0.5 g / L, Na2HPO4 0.5 g / L, NaCl 0.4 g / L, KNO3 1.0 g / L, NH4Cl 0.5 g / L, MgSO4·7H2O 1.0 g / L, CaCl2 0.2 g / L, FeSO4·7H2O 0.004 g / L, CuSO4·5H2O 0.00 4 g / L, MnSO4·H2O 0.004 g / L, ZnSO4·7H2O 0.004 g / L, NaMoO4·2H2O 0.00024 g / L.

[0027] PDA medium: potato 200 g / L, glucose 20 g / L, peptone 5 g / L, potassium dihydrogen phosphate 3 g / L, magnesium sulfate 1.5 g / L, agar 20 g / L.

[0028] Example 1 This example provides an experiment on the utilization effect of Aspergillus S8 on H2 gas, and the steps are as follows: Add 50 mL of inorganic salt culture medium to a 250 mL sealed bottle. Replace the headspace with nitrogen and add pure H₂ gas. Inoculate each bottle with a 2% suspension of isolated and purified Aspergillus S8. Add ambient air to the control bottle as a control. After 7 days of static culture at 28°C, measure the following parameters: The H₂ utilization rate of Aspergillus sp. S8 was determined using a gas chromatography-mass spectrometer (GC2014C, purchased from Shimadzu Corporation). The test conditions were: TCD detector, inlet temperature 100°C, detector temperature 100°C, oven temperature 90°C, and argon carrier gas at a flow rate of 30 mL / min. The results are shown in Table 1.

[0029] The volume average utilization rate was measured by the drainage gas collection method, and the results are shown in Table 1.

[0030] When determining the dry weight of mycelium, the filter paper was first dried to constant weight and the weight of the filter paper was accurately weighed. After the Aspergillus S8 fermentation broth was thoroughly mixed, 30 mL was accurately measured for filtration, and the filter residue was thoroughly washed several times with distilled water until the filtrate was colorless. It was then placed in a 55°C constant temperature drying oven and dried to constant weight. The total mass of the filter residue and filter paper was accurately weighed. The dry weight of mycelium was calculated as follows: dry weight of mycelium (mg) = total mass of filter residue and filter paper (mg) - mass of filter paper (mg). The determination results are shown in Table 1.

[0031] Table 1 Utilization effect of Aspergillus S8 on H2 gas

[0032] As shown in Table 1, after adding Aspergillus S8 in sealed bottle, ambient air gas utilization rate is only 2.016%, and volume utilization rate is 2.437%, and mycelia dry weight content is 28.178 mg.And Aspergillus S8 is added in pure H2gas, H2gas utilization ability is more significant, and this bacterial strain can efficiently utilize H2gas as the nutrient obtained by its growth and reproduction, and this bacterial strain is to H2gas utilization rate is 95.721%, and volume utilization rate is 51.110%, and mycelia dry weight is 41.833 mg, and Aspergillus S8 is in pure H2gas utilization rate is 47.480 times of control group, and volume utilization rate is 20.972 times for control group, and mycelia dry weight is 1.730 times for control group.Show that Aspergillus S8 provided by the present invention is to H2gas utilization effect is better, has stronger H2gas utilization rate, can utilize H2gas as the nutrient of growth and reproduction.

[0033] Example 2 This example provides an experiment on the utilization effect of Aspergillus sp. S8 on a mixture of CO2 and H2, and the steps are as follows: 50 mL of inorganic salt culture medium was added to a 250 mL sealed bottle. The headspace within the bottle was replaced with nitrogen, and pure CO2 and H2 gas were added to the bottle. The mixtures were divided into three groups: CH1 (CO2:H2 = 1:2), CH2 (CO2:H2 = 1:1), and CH3 (CO2:H2 = 2:1). The sealed bottles were refilled with the same gas mixture every 7 days, and this cycle was repeated four times. Each bottle was inoculated with a 2% suspension of isolated and purified Aspergillus S8. A control bottle was filled with ambient air as a control.

