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Different routes followed during the directed evolution of AAO. The stars indicate the newly acquired mutations, while the circles depict accumulated substitutions. The TAI (Total Activity Improvements) were determined with p-methoxybenzyl alcohol as the substrate for evolution towards secretion, with 1-(p-methoxyphenyl)-ethanol as the substrate for evolution for secondary alcohol oxidation and with 5-hydroxymethylfurfural in the evolution for FDCA synthesis.
