离子交换层析结合反向色谱纯化聚二氨基丙酸

Purification of poly(L-diaminopropionic acid)by ion-exchange and reversed-phase chromatography

ε-聚赖氨酸生产菌株Streptomyces albulus PD-1可合成一种新型非蛋白质氨基酸均聚物聚二氨基丙酸,采用离子交换层析和反向色谱,对聚二氨基丙酸的分离纯化进行研究。离子交换层析柱选用DEAE-Sepharose Fast Flow填料,50 mmol/L磷酸盐缓冲液(p H 7.5)平衡上样,含0.5 mol/L Na Cl的磷酸盐缓冲液(p H 7.5)洗脱,收集洗脱液用分子筛Sephadex G-25除去磷酸盐缓冲液。然后用C18反相色谱进一步纯化,流动相为V(甲醇)/V(0.1%磷酸)=5/95。经过离子交换层析和反向色谱,纯化得到聚二氨基丙酸纯品,回收率为39.8%,样品纯度达98.4%,为后续的聚二氨基丙酸的深入研究奠定基础。

The ε-polylysine producing strain Streptomyces albulus PD-1 could synthesize a novel non-proteinic amino acid oligomer：poly （ L-diaminopropionic acid ） （ PDAP ）-Ion-exchange and reversed-phase chromatography were used to study the purification conditions of PDAP-In the ion-exchange chromatography purification,DEAE-Sepharose Fast Flow was selected,50 mmol/L phosphate buffer was used as mobile phase and 0-5 mol/L NaCl in the phosphate buffer acted as elution-In the reversed-phase chromatography purification,ratio of methanol to 0-1% phosphoric acid was 5/95 （ V/V） and was used as the mobile phase-Pure PDAP sample was obtained by ion-exchange and reversed-phase chromatography, the recovery rate of the purification was 39-8%,and the purity of the sample was 98.4%.