Preparative Coupled Enzymatic Synthesis of L-Homophenylalanine and 2-Hydroxy-5-oxoproline with Direct In Situ Product Crystallization and Cyclization
Sven Tiedemann, Annabel Stang, Simon Last, Thierry Gefflaut, Jan von Langermann
TL;DR
Scientists developed a continuous method to efficiently produce two chemicals, L-homophenylalanine and 2-hydroxy-5-oxoproline, using an enzyme from Megasphaera elsdenii.
Contribution
A novel coupled enzymatic synthesis with in situ crystallization and cyclization for preparative-scale production of two compounds.
Findings
Fed-batch process proved most viable for the coupled synthesis.
Over 18 g of L-homophenylalanine and 9 g of 2-hydroxy-5-oxoproline were produced at gram scale.
The process improves productivity through spontaneous crystallization and cyclization.
Abstract
A continuous in situ crystallization concept is presented for the coupled preparative synthesis of L-homophenylalanine and 2-hydroxy-5-oxoproline (a cyclized form of α-ketoglutarate) using the α-transaminase from Megasphaera elsdenii. The process consists of a spontaneous reactive crystallization step of the enantiopure amino acid itself and a parallel spontaneous cyclization of the deaminated cosubstrate in solution. In parallel, these effects significantly improve the overall productivity of the biocatalytic reaction. Batch, repetitive, and fed-batch processes were investigated, and the fed-batch option proved to be the most viable option. The fed-batch process was subsequently used for a coupled synthesis approach at the gram scale. In total, >18 g of chemically pure L-homophenylalanine and >9 g of 2-hydroxy-5-oxoproline were isolated. This optimized process allows for the design of…
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Taxonomy
TopicsEnzyme Catalysis and Immobilization · Microbial Metabolic Engineering and Bioproduction · Carbohydrate Chemistry and Synthesis