Zhao Ming, Ma Yan, Pan Ying-Hong, Zhang Chun-Hua, Yuan Wen-Xia
College of Pu-erh Tea, Yunnan Agricultural University, Kunming 650201, China.
Biomed Chromatogr. 2011 Aug;25(8):908-12. doi: 10.1002/bmc.1543. Epub 2011 Jan 26.
Paper mulberry (Broussonetia papyrifera, syn. Morus papyrifera L.) is a Chinese traditional medicine and its low-molecular-weight extracts are reported to have antifungal activity. In this study, two proteins (PMAPI and PMAPII) with activity against Trichoderma viride were obtained from paper mulberry leaves with a fast protein liquid chromatography (FPLC) unit. The purification protocol employed (NH(4))(2)SO(4) precipitation, ion-exchange chromatography and hydrophobic-interaction chromatography on FPLC. Molecular masses were 18,798 Da for PMAPI, and 31,178 Da for PMAPII determined by Matrix-assisted laser desorption ionization time-of-flight mass spectrometry. Peptide mapping fingerprint analysis showed that PMAPI has no peptides similar to PMAPII. N-terminal amino acid sequencing revealed that PMAPI is a hevein-like protein, and PMAPII is a class I chitinase. They both had a half-maximal inhibitory concentration (IC50) of 0.1 µg/µL against T. viride. This is the first report of high-molecular-weight extracts with antifungal activity from paper mulberry.
构树（Broussonetia papyrifera，同物异名Morus papyrifera L.）是一种传统中药，据报道其低分子量提取物具有抗真菌活性。在本研究中，使用快速蛋白质液相色谱（FPLC）装置从构树叶中获得了两种对绿色木霉具有活性的蛋白质（PMAPI和PMAPII）。所采用的纯化方案包括硫酸铵沉淀、离子交换色谱和FPLC上的疏水相互作用色谱。通过基质辅助激光解吸电离飞行时间质谱测定，PMAPI的分子量为18,798 Da，PMAPII的分子量为31,178 Da。肽图谱指纹分析表明，PMAPI没有与PMAPII相似的肽段。N端氨基酸测序显示，PMAPI是一种类橡胶素蛋白，PMAPII是一种I类几丁质酶。它们对绿色木霉的半数抑制浓度（IC50）均为0.1 µg/µL。这是关于构树中具有抗真菌活性的高分子量提取物的首次报道。
