Zaitseva A S, Arlyapov V A, Yudina N Yu, Alferov S V, Reshetilov A N
Tula State University, 92 Lenin Prosp., Tula, 300012, Russia.
Tula State University, 92 Lenin Prosp., Tula, 300012, Russia; G.K. Skryabin Institute of Biochemistry and Physiology of Microorganisms, Russian Academy of Sciences, 5 Prosp. Nauki, Pushchino, Moscow Region, 142290, Russia.
Enzyme Microb Technol. 2017 Mar;98:43-51. doi: 10.1016/j.enzmictec.2016.12.005. Epub 2016 Dec 26.
We investigated the use of one- and two-mediator systems in amperometric BOD biosensors (BOD, biochemical oxygen demand) based on the yeast Debaryomyces hansenii. Screening of nine mediators potentially capable of electron transfer - ferrocene, 1,1'-dimethylferrocene, ferrocenecarboxaldehyde, ferroceneacetonitrile, neutral red, 2,6-dichlorophenolindophenol, thionine, methylene blue and potassium ferricyanide - showed only ferrocene and neutral red to be efficient electron carriers for the eukaryotes studied. Two-mediator systems based on combinations of the investigated compounds were used to increase the efficiency of electron transfer. The developed two-mediator biosensors exceeded their one-mediator analogs by their characteristics. The most preferable two-mediator system for developing a BOD biosensor was a ferrocene-methylene blue combination that ensured a satisfactory long-time stability (43 days), selectivity, sensitivity (the lower limit of the determined BOD concentrations, 2.5mg О/dm) and speed (assay time for one sample, not greater than 10min) of BOD determination. Analysis of water samples showed that the use of a ferrocene-methylene blue two-mediator system and the yeast D. hansenii enabled registration of data that highly correlated with the results of the standard method (R=0.9913).
我们研究了基于汉逊德巴利酵母的安培型生化需氧量（BOD）生物传感器中一介质和双介质系统的应用。对九种可能具有电子转移能力的介质进行筛选，包括二茂铁、1,1'-二甲基二茂铁、二茂铁甲醛、二茂铁乙腈、中性红、2,6-二氯酚靛酚、硫堇、亚甲基蓝和铁氰化钾，结果表明只有二茂铁和中性红是所研究真核生物的有效电子载体。基于所研究化合物组合的双介质系统用于提高电子转移效率。所开发的双介质生物传感器在性能上超过了其一介质类似物。用于开发BOD生物传感器的最优选双介质系统是二茂铁 - 亚甲基蓝组合，该组合确保了令人满意的长期稳定性（43天）、选择性、灵敏度（所测定BOD浓度的下限为2.5mg О/dm）和BOD测定速度（一个样品的分析时间不超过10分钟）。水样分析表明，使用二茂铁 - 亚甲基蓝双介质系统和汉逊德巴利酵母能够记录与标准方法结果高度相关的数据（R = 0.9913）。
