LT4792B - The strain pseudomonas fluorescens ign57 destroying oil and oil products - Google Patents
The strain pseudomonas fluorescens ign57 destroying oil and oil products Download PDFInfo
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- LT4792B LT4792B LT2000058A LT2000058A LT4792B LT 4792 B LT4792 B LT 4792B LT 2000058 A LT2000058 A LT 2000058A LT 2000058 A LT2000058 A LT 2000058A LT 4792 B LT4792 B LT 4792B
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- 241000589540 Pseudomonas fluorescens Species 0.000 title claims abstract description 25
- 239000002689 soil Substances 0.000 claims abstract description 45
- 230000003042 antagnostic effect Effects 0.000 claims abstract description 4
- 244000005700 microbiome Species 0.000 claims abstract 3
- 239000000758 substrate Substances 0.000 claims description 12
- 230000015556 catabolic process Effects 0.000 claims description 11
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 10
- 239000007003 mineral medium Substances 0.000 claims description 7
- 238000000034 method Methods 0.000 claims description 5
- 150000001875 compounds Chemical class 0.000 claims description 4
- 230000001804 emulsifying effect Effects 0.000 claims description 3
- 241000894006 Bacteria Species 0.000 claims 1
- 239000003921 oil Substances 0.000 abstract description 41
- 239000000295 fuel oil Substances 0.000 abstract description 11
- 239000000203 mixture Substances 0.000 abstract description 7
- 238000006731 degradation reaction Methods 0.000 description 10
- UFWIBTONFRDIAS-UHFFFAOYSA-N Naphthalene Chemical compound C1=CC=CC2=CC=CC=C21 UFWIBTONFRDIAS-UHFFFAOYSA-N 0.000 description 8
- 239000003209 petroleum derivative Substances 0.000 description 8
- 239000000654 additive Substances 0.000 description 6
- 230000001580 bacterial effect Effects 0.000 description 6
- 239000003208 petroleum Substances 0.000 description 6
- 241000589776 Pseudomonas putida Species 0.000 description 5
- 229930195733 hydrocarbon Natural products 0.000 description 5
- 150000002430 hydrocarbons Chemical class 0.000 description 5
- 239000002609 medium Substances 0.000 description 5
- 150000003839 salts Chemical class 0.000 description 5
- 239000000725 suspension Substances 0.000 description 5
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- 239000002028 Biomass Substances 0.000 description 4
- 239000004215 Carbon black (E152) Substances 0.000 description 4
- 238000004140 cleaning Methods 0.000 description 4
- 239000003344 environmental pollutant Substances 0.000 description 4
- 229910052757 nitrogen Inorganic materials 0.000 description 4
- 231100000719 pollutant Toxicity 0.000 description 4
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- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 description 3
- 238000000354 decomposition reaction Methods 0.000 description 3
- 230000002538 fungal effect Effects 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 3
- 239000002002 slurry Substances 0.000 description 3
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 2
- FERIUCNNQQJTOY-UHFFFAOYSA-N Butyric acid Chemical compound CCCC(O)=O FERIUCNNQQJTOY-UHFFFAOYSA-N 0.000 description 2
- SRBFZHDQGSBBOR-IOVATXLUSA-N D-xylopyranose Chemical compound O[C@@H]1COC(O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-IOVATXLUSA-N 0.000 description 2
- 241000192125 Firmicutes Species 0.000 description 2
- 239000001888 Peptone Substances 0.000 description 2
- 108010080698 Peptones Proteins 0.000 description 2
- 241000187561 Rhodococcus erythropolis Species 0.000 description 2
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 150000007513 acids Chemical class 0.000 description 2
- 239000010692 aromatic oil Substances 0.000 description 2
- 238000006065 biodegradation reaction Methods 0.000 description 2
- 238000009264 composting Methods 0.000 description 2
- NNBZCPXTIHJBJL-UHFFFAOYSA-N decalin Chemical compound C1CCCC2CCCCC21 NNBZCPXTIHJBJL-UHFFFAOYSA-N 0.000 description 2
- DIOQZVSQGTUSAI-UHFFFAOYSA-N decane Chemical compound CCCCCCCCCC DIOQZVSQGTUSAI-UHFFFAOYSA-N 0.000 description 2
- 238000007865 diluting Methods 0.000 description 2
- 239000003337 fertilizer Substances 0.000 description 2
- DCAYPVUWAIABOU-UHFFFAOYSA-N hexadecane Chemical compound CCCCCCCCCCCCCCCC DCAYPVUWAIABOU-UHFFFAOYSA-N 0.000 description 2
- 230000002209 hydrophobic effect Effects 0.000 description 2
- 235000013372 meat Nutrition 0.000 description 2
- RZJRJXONCZWCBN-UHFFFAOYSA-N octadecane Chemical compound CCCCCCCCCCCCCCCCCC RZJRJXONCZWCBN-UHFFFAOYSA-N 0.000 description 2
- 235000019319 peptone Nutrition 0.000 description 2
- 229910052698 phosphorus Inorganic materials 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
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- 230000009897 systematic effect Effects 0.000 description 2
- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-N 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- 235000001674 Agaricus brunnescens Nutrition 0.000 description 1
