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Xiaofang Wei 1 Xingli Li 2 Keyu Liu 3 Yuehui She 4 Jing Wang 1

MEOR Using Facultative Anaerobic Indigenous Consortia and Bio-surfactants. Xiaofang Wei 1 Xingli Li 2 Keyu Liu 3 Yuehui She 4 Jing Wang 1 1 China University of Petroleum 2 GRI, Jiangsu Oilfield 3 CSIRO Petroleum 4 Yangtz University. Acknowledgements. CSIRO WfO Flagship

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Xiaofang Wei 1 Xingli Li 2 Keyu Liu 3 Yuehui She 4 Jing Wang 1

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  1. MEOR Using Facultative Anaerobic Indigenous Consortia and Bio-surfactants Xiaofang Wei1 Xingli Li2Keyu Liu3 Yuehui She4 Jing Wang1 1 China University of Petroleum 2 GRI, Jiangsu Oilfield 3 CSIRO Petroleum 4 Yangtz University

  2. Acknowledgements • CSIRO WfO Flagship • Biochemical Engineering Laboratory, CUP, Beijing • Yijng Luo, Mang Lu, Peiwen Zheng, Zhongzhi Zhang • The Langfang Research Institute of Petroleum Exploration and Development, PetroChina • Core-flooding technical support 30th IEA Workshop and Symposium on EOR Sept. 21-23, 2009, Canberra, Australia

  3. Presentation Outline • Background of the MEOR study • Nutrient stimulation • MEOR screening and assessment • Bacteria selection and metabolite analysis • Oil spreading and blood agar lysis methods • Emulsification • Interfacial tension (IFT) • Core flooding experiments • Experimental procedure • Results • Conclusions and Future work 30th IEA Workshop and Symposium on EOR Sept. 21-23, 2009, Canberra, Australia

  4. Unrecovered oil range from 1-2 trillion barrels Enhanced Oil Recovery 30th IEA Workshop and Symposium on EOR Sept. 21-23, 2009, Canberra, Australia

  5. World Average Oil Price 30th IEA Workshop and Symposium on EOR Sept. 21-23, 2009, Canberra, Australia

  6. Oil Consumption (BP Statistical Review, 2006) 1 km l l Australia: 0.9 mbbl/day China: 7.0 US: 20.6 World: 82.5 US 1 cubic km oil / year China Australia United States 30th IEA Workshop and Symposium on EOR Sept. 21-23, 2009, Canberra, Australia

  7. EOR Portfolio NON-THERMAL Imm. Gas Drives Other Chemical Miscible Slug Process Polymer CO2 MEOR Enriched Gas Drive Surfactant Flue Gas FOAM Vaporizing Gas Drive Alkaline Inert Gas CO2 Miscible Micellar MEOR is a rapidly growing EOR technique currently under laboratory research or field investigation N2 Miscible ASP Alcohol Emulsion 30th IEA Workshop and Symposium on EOR Sept. 21-23, 2009, Canberra, Australia

  8. MEOR Mechanisms and Strategies • Acids, gas, solvents production • Commonly made by carbohydrate fermentation • Inject carbohydrates (molasses, etc.) • Selective plugging • Create biofilms, cell mass and/or mineral precipitates • Any nutrient could work: use nitrate as electron acceptor • If reservoir has high divalent cation concentration, CO2 production may stimulate carbonate formation • Hydrocarbon degradation • Inject electron acceptor and/or limiting nutrients (N, S, P, metals) • Biosurfactant production • Emulsifiers • Periodic cycles of nutrient-excess and nutrient limitation 30th IEA Workshop and Symposium on EOR Sept. 21-23, 2009, Canberra, Australia

  9. Microbially Converting CO2 to CH4 in Depleted Reservoirs (from Maeda, 2009) 30th IEA Workshop and Symposium on EOR Sept. 21-23, 2009, Canberra, Australia

