压电体声波微生物传感及其应用研究

【作者】 张进忠；

【导师】 魏万之；

【作者基本信息】 湖南大学 ， 分析化学， 2001， 博士

【摘要】 压电体声波传感器具有响应谱广、灵敏度高、结构简单和易实现数字化等独特优点，其应用已涉及到分析化学、生命科学、环境监测和表面科学等众多领域。传统的微生物测定方法操作繁琐并且耗时，20世纪90年代提出了两种快速而简单的压电体声波传感器微生物测定方法。本文充分利用串联式压电传感器（SPQC）对溶液电导率的响应，以及单面触液型压电体声波传感器对电极上附加质量、溶液的粘度和密度的响应，并和微生物的生长代谢特征相结合，提出了几种新型体声波微生物传感方法，拓宽了压电体声波传感器在环境监测中的应用，信息的提取更加准确容易，测定更加快速方便。本论文还将SPQC检测器离子色谱法用于乳酸菌发酵过程的监测，获得了最优的发酵条件。采用阻抗分析技术，实时监测了微生物过程，在微生物生长代谢和酶催化溶菌响应模型方面开展了一些工作，从而可以充分利用响应曲线提供的信息，准确地描述微生物的生理生化过程，获得微生物生长和溶解过程的各动力学参数。本论文的创新性工作可概括如下： 1.提出了两种串联式压电微生物传感方法。根据SPQC传感器对溶液电导率的灵敏响应，建立了快速测定生化需氧量和致突变测试的新方法。论文中还采用分光光度法，实现了致突变物的测试。 2.将SPQC检测器离子色谱法用于乳酸菌发酵过程研究，考察了发酵过程中有机酸浓度的变化，对SPQC检测器和发酵条件进行了优化。 3.提出了几种单面触液型体声波微生物传感方法。利用单面触液型压电传感器对溶液粘-密度变化和电极上附加质量的响应，采用阻抗分析技术，实时监测了溶菌酶对溶壁微球菌（M.lysodeiteicus）的溶解过程、硫酸二甲酯（DMS）对鼠伤寒沙门氏菌（S.typhimurium）的致突变过程和氧化亚铁硫杆菌（T.ferrooxidans）存在下Fe²⁺的生物氧化过程，获得了反映这些过程的多维信息。 4.充分利用阻抗响应曲线提供的信息，结合Gompertz细菌生长模型，获得了致突变过程和生物氧化过程中微生物生长的动力学参数（μ_m、A和λ）。 5.建立了溶菌酶催化微球菌溶解的阻抗响应模型。根据溶菌速率与加入溶菌酶的量和初始菌液浓度的关系，结合动态电阻与溶液粘度和密度变化的关系，建立了反映溶菌过程的阻抗响应模型，并获得了该过程的动力学参数（自发溶菌速率K₁和酶催化溶菌速率K₂）。更多还原

【Abstract】 Piezoelectric bulk acoustic wave (PBAW) sensor has been applied to numerous fields, including analytical chemistry, life sciences, environmental monitoring and surface science because of its advantages such as broad sensing- spectrum, high sensitivity, simple construction and easy to realize numeralization. Conventional methods for microorganism determination are cumbersome and time-consuming. In the 1990抯 two types of PBAW sensors for microorganism detection were proposed, which had advantages over conventional methods and were more rapid and simpler. In this thesis, combined with some characteristics of microbial growth and metabolism, several new types of PBAW microbial sensing methods were presented based on the response of SPQC sensor to the conductivity of solution, and the response of one-face-contacting liquid PQC sensor to the loading mass on gold electrode and the viscosity and density of solution. These new PBAW methods are easier and broaden their application in environmental monitoring. Moreover, the detection is more rapid and simpler and the accurate extract of information is easier. Also in this thesis, ion chromatography with SPQC detector was utilized to monitor Lactobacillus fermentation process, and optimum fermentation conditions were obtained. Some microbial processes were real time monitored by impedance analysis technique. Some work has been done on the response models of microbial growth and metabolism and bacteriolysis catalyzed by enzyme, so the information provided by response curves can be utilized sufficiently. Thus physiological and biochemical processes can be described accurately, and the dynamics parameters of microbial growth and bacteriolytic process can be obtained. The creative works are summarized as follows: 1. Two types of SPQC microbial sensing methods were developed. New methods for the rapid determination of biochemical oxygen demand and mutagenicity test were established based on the sensitive response of SPQC sensor to the conductivity of solution. Also in this thesis, mutagenicity test was realized by spectrophotometry. ii 2. Ion chromatography with SPQC detector was utilized to study Lactobacillus fermentation process. The changes of concentrations of organic acids were examined during the fermentation process, and SPQC detector and fermentation conditions were optimized. 3. Several one-face-contacting liquid PBAW microbial sensing methods were proposed. The bacteriolytic process of lysozyme to M lysodeiteicus, mutagenic process of dimethyl sulfate to S. typhimurium, bio-oxidation process of ferrous ion in the presence of 7? ferrooxidans were real time monitored by using impedance analysis technique, which is based on the response of PQC to the viscosity and density changes and the loading mass on gold electrode. Multidimensional information reflecting these processes was acquired. 4. The information provided by impedance response curves was utilized sufficiently, combined with Gompertz bacterial growth model, and thus the dynamics parameters (pin, A and 2) of microbial growth were obtained during the mutagenic process and the bio-oxidation process. 5. Impedance response model of the bacteriolysis of M lysodeiteicus catalyzed by lysozyme was established. The model was based on the relationship between the motional resistance and the viscosity and density changes of solution, combined with the更多还原