RU2154103C1 - Strain pseudomonas species 78g reserved for degradation of organophosphorus toxin destruction products - Google Patents
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- RU2154103C1 RU2154103C1 RU99100570A RU99100570A RU2154103C1 RU 2154103 C1 RU2154103 C1 RU 2154103C1 RU 99100570 A RU99100570 A RU 99100570A RU 99100570 A RU99100570 A RU 99100570A RU 2154103 C1 RU2154103 C1 RU 2154103C1
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- organophosphorus
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- 241000589516 Pseudomonas Species 0.000 title claims description 5
- 230000015556 catabolic process Effects 0.000 title claims description 5
- 238000006731 degradation reaction Methods 0.000 title claims description 5
- 230000006378 damage Effects 0.000 title abstract description 16
- 239000003053 toxin Substances 0.000 title abstract 2
- 231100000765 toxin Toxicity 0.000 title abstract 2
- 230000001580 bacterial effect Effects 0.000 claims abstract description 4
- 239000007857 degradation product Substances 0.000 claims description 4
- 230000007123 defense Effects 0.000 claims description 2
- 231100000614 poison Toxicity 0.000 claims description 2
- 239000003440 toxic substance Substances 0.000 claims description 2
- 230000000694 effects Effects 0.000 abstract description 6
- 238000005516 engineering process Methods 0.000 abstract description 3
- 230000002906 microbiologic effect Effects 0.000 abstract description 3
- 229920001817 Agar Polymers 0.000 abstract description 2
- 239000008272 agar Substances 0.000 abstract description 2
- 239000000126 substance Substances 0.000 abstract 3
- 241000589774 Pseudomonas sp. Species 0.000 abstract 1
- 230000007918 pathogenicity Effects 0.000 abstract 1
- 108700012359 toxins Proteins 0.000 abstract 1
- 239000000047 product Substances 0.000 description 10
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 3
- 238000000354 decomposition reaction Methods 0.000 description 3
- YACKEPLHDIMKIO-UHFFFAOYSA-N methylphosphonic acid Chemical compound CP(O)(O)=O YACKEPLHDIMKIO-UHFFFAOYSA-N 0.000 description 3
- 244000005700 microbiome Species 0.000 description 3
- 239000000575 pesticide Substances 0.000 description 3
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 2
- 241000194107 Bacillus megaterium Species 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 2
- 108010053446 Carbon-phosphorus lyase Proteins 0.000 description 2
- 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 2
- SIKJAQJRHWYJAI-UHFFFAOYSA-N Indole Chemical compound C1=CC=C2NC=CC2=C1 SIKJAQJRHWYJAI-UHFFFAOYSA-N 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 239000008103 glucose Substances 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 230000000877 morphologic effect Effects 0.000 description 2
- 235000015097 nutrients Nutrition 0.000 description 2
- 230000001717 pathogenic effect Effects 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- BLALDUPQYCGKAG-UHFFFAOYSA-N 3,3-dimethylbutan-2-yloxy(methyl)phosphinic acid Chemical compound CC(C)(C)C(C)OP(C)(O)=O BLALDUPQYCGKAG-UHFFFAOYSA-N 0.000 description 1
- 102000016938 Catalase Human genes 0.000 description 1
- 108010053835 Catalase Proteins 0.000 description 1
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- 229930195725 Mannitol Natural products 0.000 description 1
- 229910002651 NO3 Inorganic materials 0.000 description 1
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 description 1
- 102000004316 Oxidoreductases Human genes 0.000 description 1
- 108090000854 Oxidoreductases Proteins 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 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
- 229930006000 Sucrose Natural products 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 229910021529 ammonia Inorganic materials 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
- 238000004587 chromatography analysis Methods 0.000 description 1
- WOAFDHWYKSOANX-UHFFFAOYSA-N diisopropyl methylphosphonate Chemical compound CC(C)OP(C)(=O)OC(C)C WOAFDHWYKSOANX-UHFFFAOYSA-N 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- 239000007789 gas Substances 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
- PZOUSPYUWWUPPK-UHFFFAOYSA-N indole Natural products CC1=CC=CC2=C1C=CN2 PZOUSPYUWWUPPK-UHFFFAOYSA-N 0.000 description 1
- RKJUIXBNRJVNHR-UHFFFAOYSA-N indolenine Natural products C1=CC=C2CC=NC2=C1 RKJUIXBNRJVNHR-UHFFFAOYSA-N 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 125000001477 organic nitrogen group Chemical group 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- 229910052698 phosphorus Inorganic materials 0.000 description 1
- 239000011574 phosphorus Substances 0.000 description 1
- 239000001509 sodium citrate Substances 0.000 description 1
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical class O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 239000008399 tap water Substances 0.000 description 1
- 235000020679 tap water Nutrition 0.000 description 1
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 1
- 239000002676 xenobiotic agent Substances 0.000 description 1
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- Enzymes And Modification Thereof (AREA)
Abstract
Description
Изобретение относится к биотехнологии, в частности микробиологической очистке окружающей среды от продуктов деструкции ФОВ, может быть использовано для деградации остаточных количеств продуктов деструкции ФОВ при различных технологиях их уничтожения. The invention relates to biotechnology, in particular microbiological purification of the environment from the products of the destruction of the FOV, can be used to degrade the residual products of the destruction of the FOV with various technologies for their destruction.
