Enzymatic conversion of proteins using immobilized...

C - Chemistry – Metallurgy – 12 – P

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C12P 21/02 (2006.01) C07K 14/62 (2006.01) C12N 9/58 (2006.01) C12P 21/06 (2006.01)

Patent

CA 2138099

The present invention concerns a novel process for using a recently discovered dipeptidylaminopeptidase isolated from the cellular slime mold, Dictyostelium discodeum (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 reation 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).

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