TY - JOUR

T1 - Insights into a dual function amide oxidase/macrocyclase from lankacidin biosynthesis

AU - Dorival, Jonathan

AU - Risser, Fanny

AU - Jacob, Christophe

AU - Collin, Sabrina

AU - Dräger, Gerald

AU - Paris, Cédric

AU - Chagot, Benjamin

AU - Kirschning, Andreas

AU - Gruez, Arnaud

AU - Weissman, Kira J.

N1 - Funding information: Funding for this work was provided by the Agence Nationale de la Recherche (ANR-11-JSV8-003-01 and ANR-16-CE92-0006-01 to K.J.W.), the Centre National de la Recherche Scientifique (CNRS), the ‘IMPACT Biomolecules’ project of the Lorraine Université d’Excellence (Investissements d’avenir—ANR 15-004), the Lorraine Region (Bonus Qualité Recherche (BQR) grants to K.J.W. and B.C.) and the Deutsche For-schungsgemeinschaft (Cluster of Excellence REBIRTH, “From Regenerative Biology to Reconstructive Therapy” EXC 62 (to G.D. and A.K.), and grant Ki 397/20-1 (to A.K.)). Jean-Christophe Lec is thanked for invaluable assistance with the kinetics analysis, Russell Cox for helpful discussions, and Peter Leadlay for suggesting the use of EMAA and DATD and for editing of the manuscript. We also acknowledge the following for help in data acquisition: Pierre Legrand and Andrew Thompson (Soleil Synchrotron, Beamline Proxima1), William Shepard and Martin Savko (Soleil Synchrotron, Proxima2), and Javier Perez and Aurelien Thureau (Soleil Synchrotron, Swing). A portion of the NMR data was recorded on the NMR spectrometer of the Plateforme de Bio-physique et Biologie Structurale (B2S) (IBSLor, UMS2008, CNRS-UL-INSERM).

PY - 2018/9/28

Y1 - 2018/9/28

N2 - Acquisition of new catalytic activity is a relatively rare evolutionary event. A striking example appears in the pathway to the antibiotic lankacidin, as a monoamine oxidase (MAO) family member, LkcE, catalyzes both an unusual amide oxidation, and a subsequent intramolecular Mannich reaction to form the polyketide macrocycle. We report evidence here for the molecular basis for this dual activity. The reaction sequence involves several essential active site residues and a conformational change likely comprising an interdomain hinge movement. These features, which have not previously been described in the MAO family, both depend on a unique dimerization mode relative to all structurally characterized members. Taken together, these data add weight to the idea that designing new multifunctional enzymes may require changes in both architecture and catalytic machinery. Encouragingly, however, our data also show LkcE to bind alternative substrates, supporting its potential utility as a general cyclization catalyst in synthetic biology.

AB - Acquisition of new catalytic activity is a relatively rare evolutionary event. A striking example appears in the pathway to the antibiotic lankacidin, as a monoamine oxidase (MAO) family member, LkcE, catalyzes both an unusual amide oxidation, and a subsequent intramolecular Mannich reaction to form the polyketide macrocycle. We report evidence here for the molecular basis for this dual activity. The reaction sequence involves several essential active site residues and a conformational change likely comprising an interdomain hinge movement. These features, which have not previously been described in the MAO family, both depend on a unique dimerization mode relative to all structurally characterized members. Taken together, these data add weight to the idea that designing new multifunctional enzymes may require changes in both architecture and catalytic machinery. Encouragingly, however, our data also show LkcE to bind alternative substrates, supporting its potential utility as a general cyclization catalyst in synthetic biology.

UR - http://www.scopus.com/inward/record.url?scp=85054086513&partnerID=8YFLogxK

U2 - 10.1038/s41467-018-06323-w

DO - 10.1038/s41467-018-06323-w

M3 - Article

C2 - 30266997

AN - SCOPUS:85054086513

VL - 9

JO - Nature Communications

JF - Nature Communications

SN - 2041-1723

M1 - 3998

ER -