CN103232943B - Instant premixed dry powder for separating polycyclic aromatic hydrocarbons and degrading fungi - Google Patents
Instant premixed dry powder for separating polycyclic aromatic hydrocarbons and degrading fungi Download PDFInfo
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- CN103232943B CN103232943B CN201310143989.7A CN201310143989A CN103232943B CN 103232943 B CN103232943 B CN 103232943B CN 201310143989 A CN201310143989 A CN 201310143989A CN 103232943 B CN103232943 B CN 103232943B
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Abstract
The invention discloses instant premixed dry powder for separating polycyclic aromatic hydrocarbons and degrading fungi. The instant premixed dry powder comprises ultrafine polycyclic aromatic hydrocarbon particles, a bacterial growth inhibitor, a soluble inorganic salt and agar according to the weight ratio of 0.1-0.5: 0.01-0.05: 2-5: 10-30. The instant premixed dry powder disclosed by the invention has the advantages of simplicity and convenience in use, short consumed time, capability of dissolving the polycyclic aromatic hydrocarbons without an organic solvent, capability of greatly reducing toxicity and harm to experimental staff and an environment, little growth of mixed bacteria, high separation efficiency and good separation effect.
Description
Technical field
The invention belongs to environmental microorganism technical field, relate to a kind of instant premix dry powder for separating of degrading polycyclic aromatic hydrocarbons fungi.
Background technology
Along with the develop rapidly of China's oil petrochemical industry, fragile ecotope is also faced with more and more acid test.Wherein, petroleum hydrocarbon contaminant often the number of ways such as is revealed by accident and is entered the environment such as soil and water, causes environmental pollution.Particularly in petroleum hydrocarbon contaminant, a large amount of polycyclic aromatic hydrocarbonss (Polycyclic Aromatic Hydrocarbons, PAHs) is difficult to natural degradation, and environment has been caused to routinely serious threat.And because it has carcinogenic teratogenesis mutagenic " three cause " effect, serious threat HUMAN HEALTH and the ecosystem are stable, by a plurality of countries, are listed in priority pollutants Black List.Adopt biological treating, particularly the polycyclic aromatic hydrocarbons in fungus degrading petroleum pollution is to control and even eliminate the effective measure that Polycyclic Aromatic Hydrocarbonat Existing in Environment pollutes, thereby being subject to domestic and international researchist's extensive attention, a series of patents are authorized by China and international mechanism of related tissue.Such as: Chinese patent (200810228377.7) discloses a kind of microbiobacterial agent of repairing for polycyclic aromatic hydrocarbon pollution being comprised of Mucor, little gram of Mildy Way, sickle-like bacteria and Penicllium chrysogenum etc.; Chinese patent (200910234389.5) discloses a kind of method that adopts the arbuscular mycorrhizas such as Glomus mosseae to repair polycyclic aromatic hydrocarbon pollution; United States Patent (USP) (US20040023362A1) discloses a kind of method that adopts the degrading polycyclic aromatic hydrocarbons such as penicillium janthinellum penicillium janthinellum; United States Patent (USP) (7118906B2) discloses a kind of method that adopts pale yellow bag hole bacterium Flavodon flavus to administer waste water polluted by polycyclic aromatic hydrocarbon.
Traditional enrichment culture-flat board coating separation method is mainly followed in the separation of degrading polycyclic aromatic hydrocarbons fungi.But, traditional preparation flow of degrading polycyclic aromatic hydrocarbons fungi culture medium is quite complicated, consuming time longer, and owing to will guaranteeing being evenly distributed of polycyclic aromatic hydrocarbons particle, operating process needs the organic solvents of a large amount of severe toxicity, not only increased reagent cost but also to environment.Meanwhile, owing to lacking bacterial growth inhibitor, often cause varied bacteria growing too vigorous, to such an extent as to the separation efficiency of target degrading polycyclic aromatic hydrocarbons fungi is low, separating effect is poor.These defects have seriously limited the separated and even further oil polluted environment of degrading polycyclic aromatic hydrocarbons fungi and have administered process.The preparation flow of the substratum of Innovation degrading polycyclic aromatic hydrocarbons fungi, becomes mission critical urgently to be resolved hurrily.
Adopting instant premix dry powder technology of preparing is the available strategy addressing this problem.For example: Chinese patent (200710121709.7) discloses a kind of powder mixture for animal cell culture and preparation method thereof; Chinese patent (200610078090.1) discloses a kind of dry-powder-type culturing substrate for plant breeding, tissue culture and molecular biology research and preparation method thereof; United States Patent (USP) (US20120276630A1) discloses a kind of dehydrated medium for cell cultures.But, the relevant report of still not preparing for the instant premix dry powder of degrading polycyclic aromatic hydrocarbons fungi so far both at home and abroad.
