C - Chemistry – Metallurgy – 12 – N
Patent
C - Chemistry, Metallurgy
12
N
C12N 15/57 (2006.01) C12N 9/48 (2006.01) C12N 9/64 (2006.01) C12N 15/62 (2006.01) C12N 15/70 (2006.01) C12P 21/06 (2006.01)
Patent
CA 2403330
The invention relates to a method for producing biologically active serine proteases, isolated serine protease domains and their amino-acid variants in a prokaryotic host. The method is characterised by an N-terminal addition of a helper sequence consisting of a dipeptide which is suitable for degradation by a dipeptidyl amino peptidase, by the expression of the serine protease(s) and/or their fragments containing the N-terminal dipeptide helper sequence, optionally as inclusion bodies, and by the renaturation of the expressed proteins and the activation of the serine protease(s) and/or serine protease domains by splitting-off the helper sequence using an exopeptidase.
L'invention concerne un procédé de fabrication de sérine-protéases biologiquement actives, de domaines isolés de sérine-protéases et de leurs variantes aminoacides chez un hôte procaryote, caractérisé par addition N terminal d'une séquence auxiliaire d'un dipeptide appropriée pour la dégradation par une dipeptidylaminopeptidase, l'expression de la/ ou des sérine-protéases et/ou de leurs fragments avec des séquences auxiliaires dipeptidiques N terminal, éventuellement comme corps d'inclusion, et par renaturation de la protéine exprimée et activation de la/ ou des sérine-protéases et/ou des domaines de sérine-protéases par dédoublement de la séquence auxiliaire par une exopeptidase.
Bode Wolfram
Huber Robert
Jenne Dieter
Parry Marina A. A.
Wilharm Elke
Bode Wolfram
Huber Robert
Jenne Dieter
Max-Planck-Gesellschaft E.v.
Parry Marina A. A.
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