Isolation of individual amino acids from various microbiological sources using reversed-phase high-performance liquid chromatography.
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 (Z-Cl) 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.
Egorova
T A
TA
Department of Biotechnology, Lomonosov State Academy of Fine Chemical Technology, Moscow, Russian Federation.
Eremin
S V
SV
Mitsner
B I
BI
Zvonkova
E N
EN
Shvets
V I
VI
D000596:Amino Acids / Q000302:isolation & purification*; D000460:Chlorophyta / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D000458:Cyanobacteria / Q000737:chemistry; D007887:Fabaceae / Q000201:enzymology; D006088:Gram-Negative Aerobic Bacteria / Q000737:chemistry; D006216:Halobacteriaceae / Q000737:chemistry; D006868:Hydrolysis; D010946:Plants, Medicinal; D013056:Spectrophotometry, Ultraviolet; D014510:Urease / Q000737:chemistry
D000596:Amino Acids; D014510:Urease