  10. Implementing MEOR (from McInerney,2009) • Not all reservoirs are candidates for MEOR • Environmental conditions limit microbial growth • Recommendations: • Temperature: <80oC • pH: 5-9 • Salinity: <10% • Depth: <2500 m • API gravity: > 15o • Residual oil saturation: >25% 30th IEA Workshop and Symposium on EOR Sept. 21-23, 2009, Canberra, Australia

  11. Background of the Reservoir Investigated • Characteristics of the Huatugou Oil Reservoir, Qinghai Oilfield, China • Production history: 1958-present • EOR: Water-flooding • Formation Water Salinity <2x104 ppm • Reservoir P/T: 45℃ /22 MPa • Fm water pH≤7 • Low sulfate concentration 30th IEA Workshop and Symposium on EOR Sept. 21-23, 2009, Canberra, Australia

  12. Microbial Screening • Hydrocarbon Degrading Bacteria (HBD) • Microbes population>105 cells/ml(Bottle test) • Sulphate Reducing Bacteria (SRB) • Not Detected (PCR) 30th IEA Workshop and Symposium on EOR Sept. 21-23, 2009, Canberra, Australia

  13. MEOR Screening The Huatugou reservoir is a good candidate for MEOR • Relatively high microbe population: >105 (Cell counting) • Low formation Water Salinity: <2x104 ppm • Low reservoir T: 45℃ • Suitable formation water chemistry: pH≤7 • Low corrosion risk: no SRB detected 30th IEA Workshop and Symposium on EOR Sept. 21-23, 2009, Canberra, Australia

  14. Nutrient Stimulation • Proper nutrient (Carbon Source: Reservoir oil) • Target facultative anaerobic indigenous consortia • Cell Population is up to 108 cells/ml • Minor IFT and Viscosity changes • Gas production observed 30th IEA Workshop and Symposium on EOR Sept. 21-23, 2009, Canberra, Australia

  15. Bio-surfactant: species selection and identification • Two strains were selected and named as BIOS682-1and BIOS682-2 • Initial screening by the blood agar lysis method: positive • Screening using the oil spreading method • Pure culture clear zone diameters : 4.5 cm and 4.8 cm • Mixed culture clear zone diameter: 5.0 cm • Species identification using 16s RNA • BIOS682-1: Brevibacillus agri (99% similarity) • BIOS682-2: Brevibacillus levicki (95%, a possible new species) 30th IEA Workshop and Symposium on EOR Sept. 21-23, 2009, Canberra, Australia

  16. Bio-surfactants: speciesculture and assessment • Mixed culture of two selected strains • 5ml oil/50ml-medium cultured for 3 weeks at 45℃ • Under facultative anaerobic condition (headspace filled with N2) • Oil-water separation • Oil was centrifuged at 5000 rpm for 10 min at room temperature • Oil was extracted by using CCl4 (oil in water) • The surface tension • Equipment: Contact Angle analyzer (DCA322) • Emulsification (Culture and reservoir oil 1:1 v/v) • Emulsification maintained for 100 hrs after 15 min u/s bathing • The ratio between emulsified oil and water measured 30th IEA Workshop and Symposium on EOR Sept. 21-23, 2009, Canberra, Australia

  17. Bio-surfactant producing species:culture and assessment • Surface tension reduction: 24% • Emulsification improvement: 14% 30th IEA Workshop and Symposium on EOR Sept. 21-23, 2009, Canberra, Australia

  18. Bio-surfactant analysis • Bio-surfactant of lipopeptide extraction protocol • centrifuged twice at 4°C @10,000 rpm for 20 min • the supernatant was mixed with 6mol HCL @ pH=2 (24 hrs 4°C) • The floccules were collected using centrifuge (4°C @10,000 rpm for 20 min) • The settlings were washed using acidic solution (pH=2), then dissolved in weak alkali solution (pH=8) • Extracted with DCM and methanol solution (v/v 3:1) • Dried under vacuum at low temperature • The primary products were dissolved in alkali solution (pH=8) • The extraction was then dried under vacuum 30th IEA Workshop and Symposium on EOR Sept. 21-23, 2009, Canberra, Australia

