6RX8

EDDS lyase variant D290M/Y320M with bound fumarate

Summary for 6RX8

• Entry DOI: 10.2210/pdb6rx8/pdb
• Descriptor: Argininosuccinate lyase, FUMARIC ACID, SODIUM ION, ... (4 entities in total)
• Functional Keywords: edds lyase, c-n lyase, aspartase/fumarase, protein engineering, lyase
• Biological source: Chelativorans sp. (strain BNC1)
• Total number of polymer chains: 1
• Total formula weight: 56020.50

Authors

Grandi, E.,Poelarends, G.J.,Thunnissen, A.M.W.H. (deposition date: 2019-06-07, release date: 2019-10-30, Last modification date: 2024-01-24)

Primary citation

Zhang, J.,Grandi, E.,Fu, H.,Saravanan, T.,Bothof, L.,Tepper, P.G.,Thunnissen, A.W.H.,Poelarends, G.J.
Engineered C-N Lyase: Enantioselective Synthesis of Chiral Synthons for Artificial Dipeptide Sweeteners.
Angew.Chem.Int.Ed.Engl., 59:429-435, 2020

PubMed Abstract: Aspartic acid derivatives with branched N-alkyl or N-arylalkyl substituents are valuable precursors to artificial dipeptide sweeteners such as neotame and advantame. The development of a biocatalyst to synthesize these compounds in a single asymmetric step is an as yet unmet challenge. Reported here is an enantioselective biocatalytic synthesis of various difficult N-substituted aspartic acids, including N-(3,3-dimethylbutyl)-l-aspartic acid and N-[3-(3-hydroxy-4-methoxyphenyl)propyl]-l-aspartic acid, precursors to neotame and advantame, respectively, using an engineered variant of ethylenediamine-N,N'-disuccinic acid (EDDS) lyase from Chelativorans sp. BNC1. This engineered C-N lyase (mutant D290M/Y320M) displayed a remarkable 1140-fold increase in activity for the selective hydroamination of fumarate compared to that of the wild-type enzyme. These results present new opportunities to develop practical multienzymatic processes for the more sustainable and step-economic synthesis of an important class of food additives.

PubMed: 31625664
DOI: 10.1002/anie.201910704

Experimental method

X-RAY DIFFRACTION (1.92 Å)