ISMOS-2, Aarhus, June 17-19, 2009 Winogradsky Institute of Microbiology, Russian Academy of Sciences INMI Syntrophic Syntrophic Acetate Degradation to Acetate Degradation to Methane in a High Methane in a High-Temperature Temperature Petroleum Reservoir Petroleum Reservoir Petroleum Reservoir Petroleum Reservoir Natalya M. Shestakova 1 , Tamara N. Nazina 1 , Qingxian Feng 2 , Fangtian Ni 2 , Tatiana P T 1 Ad iB P lt 3 S S Bl 1 Mikh il V I 1 P . T ourova 1 , Andrei B. Poltaraus 3 , Sergey S. Belyaev 1 , Mikhail V . Ivanov 1 1 Winogradsky Institute of Microbiology, Russian Academy of Sciences, Russia 2 Dagang Oilfield Company, China 3 Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Russia *E-mail: [email protected]
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ISMOS-2, Aarhus, June 17-19, 2009Winogradsky Institute of Microbiology, Russian Academy of SciencesINMI
SyntrophicSyntrophic Acetate Degradation to Acetate Degradation to Methane in a HighMethane in a High--Temperature Temperature
Petroleum ReservoirPetroleum ReservoirPetroleum ReservoirPetroleum ReservoirNatalya M. Shestakova1, Tamara N. Nazina1, Qingxian Feng2, Fangtian Ni2, Tatiana
P T 1 A d i B P lt 3 S S B l 1 Mikh il V I 1P. Tourova1, Andrei B. Poltaraus3, Sergey S. Belyaev1, Mikhail V. Ivanov1
1Winogradsky Institute of Microbiology, Russian Academy of Sciences, Russia
2Dagang Oilfield Company, China
3Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Russia
Our aim was to examine the distribution,biodiversity and geochemical activity of themicroorganisms in a high-temperaturemicroorganisms in a high temperaturepetroleum reservoir1. Characterization of a microbial community
combining culture-based, radioisotope andl l th dmolecular methods
2. Investigation of microbial interactions duringacetate degradation to methane under thermophiliccondition
Dagang Oilfield
Horizon 1200 –Horizon depth
1200 1400 m
T t 59oCTemperature 59oC
pH 7.1-7.6
Salinity 5.8-6.6 g/l
The biotech trialThe biotech trial
G lGoal:Enhancement of the oil recovery
Proposal:Production of oil-displacing compounds by microbes in situp g p y
Method:I j t t + i i t t ti l t th itInject water+air mixture to stimulate the community
We will study the water from production wellsWe will study the water from production wells
Near-bottom zone of the injection wellProduction wells
35
40
45
1200
1400
ion
20
25
30
800
1000
e pr
oduc
t
10
15
20
400
600
Met
hane
0
5
1
1002
-1
1008
-1
1012
-1
1017
1017
-3
1017
-5
1032
1050
-1
1050
-3
1094
0
200
8 m3 25 m3 1094-1
Methanogenesis was registered from both labeled bicarbonate and acetate
Increase of Acetate in Formation Waters
160
180
120
140
160
80
100
20
40
60
1
1002
-1
008-
1
12-1
17 -3
Jan. 2003
May 20050
20
10 101
10
1017
-
1017
-5
1032
1050
-1
1050
-3
1094
July 2001
Production wells
Methanogenesis and Acetate contentMethanogenesis and Acetate contentcorrelate
101750 200
1008-10 25 60
40
50
160
200
0,2
0,25
48
60
y
30 1200,15 36
, μg/
l day
ate,
mg/
l
10
20
40
80
0,05
0,1
12
24
Met
hane
,
Acet
a
0
10
VII, 2001 I, 2002 VI, 2002 I, 2003 IX, 2003 XII, 20040
40
0
0,05
VII, 2001 I, 2002 VI, 2002 I, 2003 IX, 2003 XII, 20040
M
VII, 2001 I, 2002 VI, 2002 I, 2003 IX, 2003 XII, 2004
- Acetate content - From NaH14CO3 - From 14CH3COONa
16S rRNA analysis
methanogenic enrichmentsmethanogenic enrichmentsnatural formation water formation water from the zoneformation water from the zone of the pilot trialmethanogenic enrichmentmethanogenic enrichment culture on acetate
Ph i l f I l t d M thPhysiology of Isolated Methanogens
Property KZ3 KZ24aSalinity, % NaCl 0 - 1 0 - 1
Temperature range,0C 40-65-60 40-65- 65
S b t tSubstrates:
H2+CO2 + +
Foramate +/Foramate +/- -
Acetate - -
Methane Production from Acetate by Pure Cultures and Co-cultures of Methanothermobacter -
Thermoanaerobacter
3,5
4
140
160
day
2
2,5
3
100
120
140
or µ
g/l d
1
1,5
2
40
60
80
hane
, %
0
0,5
Pure cultures Co culturesPure cultures Co-cultures0
20
40
Met
h
Pure cultures Co culturesPure cultures Co-cultures Pure cultures Co-cultures
Conclusions
1. The processes of methanogenesis from both bicarbonate and acetatewere registered in the formation waters by culture-based andradioisotope methods.
2 By 16S rRNA gene analysis was revealed that H utilizing2. By 16S rRNA gene analysis was revealed that H2-utilizingmethanogens of the genus Methanothermobacter predominated in themicrobial community. Phylotypes of acetoclastic methanogens were notf d i th i i l f ti tfound in the original formation water.
3.Phylotypes of fermentative bacteria and archaea, sulfate-reducing andsyntrophic (Thermacetogenium) bacteria were revealed in the high-syntrophic (Thermacetogenium) bacteria were revealed in the hightemperature Dagang oil field.
4. It was shown that a syntrophic association of T. ethanolicus and M.thermautotrophicus carried out the reaction of acetate degradation tomethane.
AcknowledgementsAcknowledgements
Dagang Oilfield Company, China
Winogradsky Institute of Microbiology, Russia
Engelhardt Institute of Molecular Biology, Russia
Thank you!
AcknowledgementsAcknowledgements
The work was supported by the Dagang OilfieldCompany (DFT04-122-IM-18-20RU), Russian Ministry ofEducation and Science (4174.2008.4) and the RussianA d f S i (P "M l l d C ll lAcademy of Sciences (Program "Molecular and CellularBiology").