[0034] After 7 days of static cultivation at 28°C, the average utilization rate of a mixture of CO₂ and H₂ by Aspergillus S8 over four cycles (i.e., measurement was performed once per cycle, and the average value was taken; the same applies to the following examples) was determined using a gas chromatography-mass spectrometer (GC2014C, purchased from Shimadzu Corporation). The results are shown in Table 2. The testing conditions were the same as in Example 1. The average volumetric utilization rate over four cycles was determined using the water-displacement gas collection method (i.e., measurement was performed once per cycle, and the average value was taken; the same applies to the following examples). The results are shown in Table 2. Mycelial dry weight was determined in the fourth cycle as in Example 1, and the results are shown in Table 2.

[0035] Table 2 Utilization effect of Aspergillus S8 on CO2 and H2 mixed gas in 4 cycles

[0036] As shown in Table 2, after adding Aspergillus S8 to a sealed bottle, its utilization rate of ambient air was only 3.124%, its volume utilization rate was 3.560%, and its mycelial dry weight was 30.561 mg. However, when Aspergillus S8 was added to a pure CO2 and H2 mixture, the CH3 group showed the most significant utilization, followed by the CH2 group and the CH1 group. This strain efficiently utilized the pure CO2 and H2 mixture as a carbon and energy source for its growth and reproduction. The CH3 group had a utilization rate of 96.524%, a volume utilization rate of 63.404%, and a mycelial dry weight of 48.03 mg. In the pure CO2 and H2 mixture, Aspergillus S8's gas utilization rate was 30.897 times that of the control group, its volume utilization rate was 17.810 times that of the control group, and its mycelial dry weight was 1.571 times that of the control group. It shows that the Aspergillus S8 provided by the present invention has a good utilization effect on the pure CO2 and H2 mixed gas, has a strong pure CO2 and H2 gas utilization rate, and can use the pure CO2 and H2 mixed gas as a carbon source and energy source for growth and reproduction.

[0037] Example 3 This example provides an experiment on the utilization effect of Aspergillus S8 on a mixture of CH4 and H2, and the steps are as follows: 50 mL of inorganic salt culture medium was added to a 250 mL sealed bottle. The headspace within the bottle was replaced with nitrogen, and pure CH₄ and H₂ gases were added to the bottle. The mixtures were divided into three groups: MH1 (CH₄:H₂ = 1:2), MH2 (CH₄:H₂ = 1:1), and MH3 (CH₄:H₂ = 2:1). The sealed bottles were refilled with the same gas mixture every 7 days, and this cycle was repeated four times. Each bottle was inoculated with a 2% suspension of isolated and purified Aspergillus S8. A control bottle was filled with ambient air as a control.

[0038] After 7 days of static cultivation at 28°C, the average utilization rate of a mixture of CH₄ and H₂ by Aspergillus sp. S8 over four cycles was measured using a gas chromatography-mass spectrometer (GC2014C, purchased from Shimadzu Corporation). The results are shown in Table 3. The test conditions were the same as in Example 1. The average volumetric utilization rate over four cycles was determined using the water-discharge gas collection method, and the results are shown in Table 3. Mycelial dry weight was determined in the fourth cycle as in Example 2, and the results are shown in Table 3.

[0039] Table 3 Utilization effect of Aspergillus S8 on CH4 and H2 mixed gas in 4 cycles

[0040] As shown in Table 3, after adding Aspergillus S8 to the sealed bottle, the gas utilization rate of ambient air was only 3.124%, the volume utilization rate was 3.560%, and the dry weight of mycelium was 30.561 mg. However, when Aspergillus S8 was added to a pure CH4 and H2 mixture, the MH3 group showed the most significant utilization, followed by the MH2 and MH1 groups. This strain efficiently utilized the pure CH4 and H2 mixture as a carbon and energy source for its growth and reproduction. The MH3 group had a CH4 and H2 mixture utilization rate of 94.620%, a volume utilization rate of 54.958%, and a dry weight of 89.350 mg. In the pure CH4 and H2 mixture, the gas utilization rate of Aspergillus S8 was 30.288 times that of the control group, the volume utilization rate was 15.437 times that of the control group, and the dry weight of mycelium was 2.923 times that of the control group. It shows that the Aspergillus S8 provided by the present invention has a good utilization effect on pure CH4 and H2 gases, has a strong pure CH4 and H2 gas utilization rate, and can use pure CH4 and H2 gases as carbon sources and energy sources for growth and reproduction.

