CN102242071B - Application of Pichia guilliermondii 510-6jm in treatment of petroleum hydrocarbon contaminations - Google Patents
Application of Pichia guilliermondii 510-6jm in treatment of petroleum hydrocarbon contaminations Download PDFInfo
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- CN102242071B CN102242071B CN 201110074688 CN201110074688A CN102242071B CN 102242071 B CN102242071 B CN 102242071B CN 201110074688 CN201110074688 CN 201110074688 CN 201110074688 A CN201110074688 A CN 201110074688A CN 102242071 B CN102242071 B CN 102242071B
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Abstract
The invention discloses a novel microorganism, and in particular relates to a novel microorganism Pichia guilliermondii which is capable of degrading 12C-36C linear chain alkanes. The number of the strain of the Pichia guilliermondii is 510-6jm (Pichia guillierondii sp. 510-6jm) CCTCC (China Center for Type Culture Collection) M 2010173. The novel strain Pichia guilliermondii is derived from deep sea, can be used for degrading petroleum hydrocarbon substances, and is mainly applied to bioremediation and industrial washing industries. The invention discloses raw materials required for cultivating the bacterium. Bacteria cells can be cultivated by utilizing the raw materials and are used for degrading petroleum hydrocarbons.
Description
Technical field
The present invention relates to microbial technology field, specifically relate to a kind of Pichia guilliermondii (Pichia guilliermondii sp.) 510-6jm of new separation, can utilize it at the petroleum hydrocarbon class pollutant degradation function, be used for polluting the use that reparation and industry are cleaned.
Background technology
Biological restoration is the biotechnology of a class low consumption, efficient and environmental safety, mainly relies on the natural metabolism process degraded of bacterium, fungi even higher plant and cell free enzyme, removes out the pollutent in the environment.Up to now, genus surplus the microorganism of known energy degraded oil hydro carbons has 100, kind more than 200, they belong to bacterium, actinomycetes, mould and algae.The bacterium of degraded oil has Rhodopseudomonas, some bacterial strain during Pichia guilliermondii genus, Mycosphaerella, Vibrio etc. belong to.Wherein the most common is Rhodopseudomonas, has found that it contains the plasmid of multiple degradation of hydrocarbon, and once is used for genetically engineered, to the degraded of short chain alkanes such as ethane, propane, butane or long chain alkane and aromatic hydrocarbons.Common degraded is the bacterial strain during the mould that has has that aspergillus, mould, a spore are mould etc. and belongs to, and white-rot fungi is very high to organic degradation efficiencies such as oil.The yeast protein that often decomposes oil with candiyeast in the yeast and produce.Find again that in recent years as if cyanobacteria and green alga degradable aromatic hydrocarbons, especially blue-green algae have the ability of the multiple aromatic hydrocarbons of oxidation.
Each state-owned nearly 80,000 ocean-going merchantman of the whole world shuttle back and forth in each harbour, the whole world at present, and they can reach more than 1,000,000 tons at the greasy dirt of term of voyage to ocean disposal every year.In addition, the marine oil overflow accident also frequently takes place in various places in recent years, marine oil pollution serious threat to the safety of the marine eco-environment.
Marine oil spill will experience dissolving, volatilization, photochemical oxidation decomposition, absorption and a series of processes such as sedimentation and biological degradation.Wherein, biological degradation is the most important approach of thoroughly eliminating greasy dirt, and the bioremediation technology that utilizes microbiological deterioration initiatively to remove oil spilling is current a kind of very important Environmental Biotechnology.Wherein, Pichia guilliermondii is one of oil degradation dominant bacteria important in the ocean.The present invention will provide important bacterial classification for petroleum hydrocarbon pollutes biological restoration.
Summary of the invention
The invention provides a kind of new microorganism-Pichia guilliermondii that petroleum hydrocarbon pollutes biological restoration that can be used in.Its called after Pichia guilliermondii Pichia guilliermondii sp.510-6jm, be preserved in Chinese typical culture collection center (CCTCC) on July 10th, 2010, the address: Chinese Wuhan Wuhan University, its preservation registration number is CCTCC NO:M2010173.Pichia guilliermondii of the present invention separates from the surface seawater sample, and this sample collecting is from the Indian Ocean.
