Affi Mahmoud, Solliec Camille, Legentilhomme Patrick, Comiti Jacques, Legrand Jack, Jouanneau Sulivan, Thouand Gérald
University of Nantes, UMR CNRS 6144 GEPEA, CBAC group, 18 bd. Gaston Defferre, La Roche sur Yon, 85035, France.
Ecole des Mines de Nantes, UMR CNRS 6144 GEPEA, 4, rue Alfred-Kastler, Nantes, CEDEX, 44307, France.
Anal Bioanal Chem. 2016 Dec;408(30):8761-8770. doi: 10.1007/s00216-016-9490-3. Epub 2016 Apr 4.
Water quality and water management are worldwide issues. The analysis of pollutants and in particular, heavy metals, is generally conducted by sensitive but expensive physicochemical methods. Other alternative methods of analysis, such as microbial biosensors, have been developed for their potential simplicity and expected moderate cost. Using a biosensor for a long time generates many changes in the growth of the immobilized bacteria and consequently alters the robustness of the detection. This work simulated the operation of a biosensor for the long-term detection of cadmium and improved our understanding of the bioluminescence reaction dynamics of bioreporter bacteria inside an agarose matrix. The choice of the numerical tools is justified by the difficulty to measure experimentally in every condition the biosensor functioning during a long time (several days). The numerical simulation of a biomass profile is made by coupling the diffusion equation and the consumption/reaction of the nutrients by the bacteria. The numerical results show very good agreement with the experimental profiles. The growth model verified that the bacterial growth is conditioned by both the diffusion and the consumption of the nutrients. Thus, there is a high bacterial density in the first millimeter of the immobilization matrix. The growth model has been very useful for the development of the bioluminescence model inside the gel and shows that a concentration of oxygen greater than or equal to 22 % of saturation is required to maintain a significant level of bioluminescence. A continuous feeding of nutrients during the process of detection of cadmium leads to a biofilm which reduces the diffusion of nutrients and restricts the presence of oxygen from the first layer of the agarose (1 mm) and affects the intensity of the bioluminescent reaction. The main advantage of this work is to link experimental works with numerical models of growth and bioluminescence in order to provide a general purpose model to understand, anticipate, or predict the dysfunction of a biosensor using immobilized bioluminescent bioreporter in a matrix.
水质和水管理是全球性问题。对污染物尤其是重金属的分析,通常采用灵敏但昂贵的物理化学方法。已开发出其他分析方法,如微生物生物传感器,因其潜在的简易性和预期的适中成本。长时间使用生物传感器会使固定化细菌的生长产生许多变化,从而改变检测的稳健性。这项工作模拟了用于长期检测镉的生物传感器的运行,并增进了我们对琼脂糖基质内生物报告细菌生物发光反应动力学的理解。数值工具的选择是合理的,因为在每种条件下都难以通过实验测量生物传感器长时间(数天)的功能。通过将扩散方程与细菌对营养物质的消耗/反应相耦合,对生物量分布进行了数值模拟。数值结果与实验分布显示出非常好的一致性。生长模型证实细菌生长受营养物质扩散和消耗的制约。因此,在固定化基质的第一毫米处细菌密度很高。该生长模型对凝胶内生物发光模型的开发非常有用,并且表明需要氧气浓度大于或等于饱和度的22%才能维持显著水平的生物发光。在镉检测过程中持续供应营养物质会导致形成生物膜,这会减少营养物质的扩散并限制琼脂糖第一层(1毫米)中氧气的存在,从而影响生物发光反应的强度。这项工作的主要优点是将实验工作与生长和生物发光的数值模型联系起来,以便提供一个通用模型来理解、预测或预知使用固定在基质中的生物发光生物报告器的生物传感器的功能障碍。