[0041] Example 4 This example provides an experiment on the utilization effect of Aspergillus S8 on a mixed gas of CO2 and CH4, and the steps are as follows: 50 mL of inorganic salt culture medium was added to a 250 mL sealed bottle. The headspace in the sealed bottle was replaced with nitrogen, and pure CO₂ and CH₄ gases were added to the bottle. The mixture was divided into three groups according to the volume ratio: CM1 (CO₂: CH₄ = 1:2), CM2 (CO₂: CH₄ = 1:1), and CM3 (CO₂: CH₄ = 2:1). The sealed bottle was refilled with the same gas mixture every 7 days, and each cycle was repeated four times. Each bottle was inoculated with a 2% suspension of isolated and purified Aspergillus S8. A control bottle was filled with ambient air as a control.

[0042] After 7 days of static cultivation at 28°C, the average utilization rate of a mixture of CO₂ and CH₄ by Aspergillus sp. S8 over four cycles was measured using a gas chromatography-mass spectrometer (GC2014C, purchased from Shimadzu Corporation). The results are shown in Table 4. The test conditions were the same as in Example 1. The average volumetric utilization rate over four cycles was determined using the water-discharge gas collection method, and the results are shown in Table 4. Mycelial dry weight in the fourth cycle was determined as in Example 1, and the results are shown in Table 4.

[0043] Table 4 Utilization effect of Aspergillus S8 on CO2 and CH4 mixed gas in 4 cycles

[0044] As shown in Table 4, after adding Aspergillus S8 to the sealed bottle, the gas utilization rate of ambient air was only 3.124%, the volume utilization rate was 3.560%, and the dry weight of mycelium was 30.561 mg. However, when Aspergillus S8 was added to a pure CO2 and CH4 mixture, the CM3 group showed the most significant utilization, followed by the CM2 and CM1 groups. This strain efficiently utilized the pure CO2 and CH4 mixture as a carbon and energy source for its growth and reproduction. The CM3 group had a utilization rate of 97.014%, a volume utilization rate of 73.632%, and a dry weight of 91.771 mg. In the pure CO2 and CH4 mixture, the gas utilization rate of Aspergillus S8 was 31.054 times that of the control group, the volume utilization rate was 20.683 times that of the control group, and the dry weight of mycelium was 3.002 times that of the control group. It shows that the Aspergillus S8 provided by the present invention has a good utilization effect on the pure CO2 and CH4 mixed gas, has a strong pure CO2 and CH4 mixed gas utilization rate, and can use the pure CO2 and CH4 mixed gas as a carbon source and energy source for growth and reproduction.

[0045] Example 5 This example provides an experiment on the utilization effect of Aspergillus sp. S8 on a mixed gas of CO2, CH4, and H2, and the steps are as follows: To a 250 mL sealed bottle, add 50 mL of inorganic salt culture medium. Replace the headspace with nitrogen and then add pure CO₂, CH₄, and H₂ gases at a 1:1:1 volume ratio. Refill the bottle with the same gas mixture every 7 days, repeating this cycle four times. Each bottle was inoculated with a 2% suspension of isolated and purified Aspergillus S8. A control bottle was filled with ambient air as a control.

[0046] After 7 days of static cultivation at 28°C, the average utilization rate of a mixture of CO₂, CH₄, and H₂ by Aspergillus S8 over four cycles was measured using a gas chromatography-mass spectrometer (GC2014C, purchased from Shimadzu Corporation). The results are shown in Table 5. The testing conditions were the same as in Example 1. The average volumetric utilization rate over four cycles was determined using the water-discharge gas collection method, and the results are shown in Table 5. Mycelial dry weight was determined in the fourth cycle as in Example 1, and the results are shown in Table 5.