- 241000589149 Azotobacter vinelandii Species 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 102000016938 Catalase Human genes 0.000 description 1
- 108010053835 Catalase Proteins 0.000 description 1
- FBPFZTCFMRRESA-FSIIMWSLSA-N D-Glucitol Natural products OC[C@H](O)[C@H](O)[C@@H](O)[C@H](O)CO FBPFZTCFMRRESA-FSIIMWSLSA-N 0.000 description 1
- FBPFZTCFMRRESA-JGWLITMVSA-N D-glucitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-JGWLITMVSA-N 0.000 description 1
- RWSOTUBLDIXVET-UHFFFAOYSA-N Dihydrogen sulfide Chemical compound S RWSOTUBLDIXVET-UHFFFAOYSA-N 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- QIVBCDIJIAJPQS-VIFPVBQESA-N L-tryptophane Chemical compound C1=CC=C2C(C[C@H](N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-VIFPVBQESA-N 0.000 description 1
- GUBGYTABKSRVRQ-PICCSMPSSA-N Maltose Natural products O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-PICCSMPSSA-N 0.000 description 1
- 241000187486 Mycobacterium flavescens Species 0.000 description 1
- CTQNGGLPUBDAKN-UHFFFAOYSA-N O-Xylene Chemical compound CC1=CC=CC=C1C CTQNGGLPUBDAKN-UHFFFAOYSA-N 0.000 description 1
- 239000005662 Paraffin oil Substances 0.000 description 1
- 241000589517 Pseudomonas aeruginosa Species 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 241000223260 Trichoderma harzianum Species 0.000 description 1
- QIVBCDIJIAJPQS-UHFFFAOYSA-N Tryptophan Natural products C1=CC=C2C(CC(N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-UHFFFAOYSA-N 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- WQZGKKKJIJFFOK-PHYPRBDBSA-N alpha-D-galactose Chemical compound OC[C@H]1O[C@H](O)[C@H](O)[C@@H](O)[C@H]1O WQZGKKKJIJFFOK-PHYPRBDBSA-N 0.000 description 1
- 229910021529 ammonia Inorganic materials 0.000 description 1
- 230000008485 antagonism Effects 0.000 description 1
- PYMYPHUHKUWMLA-UHFFFAOYSA-N arabinose Natural products OCC(O)C(O)C(O)C=O PYMYPHUHKUWMLA-UHFFFAOYSA-N 0.000 description 1
- 125000003118 aryl group Chemical group 0.000 description 1
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 238000005422 blasting Methods 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 239000005018 casein Substances 0.000 description 1
- BECPQYXYKAMYBN-UHFFFAOYSA-N casein, tech. Chemical compound NCCCCC(C(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(CC(C)C)N=C(O)C(CCC(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(C(C)O)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(COP(O)(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(N)CC1=CC=CC=C1 BECPQYXYKAMYBN-UHFFFAOYSA-N 0.000 description 1
- 235000021240 caseins Nutrition 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 239000013505 freshwater Substances 0.000 description 1
- 229930182830 galactose Natural products 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 230000002906 microbiologic effect Effects 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 230000000877 morphologic effect Effects 0.000 description 1
- 150000002823 nitrates Chemical class 0.000 description 1
- 229940038384 octadecane Drugs 0.000 description 1
- 238000006395 oxidase reaction Methods 0.000 description 1
- 239000012188 paraffin wax Substances 0.000 description 1
- 239000010690 paraffinic oil Substances 0.000 description 1
- 229960005382 phenolphthalein Drugs 0.000 description 1
- KJFMBFZCATUALV-UHFFFAOYSA-N phenolphthalein Chemical compound C1=CC(O)=CC=C1C1(C=2C=CC(O)=CC=2)C2=CC=CC=C2C(=O)O1 KJFMBFZCATUALV-UHFFFAOYSA-N 0.000 description 1
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- 108090000623 proteins and genes Proteins 0.000 description 1
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- 102220201851 rs143406017 Human genes 0.000 description 1
- 239000013049 sediment Substances 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000000600 sorbitol Substances 0.000 description 1
- 239000007921 spray Substances 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 239000010902 straw Substances 0.000 description 1
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- 239000010409 thin film Substances 0.000 description 1
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- PXXNTAGJWPJAGM-UHFFFAOYSA-N vertaline Natural products C1C2C=3C=C(OC)C(OC)=CC=3OC(C=C3)=CC=C3CCC(=O)OC1CC1N2CCCC1 PXXNTAGJWPJAGM-UHFFFAOYSA-N 0.000 description 1
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
Description
Išradimas priklauso mikrobiologijos, biotechnologijos bei aplinkos apsaugos sritims ir gali būti panaudotas vandens, dirvožemio bei grunto, užterštų naftos produktais, ypač sunkiosiomis jos frakcijomis, biologiniam valymui suintensyvinti.The invention relates to the fields of microbiology, biotechnology and environmental protection and can be used to intensify biological treatment of water, soil and soil contaminated by petroleum products, especially its heavy fractions.
