Easy detection of siderophore production in diluted growth media using an improved CAS reagent
Easy detection of siderophore production in diluted growth media using an improved CAS reagent
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DOI：
10.1016/j.mimet.2021.106310
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发表时间：
2021-08-26
影响因子：
2.2
通讯作者：
Morimoto, Kinjiro
中科院分区：
文献类型：
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作者：
Murakami, Chiho;Tanaka, Arowu R.;Morimoto, Kinjiro
铁载体是由各种微生物，特别是包括根际细菌在内的病原菌产生的低分子量有机化合物，对铁具有高亲和力。尽管大多数微生物被认为在缺铁条件下分泌铁载体，但尚不清楚有多少微生物在自然环境中产生铁载体。此外，广泛用于检测铁载体的铬天青磺酸盐（CAS）测定法需要改进以获得更广泛的适用性。我们在 96 孔微孔板中开发了一种简单、高通量的 CAS 测定，其中使用浓缩的 CAS 试剂和常用的稀释生长培养基，且没有人工铁消耗。改进的微孔板 CAS 穿梭试验表明，它可以轻松检测假单胞菌 (P.) 荧光、恶臭假单胞菌、稳定伯克霍尔德菌和米酵母释放的铁载体，作为产生铁载体的细菌模型。这种 CAS 穿梭试验与稀释的生长培养基一起使用是筛选新的产生铁载体的细菌的有前途的工具。
Siderophores are low molecular weight organic compounds produced by various microorganisms, especially pathogenic bacteria including rhizobacteria, and have a high affinity for iron. Although most microorganisms are thought to secrete siderophores under iron-depleted conditions, it is unclear how many microorganisms produce siderophores in the natural environment. Also, the chrome azurol sulfonate (CAS) assay, which is widely used for the detection of siderophores, needs to be improved for wider applicability. We developed a simple, highthroughput CAS assay in a 96-well microplate with a concentrated CAS reagent and commonly used diluted growth media in the absence of artificial iron depletion. The improved microplate CAS shuttle assay revealed that it could easily detect siderophores released from Pseudomonas (P.) fluorescence, P. putida, Burlkholderia stabilis, and Ottowia oryzae, as models of siderophore-producing bacteria. This CAS shuttle assay employed along with diluted growth media is a promising tool to screen new siderophore-producing bacteria.