Summary of the invention
The object of the invention is to overcome the problems such as that prior art exists is slow for current degrading polycyclic aromatic hydrocarbons fungi separation and Culture speed, inferior separating effect, a kind of instant premix dry powder for separating of degrading polycyclic aromatic hydrocarbons fungi is provided.
Technical scheme of the present invention is summarized as follows:
For separating of an instant premix dry powder for degrading polycyclic aromatic hydrocarbons fungi, be that the ratio of 0.1～0.5:0.01～0.05:2～5:10～30 is comprised of ultra micro polycyclic aromatic hydrocarbons particle, bacterial growth inhibitor, soluble inorganic salt and agar by weight.
The mass ratio of described ultra micro polycyclic aromatic hydrocarbons particle, bacterial growth inhibitor, soluble inorganic salt and agar is preferably 0.2:0.03:4:20.
Phenanthrene and pyrene that described ultra micro polycyclic aromatic hydrocarbons particle is particle diameter is 1:0.01～10 lower than the mass ratio of 150 μ m form.
The mass ratio of described phenanthrene and pyrene is preferably 1:0.1.
Described bacterial growth inhibitor is preferably rose-bengal or Rifampin.
The K that described soluble inorganic salt is 0.5～2:0.5～2:0.5～2:0.5～2:0.1～0.5:0.01～0.05:0.02～0.1 by mass ratio
2hPO
4, KH
2pO
4, NH
4nO
3, NaCl, MgSO
4, CaCl
2and FeCl
3form.
Described K
2hPO
4, KH
2pO
4, NH
4nO
3, NaCl, MgSO
4, CaCl
2and FeCl
3mass ratio be preferably 1:1:1:1:0.2:0.02:0.05.
The present invention has the following advantages:
1. easy to use, consuming time extremely short;
2. without with an organic solvent dissolving polycyclic aromatic hydrocarbons, significantly reduce the murder by poisoning to experimenter and environment;
3. varied bacteria growing is few, separation efficiency is high, good separating effect.
Embodiment
By specific embodiment, set forth the present invention below, object is to make those skilled in the art can understand better the present invention, but not as the restriction to the scope of the present invention.
Embodiment 1
For separating of an instant premix dry powder for degrading polycyclic aromatic hydrocarbons fungi, by weight being formed by ultra micro polycyclic aromatic hydrocarbons particle, bacterial growth inhibitor, soluble inorganic salt and agar for the ratio of 0.2:0.03:4:20.
Phenanthrene and pyrene that ultra micro polycyclic aromatic hydrocarbons particle is particle diameter is 1:0.1 lower than the mass ratio of 150 μ m form.
Bacterial growth inhibitor is rose-bengal.
The K that soluble inorganic salt is 1:1:1:1:0.2:0.02:0.05 by mass ratio
2hPO
4, KH
2pO
4, NH
4nO
3, NaCl, MgSO
4, CaCl
2and FeCl
3form.
Embodiment 2
For separating of an instant premix dry powder for degrading polycyclic aromatic hydrocarbons fungi, by weight being formed by ultra micro polycyclic aromatic hydrocarbons particle, bacterial growth inhibitor, soluble inorganic salt and agar for 0.1:0.05:3:10.
Phenanthrene and pyrene that ultra micro polycyclic aromatic hydrocarbons particle is particle diameter is 1:0.01 lower than the mass ratio of 150 μ m form.
Bacterial growth inhibitor is rose-bengal.
The K that soluble inorganic salt is 0.5:2:1:0.5:0.5:0.03:0.02 by mass ratio
2hPO
4, KH
2pO
4, NH
4nO
3, NaCl, MgSO
4, CaCl
2and FeCl
3form.
Embodiment 3
For separating of an instant premix dry powder for degrading polycyclic aromatic hydrocarbons fungi, by weight being formed by ultra micro polycyclic aromatic hydrocarbons particle, bacterial growth inhibitor, soluble inorganic salt and agar for 0.3:0.01:5:20.
Phenanthrene and pyrene that ultra micro polycyclic aromatic hydrocarbons particle is particle diameter is 1:1 lower than the mass ratio of 150 μ m form.
Bacterial growth inhibitor is rose-bengal.
The K that soluble inorganic salt is 1:0.5:2:1:0.1:0.05:0.08 by mass ratio
2hPO
4, KH
2pO
4, NH
4nO
3, NaCl, MgSO
4, CaCl
2and FeCl
3form.
