Cheng E, Miranda M T, Tominaga M
Department of Biochemistry, University of São Paulo, Brazil.
Int J Pept Protein Res. 1988 Feb;31(2):116-25. doi: 10.1111/j.1399-3011.1988.tb00013.x.
The tripeptides Z-Pro-Leu-Gly-OEt, Z-Pro-Leu-Gly-NH2 and Z-Pro-Leu-Gly-OBzl were synthesized by thermolysin and alpha-chymotrypsin catalysis. The optimum conditions for the couplings between Z-Pro-OH and H-Leu-OEt or H-Leu-Gly-OEt catalysed by thermolysin were determined by a systematic study involving analysis of pH effect, ammonium sulfate concentration, reaction time, enzyme concentration, and relative proportion of the carboxyl and amine components. The best yield obtained for Z-Pro-Leu-OEt was 77% and for Z-Pro-Leu-Gly-OEt, 100%. Z-Pro-Leu-OEt was coupled to H-Gly-OEt, H-Gly-NH2 and H-Gly-OBzl. The best conditions to obtain Z-Pro-Leu-Gly-OEt and Z-Pro-Leu-Gly-NH2 were determined by the study of some factors that affect the reaction yield, such as organic solvent presence, substrate ratio and aqueous and organic solvent ratio. The yield obtained under optimum synthesis conditions was 55% for Z-Pro-Leu-Gly-OEt and 61% for Z-Pro-Leu-Gly-NH2. Z-Pro-Leu-Gly-OBzl was synthesized with 42% yield.
通过嗜热菌蛋白酶和α-胰凝乳蛋白酶催化合成了三肽Z-脯氨酸-亮氨酸-甘氨酸乙酯、Z-脯氨酸-亮氨酸-甘氨酸酰胺和Z-脯氨酸-亮氨酸-甘氨酸苄酯。通过系统研究确定了嗜热菌蛋白酶催化Z-脯氨酸-OH与H-亮氨酸乙酯或H-亮氨酸-甘氨酸乙酯偶联的最佳条件，该研究涉及分析pH效应、硫酸铵浓度、反应时间、酶浓度以及羧基和胺组分的相对比例。Z-脯氨酸-亮氨酸乙酯的最佳产率为77%，Z-脯氨酸-亮氨酸-甘氨酸乙酯的最佳产率为100%。Z-脯氨酸-亮氨酸乙酯与H-甘氨酸乙酯、H-甘氨酸酰胺和H-甘氨酸苄酯偶联。通过研究一些影响反应产率的因素，如有机溶剂的存在、底物比例以及水相和有机相比例，确定了获得Z-脯氨酸-亮氨酸-甘氨酸乙酯和Z-脯氨酸-亮氨酸-甘氨酸酰胺的最佳条件。在最佳合成条件下，Z-脯氨酸-亮氨酸-甘氨酸乙酯的产率为55%，Z-脯氨酸-亮氨酸-甘氨酸酰胺的产率为61%。Z-脯氨酸-亮氨酸-甘氨酸苄酯的合成产率为42%。
