Mutagenesis of the l-Amino Acid Ligase RizA Increased the Production of Bioactive Dipeptides

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dc.identifier.uri Bordewick, Sven Berger, Ralf G. Ersoy, Franziska 2022-02-07T06:23:13Z 2022-02-07T06:23:13Z 2021
dc.identifier.citation Bordewick, S.; Berger, R.G.; Ersoy, F.: Mutagenesis of the l-Amino Acid Ligase RizA Increased the Production of Bioactive Dipeptides. In: Catalysts : open access journal 11 (2021), Nr. 11, 1385. DOI:
dc.description.abstract The l-amino acid ligase RizA from B. subtilis selectively synthesizes dipeptides containing an N-terminal arginine. Many arginyl dipeptides have salt-taste enhancing properties while Arg-Phe has been found to have an antihypertensive effect. A total of 21 RizA variants were created by site-directed mutagenesis of eight amino acids in the substrate binding pocket. The variants were recombinantly produced in E. coli and purified by affinity chromatography. Biocatalytic reactions were set up with arginine and four amino acids differing in size and polarity (aspartic acid, serine, alanine, and phenylalanine) and were analyzed by RP-HPLC with fluorescence detection. Variant T81F significantly improved the yield in comparison to wild type RizA for aspartic acid (7 to 17%), serine (33 to 47%) and alanine (12 to 17%). S84F increased product yield similarly for aspartic acid (7 to 17%) and serine (33 to 42%). D376E increased the yield with alanine (12 to 19%) and phenylalanine (11 to 26%). The largest change was observed for S156A, which showed a yield for Arg-Phe of 40% corresponding to a 270% increase in product concentration. This study expands the knowledge about positions governing the substrate specificity of RizA and may help to inform future protein engineering endeavors. eng
dc.language.iso eng
dc.publisher Basel : MDPI
dc.relation.ispartofseries Catalysts : open access journal 11 (2021), Nr. 11
dc.rights CC BY 4.0 Unported
dc.subject l-amino acid ligase eng
dc.subject biocatalysis eng
dc.subject mutagenesis eng
dc.subject coupled catalysis eng
dc.subject arginyl dipeptides eng
dc.subject salt taste eng
dc.subject protein engineering eng
dc.subject substrate specificity eng
dc.subject.ddc 540 | Chemie ger
dc.title Mutagenesis of the l-Amino Acid Ligase RizA Increased the Production of Bioactive Dipeptides
dc.type Article
dc.type Text
dc.relation.essn 2073-4344
dc.relation.doi 10.3390/catal11111385
dc.bibliographicCitation.issue 11
dc.bibliographicCitation.volume 11
dc.bibliographicCitation.firstPage 1385
dc.description.version publishedVersion
tib.accessRights frei zug�nglich

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