Figure 1

Deletion analysis of the GLuc signal sequence. a) The pKM152 plasmid structure is shown. Non-homologous end joining in K. marxianus produces circular plasmid efficiently, that makes deletion and mutagenized sequence addition experiments easier and faster. b) Deletion series of GLuc signal sequences and associated levels of secretory GLuc activity. The N-terminal signal sequence of GLuc has a hydrophobic amino acid stretch (bold letters in wild-type sequence (WT)) between the 4th Lys (K) and the 16th Glu (E). Positively and negatively charged amino acids are underlined and italicized, respectively. RLU; relative luminescence units (value/(μl · sec · OD₆₀₀)), Fold; ratio of mutagenized GLuc activity per wild type activity. n; number of transformants used for calculation, N; number of transformants measured.