Recombinant Production of Arginyl Dipeptides by l-Amino Acid Ligase RizA Coupled with ATP Regeneration

Arginyl dipeptides like Arg-Ser, Arg-Ala, and Arg-Gly are salt-taste enhancers and can potentially be used to reduce the salt content of food. The l-amino acid ligase RizA from B. subtilis selectively synthesizes arginyl dipeptides. However, industrial application is prevented by the high cost of th...

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Bibliographic Details
Published inCatalysts Vol. 11; no. 11; p. 1290
Main Authors Bordewick, Sven, Mast, Tim A., Berger, Ralf G., Ersoy, Franziska
Format Journal Article
LanguageEnglish
Published Basel MDPI AG 01.11.2021
Subjects
acetate kinase
Adenosine triphosphate
Alanine
Amino acids
arginyl dipeptides
ATP regeneration
Blood pressure
Catalysts
Chemical reactions
coupled catalysis
E coli
Enzymes
Experiments
Fluorescence
Glycine
High performance liquid chromatography
Industrial applications
Kinases
l -amino acid ligase
Regeneration
salt taste
Taste
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Summary:Arginyl dipeptides like Arg-Ser, Arg-Ala, and Arg-Gly are salt-taste enhancers and can potentially be used to reduce the salt content of food. The l-amino acid ligase RizA from B. subtilis selectively synthesizes arginyl dipeptides. However, industrial application is prevented by the high cost of the cofactor adenosine triphosphate (ATP). Thus, a coupled reaction system was created consisting of RizA and acetate kinase (AckA) from E. coli providing ATP regeneration from acetyl phosphate. Both enzymes were recombinantly produced in E. coli and purified by affinity chromatography. Biocatalytic reactions were varied and analyzed by RP-HPLC with fluorescence detection. Under optimal conditions the system produced up to 5.9 g/L Arg-Ser corresponding to an ATP efficiency of 23 g Arg-Ser per gram ATP. Using similar conditions with alanine or glycine as second amino acid, 2.6 g/L Arg-Ala or 2.4 g/L Arg Gly were produced. The RizA/AckA system selectively produced substantial amounts of arginyl dipeptides while minimizing the usage of the expensive ATP.
ISSN:2073-4344
2073-4344
DOI:10.3390/catal11111290