Application of reductive amination by heterologously expressed Thermomicrobium roseum L-alanine dehydrogenase to synthesize L-alanine derivatives

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dc.authoridValjakka, Jarkko/0000-0002-8582-8581
dc.contributor.authorDedeakayogullari, Huri
dc.contributor.authorValjakka, Jarkko
dc.contributor.authorTurunen, Ossi
dc.contributor.authorYilmazer, Berin
dc.contributor.authorDemir, Garip
dc.contributor.authorJanis, Janne
dc.contributor.authorBinay, Baris
dc.date.accessioned2024-05-19T14:50:19Z
dc.date.available2024-05-19T14:50:19Z
dc.date.issued2023
dc.departmentİstinye Üniversitesien_US
dc.description.abstractUnnatural amino acids are unique building blocks in modern medicinal chemistry as they contain an amino and a carboxylic acid functional group, and a variable side chain. Synthesis of pure unnatural amino acids can be made through chemical modification of natural amino acids or by employing enzymes that can lead to novel molecules used in the manufacture of various pharmaceuticals. The NAD+ -dependent alanine dehydrogenase (AlaDH) enzyme catalyzes the conversion of pyruvate to L-alanine by transferring ammonium in a reversible reductive amination activity. Although AlaDH enzymes have been widely studied in terms of oxidative deamination activity, reductive amination activity studies have been limited to the use of pyruvate as a substrate. The reductive amination potential of heterologously expressed, highly pure Thermomicrobium roseum alanine dehydrogenase (TrAlaDH) was examined with regard to pyruvate, a-ketobutyrate, a-ketovalerate and a-ketocaproate. The biochemical properties were studied, which included the effects of 11 metal ions on enzymatic activity for both reactions. The enzyme accepted both derivatives of L-alanine (in oxidative deamination) and pyruvate (in reductive amination) as substrates. While the kinetic KM values associated with the pyruvate derivatives were similar to pyruvate values, the kinetic k(cat) values were significantly affected by the side chain increase. In contrast, K-M values associated with the derivatives of L-alanine (L-a-aminobutyrate, L-norvaline, and L-norleucine) were approximately two orders of magnitude greater, which would indicate that they bind very poorly in a reactive way to the active site. The modeled enzyme structure revealed differences in the molecular orientation between L-alanine/pyruvate and L-norleucine/a-ketocaproate. The reductive activity observed would indicate that TrAlaDH has potential for the synthesis of pharmaceutically relevant amino acids.en_US
dc.description.sponsorshipTUBITAK [120Z501]en_US
dc.description.sponsorshipAcknowledgement This work was supported partially by TUBITAK (Project number: 120Z501) . We thank Mr. Jouni Oljakka (Master of Science in Mathe-matics) for the statistical analysis of the data.en_US
dc.identifier.doi10.1016/j.enzmictec.2023.110265
dc.identifier.issn0141-0229
dc.identifier.issn1879-0909
dc.identifier.pmid37269617en_US
dc.identifier.scopus2-s2.0-85160710921en_US
dc.identifier.scopusqualityQ2en_US
dc.identifier.urihttps://doi.org10.1016/j.enzmictec.2023.110265
dc.identifier.urihttps://hdl.handle.net/20.500.12713/5665
dc.identifier.volume169en_US
dc.identifier.wosWOS:001017261700001en_US
dc.identifier.wosqualityN/Aen_US
dc.indekslendigikaynakWeb of Scienceen_US
dc.indekslendigikaynakScopusen_US
dc.language.isoenen_US
dc.publisherElsevier Science Incen_US
dc.relation.ispartofEnzyme and Microbial Technologyen_US
dc.relation.publicationcategoryMakale - Uluslararası Hakemli Dergi - Kurum Öğretim Elemanıen_US
dc.rightsinfo:eu-repo/semantics/closedAccessen_US
dc.snmz20240519_kaen_US
dc.subjectUnnatural Amino Acidsen_US
dc.subjectMedicinal Chemistryen_US
dc.subjectAlanine Dehydrogenaseen_US
dc.subjectThermomicrobium Roseumen_US
dc.subjectReductive Amination Mechanismen_US
dc.subject& Alpha;-Keto Acidsen_US
dc.subjectMolecular Modelingen_US
dc.titleApplication of reductive amination by heterologously expressed Thermomicrobium roseum L-alanine dehydrogenase to synthesize L-alanine derivativesen_US
dc.typeArticleen_US

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