Microorganism of the present invention is that 0.8-1.5 μ m is long, 0.6-1.2 μ m is wide, does not give birth to flagellum, does not produce gemma.In NaCl concentration be on 3% the LB nutrient agar 28 ℃ cultivate and can form distinctive colonial morphology in 3 days: small, oyster white, translucent, central protrusion, surface wettability, edge have irregular and transparent halo.Can utilize following carbon source: polysorbate40, tween 80, monomethyl succsinic acid, succsinic acid, γ-An Jidingsuan, seminose, melibiose.Can utilize the straight-chain paraffin of carbon chain lengths 12C～36C, growth rapidly in 16C～32C.
Table 1 is Pichia guilliermondii physiological and biochemical property of the present invention.
Diesel oil Pichia guilliermondii of the present invention is compared and is had following feature with existing other kind that belong to together:
Table 1 physiological and biochemical property
SEQ ID No.1 is the 26S rDNA sequence D 1/D2 district of Pichia guilliermondii of the present invention, and SEQ ID No.2 is the ITS sequence of Pichia guilliermondii of the present invention.
Description of drawings
The transmission electron microscope photo of Fig. 1 diesel oil Pichia guilliermondii of the present invention.
Embodiment
Embodiment
The separation of Pichia guilliermondii 510-6jm
Gather the top layer seawater sample from the Indian Ocean, in the laboratory, can obtain Pichia guilliermondii 510-6jm of the present invention by following condition cultivation.
Artificial seawater substratum: NaCl=24g/L; MgSO
47H
2O=7.0g/L; NH
4NO
3=1g/L; KCl=0.7g/L; KH
2PO
4=2.0g/L; Na
2HPO
4=3.0g/L, pH7.4 adds an amount of micro-mixed solution after the sterilization, and mends with diesel oil as sole carbon source.Diesel oil and trace element are through 0.22 μ M membrane filtration degerming.The 100mL artificial seawater that sample is inoculated in diesel oil content 1% (v/v) is cultivated concentrated.28 ℃, 200r/min shaking table were cultivated after 4～5 days, take out 1mL and change in the 100mL fresh culture from nutrient solution.After the enrichment like this 6 times, with final enriched substance gradient dilution, spread plate, cultivate separation, purifying list bacterium colony after 48 hours for 28 ℃.
Pichia guilliermondii 510-6jm removes the oil pollution water body
Petroleum pollution water body (pollute oil 0.5%～5%) adds inorganic nitrogen-sourced, inorganic phosphorus and bacterium liquid (inoculum size 1/10000～1/100), 1～200 liter of/ton sewage of air flow (air), and the hydrocarbon contamination clearance rate reaches 35%～40% after 7 days
Pichia guilliermondii 510-6jm removes oil pollution soil
Oil-polluted soils (polluting oil 0.5%～2%) adds nitrogenous fertilizer, phosphate fertilizer and bacterium liquid (inoculum size 1/10000～1/100), stirs ventilative 1 time in 3 days, and the after stain clearance rate reached 40%～50% in 9～12 days.
Claims (9)
1. a Pichia guilliermondii (Pichia guilliermondii sp.) 510-6jm, it is characterized in that: its preservation registration number is CCTCC M2010173.
2. the application of Pichia guilliermondii 510-6jm as claimed in claim 1 is characterized in that, is applied to repair the pollution of oil and petroleum products generation as one of the single component in the bioremediation agents or composition.
3. the application of the described Pichia guilliermondii 510-6jm of claim 1 aspect the greasy dirt cleaning of metal, glass and frosting.
4. the application of Pichia guilliermondii 510-6jm as claimed in claim 2, it is characterized in that: described reparation is to add inorganic nitrogen-sourced, inorganic phosphorus and bacterium liquid in the petroleum pollution water body, air flow 1-200 liter/ton sewage, the hydrocarbon contamination clearance rate reaches 35%-40% after 7 days.
