7QX0

Transaminase Structure of Plurienzyme (Tr2E2) in complex with PLP

Summary for 7QX0

• Entry DOI: 10.2210/pdb7qx0/pdb
• Descriptor: Aminotransferase TR2, PYRIDOXAL-5'-PHOSPHATE (2 entities in total)
• Functional Keywords: transaminase, hydrolase
• Biological source: Acidihalobacter
• Total number of polymer chains: 4
• Total formula weight: 207623.24

Authors

Roda, S.,Fernandez-Lopez, L.,Benedens, M.,Bollinger, A.,Thies, S.,Schumacher, J.,Coscolin, C.,Kazemi, M.,Santiago, G.,Gertzen, C.G.,Gonzalez-Alfonso, J.,Plou, F.J.,Jaeger, K.E.,Smits, S.H.,Ferrer, M.,Guallar, V. (deposition date: 2022-01-26, release date: 2023-08-16)

Primary citation

Roda, S.,Fernandez-Lopez, L.,Benedens, M.,Bollinger, A.,Thies, S.,Schumacher, J.,Coscolin, 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.,Guallar, V.
A Plurizyme with Transaminase and Hydrolase Activity Catalyzes Cascade Reactions.
Angew Chem Int Ed Engl, 61:e202207344-, 2022

PubMed 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, TR E , with efficient native transaminase (k : 69.49±1.77 min ) and artificial esterase (k : 3908-0.41 min ) activities integrated into a single scaffold, and evaluate its utility in a cascade reaction. TR E (pH : 8.0-9.5; T : 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 TR E PluriZyme was proven with a set of β-keto esters, demonstrating the potential of such designs to address bioinspired cascade reactions.

PubMed: 35734849
DOI: 10.1002/anie.202207344

Experimental method

X-RAY DIFFRACTION (3.5 Å)