a State Key Laboratory of Food Science and Technology, Jiangnan University , Wuxi , People's Republic of China.
Prep Biochem Biotechnol. 2014;44(6):545-57. doi: 10.1080/10826068.2013.835733.
A novel trypsin inhibitor with thermal and pH stability, designated Merrtine, was isolated from Glycine max L. merr. The procedure involved ammonium sulfate precipitation, ion-exchange chromatography on CM-Sephadex C-50, and affinity chromatography on Affi-gel blue gel. The 20 N-terminal amino acid sequences were determined to be DEYSKPCCDLCMCTRRCPPQ, demonstrating high homology with the sequence of Bowman-Birk type trypsin inhibitors. The molecular mass and isoelectric point of the inhibitor were estimated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and isoelectric focusing to be 20.0 kD and 5.8, respectively. Trypsin could be completely inhibited by Merrtine when the molar ratio was 8.1. The inhibitory activity of Merrtine was unaffected after exposure to temperatures up to 85 °C, as well as within the pH range 2-12. Besides inhibiting trypsin-chymotrypsin, the inhibitor demonstrated additional antifungal activity against the species of Alternaria alternate, Fusarium oxysporum, Pythium aphanidermatum, Physalospora piricola, Botrytis cinerea, and Fusarium solani. We herein report not only the trypsin inhibitor's extraction and isolation for the first time, but also its physiochemical and antifungal properties.
从 Glycine max L. merr 中分离到一种具有热稳定性和 pH 稳定性的新型胰蛋白酶抑制剂，命名为 Merrtine。该过程涉及硫酸铵沉淀、CM-Sephadex C-50 离子交换层析和 Affi-gel blue 凝胶亲和层析。20 个 N 端氨基酸序列被确定为 DEYSKPCCDLCMCTRRCPPQ，与 Bowman-Birk 型胰蛋白酶抑制剂的序列具有高度同源性。抑制剂的分子量和等电点通过十二烷基硫酸钠聚丙烯酰胺凝胶电泳（SDS-PAGE）和等电聚焦分别估计为 20.0 kD 和 5.8。当摩尔比为 8.1 时，Merrtine 可完全抑制胰蛋白酶。Merrtine 的抑制活性在高达 85°C 的温度下以及在 pH 2-12 的范围内不受影响。除了抑制胰蛋白酶-糜蛋白酶外，该抑制剂还对Alternaria alternate、Fusarium oxysporum、Pythium aphanidermatum、Physalospora piricola、Botrytis cinerea 和 Fusarium solani 等物种具有额外的抗真菌活性。我们不仅首次报道了该胰蛋白酶抑制剂的提取和分离，还报道了其理化性质和抗真菌特性。
