Molecules, Vol. 26, Pages 5636: Application of Capillary and Free-Flow Zone Electrophoresis for Analysis and Purification of Antimicrobial β-alanyl-tyrosine from Hemolymph of Fleshfly Neobellieria bullata (Molecules)

The problem of a growing resistance of bacteria and other microorganisms to conventional antibiotics gave rise to a search for new potent antimicrobial agents. Insect antimicrobial peptides (AMPs) seem to be promising novel potential anti-infective therapeutics. The dipeptide β-alanyl-tyrosine (β-Ala-Tyr) is one of the endogenous insect toxins exhibiting antibacterial activity against both Gram-negative and Gram-positive bacteria. Prior to testing its other antimicrobial activities, it has to be prepared in a pure form. In this study, we have developed a capillary zone electrophoresis (CZE) method for analysis of βâ€`Alaâ€`Tyr isolated from the extract of the hemolymph of larvae of the fleshfly Neobellieria bullata by reversed-phase high-performance liquid chromatography (RP-HPLC). Based on our previously described correlation between CZE and free-flow zone electrophoresis (FFZE), analytical CZE separation of βâ€`Alaâ€`Tyr and its admixtures have been converted into preparative purification of βâ€`Alaâ€`Tyr by FFZE with preparative capacity of 45.5 mg per hour. The high purity degree of the βâ€`Alaâ€`Tyr obtained by FFZE fractionation was confirmed by its subsequent CZE analysis.