Egorova-Zachernyuk T A, Shvets V I, Versluis K, Heerma W, Creemers A F, Nieuwenhuis S A, Lugtenburg J, Raap J
M. V. Lomonosov Moscow State Academy of Fine Chemical Technology, Russia.
J Pept Sci. 1996 Nov-Dec;2(6):341-50. doi: 10.1002/psc.72.
A simple procedure for the preparation of the specifically labelled peptide antibiotic zervamicins IC, IIA and IIB has been developed. The zervamicin molecules are labelled with stable isotopes by culturing the Emericellopsis salmosynnemata on a well-defined synthetic medium containing the highly isotopically enriched amino acid. To obtain the peptide with the specifically and highly enriched amino acid residue, precautions have been taken to prevent any de novo biosynthesis of the particular amino acid from unlabelled precursors. The enrichment of the labelled peptide is determined by mass spectrometric analysis. Following this method we have incorporated [2',4',5',6',7'-2H5]-L-Trp-1, [1'-15N]-L-Trp-1 and [2',3',4',5',6'-2H5]-L-Phl-16 into zervamicins IC, IIA and IIB on the preparative scale and without scrambling of the label. Thus, using the procedures described, isotopically labelled zervamicins can be prepared, allowing them to be studied by solid-state NMR.
已开发出一种制备特异性标记的肽抗生素泽尔瓦霉素IC、IIA和IIB的简单方法。通过在含有高度同位素富集氨基酸的明确合成培养基上培养鲑鱼短梗霉，使泽尔瓦霉素分子用稳定同位素进行标记。为了获得具有特异性和高度富集氨基酸残基的肽，已采取预防措施以防止特定氨基酸从未标记前体进行任何从头生物合成。标记肽的富集通过质谱分析来确定。按照此方法，我们已在制备规模上且无标记混乱的情况下，将[2',4',5',6',7'-2H5]-L-色氨酸-1、[1'-15N]-L-色氨酸-1和[2',3',4',5',6'-2H5]-L-苯丙氨酸-16掺入泽尔瓦霉素IC、IIA和IIB中。因此，使用所述方法，可以制备同位素标记的泽尔瓦霉素，使其能够通过固态核磁共振进行研究。
