Muhammad Saalim, Ya Lu, Kenneth J. Woycechowsky, Benjamin R. Clark
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引用次数: 0
摘要
长期以来,镍基树脂一直被用于生化研究,以纯化 His 标记的蛋白质,但迄今为止,尚未研究过镍基树脂在分离组氨酸衍生小分子方面的潜力。许多对农业具有重要意义的霉菌毒素都含有组氨酸残基,植物和细菌中的天然产物也是如此。本文介绍了一种利用不溶性镍盐 Ni3(PO4)2 纯化组氨酸天然产物的高选择性固相萃取方法。该方法分别从两种真菌菌株(金黄青霉和硬曲霉)中高选择性地结合和洗脱了两种天然产物:meleagrin 和 gartryprostatin C。结合、水洗和甲醇洗,再用甲醇醋酸盐缓冲液洗脱,这样一个简单的程序就能直接从两种真菌的粗提取物中回收 65-75% 的两种化合物。该方法可作为多步分离过程的一部分,与高效液相色谱法结合使用,很容易就能得到纯化的生物碱。
A cheap and straightforward method for the selective isolation of histidine-derived natural products using nickel(II) phosphate
Nickel-based resins have long been used in biochemical studies for the purification of His-tagged proteins, but their potential in the isolation of histidine-derived small molecules has not been investigated to date. Many agriculturally-important mycotoxins incorporate histidine residues, as do natural products from both plants and bacteria. Here, a highly-selective solid-phase extraction method is described for the purification of histidine-derived natural products using the insoluble nickel salt Ni3(PO4)2. This led to the highly-selective binding and elution of two natural products, meleagrin and gartryprostatin C, from two fungal strains, Penicillium chrysogenum and Aspergillus sclerotiorum, respectively. A simple protocol involving binding, washing with water and methanol, and elution with methanolic acetate buffer, gave 65–75% recovery of both compounds directly from crude extracts of the two fungi. The procedure can easily be used as part of a multi-step isolation process in combination with HPLC to yield the purified alkaloids.