BE/IDEA: Development of a Instrument for in situ Measurement of Microbial Enzyme Activities in Aquatic Ecosystems

微生物细胞表面酶（胞外酶）是水环境中聚合物水解的重要因子，也是微生物碳、氮、磷营养状态的指示剂。然而，像大多数其他微生物代谢率在水生环境中的测量，胞外酶活性测量通常被限制到手动测定与离散的水样。最近开发了一种使用高灵敏度荧光底物连续进行微生物酶活性测量的仪器。该仪器已被用于船上绘制密西西比河羽流表面沃茨中碱性磷酸酶和亮氨酸氨肽酶的胞外酶活性。有了这种仪器，这些酶的活动可以映射在很大程度上相同的方式，温度，盐度，浮游植物荧光，和其他参数的映射从一个研究船，而航行。然而，该系统仅限于对从地表连续泵送的水样进行水平测绘，并且需要频繁的操作员干预。 这个生物复杂性环境活动仪器开发（IDEA）项目将迈出更大的一步。研究和教育团队包括一名微生物生态学家（J. Ammerman博士）和一名地球化学家（G. Klinkhammer博士），他们都对传感器开发感兴趣并拥有专业知识，他是一位沿海物理海洋学家和建模师（R。圣歌），和非教师教育专家（先生E。Simms）。 该项目的目标是开发一种用于测量微生物酶活动的遥控仪器，并将其部署在罗格斯大学长期生态系统观测站15米深处，称为LEO-15，为期数周至数月。这种仪器被称为多酶分析仪（MEA），应该能够同时测量多达四种不同的酶活性。将在酶分析仪附近部署一个带有生物、化学和物理传感器的简单系泊装置，以帮助提供微生物速率测量的背景（沿着来自LEO-15的额外数据）。 酶和相关数据将通过时间序列方法、光谱分析技术、统计建模和其他技术进行分析，以深入了解沿海地区的微生物过程。第二次部署多边环境协定将在新的玛莎葡萄园岛海岸观测站（伍兹霍尔海洋研究所）进行，沿着有一台自动流式细胞仪和图像分析仪，用于浮游生物细胞计数和分类。 该仪器由R博士开发。Olson和同事在Biocomplexity IDEA的支持下。该酶分析仪将提供当前不可用的时间尺度上的连续真实的时间微生物代谢率测量。这种测量将为海洋观测站可以进行的广泛的化学和物理测量增加一个重要的生物组成部分。此外，水生微生物学方面的教育工作将包括为中小学教师举办培训讲习班，然后编写和实施课程，以及大学本科生和研究生的研究和教育

Microbial cell-surface enzymes (ectoenzymes) are important agents of polymer hydrolysis in aquatic environments and indicators of the state of microbial carbon, nitrogen, or phosphorus nutrition. However, like most other microbial metabolic rate measurements in aquatic environments, ectoenzyme activity measurements have usually been limited to manual assays with discrete water samples. An instrument for continuous underway measurements of microbial enzyme activities using high-sensitivity fluorescent substrates has recently been developed. This instrument has been used aboard ship to map the ectoenzyme activities alkaline phosphatase and leucine aminopeptidase in the surface waters of the Mississippi River plume. With this instrument these enzyme activities can be mapped in much the same way that temperature, salinity, phytoplankton fluorescence, and other parameters are mapped from a research ship while underway. However, this system is limited to horizontal mapping of water samples continuously pumped from the surface and requires frequent operator intervention. This Biocomplexity Instrumentation Development for Environmental Activities (IDEA) project will take the next much larger step. The research and education team includes a microbial ecologist (Dr. J. Ammerman) and a geochemist (Dr. G. Klinkhammer), both of whom have interests and expertise in sensor development, a coastal physical oceanographer and modeler (Dr. R. Chant), and a non-faculty education specialist (Mr. E. Simms). The goal of this project is to develop a remotely operated instrument for measurement of microbial enzyme activities and to deploy it at the Rutgers University Long-term Ecosystem Observatory at 15 meters depth, called LEO-15, for periods of weeks to months. This instrument is called the Multiple Enzyme Analyzer (MEA), and should be capable of the simultaneous measurement of up to four different enzyme activities. A simple mooring with biological, chemical, and physical sensors will be deployed near the enzyme analyzer to help provide a context for the microbial rate measurements (along with additional data from LEO-15). The enzyme and related data will be analyzed with time-series methods, spectral analysis techniques, statistical modeling, and other techniques to gain insight into microbial processes in the coastal zone. A second deployment of the MEA will take place at the new Martha's Vineyard Coastal Observatory (Woods Hole Oceanographic Institution), along with an automated flow cytometer and image analyzer used for plankton cell counting and sorting. This instrument is under development by Dr. R. Olson and colleagues with Biocomplexity IDEA support. This enzyme analyzer will provide continuous real time microbial metabolic rate measurements on time scales that are currently unavailable. Such measurements will add an important biological component to the extensive chemical and physical measurements that can be made at ocean observatories. In addition, education efforts in aquatic microbiology will include training workshops for primary and secondary school teachers followed by lesson development and implementation, as well as university undergraduate and graduate student research and education