Miranda M T, Tominaga M
Department of Biochemistry, University of São Paulo, Brazil.
Int J Pept Protein Res. 1991 Feb;37(2):128-33. doi: 10.1111/j.1399-3011.1991.tb00092.x.
Thermolysin was used as a catalyst to obtain the following protected di- and tripeptide esters: Z-Asn-Leu-OEt, Z-Asn-Phe-OEt, Moz-Asn-Leu-Gly-OEt, Boc-Asn-Leu-Gly-OEt, Z-Asn-Leu-Gly-OEt, Moz-Asn-Leu-Gly-OBzl, Moz-Asn-Leu-Gly-OtBu, Moz-Gln-Leu-Gly-OEt, Moz-Asn-Ile-Gly-OEt, and Moz-Asn-Leu-Ala-OEt. These compounds were obtained in pure form and the yields exceeded 50%, except for Moz-Asn-Leu-Gly-OtBu and Boc-Asn-Leu-Gly-OEt. H-Cys(Bzl)-OtBu and H-Cys(Bzl)-Pro-Leu-Gly-NH2 were both inadequate as amino components for obtaining Moz-Asn-Cys(Bzl)-OtBu, Z-Asn-Cys(Bzl)-OtBu and Moz-Asn-Cys(Bzl)-Pro-Leu-Gly-NH2 in the thermolysin-catalyzed reactions. In the attempted synthesis of the protected pentapeptide amide, this protease cleaved the Pro-Leu bond of the amino component H-Cys(Bzl)-Pro-Leu-Gly-NH2 and catalyzed the coupling between the resulting dipeptide amide and Moz-Asn-OH, thus yielding Moz-Asn-Leu-Gly-NH2 as the main product.
Z-天冬酰胺-亮氨酸乙酯、Z-天冬酰胺-苯丙氨酸乙酯、Moz-天冬酰胺-亮氨酸-甘氨酸乙酯、Boc-天冬酰胺-亮氨酸-甘氨酸乙酯、Z-天冬酰胺-亮氨酸-甘氨酸乙酯、Moz-天冬酰胺-亮氨酸-甘氨酸苄酯、Moz-天冬酰胺-亮氨酸-甘氨酸叔丁酯、Moz-谷氨酰胺-亮氨酸-甘氨酸乙酯、Moz-天冬酰胺-异亮氨酸-甘氨酸乙酯和Moz-天冬酰胺-亮氨酸-丙氨酸乙酯。这些化合物以纯形式获得，除了Moz-天冬酰胺-亮氨酸-甘氨酸叔丁酯和Boc-天冬酰胺-亮氨酸-甘氨酸乙酯外，产率均超过50%。在嗜热菌蛋白酶催化的反应中，H-Cys(Bzl)-OtBu和H-Cys(Bzl)-Pro-Leu-Gly-NH2作为氨基组分都不足以制备Moz-天冬酰胺-Cys(Bzl)-OtBu、Z-天冬酰胺-Cys(Bzl)-OtBu和Moz-天冬酰胺-Cys(Bzl)-Pro-Leu-Gly-NH2。在尝试合成受保护的五肽酰胺时，这种蛋白酶切割了氨基组分H-Cys(Bzl)-Pro-Leu-Gly-NH2的Pro-Leu键，并催化了所得二肽酰胺与Moz-天冬酰胺-OH之间的偶联反应，从而以Moz-天冬酰胺-亮氨酸-甘氨酸-NH2作为主要产物。