5. the application of Pichia guilliermondii 510-6jm as claimed in claim 4 is characterized in that: petroleum pollution water body PetroChina Company Limited. content is 0.5%-5%.
6. as the application of claim 4 or 5 described Pichia guilliermondii 510-6jm, it is characterized in that: bacterium liquid inoculum size is the 1/10000-1/100 of petroleum pollution water body.
7. the application of Pichia guilliermondii 510-6jm as claimed in claim 2: it is characterized in that: described reparation is to add nitrogenous fertilizer, phosphate fertilizer and bacterium liquid in oil-polluted soils, stirs ventilative 1 time in 3 days, and the after stain clearance rate reached 40%-50% in 9-12 days.
8. the application of Pichia guilliermondii 510-6jm as claimed in claim 7 is characterized in that: oil-polluted soils PetroChina Company Limited. content is 0.5%-2%.
9. as the application of claim 7 or 8 described Pichia guilliermondii 510-6jm, it is characterized in that: bacterium liquid inoculum size is the 1/10000-1/100 of oil-polluted soils.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110257268A (en) * | 2019-07-12 | 2019-09-20 | 东北林业大学 | One plant of degrading polyethylene generates the Pichia guilliermondii of alkane simultaneously |
| CN110257267A (en) * | 2019-07-12 | 2019-09-20 | 东北林业大学 | The Pichia guilliermondii of one plant of degradable polyethylene |
Families Citing this family (6)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103122320B (en) * | 2013-01-22 | 2014-07-02 | 浙江工业大学 | Tert-butyl 6-cyano-(3R,5R)-dihydroxyhexanoate prepared by biological catalysis and strain |
| CN106047733B (en) * | 2016-07-13 | 2020-06-30 | 华中农业大学 | Yeast strain for food fermentation, leaven and application thereof |
| CN106520580B (en) * | 2016-09-05 | 2019-08-20 | 华南理工大学 | One saccharomycete and its application in catalysis 2,5- dihydroxymethyl furans synthesis |
| CN106591204A (en) * | 2017-01-20 | 2017-04-26 | 国家海洋局第三海洋研究所 | Microecological preparation for promoting algae growth in litopenaeus vannamei aquaculture and preparation |
| CN110317738B (en) * | 2019-07-12 | 2022-10-28 | 东北林业大学 | Method for biodegradation of polyethylene by using pichia guilliermondii |
| CN110283853B (en) * | 2019-07-12 | 2023-03-31 | 东北林业大学 | Method for producing alkane by degrading polyethylene by using pichia guilliermondii |
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| CN101270340A (en) * | 2008-05-21 | 2008-09-24 | 中国农业大学 | Paris polyphylla var.yunnanensis endogenetic pichia for preparing volatile oil and antibiotic activity |
| EP1332207B1 (en) * | 2000-07-14 | 2008-10-29 | Archer-Daniels-Midland Company | Transformation systems for flavinogenic yeast |
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| EP1332207B1 (en) * | 2000-07-14 | 2008-10-29 | Archer-Daniels-Midland Company | Transformation systems for flavinogenic yeast |
| CN101270340A (en) * | 2008-05-21 | 2008-09-24 | 中国农业大学 | Paris polyphylla var.yunnanensis endogenetic pichia for preparing volatile oil and antibiotic activity |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110257268A (en) * | 2019-07-12 | 2019-09-20 | 东北林业大学 | One plant of degrading polyethylene generates the Pichia guilliermondii of alkane simultaneously |
| CN110257267A (en) * | 2019-07-12 | 2019-09-20 | 东北林业大学 | The Pichia guilliermondii of one plant of degradable polyethylene |
| CN110257268B (en) * | 2019-07-12 | 2022-05-03 | 东北林业大学 | Pichia guilliermondii strain for degrading polyethylene and producing alkane simultaneously |
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