Žinomas US patente Nr. 3 813 316 aprašytas štamas Pseudomonas aeruginosa PAO, kuris turi plazmidės CAM, OCT, NAH ir SAL su angliavandenilių utilizacijos genais ir yra panaudotas naftos produktų skaidymui.Known in U.S. Pat. No. 3,813,316 describes a strain of Pseudomonas aeruginosa PAO which has a plasmid CAM, OCT, NAH and SAL with hydrocarbon utilization genes and is used for the degradation of petroleum products.
Šio štamo trūkumas - per daug lėtas naftos degradacijos procesas.The disadvantage of this strain is that the process of oil degradation is too slow.
Žinomas pagal aut. 1. SU 1 805 097, C02F 3/34, štamas Rhodococcus erythropolis No 2 BKM AC-1339D, skaidantis naftą dirvoje.Known for aut. 1. SU 1 805 097, C02F 3/34, strain Rhodococcus erythropolis No 2 BKM AC-1339D, decomposing oil in soil.
Šio štamo trūkumas yra lėtas naftos skaidymas. Skystoje mitybinėje terpėje su 1 % naftos maksimaliai ją biodegraduoja iki 78 %, o grunte per 40 parų - iki 76 %.The disadvantage of this strain is the slow decomposition of oil. Maximum biodegradation in liquid media with 1% oil is up to 78% and in soil over 40 days up to 76%.
Žinomas pagal patentą RU 2 019 527, C02F 3/34, E02B 15/04, C09K 17/00, štamas Rhodococcus erythropolis E-15 skaido naftą ir jos produktus tundros dirvose esant žemoms temperatūroms (8-10 °C).Known according to the patent RU 2 019 527, C02F 3/34, E02B 15/04, C09K 17/00, the strain Rhodococcus erythropolis E-15 decomposes oil and its products in tundra soils at low temperatures (8-10 ° C).
Šio štamo ypatybė - geriausiai tinka naudoti geografiniuose rajonuose su šaltu klimatu.A special feature of this strain is that it is best suited for use in geographical areas with cold climates.
Žinomas pagal patentą RU 2 053 296, C12N 1/20, C02F 3/34, štamas Mycobacterium flavescens ΕΧ-91, naudojamas vandens ir dirvos valymui nuo naftos ir naftos produktų.Known according to patent RU 2 053 296, C12N 1/20, C02F 3/34, Mycobacterium flavescens ΕΧ-91, used for cleaning water and soil from oil and petroleum products.
Šio štamo trūkumas - lėtas skaidymas ir aukšta funkcionavimo temperatūra. Esant 28°C temperatūrai per 7 paras jis suskaido 74% naftos ir 55% mazuto.The disadvantage of this strain is its slow degradation and high functioning temperature. At 28 ° C it breaks down 74% of oil and 55% of fuel oil in 7 days.
Žinomas azotfiksuojantis štamas Azotobacter vinelandii 21, aprašytas patento LR 3111, C12N 15/00, aprašyme, išskirtas iš užteršto sunkiosiomis naftos frakcijomis vandens poA known azotyping strain Azotobacter vinelandii 21, described in patent LR 3111, C12N 15/00, isolated from water contaminated by heavy petroleum fractions after
τ. LT 4792 B tanklaivio avarijos ir naudojamas gėlo ir sūraus vandens bei dirvos valymui nuo naftos ir jos produktų.τ. EN 4792 B is a tanker accident and is used to clean fresh and salt water and soil from oil and oil products.
Šio štamo trūkumas - lėtas naftos ir sunkiųjų jos frakcijų skaidymas ir aukšta 30°C optimali vystymosi temperatūra. Per 21 parą šis štamas suskaidė 75,6 % parafininės naftos ir 80,2 % aromatinės naftos.The disadvantage of this strain is the slow decomposition of oil and its heavy fractions and the high optimum temperature of development of 30 ° C. Within 21 days, this strain decomposed 75.6% of paraffin oil and 80.2% of aromatic oil.
Artimiausias siūlomam štamui pagal savo sistematinę padėtį ir biodegradacines savybes štamas Pseudomonas putida 36, skaidantis naftą ir jos produktus vandenyje ir dirvožemyje (žiūrėti a. 1. SU 1 076 446, C12N 15/00, C02F 3/34, C12R 1/40).Closest to the proposed strain, according to its systematic position and biodegradable properties, is Pseudomonas putida 36, which decomposes oil and oil products in water and soil (see a. 1. SU 1 076 446, C12N 15/00, C02F 3/34, C12R 1/40).