Цель изобретения - получение штамма, способного активно разлагать широкий спектр продуктов деструкции ФОВ. Штамм непатогенен, хранится и поддерживается на МПА при температуре (5±1)oC с пересевами через 6 мес.The purpose of the invention is to obtain a strain capable of actively decomposing a wide range of products of destruction of FOV. The strain is non-pathogenic, stored and maintained on MPA at a temperature of (5 ± 1) o C with reseeding after 6 months.
Уровень техники. Изобретение относится к области биотехнологии, в частности микробиологической очистке продуктов деструкции ФОВ при различных технологиях их уничтожения, и представляет собой штамм бактерий, который имеет высокую C-P-лиазную активность и может найти применение для разложения остаточных количеств продуктов деструкции ФОВ, таких как метилфосфоновая кислота, эфиры метилфосфоновой кислоты и другие. The prior art. The invention relates to the field of biotechnology, in particular microbiological purification of products of destruction of FOV with various technologies for their destruction, and is a strain of bacteria that has a high CP-lyase activity and can be used to decompose residues of degradation products of FOV, such as methylphosphonic acid, esters methylphosphonic acid and others.
Аналогом изобретения являются штаммы микроорганизмов, предназначенные для деструкции различных ксенобиотиков. Наиболее близок изобретению по совокупности существенных признаков штамм бактерий Bacillus megaterium 501 из коллекции Всесоюзного научно-исследовательского института сельскохозяйственной микробиологии, предназначенный для деструкции фосфорорганических пестицидов [1]. Штамм 501 способен разлагать широкий спектр фосфорорганических пестицидов, но нет сведений о наличии у него C-P-лиазной активности, и отсутствуют данные о возможности его использования для деградации продуктов разложения ФОВ. An analogue of the invention are strains of microorganisms intended for the destruction of various xenobiotics. The closest to the invention in terms of essential features is the bacterial strain Bacillus megaterium 501 from the collection of the All-Union Research Institute of Agricultural Microbiology, intended for the destruction of organophosphorus pesticides [1]. Strain 501 is capable of decomposing a wide range of organophosphorus pesticides, but there is no evidence of its C-P lyase activity, and there is no evidence of the possibility of its use for the degradation of decomposition products of FOV.
Сущность изобретения. Сущность изобретения заключается в получении штамма, обладающего C-P-лиазной активностью и способного активно разлагать широкий спектр продуктов деструкции ФОВ. В качестве такого штамма предлагается штамм Pseudomonas species 78Г из коллекции Центра военно-технических проблем биологической защиты научно-исследовательского института микробиологии Министерства oбороны РФ. SUMMARY OF THE INVENTION The essence of the invention is to obtain a strain having C-P-lyase activity and is able to actively decompose a wide range of products of the destruction of FOV. As such a strain, the strain Pseudomonas species 78G from the collection of the Center for Military-Technical Problems of Biological Protection of the Research Institute of Microbiology of the Ministry of Defense of the Russian Federation is proposed.
Штамм имеет следующие морфологические и физиолого-биохимические характеристики. The strain has the following morphological and physiological and biochemical characteristics.
Морфологические признаки. Бактерии представляют собой подвижные палочки с закругленными концами размером 0,7-1,0х3,0-4,0 мкм. Неспорообразующие. Грамотрицательные. Колонии на МПА размером 2-4 мм, круглые, куполообразные, блестящие, серо-белого цвета. Morphological signs. Bacteria are motile rods with rounded ends measuring 0.7-1.0x3.0-4.0 microns. Non-spore forming. Gram-negative. Colonies on MPA 2-4 mm in size, round, domed, shiny, gray-white.