Embodiment 4
For separating of an instant premix dry powder for degrading polycyclic aromatic hydrocarbons fungi, by weight being formed by ultra micro polycyclic aromatic hydrocarbons particle, bacterial growth inhibitor, soluble inorganic salt and agar for 0.5:0.03:2:30.
Phenanthrene and pyrene that ultra micro polycyclic aromatic hydrocarbons particle is particle diameter is 1:10 lower than the mass ratio of 150 μ m form.
Bacterial growth inhibitor is Rifampin.
The K that soluble inorganic salt is 2:1:0.5:2:0.3:0.01:0.1 by mass ratio
2hPO
4, KH
2pO
4, NH
4nO
3, NaCl, MgSO
4, CaCl
2and FeCl
3form.
Embodiment 5
A kind of preparation of the instant premix dry powder for separating of degrading polycyclic aromatic hydrocarbons fungi:
In the ratio of embodiment 1,2,3 or 4, Phenanthrene and pyrene particle are ground to particle diameter lower than 150 μ m, mix, obtain the ultra micro polycyclic aromatic hydrocarbons particle mixing;
By K
2hPO
4, KH
2pO
4, NH
4nO
3, NaCl, MgSO
4, CaCl
2and FeCl
3according to the ratio of embodiment 1,2,3 or 4, mix, obtain mixing soluble inorganic salt;
Ultra micro polycyclic aromatic hydrocarbons particle, bacterial growth inhibitor, soluble inorganic salt and agar are mixed in proportion, obtain respectively a kind of instant premix dry powder for separating of degrading polycyclic aromatic hydrocarbons fungi.
The present invention be take environmental microbiology as theoretical foundation, the correlation theories knowledge of integrated environment science, preparation problem for degrading polycyclic aromatic hydrocarbons fungi culture medium, on the basis of traditional substratum, add ultra micro polycyclic aromatic hydrocarbons particle and bacterial growth inhibitor, and adopt mechanical mill mode to prepare instant premix dry powder.
Embodiment 6
Take a kind of instant premix dry powder 2.5g for separating of degrading polycyclic aromatic hydrocarbons fungi of embodiment 1, add 100ml distilled water, after fully mixing, 121 ℃ of high-temperature sterilizations 20 minutes, prepare solid plate.Expend altogether 1.3 hours, than traditional method time decreased 44.3%, efficiency improves 79.4%.Adopt gradient dilution method that oil-polluted soils sample 1# is applied on solid plate, 28 ℃ of constant temperature culture 7 days.Obtain altogether 7 degrading polycyclic aromatic hydrocarbons fungal colonies, through identifying, be Mucor.Degrading polycyclic aromatic hydrocarbons fungi accounts for 17.4% of total colony number, higher by 15.1% than traditional method.
Embodiment 7
Take a kind of instant premix dry powder 30g for separating of degrading polycyclic aromatic hydrocarbons fungi of embodiment 2, add 1000ml distilled water, after fully mixing, 121 ℃ of high-temperature sterilizations 20 minutes, prepare solid plate.Expend altogether 1.4 hours, than traditional method time decreased 47.5%, efficiency improves 90.5%.Adopt gradient dilution method that oil-polluted soils sample 2# is applied on solid plate, 28 ℃ of constant temperature culture 10 days.Obtain altogether 38 degrading polycyclic aromatic hydrocarbons fungal colonies, through being accredited as Mucor and mould.Degrading polycyclic aromatic hydrocarbons fungi accounts for 26.6% of total colony number, higher by 26.0% than traditional method.
Embodiment 8
Take a kind of instant premix dry powder 50g for separating of degrading polycyclic aromatic hydrocarbons fungi of embodiment 3, add 2000ml distilled water, after fully mixing, 121 ℃ of high-temperature sterilizations 20 minutes, prepare solid plate.Expend altogether 1.3 hours, than traditional method time decreased 51.9%, efficiency improves 107.7%.Adopt gradient dilution method that oil-polluted soils sample 3# is applied on solid plate, 28 ℃ of constant temperature culture 10 days.Obtain altogether 19 degrading polycyclic aromatic hydrocarbons fungal colonies.Degrading polycyclic aromatic hydrocarbons fungi accounts for 28.8% of total colony number, higher by 26.3% than traditional method.
Embodiment 9
Take a kind of instant premix dry powder 60g for separating of degrading polycyclic aromatic hydrocarbons fungi of embodiment 4, add 2000ml distilled water, after fully mixing, 121 ℃ of high-temperature sterilizations 20 minutes, prepare solid plate.Expend altogether 1.4 hours, than traditional method time decreased 53.3%, efficiency improves 114.3%.Adopt gradient dilution method that oil-polluted soils sample 4# is applied on solid plate, 28 ℃ of constant temperature culture 10 days.Obtain altogether 28 degrading polycyclic aromatic hydrocarbons fungal colonies.Degrading polycyclic aromatic hydrocarbons fungi accounts for 44.1% of total colony number, higher by 38.9% than traditional method.
