Schematic of the screening protocol for improved methanol tolerance by directed evolution of PML. (A) Steps for colony lift assay and staining protocol. (B) Hydrolysis of 1-napthyl palmitate and detection of 1-naphthol by fast blue to assay colonies for residual activity after exposure to methanol and heat. (C) A representative filter following step 9 in (A). Black arrows point to potential positives identified on the filter.