Šio štamo trūkumas - lėtas naftos skaidymas ir aukštoje 28 - 40 °C temperatūroje. Per 30 parų suskaido 74,5 % parafininės naftos ir 68 % aromatinės. Lėtai skaido ir sunkiausius aromatinius naftos produktus - per 21 parą suskaido iki 57,2 %.The disadvantage of this strain is the slow decomposition of oil and high temperatures of 28 - 40 ° C. Within 30 days, 74.5% of paraffinic oil and 68% of aromatic oil are broken down. Slowest degradation of the heaviest aromatic petroleum products - up to 57.2% within 21 days.
Išradimo tikslas - pagreitinti sunkiųjų naftos frakcijų degradavimą žemesnėse temperatūrose (20 - 25 °C), naudojant atrinktą Pseudomonas fluorescens IGN 57 štamą.The object of the invention is to accelerate the degradation of heavy petroleum fractions at lower temperatures (20-25 ° C) using a selected Pseudomonas fluorescens IGN 57 strain.
Siūlomas Pseudomonas fluorescens IGN 57 štamas, išskirtas iš mazutu užteršto naftos bazės dirvožemio. Štamas deponuotas Botanikos instituto Biodestruktorių tyrimo laboratorijoje, kolekcinis numeris IBL: B-57.Proposed strain Pseudomonas fluorescens IGN 57 isolated from fuel oil contaminated petroleum base soil. The strain is deposited in the Biodestructors Research Laboratory of the Botanical Institute, Collecting Number IBL: B-57.
vv
Štamas Pseudomonas fluorescens IGN 57 išskirtas panaudojant klasikinę mikrobiologinę grynų kultūrų išskyrimo metodiką (P. Gerhardt (ed.), 1981: Manual of Methods for General Bacteriology. - W?.shington). Naftuoto dirvožemio mėginys įberiamas į mineralinę terpę su skaidomu substratu ir kultivuojamas 20°C temperatūroje. Iš gautos mišrios kultūros išskiriamos grynos kultūros. Iš daugelio tokių izoliatų atrenkamas štamas, turintis didžiausią aktyvumą skaidant naftos produktus.The strain Pseudomonas fluorescens IGN 57 was isolated using the classical microbiological purification method of pure cultures (P. Gerhardt (ed.), 1981: Manual of Methods for General Bacteriology. - W. Shington). The oily soil sample is poured into mineral medium with degradable substrate and cultured at 20 ° C. Pure cultures are isolated from the resulting mixed culture. From many such isolates, the strain having the highest activity in the degradation of petroleum products is selected.
Atrinkta aktyviausia kultūra vadovaujantis 9-to leidimo Bergey apibūdintoju (R. G. F. Murray et. ai. (eds. board), 1984-1989: Bergeys Manual of Systematic Bacteriology. 9th ed., vol I-IV - Williams and Wilkins Co., Baltimore) apibūdinta kaip Pseudomonas fluorescens Migula 1895,29AL , štamo numeris - IGN 57.Selected intense culture according to the 9th permit Bergey apibūdintoju (RGF Murray et. Al. (Eds. Board), 1984-1989: Bergey Manual of Systematic Bacteriology. 9 th ed., Vol I-IV - The Williams and Wilkins Co., Baltimore ) described as Pseudomonas fluorescens Migula 1895,29 AL , strain number IGN 57.
Siūlomas mikroorganizmo štamas pasižymi šiomis savybėmis:The proposed strain has the following properties:
Morfologinės savybės. Ląstelės plonų lazdelių formos 0,5 x 2-3 ųm dydžio, sudaro binarinius junginius. Gramneigiamos. Judrios - juda kelių poliarinių žiuželių pagalba. Ląstelių viduje nekaupia ροΐι-β-oksibutirato. Nesudaro endosporų, neturi kapsulių.Morphological properties. The cells are in the form of thin rods 0.5 x 2-3 µm in size, forming binary compounds. Gram negative. Moving - Moves with the help of several polar bells. It does not accumulate ροΐι-β-oxybutyrate inside the cells. Does not form endospores, does not contain capsules.
