Isolation of individual amino acids from various microbiological sources using reversed-phase high-performance liquid chromatography |
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| Institution: | 1. Université de Carthage, INSAT (Institut National des Sciences Appliquées et de Technologie), Centre Urbain Nord, B.P. 676, 1080 Tunis, Tunisia;2. Université de Sfax, ENIS, LAVASA (Laboratoire Valorisation, Analyse et Sécurité des Aliments), BPW 3038, Sfax, Tunisia;3. Université de Lyon, Université Claude Bernard Lyon 1, BioDyMIA (Bioingénierie et Dynamique Microbienne aux Interfaces Alimentaires), Equipe Mixte d''Accueil Université Lyon 1 - ISARA Lyon n°3733, Technopole Alimentec, Rue Henri de Boissieu, 01000 Bourg en Bresse, France;2. Agricultural University of Athens, Athens, Greece;3. KU Leuven, Kortrijk, Belgium |
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| Abstract: | A new method for the preparative isolation of individual amino acids on a milligram scale based on reversed-phase high-performance liquid chromatography (RP-HPLC) after pre-column derivatization with carbobenzoxychloride (ZCl) has been developed. The chromatographic procedure was tested by the investigation of jack bean urease hydrolysate. The method has been applied to the preparative separation of Z-amino acids (from 10 up to 16) obtained from protein hydrolysates of various sources (green microalgae, blue-green algae, halophilic and methylotrophic microorganisms) and was proved to be reliable by the separation of deuterated amino acids (enrichment 97–99%) from Methylobacillus flagellatum (due to the bioconversion of CD3OD and D2O). Independent of the biological source of the protein, the amino acids were isolated with high recovery (from 68% up to 89%) and chromatographic purity (from 96% up to 99%). The method was also applied for the isolation of phenylalanine and leucine excreted by amino-acid overproducing microorganisms. |
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