Search for Biosurfactant-Producing Microorganisms in Oil-Contaminated Soils for Use in Enhanced Oil Recovery Technology

Albert Kh. Safarov, Gulnara G. Kurbanova, Aigul A. Garipova, Daniya A. Kurbanova

Author details

Albert Kh. Safarov
TatNIPIneft R&D Institute of the PJSC TATNEFT named after V.D. Shashin, Almetyevsk, Russian Federation
Gulnara G. Kurbanova
TatNIPIneft R&D Institute of the PJSC TATNEFT named after V.D. Shashin, Almetyevsk, Russian Federation
Aigul A. Garipova
TatNIPIneft R&D Institute of the PJSC TATNEFT named after V.D. Shashin, Almetyevsk, Russian Federation
Daniya A. Kurbanova
TatNIPIneft R&D Institute of the PJSC TATNEFT named after V.D. Shashin, Almetyevsk, Russian Federation

https://doi.org/10.15593/2712-8008/2025.4.16

This study focuses on the identification and isolation of microorganisms producing biological surfactants (biosurfactants) from oil-contaminated soils for subsequent use in environmentally friendly enhanced oil recovery (EOR) technologies. The relevance of this work stems from the need to develop effective and environmentally friendly methods for extracting hard-to-recover reserves (HRR) that minimize environmental risks compared to traditional physical and chemical EOR methods. Twenty strains of hydrocarbon-oxidizing microorganisms were isolated and identified from oil-contaminated soil samples in accordance with GOST 17.4.4.02-2017. Microbiological analysis (MUK 4.2.3695-21) confirmed the absence of pathogenic microflora. Six isolated strains (Bacillus spp. 10RE, Citrobacter spp. 14RE, Bacillus spp. 17RE, Pseudomonas spp. 18RE, Bacillus spp. 21RE, Rhodococcus spp. 24RE) demonstrated a pronounced capacity for surfactant biosynthesis. Key evidence was a significant decrease in the interfacial tension (IFT) of the culture fluid during its development on oil used as the sole carbon source. The values of the IFT in the medium with oil varied from 2.5 to 4.5 mN/m, which is significantly lower than the values in the medium with sucrose 3.5–13.5 mN/m, and indicates the high efficiency of the produced biosurfactants. The adaptability of microorganisms to reservoir conditions is important for application in real oil reservoirs. A study of their physiological characteristics revealed that the selected strains showed wide tolerance ranges: temperature (from 5°C to ≥ 50°C) and salinity (from 0 to 80–240 g NaCl/L). For example, Bacillus spp. 21RE grew at salinities up to 240 g/L, while Pseudomonas spp. 18RE and Citrobacter spp. 14RE exhibited thermotolerance. The obtained results demonstrate the high potential of the isolated strains for the development of innovative biotechnologies. Their ability to efficiently produce biosurfactants directly on the oil substrate and survive in extreme reservoir conditions (high salinity, wide temperature range) makes them promising agents for microbiologically enhanced oil recovery (MEOR). The use of these microorganisms and their metabolites will reduce the IFT at the oil-water interface, mobilize capillary-trapped oil, and increase the oil recovery factor, while simultaneously facilitating the bioremediation of contaminated sites.

Keywords

enhanced oil recoverybiological surfactantsmicrobiological methods of enhanced oil recoveryinterfacial tensionoil-contaminated soilsbioremediationhydrocarbon-oxidizing microorganismsBacillusPseudomonasRhodococcus.

Поиск биологических поверхностно-активных продуцирующих микроорганизмов в нефтезагрязненных грунтах с целью применения в технологии повышения нефтеотдачи пласта

Настоящее исследование посвящено поиску и выделению микроорганизмов-продуцентов биологических поверхностно-активных веществ (биоПАВ) из нефтезагрязненных грунтов для последующего применения в экологически безопасных технологиях повышения нефтеотдачи пластов (ПНП). Актуальность работы обусловлена необходимостью разработки эффективных и экологичных методов извлечения трудноизвлекаемых запасов (ТРИЗ), минимизирующих риски для окружающей среды в сравнении с традиционными физико-химическими методами ПНП. Из нефтезагрязненных почвенных проб, в соответствии с ГОСТ 17.4.4.02-2017, было выделено и идентифицировано 20 штаммов углеводородокисляющих микроорганизмов. Микробиологический анализ (МУК 4.2.3695-21) подтвердил отсутствие патогенной микрофлоры. Шесть выделенных штаммов (Bacillus spp. 10RE, Citrobacter spp. 14RE, Bacillus spp. 17RE, Pseudomonas spp. 18RE, Bacillus spp. 21RE, Rhodococcus spp. 24RE) показали выраженную способность к биосинтезу ПАВ. Ключевым доказательством послужило значительное снижение межфазного натяжения (МФН) культуральной жидкости при ее развитии на нефти, использующейся в качестве единственного источника углерода. Значения МФН на среде с нефтью варьировались от 2,5 до 4,5 мН/м, что существенно ниже показателей на среде с сахарозой 3,5–13,5 мН/м, и свидетельствует о высокой эффективности продуцируемых биоПАВ. Важным для применения в реальных нефтяных пластах является адаптивность микроорганизмов к пластовым условиям. Исследование физиологических характеристик показало, что отобранные штаммы обладают широкими диапазонами толерантности: к температуре (от 5°C до ≥ 50 °C) и солености (от 0 до 80–240 г NaCl/л). Например, Bacillus spp. 21RE рос при солености до 240 г/л, а Pseudomonas spp. 18RE и Citrobacter spp.14RE проявляли термотолерантность. Полученные результаты демонстрируют высокий потенциал выделенных штаммов для разработки инновационных биотехнологий. Их способность эффективно продуцировать биоПАВ непосредственно на нефтяном субстрате и выживать в экстремальных условиях пласта (высокая минерализация, широкий температурный диапазон) делает их перспективными агентами для микробиологических методов увеличения нефтеотдачи (ММУН). Применение таких микроорганизмов и их метаболитов позволит снизить МФН на границе «нефть – вода», мобилизовать капиллярно-удержанную нефть и повысить коэффициент извлечения нефти (КИН), одновременно способствуя биоремедиации загрязненных объектов.

Ключевые слова

повышение нефтеотдачи пластов (ПНП)биологические поверхностно-активные вещества (биоПАВбиосурфактанты)микробиологические методы увеличения нефтеотдачимежфазное натяжение (МФН)нефтезагрязненные грунтыбиоремедиацияуглеводородокисляющие микроорганизмыBacillusPseudomonasRhodococcus.

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Published: 21 November 2025
Issue date: 21 November 2025
Issue: Vol. 25, No. 4 (2025)

Search for Biosurfactant-Producing Microorganisms in Oil-Contaminated Soils for Use in Enhanced Oil Recovery Technology

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