Lam S K, Ng T B
Department of Biochemistry, Faculty of Medicine, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong, China.
Planta Med. 2002 Nov;68(11):1024-8. doi: 10.1055/s-2002-35649.
The roots of the sanchi ginseng, Panax notoginseng, were extracted with an aqueous buffer. The extract was chromatographed on a CM-cellulose column to remove extraneous unadsorbed proteins. The adsorbed fraction was dialyzed and chromatographed on Affi-gel blue gel. The adsorbed fraction was again collected, dialyzed and applied on a column of Mono S. The second peak was dialyzed and chromatographed on an FPLC-gel filtration Superdex 75 column. An antifungal protein with an N-terminal sequence similar to those of chitinases was isolated from the first peak which had a molecular mass of 35 kDa. The sequence was distinctive in that the third and ninth highly conserved N-terminal residues (C and G) were replaced by H and M, respectively. The protein inhibited mycelial growth in Coprinus comatus, Physalospora piricola, Botrytis cinerea, and Fusarium oxysporum with an IC 50 of 100 nM, 1 microM, 630 nM and 560 nM, respectively. It inhibited cell-free translation with an IC 50 of 630 nM. Its antifungal and translation-inhibitory activities were more potent than those of previously reported antifungal proteins. It inhibited human immunodeficiency virus-1 reverse transcriptase by 35.8 % at 12.6 microM and 24.7 % at 1.26 microM.
三七(Panax notoginseng)的根用缓冲液提取。提取物在CM - 纤维素柱上进行色谱分离以去除多余的未吸附蛋白质。吸附部分进行透析,然后在Affi - 凝胶蓝胶上进行色谱分离。再次收集吸附部分,透析后上样到Mono S柱上。第二个峰经透析后在FPLC凝胶过滤Superdex 75柱上进行色谱分离。从分子量为35 kDa的第一个峰中分离出一种N端序列与几丁质酶相似的抗真菌蛋白。该序列的独特之处在于,第三个和第九个高度保守的N端残基(C和G)分别被H和M取代。该蛋白抑制毛头鬼伞、梨孢盘菌、灰葡萄孢和尖孢镰刀菌的菌丝生长,IC50分别为100 nM、1 μM、630 nM和560 nM。它以630 nM的IC50抑制无细胞翻译。其抗真菌和翻译抑制活性比先前报道的抗真菌蛋白更强。它在12.6 μM时抑制人类免疫缺陷病毒1逆转录酶35.8%,在1.26 μM时抑制24.7%。