Li Dak Sum Yip Yio Chin Kenneth Li Marine Biopharmaceutical Research Center, College of Food and Pharmaceutical Sciences, Ningbo University, Ningbo 315800, China.
School of Marine Science, Ningbo University, Ningbo 315211, China.
Mar Drugs. 2020 Jun 25;18(6):332. doi: 10.3390/md18060332.
An efficient strategy was developed for the rapid separation and enrichment of bafilomycin A1 (baf A1) from a crude extract of the marine microorganism fermentation. This strategy comprises liquid-liquid extraction (LLE) with a three-phase solvent system (-hexane-ethyl acetate-acetonitrile-water = 7:3:5:5, ///) followed by separation using high-speed counter-current chromatography (HSCCC). The results showed that a 480.2-mg fraction of baf A1-enriched extract in the middle phase of the three-phase solvent system was prepared from 4.9 g of crude extract after two consecutive one-step operations. Over 99% of soybean oil, the main hydrophobic waste in the crude extract, and the majority of hydrophilic impurities were distributed in the upper and lower phase, respectively. HSCCC was used with a two-phase solvent system composed of -hexane-acetonitrile-water (15:8:12, //) to isolate and purify baf A1 from the middle phase fraction, which yielded 77.4 mg of baf A1 with > 95% purity within 90 min. The overall recovery of baf A1 in the process was determined to be 95.7%. The use of a three-phase solvent system represents a novel strategy for the simultaneous removal of hydrophobic oil and hydrophilic impurities from a microbial fermentation extract.
开发了一种从海洋微生物发酵物粗提物中快速分离和富集巴佛洛霉素 A1（baf A1）的有效策略。该策略包括使用三相溶剂系统（-己烷-乙酸乙酯-乙腈-水=7:3:5:5，///）进行液液萃取（LLE），然后使用高速逆流色谱法（HSCCC）进行分离。结果表明，从 4.9 g 粗提物中，经过两次连续的一步操作，可以制备出 480.2 mg 富含 baf A1 的中间相萃取物。超过 99%的大豆油，粗提物中的主要疏水性废物，以及大部分亲水性杂质分别分布在上相和下相中。HSCCC 采用两相溶剂系统（-己烷-乙腈-水（15:8:12，//）），从中间相部分分离和纯化 baf A1，在 90 分钟内可得到 77.4 mg 纯度大于 95%的 baf A1。该过程中 baf A1 的总回收率确定为 95.7%。三相溶剂系统的使用代表了一种从微生物发酵物中同时去除疏水性油和亲水性杂质的新策略。