Физиолого-биохимические признаки. Аэроб. Оптимум роста (27,5±2,5)oC. Пределы роста 5-40oC. Растет при pH 5,0-8,5. Оптимум pH (7,0±0,2).Physiological and biochemical characteristics. Aerobe. The optimum growth (27.5 ± 2.5) o C. The limits of growth of 5-40 o C. It grows at pH 5.0-8.5. Optimum pH (7.0 ± 0.2).
Усваивает глюкозу, сахарозу, маннит, кислоты не образует. Absorbs glucose, sucrose, mannitol, does not form acid.
Каталазная активность положительная, оксидазная активность отрицательная. Catalase activity is positive, oxidase activity is negative.
Использует минеральные и органические источники азота. Нитраты не восстанавливает. Индол, аммиак не выделяет. Желатину не разжижает. Маркерных признаков штамм не имеет. Uses mineral and organic nitrogen sources. Nitrate does not recover. Indole, ammonia does not emit. It does not dilute gelatin. The strain has no marker features.
Признаки штамма устойчивы. Штамм непатогенен. The signs of the strain are stable. The strain is non-pathogenic.
Штамм выделен из лесного серозема Саратовской области. Хранится и поддерживается на мясопептонном агаре при (5±1)oC с пересевами через 6 мес.The strain isolated from forest serozem of the Saratov region. It is stored and maintained on meat and peptone agar at (5 ± 1) o C with reseeding after 6 months.
Штамм выращивают на питательной среде следующего состава, г/л: MgSO4•7H2O по ГОСТ 4523-77 - 0,20; (NH4)2SO4 по ГОСТ 3769-78 - 2,00; KNO3 по ГОСТ 4217-77 - 0,50; трис(оксиметил)-аминометан по ТУ 6-09-4992-76 - 6,05; натрий лимоннокислый трехзамещенный по ГОСТ 22280-76 - 10,00; лимонная кислота, 1-водная, по ГОСТ 3652-69 - 0,20; глюкоза по ГОСТ 975-88 - 0,50; вода водопроводная до 1000 мл, pH 7,2±0,05.The strain is grown on a nutrient medium of the following composition, g / l: MgSO 4 • 7H 2 O according to GOST 4523-77 - 0.20; (NH 4 ) 2 SO 4 according to GOST 3769-78 - 2.00; KNO 3 according to GOST 4217-77 - 0.50; Tris (hydroxymethyl) aminomethane according to TU 6-09-4992-76 - 6.05; tri-substituted sodium citrate according to GOST 22280-76 - 10.00; citric acid, 1-aqueous, according to GOST 3652-69 - 0.20; glucose according to GOST 975-88 - 0.50; tap water up to 1000 ml, pH 7.2 ± 0.05.
Пример. Изучалась деградация продуктов деструкции фосфорорганических отравляющих веществ под действием штамма Pseudomonas species 78Г в лабораторных условиях. Example. We studied the degradation of degradation products of organophosphorus toxic substances under the influence of a strain of Pseudomonas species 78G in laboratory conditions.
Микроорганизмы выращивали в питательной среде описанного состава в конических колбах объемом 250 см3, заполненных 50 см3 среды, на термостатируемой качалке УВМТ-12-250 при 100 об/мин-1 и температуре (30±2)oC в течение суток. Затем переносили в такую же среду, содержащую один из следующих продуктов деструкции ФОВ, г/л: метилфосфоновую кислоту 0,1; диизопропиловый эфир метилфосфоновой кислоты 1,0; пинаколиловый эфир метилфосфоновой кислоты 1,0. Культивировали в течение 7 суток. Концентрацию продуктов деструкции ФОВ определяли газохроматографическим методом и вычисляли степень их разложения.Microorganisms were grown in a nutrient medium of the described composition in 250 cm 3 conical flasks filled with 50 cm 3 of medium on a UVMT-12-250 thermostatically controlled rocking chair at 100 rpm -1 and a temperature of (30 ± 2) o C during the day. Then transferred to the same medium containing one of the following products of the destruction of FOV, g / l: methylphosphonic acid 0.1; methylphosphonic acid diisopropyl ester 1.0; methylphosphonic acid pinacolyl ester 1.0. Cultivated for 7 days. The concentration of the degradation products of DOM was determined by the gas chromatographic method and the degree of their decomposition was calculated.
Результаты опыта представлены в таблице. The results of the experiment are presented in the table.
