PURIFICATION OF YERSINIABACTIN - A SIDEROPHORE AND POSSIBLE VIRULENCE FACTOR OF YERSINIA-ENTEROCOLITICA
PURIFICATION OF YERSINIABACTIN - A SIDEROPHORE AND POSSIBLE VIRULENCE FACTOR OF YERSINIA-ENTEROCOLITICA
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DOI：
10.1099/00221287-139-9-2159
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发表时间：
1993-09-01
期刊：
影响因子：
--
通讯作者：
ZAHNER, H
中科院分区：
文献类型：
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作者：
HAAG, H;HANTKE, K;ZAHNER, H
HPLC分析表明，小肠结肠炎耶尔森氏菌WA-C在铁限制条件下产生两种物质，其中一种被鉴定为2，3-二羟基苯甲酰-L-丝氨酸。另一种化合物具有铁络合活性，被称为耶尔森菌素。毛皮突变体H1852显示以铁非依赖性方式组成性地产生耶尔森菌素。从H1852发酵液中用乙酸乙酯萃取、分级层析和制备型HPLC分离得到耶尔森菌素。邻苯二酚的功能，证明了不同的化学分析和紫外-可见光谱。经测定，耶尔森菌素的相对分子质量为482 Da，纯化后的耶尔森菌素能刺激Y.小肠结肠炎菌和大肠杆菌phi的铁限制条件下，显然作为铁载体。Fe-55-耶尔森菌素的转运是TonB依赖性的，表明受体介导的跨外膜摄取。缺失受体蛋白FyuA的pesticin-resistant突变体不能运输和使用耶尔森菌素作为铁载体。
HPLC analysis revealed that Yersinia enterocolitica WA-C produced two substances under iron-limiting conditions one of which was identified as 2,3-dihydroxybenzoyl-L-serine. The other compound had iron-complexing activity and was called yersiniabactin. The fur mutant H1852 was shown to produce yersiniabactin constitutively in an iron-independent manner. Yersiniabactin was isolated by ethyl acetate extraction from the spent medium of H1852, size-fractionation chromatography and preparative HPLC. A catechol function was demonstrated with different chemical assays and by UV-visible spectroscopy. The molecular mass of yersiniabactin was determined to be 482 Da. Purified yersiniabactin stimulated growth of Y. enterocolitica and Escherichia coli phi under iron-limiting conditions and apparently served as an iron carrier. Transport of Fe-55-yersiniabactin was TonB-dependent, indicating a receptor-mediated uptake across the outer membrane. A pesticin-resistant mutant missing the receptor protein FyuA was unable to transport and use yersiniabactin as a siderophore.