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Enzymatic Pictet–Spengler Reaction: Computational Study of the Mechanism and Enantioselectivity of Norcoclaurine Synthase

Author:
Sheng, Xiang, Himo, Fahmi
Source:
Journal of the American Chemical Society 2019 v.141 no.28 pp. 11230-11238
ISSN:
1520-5126
Subject:
(S)-norcoclaurine synthase, active sites, aldehydes, alkaloids, biocatalysis, biosynthesis, crystal structure, dopamine, enantioselectivity, energy, enzyme substrates, geometry, models, organic chemistry, quantum mechanics, reaction mechanisms, site-directed mutagenesis
Abstract:
The Pictet–Spengler (PS) reaction, i.e., the acid-catalyzed condensation between β-arylethylamine and an aldehyde or a ketone and the subsequent ring closure, is an important reaction in organic chemistry. A number of enzymes (called Pictet–Spenglerases, PSases) have been identified to catalyze this reaction, usually with very high enantioselectivity, making these enzymes of potential value in biocatalysis. PSases catalyze the key step in the biosynthesis of indole and benzylisoquinoline alkaloids of plant origin, some of which have pharmacological importance. However, the reaction mechanisms and the origins of the selectivity are not fully understood. Herein, we report a quantum chemical investigation of the mechanism and enantioselectivity of norcoclaurine synthase (NCS), an enzyme that catalyzes the PS condensation between dopamine and 4-hydroxyphenylacetaldehyde (4-HPAA). A large model of the active site is designed on the basis of a recent crystal structure, and it is used to calculate the detailed energy profile of the reaction. Good agreement is obtained between the calculated energies and available experimental information. Both the “dopamine-first” and the “HPAA-first” binding modes of the substrates reported in the literature are shown to be energetically accessible in the enzyme–substrate complex. However, it is demonstrated that only the dopamine-first pathway is associated with feasible energy barriers. Key active site residues are identified, and their roles in the catalysis are discussed and compared to site-directed mutagenesis experiments. Very importantly, the calculations are able to reproduce and rationalize the observed enantioselectivity of NCS. A detailed analysis of the geometries of the intermediates and transition states helps to pinpoint the main factors controlling the selectivity.
Agid:
6539371