Kultūriniai požymiai. Ant mėsos ir peptono agaro (MPA) užauga per 24 vai, sudarydamas apvalias taisyklingas, lygaus krašto, skaidrias, iškilo profilio, blizgančias kolonijas 1-3 mm skersmens. Sintetina geltonai žalią fluorescuojantį intraląstelinį pigmentą kuris lengvai pastebimas praeinančioje šviesoje. Auginant mėsos ir peptono buljone (MPB) atsiranda homogeniškas drumstumas, plona plėvelė ir mažas kiekis nuosėdų Ant bulvės užauga bespalvis štrichas su matiniu paviršiumi. Ant agarizuotos mineralinės terpės su nafta pradeda augti per 24 vai. sudarydamos smulkias kolonijas. Ant terpės su mazutu augimas pasirodo po 72 vai. Aplink kolonijas tiek su nafta, tiek su mazutu sudaro lipofobines substrato atstūmimo zonas.Cultural attributes. Meat and peptone agar (MPA) grows in 24 hours, forming round, regular, smooth, clear, raised, glossy colonies 1-3 mm in diameter. Synthesizes yellow-green fluorescent intracellular pigment that is easily visible in passing light. Growing meat and peptone broth (MPB) produces a homogeneous turbidity, a thin film and a small amount of sediment. On agarized mineral medium with oil begins to grow within 24 hours. forming small colonies. The growth on fuel oil medium appears after 72 hours. Lipophobic substrate repulsion zones are formed around the colonies with both oil and fuel oil.
Fiziologinės savybės. Griežtas aerobas, katalazės ir oksidazės reakcijos teigiamos. Tipiškas mezofilas, gerai auga 4-37°C temperatūrų intervale, optimali temperatūra 20-28 °C. 42 °C temperatūroje neauga. Auga ant Ešbio terpės. Gerai įsisavina mineralinius azoto šaltinius, nitratus redukuoja iki azoto. Skystina želatiną krakmolo nehidrolizuoja, kazeino proteolizė silpna. Sudaro sieros vandenilį ir amoniaką. Naudoja gliukozę, ksilozę, sacharozę bei galaktozę, sudarydamas rūgštis, su maltoze rūgščių nesudaro, gerai auga su sorbitu ir etanoliu. Auga MPB su 0,6 % NaCI. Neįsisavina sviesto rūgšties ir triptofano. Auga ant mineralines terpės su angliavandeniliais: heksanu, dekanu, heksadekanu, oktadekanu, ksilolu, toluolu, n' ftalinu, dekalinu, parafinu. Mineralinėje terpėje su angliavandeniliniais substratais sintetina ekstraceliuliarinį emulguojantį junginį, padedantį lengviau įsisavinti hidrofobinį substratąPhysiological properties. Strict aerobic, catalase and oxidase reactions positive. Typical mesophyll, growing well in the temperature range 4-37 ° C, optimum temperature 20-28 ° C. Does not grow at 42 ° C. Grows on Ashbee medium. It absorbs mineral nitrogen well, reduces nitrates to nitrogen. Liquid gelatin does not hydrolyze starch, casein proteolysis is weak. Forms hydrogen sulphide and ammonia. Uses glucose, xylose, sucrose and galactose to form acids, does not form acids with maltose, grows well with sorbitol and ethanol. Growth of MPB with 0.6% NaCl. Does not absorb butyric acid and tryptophan. It grows on mineral medium containing hydrocarbons: hexane, decane, hexadecane, octadecane, xylene, toluene, n 'phthalin, decalin, paraffin. Extracellular emulsifying compound synthesized in mineral medium with hydrocarbon substrates to facilitate hydrophobic substrate uptake
Siūlomo štamo Pseudomonas fluorescens IGN 57 biodestrukcinis pajėgumas buvo tirtas auginant skystoje mineralinėje terpėje su naftalinu, nafta ir mazutu kaip vieninteliais anglies šaltiniais. Buvo panaudota mineralinė terpė tokios sudėties (g/l): NH4NO3 -1,5; KH2PO4 1; KCI - 0,5; MgSCb χ 7H2O - 0,5; FeSCL x 7H2O - 0,001. pH 7,2-7,4. Į terpę substrato dėta 1% Kultivuota ant kratyklės 20 parų esant 20 - 25 °C temperatūroms. Likučiai matuotiThe biodestructive capacity of the proposed strain Pseudomonas fluorescens IGN 57 has been investigated in liquid mineral medium with naphthalene, petroleum and fuel oil as the sole carbon sources. A mineral medium of the following composition (g / l) was used: NH4NO3 -1.5; KH 2 PO 4 1; KCl - 0.5; MgSO 4 χ 7H 2 O - 0.5; FeSCL x 7H2O - 0.001. pH 7.2-7.4. Substrate in medium was 1% Cultured on a shaker for 20 days at 20-25 ° C. Residues were measured
7-ą ir 20-ą kultivavimo parą.7th and 20th cultivation day.
Rezultatai pateikti 1 lentelėje.The results are shown in Table 1.
lentelėtable
Štamo Pseudomonas fluorescens IGN 57 angliavandenilinių substratų suskaidymasDegradation of hydrocarbon substrates of Pseudomonas fluorescens IGN 57
Siūlomas štamas per 7 paras laboratorinėse sąlygose suskaido (lyginant su išmatuotu substrato kiekiu bandymo pradžioje) 87,9 % naftalino, 84,0 % naftos ir 60,7 % mazuto. 20-ą parą naftaiinas suskaidytas iki 99,9 %, o nafta - 99,8 %.The proposed strain decomposes under laboratory conditions (relative to the measured substrate content at the start of the test) in 87.9% of naphthalene, 84.0% of petroleum and 60.7% of fuel oil within 7 days. On day 20, naphthalene is broken down to 99.9% and oil to 99.8%.
