Fig. 4

Reverse engineering of the strain CEN.PK113-7D for improved glycerol growth performance by allelic replacement of the GUT1 (G) and UBR2 (U) allele. The replacement cassettes obtained from the evolved strains JL1, PW-1, PW-2, and the wild-type isolate CBS 6412-13A (CBS) comprise the ORF and at least 1000 bp upstream and 300 bp downstream of the start and stop codon, respectively. The growth performance on glycerol of the reverse-engineered strains is presented by showing the growth rates and lag phases and compared to the growth performance of the respective reference strain (Ref) from where the replacement cassettes were obtained. Mean values and standard deviations were obtained from at least three biological replicates