Table S1.
In situ sulfate reduction rates
(SRR) determined from a
variety of production well and
processing facility samples
from two oil fields on Alaska’s
NorthSlope.
Table S2. Alkane-to-pristane or alkane-to-phytane peak area ratios for crude oil incubated in
the presence (inoculated) or absence (sterile) of a methanogenic population enriched from a
hot oilfield production well from Field B on Alaska’s North Slope. Bolded numbers show a
decrease of the ratio in inoculated incubations relative to sterile incubations indicating that
biodegradation has occurred.
Alkane/Pristane
Alkane/Phytane
Alkane
Sterile
Inoculated
Sterile
Inoculated
C10
0.874
0.490
1.034
0.558
C11
1.621
0.820
1.621
0.976
C12
1.647
1.083
1.949
1.287
C13
2.064
1.480
2.443
1.740
C14
1.884
1.501
2.229
1.835
C15
1.629
1.402
1.928
1.561
C16
1.511
1.366
1.789
1.547
C17
1.792
1.652
2.121
2.112
C18
1.226
1.180
1.451
1.38
C19
1.128
1.072
1.335
1.207
C20
1.058
1.057
1.252
1.197
able S3. Amounts (mol) of n-alkanes present in sterile and inoculated crude oilamended incubations enriched from production well fluids from Field B on Alaska’s North
Slope and expected amounts (mol) of methane based on predicted stoichiometric
reactions.
n-alkane
C#
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
mol nalkane,
sterile
control
0.229
0.316
0.452
0.453
0.496
0.634
0.570
0.510
0.963
0.521
0.657
0.715
0.689
0.711
0.716
0.700
0.836
0.778
0.728
0.631
0.623
0.614
0.508
mol nalkane,
inoculated
1
0.102
0.161
0.242
0.256
0.295
0.379
0.338
0.283
0.547
0.224
0.216
0.203
0.188
0.185
0.217
0.272
0.348
0.374
0.391
0.395
0.394
0.386
0.321
total mol
n-alkane
consumed
mol nalkane,
inoculated 2
mol CH4 per
mol n-alkane
consumed
mol CH4
theoretically
expected
0.127
0.155
0.210
0.197
0.201
0.254
0.232
0.227
0.417
0.296
0.441
0.512
0.501
0.526
0.499
0.428
0.488
0.403
0.337
0.237
0.229
0.228
0.187
7
7.75
8.5
9.25
10
10.75
11.5
12.25
13
13.75
14.5
15.25
16
16.75
17.5
18.25
19
19.75
20.5
21.25
22
22.75
23.5
mol CH4
expected
0.886
1.201
1.783
1.824
2.010
2.735
2.662
2.783
5.416
4.074
6.397
7.815
8.017
8.811
8.740
7.818
9.263
7.969
6.912
5.027
5.048
5.198
4.399
7.33
116.79
Table S4. Summary of OTU in hydrocarbon-degrading methanogenic culture enriched from Field B.
Primers1
Bacteria
OTU
%
(Access.
Total2
No.)
BBACe11
29
(GU357467)
Most similar sequences
(Access. No.)
Uncultured G55_D25_M_B_H02 (DQ997951)
% identity
(% query
coverage)3
99 (98)
Source
Thermophilic anaerobic solid waste
bioreactor
ANS oil-water separator*
Unpublished, not stated
ANS oil processing facility coalescer*
Mesothermic ANS oil well
ANS oil well*
Thermophilic anaerobic methanogenic
reactor
ANS oil well*
Thermophilic microbial fuel cell
Anaerobaculum hydrogeniformans OS1
99 (98)
Thermotoga elfii strain G1 (DQ374393)
99 (100)
CO1SHNF443 (FJ469358)
99 (100)
20
Uncultured D004011L07 (EU721778)
99 (99)
Uncultured 2L1SGXO566 (FJ469295)
99 (87)
20
Thermacetogenium phaeum DSM 12270
98 (98)
(NR_024688)
7
Uncultured 2L6SGXO407 (FJ469312)
99 (86)
Uncultured Aminanaerobia clone SHBZ405
99 (99)
(EU638937)
Archaea BARCe03
100
Uncultured NAK-a1 (DQ867048)
99 (100)
Natural gas field
(GU357464)
PS35SGXN478 (FJ44511)
99 (99)
ANS oil-water separator*
mcrA4
BMEg02
100
Uncultured PS130SGXI591
99 (100)
ANS oil-water separator*
(GU357469)
Uncultured TUM-dMCRA-TR1-B1-5
92 (64)
biogas reactor
(FJ240435)
Uncultured MDS-m-H03 (AB353242)
92 (58)
mesophilic digested sludge
Methanobacterium aarhusense (AY386125)
81 (98)
marine sediment
5
mcrAA BMEg02AA 100
Uncultured PS1130SGXI591AA
100 (100)
ANS oil-water separator*
Uncultured TUM-dMCRA-TR1-B1-5
97 (64)
biogas reactor
(FJ240435)
Uncultured MDS-m-H03 (AB353242)
96 (58)
mesophilic digested sludge
Methanobacterium aarhusense (AY386125)
91 (100)
marine sediment
1 Primer sets used for each group are described in Experimental procedures
2 Bacteria, 41 sequences; Archaea, 46 sequences; mcrA, 23 sequences
3 % coverage of query sequence by database sequence, e.g. values < 100 indicate the database sequence being compared was shorter than the
query sequence
4 Nucleotide sequence similarity
5 Amino acid similarity from deduced sequences
* From the same ANS (Alaskan North Slope) oil field facility as the current enrichment
BBACe04
(GU357465)
BBACe02
(GU357463)
BBACe03
(GU357464)
BBACe10
(GU357466)
24
Fig. S1. Phylogenetic relationship of deduced methyl-coenzyme M reductase subunit A (mcrA) amino acid sequences from the Field B
enrichment with respect to related sequences. The tree is constructed from approximately 219 amino acid positions (deduced from the
mcrA nucleotide sequences) using the neighbor-joining algorithm. One thousand bootstrap replications were performed; only values
greater than 700 are shown. The sequence shown in bold (BME09g02) and followed by an asterix is from the enrichment. It is
representative of 23 sequences >99% similarity. Sequences in bold beginning with "PS" are from samples taken from the same field in
2006 (Duncan et al., 2009).
Reference:
Duncan, K. E., Gieg, L. M., Parisi, V. A., Tanner, R. S., Suflita, J. M., Green Tringe, S. and Bristow, J. (2009) Biocorrosive
thermophilic microbial communities in Alaskan North Slope oil facilities. Environ Sci Technol 43: 7977-7984.