improved variants screened from HRPL-library. α-factor pre-leader is represented in grey, α-factor proleader in white and mature protein in black. Suggested recombination events are indicated in different colours. New point mutations are highlighted in yellow. Squared in red and blue is highlighted the recombination event that took place between 11D11 and 11A2 to generate 7H2 mutant in the 4th cycle of evolution.