聚球藻7002嗜铁素的检测与分离

Detection and separation of the siderophores from Synechococcus sp. PCC 7002

摘要:

【目的】探究聚球藻7002嗜铁素的检测和分离方法，为深入研究海洋嗜铁素提供科学依据。【方法】在缺铁Medium A中培养聚球藻7002，利用双层平板法、混合平板法和传统铬天青S (CAS)平板法定性检测嗜铁素，用CAS蓝色液体检测液定量检测嗜铁素。采用大孔树脂XAD-2和固定化金属离子亲和层析(IMAC)对嗜铁素进行分离，IMAC采用降低pH和竞争性洗脱两种洗脱方式。【结果】混合平板定性检测法更快速、高效、便捷。缺铁培养的聚球藻7002发酵液中，嗜铁素的相对含量高达93.50%。大孔树脂分离，上样液pH调为2.0时，嗜铁素吸附充分，分离效果较好。试验发现，分离得到的聚球藻7002嗜铁素在254 nm紫外下具有明显的荧光特性。【结论】试验得到了聚球藻7002嗜铁素定性检测和分离的有效方法。

Abstract:

[Objective] In order to provide a scientific basis for further research of marine siderophores, detection and separation methods of the siderophores from Synechococcus sp. PCC 7002 were investigated. [Methods] Synechococcus sp. PCC 7002 was cultivated in iron-limited Medium A. We used two-layer plate, mixture plate and traditional chromazurol S plate to detect the siderophores qualitatively. The relative amount of siderophores was detected by the chromazurol S shuttle assay. Amberlite XAD-2 resin and immobilized metal-chelating affinity chromatography were used for siderophores separation. Two different elution methods (pH reduced elution and competitive elution) were utilized in immobilized metal-chelating affinity chromatography. [Results] Mixture plate was more rapid, efficient and convenient among the three qualitative detection methods. The relative amount of siderophores from the iron-limited culture was up to 93.50%. Siderophores adsorbed sufficiently into Amberlite XAD-2 resin and the separating efficiency was relatively high when the sample pH was adjusted to 2. Strong fluorescence of siderophores from Synechococcus sp. PCC 7002 was observed at 254 nanometer ultraviolet light. [Conclusion] In present study, we got the effective detection and separation methods of siderophores from Synechococcus sp. PCC 7002.