vavilovВавиловский журнал генетики и селекцииVavilov Journal of Genetics and Breeding2500-3259Institute of Cytology and Genetics of Siberian Branch of the RAS10.18699/VJGB-22-70vavilov-3479Research ArticleГЕНЕТИКА МИКРООРГАНИЗМОВMICROBIAL GENETICSАктивность гена алканмонооксигеназы alkB у штаммов углеводородокисляющих бактерий, выделенных из нефтепродуктовActivity of alkanmonooxygenase alkB gene in strains of hydrocarbon-oxidizing bacteria isolated from petroleum productshttps://orcid.org/0000-0002-4005-8114ШапироТ. Н.ShapiroT. N.

Москва

Moscow

floyd52@rambler.ruhttps://orcid.org/0000-0002-7304-7753МанучароваН. А.ManucharovaN. A.

Москва

Moscow

https://orcid.org/0000-0002-7054-0024ЛобаковаЕ. С.LobakovaE. S.

Москва

Moscow

Московский государственный университет им. М.В. Ломоносова, биологический факультетРоссияLomonosov Moscow State University, Faculty of BiologyRussian FederationМосковский государственный университет им. М.В. Ломоносова, факультет почвоведенияРоссияLomonosov Moscow State University, Faculty of Soil ScienceRussian Federation202209102022266575582Copyright © Шапиро Т.Н., Манучарова Н.А., Лобакова Е.С., 20222022Шапиро Т.Н., Манучарова Н.А., Лобакова Е.С.Shapiro T.N., Manucharova N.A., Lobakova E.S.This work is licensed under a Creative Commons Attribution 4.0 License.https://vavilov.elpub.ru/jour/article/view/3479

Ферменты алканмонооксигеназы AlkB и Cyp153 ответственны за аэробную деградацию н-алканов нефти и нефтепродуктов. Для доказательства использования штаммами углеводородокисляющих бактерий, выделенных из авиационного керосина ТС-1 и автомобильного бензина АИ-95, н-алканов нефти и нефтепродуктов, проведена детекция ключевых генов alkB, Alk1, Alk2, Alk3 и Cyp153, кодирующих алканмонооксигеназы AlkB и Cyp153, ответственных за окисление углеводородов с определенной длиной цепи. Установлено, что штаммы бактерий, изолированные из реактивного топлива ТС-1, за исключением Deinococcus sp. Bi7, имели как минимум один из исследованных генов деградации н-алканов. Штаммы Sphingobacterium multivorum Bi2, Alcaligenes faecalis Bi3, Rhodococcus sp. Bi4, Sphingobacterium sp. Bi5, Rhodococcus erythropolis Bi6 содержали ген alkB. У штаммов углеводородокисляющих бактерий, выделенных из бензина АИ-95, этот ген алканмонооксигеназы не был детектирован. С помощью метода ПЦР в реальном времени проанализирована активность гена alkB у всех полученных из нефтепродуктов штаммов бактерий и определено число его копий. Методом ПЦР в реальном времени с использованием праймера с другой последовательностью нуклеотидов для детекции гена alkB установлена его активность у всех штаммов бактерий, выделенных из бензина АИ-95, причем штамм Paenibacillus agaridevorans Bi11 отнесен к группе с высоким уровнем его активности (1290 копий/мл). По оценке роста исследованных углеводородокисляющих бактерий на плотной минеральной среде Эванса с модельной смесью углеводородов штаммы были разделены на три группы. Отмечены совпадения результатов по распределению штаммов углеводородокисляющих бактерий в группах по активности гена alkB и группах, сформированных на основе способности роста и использования модельной смеси углеводородов и нефтепродуктов. Полученные результаты свидетельствуют о необходимости применения комплекса молекулярногенетических и физиологических методов для всестороннего анализа распространения исследуемых генов у бактерий и оценки их активности в штаммах углеводородокисляющих бактерий, способных к биодеградации углеводородов нефтепродуктов.

Alkanmonooxygenase enzymes AlkB and Cyp153 are responsible for the aerobic degradation of n-alkanes of petroleum and petroleum products. To prove the usage of n-alkanes from oil and petroleum products by hydrocarbon- oxidizing bacteria isolated from aviation kerosene TS-1 and automobile gasoline AI-95, the detection of the key genes alkB, Alk1, Alk2, Alk3 and Cyp153 encoding alkanmonooxygenases AlkB and Cyp153 (responsible for the oxidation of hydrocarbons with a certain chain length) was carried out. It was found that bacterial strains isolated from TS-1 jet fuel, except Deinococcus sp. Bi7, had at least one of the studied n-alkane degradation genes. The strains Sphingobacterium multivorum Bi2; Alcaligenes faecalis Bi3; Rhodococcus sp. Bi4; Sphingobacterium sp. Bi5; Rhodococcus erythropolis Bi6 contained the alkB gene. In the strains of hydrocarbon-oxidizing bacteria isolated from gasoline AI- 95, this alkanmonooxygenase gene was not detected. Using the real-time PCR method, the activity of the alkB gene in all bacterial strains isolated from petroleum products was analyzed and the number of its copies was determined. By real-time PCR using a primer with a different sequence of nucleotides to detect the alkB gene, its activity was established in all bacterial strains isolated from gasoline AI-95; besides, the strain Paenibacillus agaridevorans Bi11 was assigned to the group with a high level of its activity (1290 copies/ml). According to the assessment of the growth of isolated hydrocarbon-oxidizing bacteria on a solid Evans mineral medium with the addition of the model mixture of hydrocarbons, the strains were divided into three groups. The distributions of strains of hydrocarbon-oxidizing bacteria in the groups based on the activity of the alkB gene and groups formed based on the growth ability and use of the model mixture of hydrocarbons and petroleum products were found to be consistent. The results obtained indicate that we need to use a complex of molecular and physiological methods for a comprehensive analysis of the distribution of the studied genes in bacteria and to assess their activity in the strains of hydrocarbon-oxidizing bacteria capable of biodegradation of petroleum hydrocarbons.

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The authors declare that there are no conflicts of interest present.