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Fig. 5 | Biotechnology for Biofuels

Fig. 5

From: A new laboratory evolution approach to select for constitutive acetic acid tolerance in Saccharomyces cerevisiae and identification of causal mutations

Fig. 5

Latency phase of engineered strains and CEN.PK113-7D upon inoculation of non-adapted cells in media containing acetic acid. After pre-cultivation in SMG without acetic acid (pH 6.0), cells were inoculated in 96-well plates containing SMG with acetic acid (pH 4.5, 5–9 g/L acetic acid) at an initial OD660 of 0.1. Sealed plates were incubated at 30 °C and at 400 rpm, and the OD660 was recorded every 15 min. For each acetic acid concentration, length of the latency phase was defined as the time required to reach an OD660 of 0.12. Data points represent average and standard deviation of two to eight replicates. Wells that did not show growth after 5 days (OD660 <0.12) were eliminated from the analysis (15 out of 312 for the reverse engineered strains and 44 out of 156 for CEN.PK113-7D). a Engineered strains expressing ASG1 and ADH3 alleles from MUT1A. b Engineered strains expressing ASG1, ADH3 and SKS1 alleles from MUT2B. c Engineered strain expressing the GIS4 allele from MUT3E. d Engineered strain expressing the ASG1 allele from HAT1E. e Engineered strains expressing ASG1, ADH3 and SKS1 alleles from HAT2A. f Engineered strains expressing the GIS4 allele from UV-E3

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