Kultivuojant vienoje terpėje siūlomą štamą kartu su grybu štamu Trichoderma harzianum VNB-16 po 7 parų mazuto suskaidymas siekė 71,6 %. Tuo tarpu vienas grybas suskaidė tik 15,3 % mazuto.When cultivated on a single medium, the proposed strain, together with the fungal strain Trichoderma harzianum VNB-16, showed 71.6% degradation of the fuel oil after 7 days. Meanwhile, one mushroom breaks down only 15.3% of fuel oil.
Atliekant antagonizmo tyrimus nustatyta, kad siūlomas štamas Pseudomonas fluorescens IGN 57 nėra antagonistiškas plačiausiai dirvoje paplitusioms grybų ir mielių rūšims, taip pat ir gramteigiamoms bakterijoms.Antagonism studies have shown that the proposed strain Pseudomonas fluorescens IGN 57 is not antagonistic to the most widespread fungal and yeast species, including Gram-positive bacteria.
Siūlomo štamo Pseudomonas fluorescens IGN 57 ir žinomo štamo Pseudomonas putida 36 biodegradacinių savybių palyginimas pateiktas 2 lentelėje.A comparison of the biodegradable properties of the proposed strain Pseudomonas fluorescens IGN 57 and the known strain Pseudomonas putida 36 is presented in Table 2.
lentelėtable
Štamų Pseudomonas fluorescens IGN 57 ir žinomo Pseudomonas putida 36 biodegradacinių savybių palyginimasComparison of the biodegradability of Pseudomonas fluorescens IGN 57 and known Pseudomonas putida 36
Siūlomas štamas Pseudomonas fluorescens IGN 57 palyginus su štamu Pseudomonas putida 36 pasižymi tuo, kad greičiau skaido naftą bei naftos sunkiąsias frakcijas žemesnėse temperatūrose - 20 - 25 °C. Be to, esant terpėje hidrofobiniam angliavandeniliniam substratui, išskiria į aplinką ekstraceliuliarinį emulguojantį junginį, pagreitinantį skaidymo procesą. Taip pat nėra antagonistiškas plačiausiai dirvoje paplitusioms grybų ir mielių rūšims ir gramteigiamoms bakterijoms. Minėti požymiai leidžia aprašytąjį štamą panaudoti dirvožemio ir grunto biologinio valymo nuo naftos ir jos produktų pagreitinimui.The proposed strain Pseudomonas fluorescens IGN 57, compared to Pseudomonas putida 36, is characterized by faster degradation of oil and heavy oil fractions at lower temperatures of 20-25 ° C. In addition, in the presence of a hydrophobic hydrocarbon substrate in the medium, it releases into the environment an extracellular emulsifying compound that accelerates the degradation process. It is also not antagonistic to the most widespread fungal and yeast species and Gram-positive bacteria in the soil. These features make it possible to use the strain described to accelerate the biological treatment of soil and soil from oil and oil products.
Štamo panaudojimas iliustruojamas pavyzdžiais.The use of the strain is illustrated by examples.
pavyzdysexample
Ant dirvos išsipylė nafta, užterštas plotas 200 m2, teršalų kiekis 1 m2 yra 5 kg. Bendras teršalų kiekis 1 tona. Paruošiame Pseudomonas fluorescens IGN 57 suspensiją: 40 1 fermentatoriuje išaugintos biomasės praskiedžiame 2 m'’ druskų ir kt. bakterijai reikalingų priedų tirpale. Gautą suspensiją išpurškiame ant užteršto dirvožemio sluoksnio jį perkasant ir išpurenant. Valymo proceso eigoje dirvos sluoksnio purenimą atliekame periodiškai 1 kartą per savaitę.Oil spilled on the soil, polluted area 200 m 2 , pollutants 1 m 2 is 5 kg. Total emissions per tonne. Prepare a suspension of Pseudomonas fluorescens IGN 57: dilute 2 m '' of salt, etc., in 40 L of biomass grown in the fermenter. bacterial additives in solution. The resulting slurry is sprayed onto the contaminated soil layer by digging and blasting. During the cleaning process, we loosen the soil layer periodically once a week.
