Biocatalytic asymmetric Michael addition reaction of l-arginine to fumarate for the green synthesis of N-(([(4S)-4-amino-4-carboxy-butyl]amino)iminomethyl)-l-aspartic acid lithium salt (l-argininosuccinic acid lithium salt)Electronic supplementary information (ESI) available: Analytical Data. See DOI: 10.1039/c7ra10236d

• Main Authors: Schoenenberger, B, Wszolek, A, Meier, R, Brundiek, H, Obkircher, M, Wohlgemuth, R
• Format: Journal Article
• Published: 16-10-2017
• ISSN: 2046-2069
• DOI: 10.1039/c7ra10236d

The basic natural amino acid
l
-argininosuccinate containing two chiral centers occurs in
l
-alanine,
l
-arginine,
l
-aspartate,
l
-glutamate and
l
-proline metabolic pathways and plays a role in the biosynthesis of secondary metabolites and other amino acids. It is a precursor for arginine in the urea cycle or the citrulline-NO cycle as well as a precursor to fumarate in the citric acid cycle
via
argininosuccinate lyase. We aimed to run part of the urea cycle in reverse by catalyzing not the elimination but the addition reaction of
l
-arginine to fumarate in order to synthesize
l
-argininosuccinate. Argininosuccinate lyase (ASL) from
Saccharomyces cerevisiae
has been chosen as the catalyst for this addition reaction. The selected
ARG4
gene was synthesized and homogeneously expressed in
E. coli
leading to a highly active argininosuccinate lyase. The ASL-catalyzed addition reaction of
l
-arginine to fumarate has been successfully developed at gram scale. After a standard workup procedure the pure final product
l
-argininosuccinate has been isolated in good yield and high purity.
The biocatalytic asymmetric Michael addition of
l
-arginine to fumarate using argininosuccinate lyase (ASL) has enabled the synthesis of the key metabolite
l
-argininosuccinic acid lithium salt
1
for the first time, with excellent yield and purity.