Fischer A, Schwarz A, Wandrey C, Bommarius A S, Knaup G, Drauz K
Institut für Biotechnologie, Forschungszentrum Jülich, Germany.
Biomed Biochim Acta. 1991;50(10-11):S169-74.
Several amino acid derivatives with the negatively charged N alpha-protecting groups Maleyl (Mal) and Citraconyl (Cit) were synthesized and used in enzyme-catalyzed peptide synthesis. Compared to commonly used alpha-amino protecting groups in chemical peptide synthesis (Z, Fmoc, Boc, etc.), these charged protecting groups strongly increase both water solubility of different aromatic amino acid derivatives and activities of synthesis reactions. As a consequence of the solubilizing effect, we used these groups in the kinetically controlled peptide synthesis choosing kyotorphin (tyrosyl-arginine) as a model peptide. With Mal-Tyr-OEt as substrate and Arg-OEt as nucleophile, we succeeded in the alpha-chymotrypsin-catalyzed production of about 12 kg of Tyr-Arg (50.4% overall yield) in a 300 l batch experiment.
合成了几种带有带负电荷的Nα-保护基团马来酰基（Mal）和柠康酰基（Cit）的氨基酸衍生物，并将其用于酶催化的肽合成。与化学肽合成中常用的α-氨基保护基团（Z、Fmoc、Boc等）相比，这些带电荷的保护基团极大地提高了不同芳香族氨基酸衍生物的水溶性以及合成反应的活性。作为增溶作用的结果，我们在动力学控制的肽合成中使用了这些基团，选择脑啡肽（酪氨酰-精氨酸）作为模型肽。以Mal-Tyr-OEt为底物，Arg-OEt为亲核试剂，我们在300升的分批实验中成功地通过α-胰凝乳蛋白酶催化生产了约12千克的Tyr-Arg（总产率50.4%）。
