Amperometric detection of the bacterial metabolic regulation with a microbial array chip.
Amperometric detection of the bacterial metabolic regulation with a microbial array chip.
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DOI：
10.1016/j.bios.2004.08.039
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发表时间：
2005-07
影响因子：
12.6
通讯作者：
K. Nagamine;Nobuto Matsui;T. Kaya;T. Yasukawa;H. Shiku;T. Nakayama;T. Nishino;T. Matsue
中科院分区：
文献类型：
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作者：
K. Nagamine;Nobuto Matsui;T. Kaya;T. Yasukawa;H. Shiku;T. Nakayama;T. Nishino;T. Matsue
应用胶原胶斑包埋活细菌细胞的微生物芯片研究了脱氮副球藻的代谢规律。使用扫描电化学显微镜(SECM)监测亚铁氰化物的产生，以反映位于脱氮假单胞菌内膜内的呼吸链中的电子流动。脱氮假单胞菌产生亚铁氰化物在很大程度上取决于碳源(葡萄糖或乳酸)的类型，这表明呼吸链中的电子流动速度取决于呼吸链上游代谢途径的活性。更重要的是，研究发现，在以d-(+)-葡萄糖为唯一碳源的营养琼脂培养基中，影响葡萄糖分解代谢反应的酶显著上调。利用脱氮假单胞菌的粗提物进行了酶活性测定，以确定在添加d-(+)-葡萄糖的培养物中高水平表达的酶。已证实，在含有d-(+)-葡萄糖的培养中，丙酮酸激酶和整个Entner-Doudoroff途径的酶被高度诱导。
A microbial array chip with collagen gel spots entrapping living bacterial cells has been applied to investigate the metabolic regulation in Paracoccus denitrificans. Scanning electrochemical microscopy (SECM) was used to monitor the ferrocyanide production that reflects the electron flow in the respiratory chain located within the internal membrane of P. denitrificans. The ferrocyanide production from P. denitrificans largely depends on the types of the carbon source (glucose or lactate), suggesting that the electron flow rate in the respiratory chain depends on the activity of the metabolic pathway located up-stream of the respiratory chain. More importantly, it was found that the enzymes affecting glucose catabolic reactions were significantly up-regulated in cultures with a nutrient agar medium containing d-(+)-glucose as a sole carbon source. Enzyme assays using crude extracts of P. denitrificans were carried out to identify the enzymes expressed at a higher level in cultures supplemented with d-(+)-glucose. It was confirmed that the pyruvate kinase and enzymes of the overall Entner–Doudoroff pathway were highly induced in cultures containing d-(+)-glucose.