Método para remover um dipeptídio amino-terminal de um polipeptídio precursor para produzir um polipeptídio processado e método para remover dipeptídios amino-terminais de um polipeptídio precursor para produzir um polipeptídio processado

摘要

The present invention concerns a novel process for using a recently discovered dipeptidylaminopeptidase isolated from the cellular slime mold, Dictyostelium discoideum (dDAP). The claimed process involves immobilizing dDAP onto a support surface so that even numbered, N-terminal dipeptides can be efficiently removed from precursor proteins or polypeptides to yield desired processed proteins or polypeptides. The invention provides a means for reusing dDAP in multiple conversion reaction runs, as the enzyme does not become a part of the substrate/product solution. The claimed process is particularly useful for converting a precursor human proinsulin analog (Met-Arg-Human Proinsulin having Lys at position 28 and Pro at position 29 of the corresponding insulin B-chain; MR-KPB-hPI) to a processed insulin analog (Human Proinsulin having Lys at position 28 and Pro at position 29 of the corresponding insulin B-chain, KPB-hPI).