여름정기학술대회
2022여름초록
발표자 및 발표 내용
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주저자
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공동저자
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접수자
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This
study describes the identification and quantification of peptide natural
products produced by bacteria as secondary metabolites. The target peptides are
bioactive componds from insect-pathogenic bacteria genera Photorhabdus and Xenorhabdus,
named as GameXPeptides (GXPs). The GXPs, entomopathogenic compounds, can infect
and kill several soil-dwelling insect larvae and therefore this compounds can
be used industrially for pest control in organic farming. The proposed LC-MS
based method measure four GXPs, GXP A to D, simultaneously within 10 min. As
GXPs, a class of cyclic pentapeptides composed of valine, leucine and
phenylalanine, have high similarity in both molecular weight and structure,
required finely programed separation, and higher mass resolving performance. The
GXPs are separated between pentafluorophenyl stationary phase (Kinetex F5, 100
× 2.1 mm, 2.6 µm; Phenomenex) and mobile phases, comprised 20 mM ammonium
formate, 0.1 % formic acid, and acetonitrile for gradient elution. Separated
GXPs are detected by ESI-QTOF MS (5600+ TripleTOF, ABSciex) for identification
and quantification of four GXPs. The LC-MS conditions were optimized and
validated with synthetic GXPs. The linearity, sensitivity, measurement accuracy
and precision were satisfied within acceptance criteria. The proposed method
applied to determine GXPs in bacteria extracts, genetically modified to improve
its production yield. The expression of these compounds can be drived by
regulation of the promoters, and the developed method took a role in
confirmation of the expression yield. It will be
suitable for the accurate and efficient quantification method for GXPs in
various applications.
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