Rezultatas: 1000 kg naftos, užteršiančios gruntą (500 mg naftos/cm2 arba 1 tona naftos/200 m2) ir valant šį naftingą gruntą paskleidus jį 0,5 m sluoksniu, naudojant Pseudomonas fluorescens IGN 57, suardoma 97 % naftos per parą.Result: 1000 kg of oil contaminating the soil (500 mg oil / cm 2 or 1 tonne oil / 200 m 2 ) and cleaning this oily soil by spreading it in a 0.5 m layer using Pseudomonas fluorescens IGN 57 destroys 97% of the oil daily.
pavyzdysexample
Naftos produktais užteršto dirvožemio valymas aikštelėje. Valymo aikštelėje yra 100 tonų užteršto dirvožemio. Naftos produktų 20 mg/g žemės Dirvožemis sumaišomas su trąšomis, turinčiomis N ir P bei kitų priedų, reikalingų bakterijų dauginimuisi. Patikrinamas mišinio pH ir, jei jis mažesnis už 4, mišinį reikia kalkinti. Paskui dirvožemis paskleidžiamas 0,5 m storio sluoksniu (jei teršalų daugiau, sluoksnis turi būti plonesnis). Paruošiame Pseudomonas fluorescens IGN 57 suspensiją: 100 1 fermentatoriuje išaugintos biomasės praskiedžiame 10 mJ druskų ir kt. bakterijai reikalingų priedų tirpale. Gautą suspensiją išpurškiame ant paruošto dirvožemio sluoksnio jį perkasant.On-site remediation of oil-contaminated soil. The treatment site contains 100 tonnes of contaminated soil. Petroleum products 20 mg / g of soil The soil is mixed with fertilizers containing N and P and other additives necessary for bacterial growth. The pH of the mixture is checked and, if less than 4, the mixture must be limed. The soil is then spread in a layer 0.5 m thick (if pollutants are higher, the layer should be thinner). Prepare a suspension of Pseudomonas fluorescens IGN 57 by diluting 10 mJ of salt, etc. in 100 L of biomass grown in the fermenter. bacterial additives in solution. The resulting slurry is sprayed onto the prepared soil layer by digging.
Rezultatas: 2000 kg naftos, teršiančios gruntą (20 mg naftos/1 g grunto), valant šį naftingą gruntą paskleidus jį 0,5 m sluoksniu, naudojant Pseudomonas fluorescens IGN 57, suardoma 98 % naftos per parą.Result: 2000 kg of oil polluting the soil (20 mg oil / 1 g soil), purifying this oily soil by spreading it in 0.5 m layers using Pseudomonas fluorescens IGN 57, destroys 98% of the oil daily.
pavyzdysexample
Naftos produktais užteršto dirvožemio valymas kompostavimo būdu. Vasarą aikštelėje (žiemą šildomoje patalpoje iki 20 °C) yra 100 tonų užteršto dirvožemio. Naftos produktų 20 mg/g žemės. Dirvožemis sumaišomas su trąšomis, turinčiomis N ir P bei kitų priedų, reikalingų bakterijų dauginimuisi. Į mišinį pridedama susmulkintų šiaudų. Patikrinamas mišinio pH ir, jei jis mažesnis už 4, mišinį reikia kalkinti. Paskui dirvožemis supilamas suformuojant kaupą. Jei teršalų daug ir žemės kiekis didelis, kaupe reikia įrengti oro prapūtimo sistemą (aktyvus būdas) arba išvedžioti vamzdelių sistemą natūraliai ventiliacijai paspartinti (pasyvus būdas). Paruošiame Pseudomonas fluorescens IGN 57 suspensiją: 20 1 fermentatoriuje išaugintos biomasės praskiedžiame 1 mJ druskų ir kt. bakterijai reikalingų priedų tirpale. Gautą suspensiją išpurškiame ant paruošto dirvožemio kaupo, kuo giliau jį perkasant.Composting of petroleum-contaminated soil. In summer, the site (heated in winter up to 20 ° C) contains 100 tonnes of contaminated soil. Petroleum products 20 mg / g ground. The soil is mixed with fertilizers containing N and P and other additives necessary for bacterial growth. Crushed straw is added to the mixture. The pH of the mixture is checked and, if less than 4, the mixture must be limed. The soil is then poured to form a lump. If the pollutant is high and the amount of land is large, the hood must be equipped with an air purge system (active mode) or a pipe system to accelerate natural ventilation (passive mode). Prepare a suspension of Pseudomonas fluorescens IGN 57: dilute 1 m J of salt and so on with 20 L of biomass grown in the fermenter. bacterial additives in solution. The resulting slurry is sprayed onto the prepared soil, digging as deep as possible.
