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Supporting information
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Materials and Methods
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Determination of cellular fatty acids
E. coli cells were grown at 30°C to a late-exponential phase in LB liquid medium
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with 0.5 mM IPTG.
Cells were harvested by centrifugation at 8,000 g for 10 min at
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4°C and washed three times with distilled water.
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saponified, methylated, and extracted by the procedure of the Sherlock Microbial
The cellular fatty acids were
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Identification System (MIDI, Inc., Newark, Del.).
The cellular fatty acid
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composition was then determined by gas chromatography with a HP 6890 series
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gas chromatograph (Hewlett Packard, Palo Alto, Calif.) and the Sherlock Microbial
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Identification System.
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independently.
These experiments were performed three times,
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16
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Glucose utilization
E. coli cells were cultured in LB liquid medium with 0.5 mM IPTG and with or
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without butanol (1%, vol/vol) at 30°C.
Cells were harvested from the exponential
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and stationary growth cultures (6 and 24 h after inoculation, respectively) and the
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concentration of glucose remaining in the medium was determined with a Glucose
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(HK) assay kit (Sigma, St. Louis, MO) according to the manufacturer’s protocol.
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These experiments were performed at least three times, independently.
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Supplementary Table 1. Fatty acid composition of the butanol-tolerant cells (BT).
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Cells were grown at 30°C to a late-exponential phase in LB liquid medium with 0.5
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mM IPTG. The cellular fatty acids were saponified, methylated, and extracted.
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The resulting methyl ester mixtures were separated by a Hewlett Packard 6890 gas
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chromatograph.
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system (Sherlock Microbial Identification System).
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coli transformed with a control plasmid.
Fatty acids were identified by the MIDI microbial identification
“C” indicates the wild-type E.
Fatty acid composition (% of total fatty acids)
Strain
12:0
14:0
16:0
17a
18:1
19b
Sum 1c
Sum 2d
C
3.59
6.97
45.09
22.38
2.96
9.22
7.55
2.32
BT
3.81
7.67
43.45
24.17
2.14
8.36
8.42
1.12
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a17
: 17:0 Cyclo
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b19
: 19:0 Cyclo
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cSum
1 : 14:0 3OH, 16:1 ISO I
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dSum
2 : 16:1, 15 ISO 2OH
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Supplementary Table 2. Glucose utilization by the butanol-tolerant cells (BT).
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Cells were cultured in LB liquid medium with 0.5 mM IPTG and with or without
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butanol (1%, vol/vol) at 30°C and harvested from the exponential and stationary
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growth cultures (6 and 24 h after inoculation, respectively).
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glucose remaining in the medium was analyzed enzymatically with a Glucose (HK)
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assay kit (Sigma).
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plasmid.
The concentration of
“C” indicates the wild-type E. coli transformed with a control
Glucose remaining (mg/ml)
Strain
Without butanol
1% (vol/vol) butanol
Exponential phase
Stationary phase
Exponential phase
Stationary phase
C
0.52
0
1.52
0
BT
0.3
0
1.46
0
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