Supplemental material
S. cerevisiae CICC 1300
arg1::pCpA1/G-XI
ty1:: pYIE2-Ty-XI
δ::loxP-zeo-loxPADH1p-XKS1-XKS1tTPI1p-TAL1-TAL1tPGK1p-RPE1-RPE1tFBA1p-TKL1-TKL1tPDC1p-RKI1-RKI1t
adaptive evolution &
single-colony isolation
B
45
Xylose
Xylitol
40
CIBTS0552
10
Glycerol
9
8
35
7
30
6
25
5
20
4
15
3
10
2
5
1
0
0
0
2
Ethanol
Biomass
Biomass (g DCW/l)
A
Metabolite concentration (g/l)
1
8
16
24
Hours
32
40
48
3
Figure S1, Anaerobic fermentation of CIBTS0552 on xylose. (A), Construction process of CIBTS0552. (B), Xylose fermentation. The strain
4
was cultured at 30 oC with shaking in YP medium supplemented with 40 g/l xylose in a 300 ml shake flask containing 100ml medium. The
5
shake flask was capped with a rubber stopper (a syringe needle was inserted into the rubber stopper to release CO2 during fermentation). The
6
initial OD600 was set at 1.0 (0.63 g DCW/l).
A S. cerevisiae CCTCC M94055
arg1::pCpA1/G-XI
ty1:: pYIE2-Ty-XI
δ::loxP-zeo-loxPADH1p-XKS1-XKS1tTPI1p-TAL1-TAL1tPGK1p-RPE1-RPE1tFBA1p-TKL1-TKL1tPDC1p-RKI1-RKI1t
7
CIBTS0525
gre3::pYIE2-SUT1
CIBTS0572
adaptive evolution &
single-colony isolation
CIBTS0734
B
45
Xylose
Xylitol
40
Ethanol
Biomass
10
Glycerol
9
8
35
7
30
6
25
5
20
4
15
3
10
2
5
1
0
0
0
8
16
24
Hours
32
40
48
8
Figure S2, Anaerobic fermentation of CIBTS0734 on xylose. (A), Construction process of CIBTS0734. (B), Xylose fermentation. The strain
9
was cultured at 30 oC with shaking in YP medium supplemented with 40 g/l xylose in a 300 ml shake flask containing 100ml medium. The
10
shake flask was capped with a rubber stopper (a syringe needle was inserted into the rubber stopper to release CO2 during fermentation). The
11
initial OD600 was set at 1.0 (0.63 g DCW/l).
12
Glucose
Xylose
Ethanol
12
Biomass
80
10
70
60
8
50
6
40
30
4
20
2
10
0
Biomass (g DCW/l)
Metabolite concentration (g/l)
90
0
0
24
48
72
96
120
Hours
13
14
Figure S3, Anaerobic fermentation of CIBTS0735 in undetoxified corn stover hydrolysate. The strain was cultured at 30 oC with shaking in a
15
100 ml shake flask containing 30ml undetoxified corn stover hydrolysate. The hydrolysate was only supplemented with 1 g/l urea and its pH
16
was adjusted to 6.0. The shake flask was capped with a rubber stopper (a syringe needle was inserted into the rubber stopper to release CO2
17
during fermentation) and the initial OD600 was set at 8.0 (5 g DCW/l). The hydrolysate contained 82.3 g/l glucose, 54.2 g/l xylose, 8.4 g/l acetic
18
acid and 2.6 g/l HMF.
19
20
21
Figure S4, Genetic maps for pCpA1/G-XI and pYIE2-Ty-XI
22
23
Figure S5, Genetic map for pYIE2-GXF1
24
25
Figure S6, DNA fragment used to overexpress xylulokinase and the four nonoxidative
26
enzymes in the pentose phosphate pathway at the δ locus.