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In situ detection of microbial c-type cytochrome based on intrinsic peroxidase-like activity using screen-printed carbon electrode
Biosensors and Bioelectronics ( IF 12.6 ) Pub Date : 2018-04-22 , DOI: 10.1016/j.bios.2018.04.049
Junlin Wen , Daigui He , Zhen Yu , Shungui Zhou

C-type cytochromes (c-cyts) facilitate microbial extracellular electron transfer and play critical roles in biogeochemical cycling, bioelectricity generation and bioremediation. In this study, a simple and effective method has been developed to detect microbial c-cyts by means of peroxidase mimetic reaction on screen-printed carbon electrode (SPCE). To this end, bacteria cells were immobilized onto the working electrode surface of SPCE by a simple drop casting. After introducing 3,3′,5,5′-tetramethylbenzidine (TMB) solution, microbial c-cyts with peroxidase-like activity catalyze the oxidation of TMB in the presence of hydrogen peroxide. The oxidized TMB was electrochemically determined and the current signal was employed to calculate the c-cyts content. This electrochemical method is highly sensitive for microbial c-cyts with a low detection limit of 40.78 fmol and a wide detection range between 51.70 fmol and 6.64 pmol. Moreover, the proposed technique can be universally expanded to detect c-cyts in other bacteria species such as Fontibacter ferrireducens, Pseudomonas aeruginosa, Comamonas guangdongensis and Escherichia coli. Furthermore, the proposed method confers an in situ facile and quantitative c-cyts detection without any destructive sample preparations, complex electrode modifications and expensive enzyme- or metal particle- based signal amplification. The suggested method advances an intelligent strategy for in situ quantification of microbial c-cyts and consequently holds promising application potential in microbiology and environmental science.



中文翻译:

使用丝网印刷碳电极基于固有过氧化物酶样活性原位检测微生物c型细胞色素

C型细胞色素(c-cyts)促进微生物细胞外电子转移,并在生物地球化学循环,生物电产生和生物修复中发挥关键作用。在这项研究中,已开发出一种简单有效的方法,可通过在丝网印刷碳电极(SPCE)上进行过氧化物酶模拟反应来检测微生物c-cyt。为此,通过简单的滴铸将细菌细胞固定在SPCE的工作电极表面上。引入3,3',5,5'-四甲基联苯胺(TMB)溶液后,具有过氧化物酶样活性的微生物c-cyt在过氧化氢的存在下催化TMB的氧化。电化学测定氧化的TMB,并使用电流信号计算c-cyts含量。该电化学方法对微生物c-细胞高度敏感,检测限低至40.78 fmol,检测范围介于51.70 fmol至6.64 pmol之间。此外,所提出的技术可以普遍扩展以检测其他细菌物种中的c-cyts,例如铜绿假单胞菌广东鸭毛大肠杆菌。此外,所提出的方法可以在不进行任何破坏性样品制备,复杂的电极修饰和昂贵的基于酶或金属颗粒的信号放大的情况下,进行现场,定量的c-cyts检测。所提出的方法为微生物c-cyt的原位定量分析提供了一种智能策略,因此在微生物学和环境科学中具有广阔的应用前景。

更新日期:2018-04-22
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