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用于电化学定量评估和定性表征富马酸对培养受体影响的生物传感器方法。

Biosensor approach for electrochemical quantitative assessment and qualitative characterization of the effect of fusaric acid on a culture-receptor.

作者信息

Emelyanova Elena V, Antipova Tatiana V

机构信息

Laboratory of Biosensor, G.K. Skryabin Institute of Biochemistry and Physiology of Microorganisms, Pushchino Center for Biological Research of the Russian Academy of Sciences, prosp. Nauki 5, Pushchino, Moscow Region 142290, Russian Federation.

Laboratory of Secondary Metabolites, G.K. Skryabin Institute of Biochemistry and Physiology of Microorganisms, Pushchino Center for Biological Research of the Russian Academy of Sciences, prosp. Nauki 5, Pushchino, Moscow Region 142290, Russian Federation.

出版信息

J Biotechnol. 2022 Sep 20;357:1-8. doi: 10.1016/j.jbiotec.2022.08.004. Epub 2022 Aug 11.

Abstract

Fusaric acid (FA) is a secondary fungal metabolite, which is widespread on corn and corn-based feed and food; FA has non-specific toxicity. Biosensor method is an express and easy-to-use method for quantitative and qualitative assessment of FA effect. Search for cultures has been performed for the formation of laboratory models of FA biosensor with the Clark-type oxygen electrode as transducer: respiration intensity of chosen cultures changed in the presence of FA. Resting cells of Fusarium oxysporum f. sp. vasinfectum and Bacillus subtilis were used as receptors of the amperometric biosensor for FA determination in aqueous solution. To enhance the sensitivity of detection, induction by substrate was performed for Bacillus subtilis. Response-concentration linear dependencies were obtained in a range of 0.5-500 FA mg/L. Biosensor models were applied to characterize influence of FA on microbial cells and investigate some features of FA transport. The dependences of the cells' response to FA on FA concentration were obtained; the kinetic parameters S and V were determined for each culture. Inhibition-threshold FA (S) concentrations were similar for both studied cultures. At concentrations lower than S, the process of simple diffusion governed FA transport into cells and caused the cells' response to FA for non-induced culture.

摘要

镰刀菌酸(FA)是一种真菌次生代谢产物,广泛存在于玉米及以玉米为原料的饲料和食品中;FA具有非特异性毒性。生物传感器法是一种用于定量和定性评估FA效应的快速且易于使用的方法。为构建以克拉克型氧电极作为换能器的FA生物传感器实验室模型,已开展了寻找合适培养物的工作:所选培养物的呼吸强度在FA存在时会发生变化。尖孢镰刀菌萎蔫专化型和枯草芽孢杆菌的静息细胞被用作安培型生物传感器的受体,用于测定水溶液中的FA。为提高检测灵敏度,对枯草芽孢杆菌进行了底物诱导。在0.5 - 500 FA mg/L范围内获得了响应 - 浓度线性关系。生物传感器模型被用于表征FA对微生物细胞的影响,并研究FA转运的一些特性。得到了细胞对FA的响应与FA浓度的关系;确定了每种培养物的动力学参数S和V。两种研究培养物的抑制阈值FA(S)浓度相似。在低于S的浓度下,简单扩散过程控制着FA进入细胞,并导致未诱导培养物中的细胞对FA产生响应。

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