Aprotinin and aprotinin analogues expressed in yeast

Synthetic genes encoding aprotinin and aprotinin analogues were constructed and fused in frame to the S. cerevisiae mating factor alpha 1 signal-leader (1-85) sequence. Expression in yeast resulted in secretion into the culture medium of a moderate yield of correctly processed aprotinin (1-58) together with two N-terminally extended forms. Des-Arg1, Pro2-aprotinin was expressed in a higher yield. In this case only the correct N-terminal amino acid sequence was found. Substitution of Ser42 for Arg42 in the potential internal KEX2 processing site improved the secretion yield. The aprotinins are characterized by an inhibition profile similar to that of native aprotinin. Des-Arg1, Pro2-[Arg15, Ser42] aprotinin has a strongly increased plasma kallikrein inhibition profile.