硫酸盐还原菌生物膜活性的原位快速测定研究

Sulfate-reducing bacteria (SRB) are the most corrosive and the most extensively studied corrosion related microorganisms, they are widely distributed in various marine environments. Although it has been clearly known that the metabolic activity of SRB is closely related with their roles in corrosion processes, determination of SRB metabolic activity is still a big problem. In this program, the complex SRB biofilm is set as research object, with an aim to develop a in-situ rapid method for determination of SRB biofilm metabolic activity and then evaluate its application performances. Based on the characteristic metabolic pathways of SRB, the metabolic activity of SRB biofilm is to be determined by specific analysis of the state of sulfide metabolite. Then functional modification and advanced material loading are to be introduced to fabricate the active recognition unit, which can enhance the transmission and identification efficiency of the recognition process in SRB biofilm, and thus reveal the spatial distribution characteristics of SRB biofilm activity. Finally, the validity and efficiency of SRB biofilm activity determination is to be verified and corrected, the determination time, stability, specificity, reproducibility and other performances are to be systematically evaluated, and the determination parameters and processes are to be specified. To achieve the spatial distribution status of biofilm activity timely, this program will construct a SRB biofilm activity determination approach systematically, which could provide theoretical basis and guidance for revealing the mechanism of SRB in the corrosion process and monitoring of microbial influenced corrosion status.

硫酸盐还原菌（SRB）是腐蚀性最强，也是研究最广泛的腐蚀微生物，广泛存在于海洋环境中。SRB的代谢活性与其参与的腐蚀过程密切相关，而SRB代谢活性的快速测定仍是一大难题。本研究以复杂的SRB生物膜体系为研究对象，旨在开发原位、快速的SRB生物膜代谢活性测定方法，并完成其应用性能评价。研究从SRB种群独特的代谢路径出发，以代谢产物硫化物为活性指示物，通过实时分析硫化物的代谢状态实现SRB活性的快速测定；借助功能化改性和先进材料搭载，解决活性识别单元在生物膜内的传质和识别难题，揭示生物膜内代谢活性的立体空间分布特征；验证并校正SRB生物膜活性测定的正确性和有效性，系统评价测试时间、稳定性、选择性、重现性等性能，规范测定参数和流程。本研究将系统完成SRB生物膜代谢活性测试方法的构建，实现生物膜代谢活性的实时、准确测定，为揭示SRB在腐蚀过程中的作用机理，实现微生物腐蚀状态监测提供理论依据和指导。

硫酸盐还原菌（SRB）是腐蚀性最强，也是研究最广泛的腐蚀微生物，广泛存在于海洋环境中。SRB的代谢活性与其参与的腐蚀过程密切相关，而SRB代谢活性的快速测定仍是一大难题。本研究以复杂的SRB生物膜体系为研究对象，构建了8套灵敏度高、特异性好、成本低廉的SRB 生物膜活性测定系统，分别为基于全固态接触式微电极探针的SRB生物膜代谢活性实时、连续监测方法，基于GSH-Au(I)配合物荧光探针的SRB生物膜代谢活性检测系统，生物膜内腐蚀微生物代谢活性分子ATP的双模式测定平台，基于磁控修饰抗菌肽材料的活性分子区分测定方法，基于无机纳米材料识别的高稳定性双信号响应检测平台，基于CRISPR/Cas12a集成ZIF-90@Ag3AuS2@Fe3O4纳米复合材料的ADP和ATP的超灵敏区分定量检测平台，适用于腐蚀现场的无装置化ATP纸基试剂盒，以及基于靶标竞争性结合的生物膜内活性分子和电荷传递载体同时检测平台。所开发的无装置化ATP纸基试剂盒通过设计基于ZIF-90的识别体系，解决了传统生物识别材料的稳定性难题，通过引入多糖酶解水凝胶和纳米酶催化显色特性，实现了生物膜内靶标分子的可视化测定，整个测试过程简便、快捷，且摆脱了对仪器设备的依赖，对于较高的现场应用潜力。此外，新型生物膜活性测定方法能够将活性指示分子的检测限由μM级降低至fM级，把整个测试时间控制在15分钟以内，并已用于初步揭示了不同金属材料表面生物膜活性的长期演变特征，为明确生物膜内腐蚀微生物代谢活性对金属腐蚀过程的影响，建立微生物腐蚀预测、监测模型提供可靠了重要手段。基于上述研究结果，研究团队在项目执行期间共发表关于SRB生物膜活性测定的期刊论文19篇，获山东省科学技术奖二等奖等奖励。

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