从中国板栗中提取新型抗真菌蛋白——栗壳蛋白的纯化

Purification of castamollin, a novel antifungal protein from Chinese chestnuts.

作者信息

Wang H X, Ng T B

机构信息

Department of Microbiology, College of Biological Science, China Agricultural University, Beijing, China.

出版信息

Protein Expr Purif. 2003 Nov;32(1):44-51. doi: 10.1016/S1046-5928(03)00212-2.

DOI:10.1016/S1046-5928(03)00212-2

PMID:14680938

Abstract

A novel antifungal protein, designated castamollin, was isolated from Chinese chestnut (Castanea mollisima) seeds with a procedure involving ion exchange chromatography on DEAE-cellulose, affinity chromatography on Affi-gel blue gel, ion exchange chromatography on CM-Sepharose and FPLC-gel filtration on Superdex 75. Castamollin possessed a novel N-terminal sequence demonstrating little similarity to N-terminal sequences of Castanea sativa chitinase. Castamollin exhibited a molecular mass of 37kDa in gel filtration and SDS-PAGE. It inhibited the activity of human immunodeficiency virus-1 reverse transcriptase with an IC(50) of 7microM and translation in a cell-free rabbit reticulocyte lysate system with an IC(50) of 2.7microM. Castamollin displayed antifungal activity against Botrytis cinerea, Mycosphaerella arachidicola, Physalospora piricola, and Coprinus comatus but was devoid of lectin activity.

摘要

从中国板栗（Castanea mollisima）种子中分离出一种名为栗蘑素的新型抗真菌蛋白，其分离过程包括在DEAE - 纤维素上进行离子交换色谱、在Affi - 凝胶蓝胶上进行亲和色谱、在CM - 葡聚糖凝胶上进行离子交换色谱以及在Superdex 75上进行快速蛋白质液相色谱 - 凝胶过滤。栗蘑素具有一个新的N端序列，与欧洲栗几丁质酶的N端序列几乎没有相似性。在凝胶过滤和SDS - PAGE中，栗蘑素的分子量为37kDa。它在无细胞兔网织红细胞裂解物系统中抑制人免疫缺陷病毒1逆转录酶的活性，IC（50）为7μM，抑制翻译的IC（50）为2.7μM。栗蘑素对灰葡萄孢、落花生球腔菌、梨轮纹病菌和毛头鬼伞具有抗真菌活性，但没有凝集素活性。