Claims (5)
1. the instant premix dry powder for separating of degrading polycyclic aromatic hydrocarbons fungi, it is characterized in that being that the ratio of 0.1～0.5:0.01～0.05:2～5:10～30 is comprised of ultra micro polycyclic aromatic hydrocarbons particle, bacterial growth inhibitor, soluble inorganic salt and agar by weight, phenanthrene and pyrene that described ultra micro polycyclic aromatic hydrocarbons particle is particle diameter is 1:0.01～10 lower than the mass ratio of 150 μ m form, the K that described soluble inorganic salt is 0.5～2:0.5～2:0.5～2:0.5～2:0.1～0.5:0.01～0.05:0.02～0.1 by mass ratio
2hPO
4, KH
2pO
4, NH
4nO
3, NaCl, MgSO
4, CaCl
2and FeCl
3form.
2. a kind of instant premix dry powder for separating of degrading polycyclic aromatic hydrocarbons fungi according to claim 1, is characterized in that the mass ratio of described ultra micro polycyclic aromatic hydrocarbons particle, bacterial growth inhibitor, soluble inorganic salt and agar is 0.2:0.03:4:20.
3. a kind of instant premix dry powder for separating of degrading polycyclic aromatic hydrocarbons fungi according to claim 1, the mass ratio that it is characterized in that described phenanthrene and pyrene is 1:0.1.
4. a kind of instant premix dry powder for separating of degrading polycyclic aromatic hydrocarbons fungi according to claim 1 and 2, is characterized in that described bacterial growth inhibitor is rose-bengal or Rifampin.
5. a kind of instant premix dry powder for separating of degrading polycyclic aromatic hydrocarbons fungi according to claim 1, is characterized in that described K
2hPO
4, KH
2pO
4, NH
4nO
3, NaCl, MgSO
4, CaCl
2and FeCl
3mass ratio be 1:1:1:1:0.2:0.02:0.05.
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| CN113755337B (en) * | 2021-08-30 | 2022-06-10 | 中国科学院广州地球化学研究所 | Polycyclic aromatic hydrocarbon degrading fungi GIG-1 and GIG-2 in petroleum-polluted soil, mixed microbial inoculum thereof and application thereof |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101050421A (en) * | 2007-03-30 | 2007-10-10 | 中国科学院沈阳应用生态研究所 | Method for quick filtering out bacteria having characterization of adsorption for degrading polycyclic aromatic hydrocarbon |
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Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101050421A (en) * | 2007-03-30 | 2007-10-10 | 中国科学院沈阳应用生态研究所 | Method for quick filtering out bacteria having characterization of adsorption for degrading polycyclic aromatic hydrocarbon |
Non-Patent Citations (7)
| Title |
|---|
| Bishnoi K et al.Biodegradation of polycyclic aromatic hydrocarbons by white rot fungi Phanerochaete chrysosporium in sterile and unsterile soil.《Journal of Scientific & Industrial Research》.2008,第67卷 * |
| Bishnoi K et al.Biodegradation of polycyclic aromatic hydrocarbons by white rot fungi Phanerochaete chrysosporium in sterile and unsterile soil.《Journal of Scientific & * |
| Industrial Research》.2008,第67卷 * |
| 土壤和地下水中多环芳烃生物降解研究进展;谭文捷 等;《生态环境》;20070430;第16卷(第4期);1310-1317 * |
| 多环芳烃降解菌的筛选及其对芘的降解研究;王蕾 等;《环境科学与技术》;20100131;第33卷(第1期);43-47 * |
| 王蕾 等.多环芳烃降解菌的筛选及其对芘的降解研究.《环境科学与技术》.2010,第33卷(第1期), * |
| 谭文捷 等.土壤和地下水中多环芳烃生物降解研究进展.《生态环境》.2007,第16卷(第4期), * |
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Address after: 100010 Oceanic Building, Oceanic Building, Chaoyangmen North Street, Dongcheng District, Beijing, 25 Co-patentee after: CNOOC Energy Development Co., Ltd. Patentee after: China Offshore Oil Group Co., Ltd. Co-patentee after: COES Limited Address before: 100010 Oceanic Building, Oceanic Building, Chaoyangmen North Street, Dongcheng District, Beijing, 25 Co-patentee before: CNOOC Energy Development Co., Ltd. Patentee before: China National Offshore Oil Corporation Co-patentee before: COES Limited |
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