Marco Corti, Francesca Rinaldi, Daniela Monti
May 1, 2019
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Influential Citations
3
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Journal
Journal of Pharmaceutical and Biomedical Analysis
Abstract
Highlightsω−transamimase was immobilized on epoxy monolithic silica.The activity of the IMER was studied by an integrated HPLC system.Optimization of catalytic properties was carried out by a DoE approach.The synthesis of chiral amines of pharmaceutical interest was investigated. ABSTRACT An integrated chromatographic system was developed to rapidly investigate the biocatalytic properties of ω‐transaminases useful for the synthesis of chiral amines. ATA‐117, an (R)‐selective ω‐transaminase was selected as a proof of concept. The enzyme was purified and covalently immobilized on an epoxy monolithic silica support to create an immobilized enzyme reactor (IMER). Reactor efficiency was evaluated in the conversion of a model substrate. The IMER was coupled through a switching valve to an achiral analytical column for separation and quantitation of the transamination products. The best conditions of the transaminase‐catalyzed bioconversion were optimized by a design of experiments (DoE) approach. The production of (R)‐1‐(4‐methoxyphenyl)propan‐2‐amine and (R)‐1‐methyl‐3‐phenylpropylamine, intermediates for the synthesis of the bronchodilator formoterol and the antihypertensive dilevalol respectively, was achieved in the presence of different amino donors. The enantiomeric excess (ee) was determined off‐line by developing a derivatization procedure using N&agr;‐(2,4‐dinitro‐5‐fluorophenyl)‐L‐alaninamide reagent. The most satisfactory conversion yields were 60% for (R)‐1‐(4‐methoxyphenyl)propan‐2‐amine and 29% for (R)‐1‐methyl‐3‐phenylpropylamine, using isopropylamine as amino donor. The enantiomeric excess of the reactions were 84%R and 99%R, respectively.