  19. Bio-surfactant analysis • Thin Layer Chromatography (TLC) • Initial screening • Liquid Chromatography-Mass Spectrometry (LC-MS). • The purified product was hydrolyzed for 14 hrs at 110 ℃ with 6N HCl in N2 chamber • Equipment: Agilent 1100 system a-Amino acid 30th IEA Workshop and Symposium on EOR Sept. 21-23, 2009, Canberra, Australia

  20. Bio-surfactant analysis • Thin Layer Chromatography (TLC) Metabolite Standard Sample lipopeptidebio-surfactant(Pink): Qualitative analysis Solution used is a mixture of n-butanol/alcohol/water(4:1:1) 30th IEA Workshop and Symposium on EOR Sept. 21-23, 2009, Canberra, Australia

  21. Bio-surfactant analysis Metabolite Standard Sample lipopeptidebio-surfactant-Amino Acid (Yellow) 30th IEA Workshop and Symposium on EOR Sept. 21-23, 2009, Canberra, Australia

  22. Bio-surfactant analysis Amino Acid contents in the metabolites 30th IEA Workshop and Symposium on EOR Sept. 21-23, 2009, Canberra, Australia

  23. 4. Core-flooding Experiment Oil Collector Core Holder Water Microbes Core-flooding experiment design: • T/P: 45℃/22 MPa • Water-cut: 70% • Incubation time: 3 days • Injected volume: 2.5 PV of Cultured solution 30th IEA Workshop and Symposium on EOR Sept. 21-23, 2009, Canberra, Australia

  24. Synthetic Core Plugs for Core Flooding Experiments • Material: oil zone sandstone recovered from the producing well, • DisintegratedCleaned • Sieved to 30, 80 and 120 meshes, respectively • A two-layer heterogeneous sand packs with different permeabilities was constructed • Layer 1: 2:2:1 mix of 30-80, 80-120 and >120 meshes • Layer 2: 1:1 mix of 30-80 and 80-120 meshes 30th IEA Workshop and Symposium on EOR Sept. 21-23, 2009, Canberra, Australia

  25. Core-flooding Experiment η＝(B－A)－C • B: final oil recovery % • A: oil recovery % from 0.5 pv nutrient solution injection • C: oil recovery % (Blank) from 0.5 pv synthetic formation water injection • η: incremental oil recovery (%) Core plug properties 30th IEA Workshop and Symposium on EOR Sept. 21-23, 2009, Canberra, Australia

  26. Results of core-flooding 30th IEA Workshop and Symposium on EOR Sept. 21-23, 2009, Canberra, Australia

  27. Conclusions • The Huatugou reservoir in the Qinghai Oilfield is a good candidate for MEOR • The indigenous facultative anaerobic flora from the reservoir was stimulated by adding proper nutrients to promote their activities • The two indigenous aerobic species selected can co-metabolize well and produced more bio-surfactant than from a single strain in the reservoir • The two species identified by 16s RNA are Brevibacillus agri (BIOS682-1); and a possible new species in genus of Bacillus brevis (BIOS682-2). 30th IEA Workshop and Symposium on EOR Sept. 21-23, 2009, Canberra, Australia

  28. Conclusions • The metabolites of BIOS682 contain low concentration of amino acids • An 11% incremental recovery was achieved using a combined microbial and biosurfactant solution injection. • The facultative anaerobic flora play an important role in peeling the oil form rock or breaking down the large oil drops. • The use of cultured solution with bio-surfactant and other metabolites is much more effective than the use of the indigenous microbes alone in recovering residual oil. 30th IEA Workshop and Symposium on EOR Sept. 21-23, 2009, Canberra, Australia

  29. Future work • In-situ experiments (Field Trials) • Further analysis of the metabolites 30th IEA Workshop and Symposium on EOR Sept. 21-23, 2009, Canberra, Australia

  30. Dr Xiaofang Wei Research Institute of Petroleum Exploration and Development, PetroChina weixfanf@163.com Dr Keyu Liu Research Team Leader CSIRO Petroleum Keyu.Liu@csiro.au Thank You 30th IEA Workshop and Symposium on EOR Sept. 21-23, 2009, Canberra, Australia

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