Mono-\u3cem\u3eN\u3c/em\u3e-acyl-2,6-diaminopimelic Acid Derivatives: Analysis by Electromigration and Spectroscopic Methods and Examination of Enzyme Inhibitory Activity

Thirteen mono-N-acyl derivatives of 2,6-diaminopimelic acid (DAP)—new potential inhibitors of the dapE-encoded N-succinyl-l,l-diaminopimelic acid desuccinylase (DapE; EC 3.5.1.18)—were analyzed and characterized by infrared (IR) and nuclear magnetic resonance (NMR) spectroscopies and two capillary electromigration methods: capillary zone electrophoresis (CZE) and micellar electrokinetic chromatography (MEKC). Structural features of DAP derivatives were characterized by IR and NMR spectroscopies, whereas CZE and MEKC were applied to evaluate their purity and to investigate their electromigration properties. Effective electrophoretic mobilities of these compounds were determined by CZE in acidic and alkaline background electrolytes (BGEs) and by MEKC in acidic and alkaline BGEs containing a pseudostationary phase of anionic detergent sodium dodecyl sulfate (SDS) or cationic detergent cetyltrimethylammonium bromide (CTAB). The best separation of DAP derivatives, including diastereomers of some of them, was achieved by MEKC in an acidic BGE (500 mM acetic acid [pH 2.54] and 60 mM SDS). All DAP derivatives were examined for their ability to inhibit catalytic activity of DapE from Haemophilus influenzae (HiDapE) and ArgE from Escherichia coli (EcArgE). None of these DAP derivatives worked as an effective inhibitor of HiDapE, but one derivative—N-fumaryl, Me-ester-DAP—was found to be a moderate inhibitor of EcArgE, thereby providing a promising lead structure for further studies on ArgE inhibitors

Investigation of the protein architecture of the fungal hexosaminidases

Katedra biochemieDepartment of BiochemistryFaculty of SciencePřírodovědecká fakult

Analysis, characterization and separation of 2,6-diaminopimelic acid derivatives by capillary zone electrophoresis and micellar electrokinetic chromatography

In this work, capillary zone electrophoresis (CZE) and micellar electrokinetic chromatography (MEKC) with UV-photometric detection were used for the determination of degree of chemical purity, limit of detection (LOD) and limit of quantitation (LOQ) of N-acyl derivatives of 2,6-diaminopimelic acid (DAP). The characterization of DAP derivatives was performed by determination of their effective electroforetic mobilities in several background electrolytes in acidic and alkalic pH range. The mobilities were corrected to reference temperature 25 řC. Separation of the mixture of DAP derivatives were achieved by MEKC in acid background elektrolyte (500 mmol·dm-3 acetic acid, pH 2.54) using anionic surfactant 60 mmol·dm-3 sodium dodecylsulfate (SDS) as a constituent of the micellar pseudostationary phase. Subject words: analytical chemistry, separation methods, capillary electromigration methods, derivatives of amino acids Key words: capillary zone electrophoresis, micellar electrokinetic chromatography, derivatives of 2,6-diaminopimelic aci

Analysis, characterization and separation of 2,6-diaminopimelic acid derivatives by capillary zone electrophoresis and micellar electrokinetic chromatography

V rámci diplomové práce byly metodami kapilární zónové elektroforézy (CZE) a micelární elektrokinetické chromatografie (MEKC) s UV-fotometrickou detekcí určeny stupně chemické čistoty, meze detekce (LOD) a meze stanovitelnosti (LOQ) N-acyl derivátů kyseliny 2,6-diaminopimelové (DAP). Charakterizace derivátů DAP byla provedena stanovením jejich efektivních elektroforetických pohyblivostí korigovaných na referenční teplotu 25 řC v několika základních elektrolytech, v kyselé i v alkalické oblasti pH. Separace směsi derivátů DAP byla provedena metodou MEKC v kyselém základním elektrolytu, 500 mmol·dm-3 kyselině octové, pH 2,54, s přídavkem anionického tenzidu 60 mmol·dm-3 dodecylsulfátu sodného (SDS). Předmětová slova: analytická chemie, separační metody, kapilární elektromigrační metody, deriváty aminokyselin Klíčová slova: kapilární zónová elektroforéza, micelární elektrokinetická chromatografie, deriváty kyseliny 2,6-diaminopimelovéIn this work, capillary zone electrophoresis (CZE) and micellar electrokinetic chromatography (MEKC) with UV-photometric detection were used for the determination of degree of chemical purity, limit of detection (LOD) and limit of quantitation (LOQ) of N-acyl derivatives of 2,6-diaminopimelic acid (DAP). The characterization of DAP derivatives was performed by determination of their effective electroforetic mobilities in several background electrolytes in acidic and alkalic pH range. The mobilities were corrected to reference temperature 25 řC. Separation of the mixture of DAP derivatives were achieved by MEKC in acid background elektrolyte (500 mmol·dm-3 acetic acid, pH 2.54) using anionic surfactant 60 mmol·dm-3 sodium dodecylsulfate (SDS) as a constituent of the micellar pseudostationary phase. Subject words: analytical chemistry, separation methods, capillary electromigration methods, derivatives of amino acids Key words: capillary zone electrophoresis, micellar electrokinetic chromatography, derivatives of 2,6-diaminopimelic acidDepartment of Analytical ChemistryKatedra analytické chemieFaculty of SciencePřírodovědecká fakult