Journal Article

PUBDB-2024-07777

http://join2-wiki.gsi.de/foswiki/pub/Main/Artwork/join2_logo100x88.png A Plurizyme with Transaminase and Hydrolase Activity Catalyzes Cascade Reactions

Roda, S. ; Fernandez-Lopez, L. ; Benedens, M. ; Bollinger, A. ; Thies, S. ; Schumacher, J. ; Coscolín, C. ; Kazemi, M. ; Santiago, G. ; Gertzen, C. G. W. ; Gonzalez-Alfonso, J. L. ; Plou, F. J. ; Jaeger, K.-E. ; Smits, S. H. J. ; Ferrer, M. (Corresponding author) ; Guallar, V. (Corresponding author)

2022
Wiley-VCH Weinheim

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Please use a persistent id in citations: doi:10.1002/anie.202207344  doi:10.3204/PUBDB-2024-07777

Abstract: Engineering dual-function single polypeptide catalysts with two abiotic or biotic catalytic entities (or combinations of both) supporting cascade reactions is becoming an important area of enzyme engineering and catalysis. Herein we present the development of a PluriZyme, TR2E2, with efficient native transaminase (kcat: 69.49±1.77 min−1) and artificial esterase (kcat: 3908–0.41 min−1) activities integrated into a single scaffold, and evaluate its utility in a cascade reaction. TR2E2 (pHopt: 8.0–9.5; Topt: 60–65 °C) efficiently converts methyl 3-oxo-4-(2,4,5-trifluorophenyl)butanoate into 3-(R)-amino-4-(2,4,5-trifluorophenyl)butanoic acid, a crucial intermediate for the synthesis of antidiabetic drugs. The reaction proceeds through the conversion of the β-keto ester into the β-keto acid at the hydrolytic site and subsequently into the β-amino acid (e.e. >99 %) at the transaminase site. The catalytic power of the TR2E2 PluriZyme was proven with a set of β-keto esters, demonstrating the potential of such designs to address bioinspired cascade reactions.

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Contributing Institute(s):

1. EMBL-User (EMBL-User)

Research Program(s):

1. 6G3 - PETRA III (DESY) (POF4-6G3) (POF4-6G3)
2. DFG project G:(GEPRIS)417919780 - Zentrum für strukturelle Studien (417919780) (417919780)

Experiment(s):

1. PETRA Beamline P13 (PETRA III)

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Database coverage:
; ; ; Chemical Reactions ; Clarivate Analytics Master Journal List ; Current Contents - Life Sciences ; Current Contents - Physical, Chemical and Earth Sciences ; DEAL Wiley ; Ebsco Academic Search ; Essential Science Indicators ; IF >= 15 ; Index Chemicus ; JCR ; Nationallizenz ; SCOPUS ; Science Citation Index Expanded ; Web of Science Core Collection

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