TY - PAT
T1 - Peptide fragment condensation and cyclisation using a subtilisin variant with improved synthesis over hydrolysis ratio
AU - Quaedflieg, Peter Jan Leonard Mario
AU - Nuijens, Timo
AU - van der Laan, Jan Metske
AU - Janssen, Dirk Barend
AU - Toplak, Ana
AU - Wu, Bian
PY - 2016/4/14
Y1 - 2016/4/14
N2 - The invention relates to a method for enzymatically synthesizing an (oligo)peptide, comprising coupling (a) an (oligo)peptide C-terminal ester or thioester and (b) an (oligo)peptide nucleophile having an N-terminally unprotected amine, wherein the coupling is carried out in a fluid comprising water, and wherein the coupling is catalyzed by a subtilisin BPN' variant or a homologue thereof, which comprises the following mutations compared to subtilisin BPN' represented by SEQUENCE ID NO: 2 or a homologue sequence thereof : - a deletion of the amino acids corresponding to positions 75-83; - a mutation at the amino acid position corresponding to S221, the mutation being S221C or S221selenocysteine; preferably a mutation at the amino acid position corresponding to P225 wherein the amino acid positions are defined according to the sequence of subtilisin BPN' represented by SEQUENCE ID NO: 2. Further, the invention relates to an enzyme suitable for use as a catalyst in a method of the invention.
AB - The invention relates to a method for enzymatically synthesizing an (oligo)peptide, comprising coupling (a) an (oligo)peptide C-terminal ester or thioester and (b) an (oligo)peptide nucleophile having an N-terminally unprotected amine, wherein the coupling is carried out in a fluid comprising water, and wherein the coupling is catalyzed by a subtilisin BPN' variant or a homologue thereof, which comprises the following mutations compared to subtilisin BPN' represented by SEQUENCE ID NO: 2 or a homologue sequence thereof : - a deletion of the amino acids corresponding to positions 75-83; - a mutation at the amino acid position corresponding to S221, the mutation being S221C or S221selenocysteine; preferably a mutation at the amino acid position corresponding to P225 wherein the amino acid positions are defined according to the sequence of subtilisin BPN' represented by SEQUENCE ID NO: 2. Further, the invention relates to an enzyme suitable for use as a catalyst in a method of the invention.
M3 - Patent
M1 - WO2016056913
ER -