Recombinant Production of Arginyl Dipeptides by <span style="font-variant: small-caps">l</span>-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 <span style="font-variant: small-caps;">l</span>-amino acid ligase RizA from <i>B. subtilis</i> selectively synthesizes...
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| Autores principales: | , , , |
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| Formato: | article |
| Lenguaje: | EN |
| Publicado: |
MDPI AG
2021
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| Materias: | |
| Acceso en línea: | https://doaj.org/article/4a26a1651067447abf25f428245dd837 |
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| Sumario: | 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 <span style="font-variant: small-caps;">l</span>-amino acid ligase RizA from <i>B. subtilis</i> 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 <i>E. coli</i> providing ATP regeneration from acetyl phosphate. Both enzymes were recombinantly produced in <i>E. coli</i> 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. |
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