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Protease‐catalyzed peptide synthesis using inverse substrates: The synthesis of Pro‐Xaa‐bonds by trypsin
Author(s) -
Schellenberger Volker,
Schellenberger Ute,
Jakubke HansDieter,
Zapevalova Nina P.,
Mitin Yuri V.
Publication year - 1991
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.260380314
Subject(s) - nucleophile , chemistry , aminolysis , moiety , peptide bond , trypsin , substrate (aquarium) , hydrolysis , stereochemistry , scissile bond , peptide synthesis , peptide , acyl group , catalysis , protease , leaving group , organic chemistry , enzyme , biochemistry , oceanography , alkyl , geology
Benzyloxycarbonyl‐ L ‐proline p ‐guanidinophenyl ester is an “inverse substrate” for trypsin; i.e., the cationic center is included in the leaving group instead of being in the acyl moiety. This substrate can be used in trypsin‐catalyzed acyl‐transfer reactions leading to the synthesis of Pro‐Xaa peptide bonds. The reaction proceeds about 20 times slower than reaction with similar alanine‐containing substrates, but the ratio between synthesis and hydrolysis is more favorable. The investigation of a series of nucleophiles led to information about the specificity of the process. Nucleophiles differing only in the P 1 ′‐position show an increasing acyl transfer efficiency in the order Phe < Gly < Ley < Ser < Ala < lle. C terminal elongation of the nucleophiles is of minor influence on their efficiency. The formation of an H bond between the acyl‐enzyme and the nucleophile seems to play an important role in the aminolysis of the acyl‐enzyme.