Method for separating and purifying epothilone D based on molecular imprinting

Abstract

The invention discloses a method for separating and purifying epothilone D based on molecular imprinting. The method includes the steps of soaking an EpoD molecular imprinting polymer with a small amount of pure methanol, filling a separation column with the materials, leaching the column sequentially through methanol-acetic acid, methanol and methanol-deionized water, vacuumizing and compacting the column, taking an EpoD crude product to be dissolved in a 20-80% methanol water solution, adding the materials in the assembled column, leaching the column after standing for 20-50 min to remove interferent so that impurity interference can be further reduced, eluting the column with a 10-90% methanol PBS solution as the eluant, completely dissolving EpoD in the eluant, collecting the eluant, conducting rotating and drying by distillation, and redissolving the pure methanol to obtain a purified EpoD methanol solution. Compared with an existing separating and purifying method, the method issimple and convenient to operate and low in cost; meanwhile, specific adsorption is realized in the adsorption process of the epothilone D, a better separation effect can be achieved, and the probability is provided for further separating and purifying the epothilone D.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for separating and purifying epothilone D based on molecular imprinting. Background technique [0002] Epothilone (epothilone) is a class of macrolide compounds, first reported in 1993 by G. Höfle et al. of the German National Center for Biotechnology (GBF). Epothilone can be isolated from the fermentation liquid of the Myxobacteria suborder S. cellulosus strain, and its main components are Epothilone A and B. The biological activities of epothilone A, B and D on cancer cells are equivalent to or even stronger than paclitaxel, especially epothilone B, whose activity is 10-100 times stronger than paclitaxel. The activity of epothilone will be greatly improved after the substitution of methyl on the C12 position and the replacement of the C12-C13 double bond with an epoxy group. Epothilones have superior anti-tumor properties than paclitaxel: ① Epothilones are mo...

AI Technical Summary

Problems solved by technology

② Epothilone has no cytotoxic activity of paclitaxel

However, the high cost in its production process has become a problem that people need to solve urgently.

In addition to factors such as low yield of the producing bacteria, long growth cycle, and susceptibility to infection, there are also major factors in the isolation and purification of epothilone

Because the expression product of S. cellulosus has complex and diverse components, the difficulty of separation and purification of epothilone is greatly increased.

[0004] At present, the methods for the separation and purification of epothilone mainly include: Sephadex combined with PHPLC separation and purification route, its purity can reach more than 90%, but the recovery rate is low, only about 75%, and the cost of filler is high. The operation is relatively complicated; the C18 normal pressure reverse column chromatography separation and purification route has a recovery rate of 80% and a purity of 85%, and its packing cost is high and the recovery rate is low; the ion exchange column chromatography separation and purification route , the recovery rate can reach 90%, but the purity of 60% is far from meeting the requirements of practical application