Rezultatas: 2000 kg naftos, teršiančios gruntą (20 mg naftos/1 g grunto), valant šį naftingą gruntą grunto kaupo sistemoje, naudojant Pseudomonas fluorescens IGN 57, suardoma 97 % naftos per 36 paras.Result: 2000 kg of oil polluting the soil (20 mg oil / 1 g soil), cleaning this oily soil in a soil accumulation system using Pseudomonas fluorescens IGN 57, destroys 97% of the oil in 36 days.
pavyzdysexample
Naftos produktais užteršto vandens telkinio valymas. Užterštas plotas 100 m2, teršalų kiekis 0,2 tonos. Užterštą plotą reikia izoliuoti (panaudojant spec. priemones) nuo platesnio pasiskleidimo. Paruošiame Pseudomonas fluorescens IGN 57 suspensiją: 10 1 fermentatoriuje išaugintos biomasės praskiedžiame 0,5 mJ druskų ir kt. bakterijai reikalingų priedų tirpale. Paruoštą suspensiją išpurkškiame ant vandens paviršiaus 5 1 į 1 m2 paviršiaus.Purification of an oil-polluted water body. The contaminated area is 100 m 2 and the amount of pollutants is 0.2 tons. Contaminated area should be isolated (by special measures) from widespread spread. Prepare a suspension of Pseudomonas fluorescens IGN 57 by diluting 0.5 m J of salt and so on with 10 L of biomass grown in the fermenter. bacterial additives in solution. Spray the prepared suspension on a water surface of 5 1 to 1 m 2 .
Rezultatas: kai vandens paviršius užterštas nafta (200 mg naftos/1 cm2 arba 200 kg/100 m2) ir valomas naudojant Pseudomonas fluorescens IGN 57, suardoma 99 % naftos per parą.The result: when the water surface is contaminated with oil (200 mg oil / 1 cm 2 or 200 kg / 100 m 2 ) and cleaned using Pseudomonas fluorescens IGN 57, 99% of the oil is degraded daily.
Štamas Pseudomonas fluorescens IGN 57 naudojimas biologiniam dirvožemio, vandens ir grunto valymui nuo naftos ir jos produktų. Jis gali būti panaudojamas biologiniam valymui avarinio naftos produktų išsipylimo į dirvą atveju, naftos produktams patekus į gėlus vandens telkinius ir taip pat gali būti panaudotas užteršto naftos produktais grunto valymui aikštelėse kompostavimo būdu.Use of Pseudomonas fluorescens IGN 57 for biological treatment of soil, water and soil from oil and oil products. It can be used for biological treatment in the event of an accidental spillage of petroleum products into the soil after oil spills into freshwater and can also be used for composting of petroleum contaminated soil on site.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2015056110A1 (en) | 2013-10-14 | 2015-04-23 | Uab "Biocentras" | Complex method for cleaning environment from oil pollutants |
| LT7087B (en) | 2024-02-07 | 2024-09-10 | Gamtos Tyrimų Centras | BIO PREPARATION FOR CLEANING SOIL CONTAMINATED BY PETROLEUM PRODUCTS AND CLEANING METHOD |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3813316A (en) | 1972-06-07 | 1974-05-28 | Gen Electric | Microorganisms having multiple compatible degradative energy-generating plasmids and preparation thereof |
| SU1076446A1 (en) | 1982-07-22 | 1984-02-29 | Западно-Сибирский научно-исследовательский геологоразведочный нефтяной институт | Strain pseudomonas putida 36 for use in purification of water and soil from petroleum and petroleum products |
| RU2019527C1 (en) | 1993-04-30 | 1994-09-15 | Северный государственный научно-исследовательский и проектно-конструкторский геологический центр | Method of soil treatment from the oil pollution |
| LT3111B (en) | 1992-07-15 | 1994-12-27 | Saulius Grigiskis | The oil pollutants decomposing microorganism azotobacter vinelandii 21 |
| RU2053296C1 (en) | 1992-11-11 | 1996-01-27 | Государственный научно-исследовательский институт прикладной микробиологии | Strain of mycobacterium flavescens used for water and soil treatment from the oil and petroleum products |
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Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3813316A (en) | 1972-06-07 | 1974-05-28 | Gen Electric | Microorganisms having multiple compatible degradative energy-generating plasmids and preparation thereof |
| SU1076446A1 (en) | 1982-07-22 | 1984-02-29 | Западно-Сибирский научно-исследовательский геологоразведочный нефтяной институт | Strain pseudomonas putida 36 for use in purification of water and soil from petroleum and petroleum products |
| LT3111B (en) | 1992-07-15 | 1994-12-27 | Saulius Grigiskis | The oil pollutants decomposing microorganism azotobacter vinelandii 21 |
| RU2053296C1 (en) | 1992-11-11 | 1996-01-27 | Государственный научно-исследовательский институт прикладной микробиологии | Strain of mycobacterium flavescens used for water and soil treatment from the oil and petroleum products |
| RU2019527C1 (en) | 1993-04-30 | 1994-09-15 | Северный государственный научно-исследовательский и проектно-конструкторский геологический центр | Method of soil treatment from the oil pollution |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2015056110A1 (en) | 2013-10-14 | 2015-04-23 | Uab "Biocentras" | Complex method for cleaning environment from oil pollutants |
| LT7087B (en) | 2024-02-07 | 2024-09-10 | Gamtos Tyrimų Centras | BIO PREPARATION FOR CLEANING SOIL CONTAMINATED BY PETROLEUM PRODUCTS AND CLEANING METHOD |
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