TABLE 1.

Metabolites measured for EC1118 and evolved strains after 30 days of fermentation on MS210 with 260 g/liter glucose at 28°C

Parameter	Result for strain:
	EC1118	K200.1(a)	K250.1(a)	K300.2(a)	K200.1(b)	K250.3(b)	K300.1(b)
Main compounds (g/liter)
Consumed glucose	259.9 ± 0.1	258.5 ± 1.4	250.7 ± 4.5	251.3 ± 5.0	260.0 ± 0.1	258.6 ± 1.5	259.8 ± 0.3
CO2	117 ± 0	112 ± 1	110 ± 1	109 ± 1	112 ± 0	112 ± 2	110 ± 1
Biomass (80%)a	4.0 ± 0.1	3.8 ± 0.1	4.0 ± 0.2	3.7 ± 0.2	3.6 ± 0.2	3.9 ± 0.0	4.0 ± 0.1
Ethanol	120.6 ± 0.6	116.3 ± 1.3	112.9 ± 1.5	113.2 ± 1.0	116.3 ± 0.7	115.5 ± 1.6	114.0 ± 0.8
Glycerol	8.5 ± 0.1	13.2 ± 0.1	12.7 ± 0.1	12.6 ± 0.2	13.3 ± 0.5	13.9 ± 0.2	14.2 ± 0.3
Succinate	0.9 ± 0.0	1.5 ± 0.1	1.5 ± 0.1	1.1 ± 0.1	1.5 ± 0.1	1.5 ± 0.1	1.7 ± 0.1
Pyruvate	0.22 ± 0.03	0.20 ± 0.00	0.20 ± 0.01	0.19 ± 0.00	0.20 ± 0.01	0.19 ± 0.01	0.19 ± 0.02
Acetate	0.9 ± 0.2	0.7 ± 0.0	0.9 ± 0.0	1.1 ± 0.0	1.0 ± 0.1	1.0 ± 0.2	1.0 ± 0.1
Acetaldehyde	0.011 ± 0.001	0.016 ± 0.001	0.023 ± 0.002	0.031 ± 0.007	0.025 ± 0.005	0.031 ± 0.010	0.029 ± 0.001
Acetoin	NDc	ND	ND	ND	ND	ND	ND
2,3-Butanediol	0.45 ± 0.02	1.00 ± 0.09	0.89 ± 0.30	1.08 ± 0.14	0.87 ± 0.11	1.10 ± 0.18	1.60 ± 0.06
Balance (%)b
Carbon	97.6 ± 0.6	96.9 ± 0.8	97.2 ± 2.5	97.0 ± 2.7	96.4 ± 0.8	96.7 ± 0.8	95.9 ± 0.5
Redox	97.4 ± 0.6	97.3 ± 0.7	97.5 ± 2.1	97.3 ± 2.1	96.6 ± 0.8	97.1 ± 1.7	96.4 ± 0.6
Yield
EtOH (g/g glucose consumed)	0.464 ± 0.002	0.450 ± 0.004	0.450 ± 0.010	0.450 ± 0.010	0.447 ± 0.003	0.447 ± 0.009	0.440 ± 0.003
Glycerol (g/g glucose consumed)	0.033 ± 0.000	0.051 ± 0.000	0.051 ± 0.001	0.050 ± 0.002	0.051 ± 0.002	0.054 ± 0.001	0.054 ± 0.001
Glycerol/EtOH ratio (%)	7.05 ± 0.07	11.32 ± 0.09	11.27 ± 0.11	10.09 ± 0.23	11.46 ± 0.40	12.01 ± 0.26	12.38 ± 0.27
Ethanol production (% [vol/vol])	15.29 ± 0.08	14.82 ± 0.13	14.84 ± 0.32	14.84 ± 0.34	14.74 ± 0.09	14.72 ± 0.29	14.50 ± 0.09
Glucose (g) required for 1% (vol/vol) ethanol production	17.00 ± 0.07	17.53 ± 0.09	17.52 ± 0.11	17.52 ± 0.23	17.64 ± 0.40	17.67 ± 0.26	17.97 ± 0.27
Residual sugar (g/liter) at:
15 days	0.1 ± 0.1	20.2 ± 1.4	58.7 ± 4.5	30.4 ± 5.0	0.1 ± 0.1	22.5 ± 1.5	0.2 ± 0.3
30 days	0.1 ± 0.1	1.5 ± 1.4	9.3 ± 4.5	8.7 ± 5.0	0.0 ± 0.1	1.4 ± 1.5	0.1 ± 0.3

^aBiomass measured at 80% of fermentation advancement.

^bCarbon balance represents the ratio between carbon moles of fermentation by-products and carbon moles of glucose. Redox balance represents the ratio between the reductance degree of fermentation by-products and the reductance degree of glucose.

^cND, not detected (<10 mg/ml).