Racemization-free and scalable amidation of l-proline in organic media using ammonia and a biocatalyst only. Issue 13 (9th June 2022)

• Record Type: Journal Article
• Authors: Pitzer, Julia; Steiner, Kerstin; Schmid, Christian; Schein, Viktor K.; Prause, Christoph; Kniely, Claudia; Reif, Michaela; Geier, Martina; Pietrich, Elena; Reiter, Tamara; Selig, Philipp; Stückler, Clemens; Pöchlauer, Peter; Steinkellner, Georg; Gruber, Karl; Schwab, Helmut; Glieder, Anton; Kroutil, Wolfgang
• Is Part Of: Green chemistry. Volume 24:Issue 13(2022)
• Page Start: 5171
• Page End: 5180
• Publication Date: 2022-06-09
• Subjects: Environmental chemistry -- Industrial applications -- Periodicals; Environmental management -- Periodicals; 660
• DOI: 10.1039/d2gc00783e
• Languages: English
• ISSNs: 1463-9262

Abstract

Abstract : This work describes the biocatalytic amidation of l -proline with ammonia, resulting in a process with optimized atom efficiency giving prolinamide in an optically pure form (ee >99%). Detailed enzyme and reaction engineering studies are provided. Abstract : Efficient amide formation is of high importance for the chemical and pharmaceutical industry. The direct biocatalytic one-pot transformation of acids into amides without substrate activation is a highly desirable but highly challenging reaction; this is why in general the acid is activated using additional reagents before amide formation occurs. In particular, amidation of α-amino acids is challenging and in general requires protection strategies for the amino functionality. A further challenge is the low solubility of the unprotected amino acids in organic solvents. Furthermore, the amidation process is prone to racemisation as observed for the acyl chloride derivative. These three challenges may be addressed using biocatalysis. Here the enzyme catalyzed, racemization-free amidation of unprotected l -proline with ammonia in an organic solvent is described. Comprehensive reaction, solvent and enzyme engineering allowed obtaining high l -prolinamide concentrations. For instance at 145 mM substrate concentration, 80% conversion was achieved employing an immobilized CalB variant and ammonia in 2-methyl-2-butanol at 70 °C. A two-fold increase in l -prolinamide formation was achieved employing the immobilized and engineered enzyme variant CalBopt-24 T245S compared to wild type CalB. In contrast to chemical processes, racemization, halogenated solvents and waste are avoided/minimized and atom efficiency is significantly improved from 45.5% to 86.4%. The excellent optical purity of the obtained product (ee >99%) and the stability of immobilized CalB pave the way for an innovative industrial process to produce l -prolinamide, a key intermediate in drug synthesis.