Protease-catalyzed peptide synthesis using inverse substrates: The influence of reaction conditions on the trypsin acyl transfer efficiency

Benzyloxycarbonyl‐L‐alanine p‐guanidinophenyl ester behaves as a trypsin “inverse substrate,” i.e., a cationic center is included in the leaving group instead of being in the acyl moiety. Using this substrate as an acyl donor, trypsin catalyzes the synthesis of peptide bonds that cannot be split by...

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Bibliographic Details
Published in:Biotechnology and bioengineering 1991-06, Vol.38 (1), p.104-108
Main Authors: Schellenberger, Volker, Jakubke, Hans-Dieter, Zapevalova, Nina P., Mitin, Yuri V.
Format: Article
Language:English
Subjects:
acyl transfer
artificial substrates
benzyloxycarbonyl-L-alanine p-guanidinophenyl ester
Biological and medical sciences
Biotechnology
catalysis
Enzyme engineering
Fundamental and applied biological sciences. Psychology
inverse substrates
Methods. Procedures. Technologies
Miscellaneous
nucleophile efficiency
nucleophiles
peptide synthesis
protease
proteinase
trypsin
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Summary:Benzyloxycarbonyl‐L‐alanine p‐guanidinophenyl ester behaves as a trypsin “inverse substrate,” i.e., a cationic center is included in the leaving group instead of being in the acyl moiety. Using this substrate as an acyl donor, trypsin catalyzes the synthesis of peptide bonds that cannot be split by this enzyme. An optimal acyl transfer efficiency was achieved between pH 8 and 9 at 30°C.The addition of as much as 50% cosolvent was shown to be of minor influence on the acyl transfer efficiency, whereas the reaction velocity decreases by more than one order of magnitude. The efficiency of H‐Leu‐NH2 and H‐Val‐NH2 in deacylation is almost the same for “inverse” and normal type substrates.
ISSN:0006-3592
1097-0290
DOI:10.1002/bit.260380114