13-07-2017 дата публикации
Номер: US20170198272A1
Принадлежит:
The present invention relates to a polypeptide having proline-specific endoprotease activity, wherein the polypeptide has less than 70% residual activity when the polypeptide has been kept at a temperature of 65° C. for 15 min. 1. A polypeptide having proline-specific endoprotease activity , wherein the polypeptide has less than 70% residual activity on acetyl-AlaAlaPro-paranitroaniline (Ac-AAP-pNA) as a substrate after the polypeptide has been kept at a temperature of 65° C. for 15 min.2. A polypeptide having proline-specific endoprotease activity claim 1 , optionally according to claim 1 , wherein the polypeptide is selected from the group consisting of:i. a polypeptide, which, when aligned with an amino acid sequence according to SEQ ID NO: 1 comprises an amino acid substitution at a position corresponding to position 469, and optionally at least one further amino acid substitution at position 204, 304, 377, 466, and/or 477, wherein the position is defined with reference to SEQ ID NO: 1;ii. a polypeptide, which, when aligned with an amino acid sequence according to SEQ ID NO: 1 comprises an amino acid selected from the group consisting of Ala (A), Cys (C), Asp (D), Glu (E), Phe (F), Gly (G), His (H), Ile (I), Lys (K), Leu (L), Met (M), Asn (N), Gln (Q), Arg (R), Ser (S), Thr (T), Val (V), Trp (W), and Tyr (Y) at a position corresponding to position 469, and optionally an amino acid of Phe (F) at position 204, Ser (S) at position 304, Ala (A) at position 377, Thr (T) at position 466 and/or Ala (A) at position 477, wherein the position is defined with reference to SEQ ID NO:1;iii. a polypeptide comprising an amino acid sequence according to SEQ ID NO: 1, wherein SEQ ID NO: 1 comprises at least one amino acid substitution selected from the group consisting of P469A, P469C, P469D, P469E, P469F, P469G, P469H, P469I, P469K, P469L, P469M, P469N, P469Q, P469R, P469S, P469T, P469V, P469W, and P469Y, and optionally an amino acid substitution I204F, P304S, P377A, P466T, and ...
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