Из результатов опыта, представленных в таблице, следует, что штамм Pseudomonas species 78Г весьма активно разлагает исследованные продукты деструкции ФОВ. Степень разложения исследованных продуктов составляет 90% через 7 суток. При этом наблюдается значительное увеличение оптической плотности культуры, что свидетельствует об использовании микроорганизмом изученных продуктов деструкции ФОВ в качестве единственного источника фосфора. From the results of the experiment, presented in the table, it follows that the strain Pseudomonas species 78G very actively decomposes the investigated products of the destruction of FOV. The degree of decomposition of the investigated products is 90% after 7 days. At the same time, a significant increase in the optical density of the culture is observed, which indicates the use by the microorganism of the studied products of the degradation of FOV as the sole source of phosphorus.
Источники информации
1. Патент 1735359 Россия, кл. C 12 N 1/20, C 02 F 3/34. - Штамм бактерий Bacillus megaterium, предназначенный для деструкции фосфорорганических пестицидов/Е. А. Гаврилова, Т.О. Лисина, Ю.В. Круглов; Всесоюзный научно-исследовательский институт сельскохозяйственной микробиологии. - N 4781535/13; Заявл. 11.01.90; Опубл. 23.05.92. -Бюл. N 19.Sources of information
1. Patent 1735359 Russia, cl. C 12 N 1/20, C 02 F 3/34. - The bacterial strain Bacillus megaterium, designed for the destruction of organophosphorus pesticides / E. A. Gavrilova, T.O. Lisina, Yu.V. Kruglov; All-Union Research Institute of Agricultural Microbiology. - N 4781535/13; Claim 01/11/90; Publ. 05/23/92. -Bul. N 19.
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| Application Number | Priority Date | Filing Date | Title |
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| RU99100570A RU2154103C1 (en) | 1999-01-10 | 1999-01-10 | Strain pseudomonas species 78g reserved for degradation of organophosphorus toxin destruction products |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| RU99100570A RU2154103C1 (en) | 1999-01-10 | 1999-01-10 | Strain pseudomonas species 78g reserved for degradation of organophosphorus toxin destruction products |
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| RU2154103C1 true RU2154103C1 (en) | 2000-08-10 |
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| RU99100570A RU2154103C1 (en) | 1999-01-10 | 1999-01-10 | Strain pseudomonas species 78g reserved for degradation of organophosphorus toxin destruction products |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116121112A (en) * | 2022-11-18 | 2023-05-16 | 广东工业大学 | A Strain of Pediococcus stetzeri and Its Application in Environmental Restoration |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| SU1652335A1 (en) * | 1989-06-08 | 1991-05-30 | Институт Биохимии И Физиологии Микроорганизмов Ан Ссср | Actinomyces strain streptomyces rochei, carrying out complete degradation of 2,4,6-trichlorophenol, or 2,4-dichlorophenol, or 2,6- dichlorophenol, or 2-chlorophenol |
| SU1735359A1 (en) * | 1990-01-11 | 1992-05-23 | Всесоюзный научно-исследовательский институт сельскохозяйственной микробиологии | Strain of bacteria bacillus megaterium, used for degradation of organophosphate pesticides |
| SU1775471A1 (en) * | 1990-12-07 | 1992-11-15 | Inst Biokhim I Fiziol Mikroorg | Process for producing preparation for decomposition of chlorophenols |
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1999
- 1999-01-10 RU RU99100570A patent/RU2154103C1/en not_active IP Right Cessation
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| SU1652335A1 (en) * | 1989-06-08 | 1991-05-30 | Институт Биохимии И Физиологии Микроорганизмов Ан Ссср | Actinomyces strain streptomyces rochei, carrying out complete degradation of 2,4,6-trichlorophenol, or 2,4-dichlorophenol, or 2,6- dichlorophenol, or 2-chlorophenol |
| SU1735359A1 (en) * | 1990-01-11 | 1992-05-23 | Всесоюзный научно-исследовательский институт сельскохозяйственной микробиологии | Strain of bacteria bacillus megaterium, used for degradation of organophosphate pesticides |
| SU1775471A1 (en) * | 1990-12-07 | 1992-11-15 | Inst Biokhim I Fiziol Mikroorg | Process for producing preparation for decomposition of chlorophenols |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116121112A (en) * | 2022-11-18 | 2023-05-16 | 广东工业大学 | A Strain of Pediococcus stetzeri and Its Application in Environmental Restoration |
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