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...

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Autores principales:	Sven Bordewick, Tim A. Mast, Ralf G. Berger, Franziska Ersoy
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	<span style="font-variant: small-caps">l</span>-amino acid ligase acetate kinase coupled catalysis arginyl dipeptides salt taste ATP regeneration Chemical technology TP1-1185 Chemistry QD1-999
Acceso en línea:	https://doaj.org/article/4a26a1651067447abf25f428245dd837
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spelling	oai:doaj.org-article:4a26a1651067447abf25f428245dd8372021-11-25T17:05:30ZRecombinant Production of Arginyl Dipeptides by <span style="font-variant: small-caps">l</span>-Amino Acid Ligase RizA Coupled with ATP Regeneration10.3390/catal111112902073-4344https://doaj.org/article/4a26a1651067447abf25f428245dd8372021-10-01T00:00:00Zhttps://www.mdpi.com/2073-4344/11/11/1290https://doaj.org/toc/2073-4344Arginyl 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.Sven BordewickTim A. MastRalf G. BergerFranziska ErsoyMDPI AGarticle<span style="font-variant: small-caps">l</span>-amino acid ligaseacetate kinasecoupled catalysisarginyl dipeptidessalt tasteATP regenerationChemical technologyTP1-1185ChemistryQD1-999ENCatalysts, Vol 11, Iss 1290, p 1290 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	<span style="font-variant: small-caps">l</span>-amino acid ligase acetate kinase coupled catalysis arginyl dipeptides salt taste ATP regeneration Chemical technology TP1-1185 Chemistry QD1-999
spellingShingle	<span style="font-variant: small-caps">l</span>-amino acid ligase acetate kinase coupled catalysis arginyl dipeptides salt taste ATP regeneration Chemical technology TP1-1185 Chemistry QD1-999 Sven Bordewick Tim A. Mast Ralf G. Berger Franziska Ersoy Recombinant Production of Arginyl Dipeptides by <span style="font-variant: small-caps">l</span>-Amino Acid Ligase RizA Coupled with ATP Regeneration
description	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.
format	article
author	Sven Bordewick Tim A. Mast Ralf G. Berger Franziska Ersoy
author_facet	Sven Bordewick Tim A. Mast Ralf G. Berger Franziska Ersoy
author_sort	Sven Bordewick
title	Recombinant Production of Arginyl Dipeptides by <span style="font-variant: small-caps">l</span>-Amino Acid Ligase RizA Coupled with ATP Regeneration
title_short	Recombinant Production of Arginyl Dipeptides by <span style="font-variant: small-caps">l</span>-Amino Acid Ligase RizA Coupled with ATP Regeneration
title_full	Recombinant Production of Arginyl Dipeptides by <span style="font-variant: small-caps">l</span>-Amino Acid Ligase RizA Coupled with ATP Regeneration
title_fullStr	Recombinant Production of Arginyl Dipeptides by <span style="font-variant: small-caps">l</span>-Amino Acid Ligase RizA Coupled with ATP Regeneration
title_full_unstemmed	Recombinant Production of Arginyl Dipeptides by <span style="font-variant: small-caps">l</span>-Amino Acid Ligase RizA Coupled with ATP Regeneration
title_sort	recombinant production of arginyl dipeptides by <span style="font-variant: small-caps">l</span>-amino acid ligase riza coupled with atp regeneration
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/4a26a1651067447abf25f428245dd837
work_keys_str_mv	AT svenbordewick recombinantproductionofarginyldipeptidesbyspanstylefontvariantsmallcapslspanaminoacidligaserizacoupledwithatpregeneration AT timamast recombinantproductionofarginyldipeptidesbyspanstylefontvariantsmallcapslspanaminoacidligaserizacoupledwithatpregeneration AT ralfgberger recombinantproductionofarginyldipeptidesbyspanstylefontvariantsmallcapslspanaminoacidligaserizacoupledwithatpregeneration AT franziskaersoy recombinantproductionofarginyldipeptidesbyspanstylefontvariantsmallcapslspanaminoacidligaserizacoupledwithatpregeneration
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Recombinant Production of Arginyl Dipeptides by <span style="font-variant: small-caps">l</span>-Amino Acid Ligase RizA Coupled with ATP Regeneration

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