An efficient procedure for preparing high-purity pingyangmycin and boanmycin from Streptomyces verticillus var. pingyangensis fermentation broth via macroporous cation-exchange resin and subsequent reversed-phase preparative chromatography

Abstract

Pingyangmycin (PYM) and boanmycin (BAM), two individual components of bleomycin (bleomycin A5 and bleomycin A6), are glycopeptide antitumor antibiotics. An efficient procedure for the preparation of PYM and BAM from Streptomyces verticillus var. pingyangensis fermentation broth using macroporous cation-exchange (MCE) resin followed by medium-pressure preparative liquid chromatography (MPLC) based on monodisperse poly(styrene-co-divinylbenzene) (p(st-dvb)) microspheres was investigated in this paper. Nine frequently used MCE resins were screened by static adsorption and desorption to enrich PYM and BAM fromthe fermentation broth, and D157 resin was found to be the most effective. After one run of column-based dynamic adsorption and desorption, the contents of PYM and BAM were increased by factors of 13.8 and 12.1 with recovery yields of 84.21% and 81.47%, respectively. The enriched samples were subjected to MPLC with columns prepacked with the PolyRP 10-300 microspheres. The operational parameters of the MPLC, including the stationary phase and mobile phase compositions, sample/stationary phase ratio, sample loading scale and flow rate, were screened and optimized. The results showed that the separation and purification for PYM and BAM by MPLC were dramatically improved with a mobile phase modifier of 0.15 mol/L ammonium chloride aqueoussolution, a flow rate of 10 mL/min and a sample/stationary phase ratio of 1.0:100 (m/v, g/mL), and PYM and BAM with purities of more than 98.65% and 99.12% were obtained, respectively. The total recoveries of PYM and BAM reached 75.38% and 70.31%. The separation and purification method is simple, efficient, energy-saving, environmentally friendly and suitable for the large-scale preparation of high-purity PYM and BAM from Streptomyces verticillus var. pingyangensis fermentation broth.