[0047] Table 5 Utilization effect of Aspergillus S8 on mixed gas of CO2, CH4 and H2 in 4 cycles

[0048] Table 5 shows that after adding Aspergillus S8 to the sealed bottle, the gas utilization rate of ambient air was only 3.124%, the volume utilization rate was 3.560%, and the dry weight of mycelium was 30.561 mg. However, when Aspergillus S8 was added to a pure CO2, CH4, and H2 gas mixture, the utilization capacity was significantly higher. This strain efficiently utilized the pure CO2, CH4, and H2 gas mixture as a carbon and energy source for its growth and reproduction. The gas utilization rate of the pure CO2, CH4, and H2 gas mixture in the treatment group was 94.722%, the volume utilization rate was 59.196%, and the dry weight of mycelium was 86.768 mg. The gas utilization rate of Aspergillus S8 in the pure CO2, CH4, and H2 gas mixture was 30.321 times that of the control group, the volume utilization rate was 16.628 times that of the control group, and the dry weight of mycelium was 2.839 times that of the control group. It shows that the Aspergillus S8 provided by the present invention has a good utilization effect on pure CO2, CH4 and H2 mixed gas, has a strong utilization rate of pure CO2, CH4 and H2 mixed gas, and can use pure CO2, CH4 and H2 mixed gas as a carbon source and energy source for growth and reproduction.

[0049] It can be concluded from the above examples that the Aspergillus sp. S8 of the present invention can efficiently metabolize flammable and explosive gases and mixed gases containing flammable and explosive gases.

[0050] Although the above embodiment provides a detailed description of the present invention, it is only a part of the embodiments of the present invention, not all of the embodiments. People can also obtain other embodiments based on this embodiment without creativity, and these embodiments all fall within the scope of protection of the present invention.

Claims

1. Aspergillus ( Aspergillus cejpii ) Application of S8 in gas metabolism, characterized in that, The gas includes hydrogen or a mixed gas containing flammable and explosive gases; the flammable and explosive gas includes hydrogen and / or methane; the preservation number of the Aspergillus S8 is CGMCC NO.40828.

2. Application of a bacterial agent containing Aspergillus S8 in gas metabolism, characterized in that: The gas includes hydrogen or a mixed gas containing flammable and explosive gases; the flammable and explosive gas includes hydrogen and / or methane; the preservation number of the Aspergillus S8 is CGMCC NO.40828.

3. The use according to claim 2, characterized in that The bacterial agent includes a bacterial suspension of Aspergillus S8.

4. The use according to any one of claims 1 to 3, characterized in that The mixed gas includes any one or more of the following four mixed gases: 1) Hydrogen and carbon dioxide; 2) Hydrogen and methane; 3) Methane and carbon dioxide; 4) Hydrogen, methane and carbon dioxide.

5. The use according to claim 4, characterized in that 1) In the mixed gas, the volume ratio of hydrogen to carbon dioxide is (1~2):(1~2).

6. The use according to claim 4, characterized in that 2) In the mixed gas, the volume ratio of hydrogen to methane is (1~2):(1~2).

7. The use according to claim 4, characterized in that 3) In the mixed gas, the volume ratio of methane to carbon dioxide is (1~2):(1~2).

8. The use according to claim 4, characterized in that 4) In the mixed gas, the volume ratio of hydrogen, methane and carbon dioxide is 1:1:

1.

9. A method for gas metabolism, characterized in that: Aspergillus S8 or a bacterial agent containing Aspergillus S8 is used to treat gas; the gas is hydrogen or a mixed gas containing flammable and explosive gases; the flammable and explosive gases include hydrogen and / or methane; the preservation number of Aspergillus S8 is CGMCC NO.40828.

10. The method according to claim 9, characterized in that The bacterial agent is a bacterial suspension of Aspergillus S8.

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