Abstract:Active‐learning strategies such as undergraduate research courses and course‐based undergraduate research experiences (CUREs), which engage students in the practical experiments, have been reported as efficient ways for effective teaching and training of graduating students. Recently, many practical examples have been published in various fields to develop critical skills needed by undergraduate students for graduate school or the workforce; however, very few examples have been published for bioanalytical topi… Show more
“…20 V) and the precursor ➔ product transition (254➔ 156) are then used for the MRM method. There are numerous examples of the use of such MRM-based analyses for organic compounds in various matrices [26, 27]. Fig.…”
BackgroundThe presence of a wide range of bioactive organic pollutants in wastewater and municipal water sources is raising concerns about their potential effects on humans. Not surprisingly, various approaches are being explored that can efficiently degrade these persistent organic pollutants. Use of peroxidases has recently been recognized as a novel remediation approach that may have potential advantages over conventional degradation techniques. However, testing the abilities of different peroxidases to degrade diverse emerging pollutants is tedious and cumbersome.ResultsIn the present study, we present a rapid and robust approach to easily test the degradability of 21 different emerging pollutants by five different peroxidases (soybean peroxidase, chloroperoxidase, lactoperoxidase, manganese peroxidase, and horseradish peroxidase) using an LC-MSMS approach. Furthermore, this approach was also used to examine the role of a redox mediator in these enzymatic degradation assays. Our results show that some of the organic pollutants can be easily degraded by all five of the peroxidases tested, whereas others are only degraded by a specific peroxidase (or when a redox mediator was present) and there are some that are completely resistant to degradation by any of the peroxidases tested (even in the presence of a redox mediator). The degradation of furosemide and trimethoprim by soybean peroxidase and chloroperoxidase, respectively, was investigated in detail by examining the transformation products generated during their degradation. Some of the products generated during enzymatic breakdown of these pollutants have been previously reported by others, however, we report many new transformation products.ConclusionsLC-MSMS approaches, like the one described here, can be used to rapidly evaluate the potential of different peroxidases (and redox requirements) to be used as bioremediation agents. Our preliminary result shows peroxidases hold tremendous potential for being used in a final wastewater treatment step.
“…20 V) and the precursor ➔ product transition (254➔ 156) are then used for the MRM method. There are numerous examples of the use of such MRM-based analyses for organic compounds in various matrices [26, 27]. Fig.…”
BackgroundThe presence of a wide range of bioactive organic pollutants in wastewater and municipal water sources is raising concerns about their potential effects on humans. Not surprisingly, various approaches are being explored that can efficiently degrade these persistent organic pollutants. Use of peroxidases has recently been recognized as a novel remediation approach that may have potential advantages over conventional degradation techniques. However, testing the abilities of different peroxidases to degrade diverse emerging pollutants is tedious and cumbersome.ResultsIn the present study, we present a rapid and robust approach to easily test the degradability of 21 different emerging pollutants by five different peroxidases (soybean peroxidase, chloroperoxidase, lactoperoxidase, manganese peroxidase, and horseradish peroxidase) using an LC-MSMS approach. Furthermore, this approach was also used to examine the role of a redox mediator in these enzymatic degradation assays. Our results show that some of the organic pollutants can be easily degraded by all five of the peroxidases tested, whereas others are only degraded by a specific peroxidase (or when a redox mediator was present) and there are some that are completely resistant to degradation by any of the peroxidases tested (even in the presence of a redox mediator). The degradation of furosemide and trimethoprim by soybean peroxidase and chloroperoxidase, respectively, was investigated in detail by examining the transformation products generated during their degradation. Some of the products generated during enzymatic breakdown of these pollutants have been previously reported by others, however, we report many new transformation products.ConclusionsLC-MSMS approaches, like the one described here, can be used to rapidly evaluate the potential of different peroxidases (and redox requirements) to be used as bioremediation agents. Our preliminary result shows peroxidases hold tremendous potential for being used in a final wastewater treatment step.
“…An integral part of most science and engineering‐based undergraduate curricula have some sort of an “Independent Study” component, where third‐ or fourth‐year students work with faculty members on a mini‐research project 1,2 . These independent study research projects are primarily lab based and require students to submit a detailed written report and/or oral presentation.…”
Section: Methodsmentioning
confidence: 99%
“…critical thinking, enzyme thermostability, senior research project, undergraduate research An integral part of most science and engineering-based undergraduate curricula have some sort of an "Independent Study" component, where third-or fourth-year students work with faculty members on a mini-research project. 1,2 These independent study research projects are primarily lab based and require students to submit a detailed written report and/or oral presentation. In some cases, these result-oriented projects can lead to eventual student publications.…”
The recent lockdown of laboratories in the COVID-19 era has negatively impacted many lab-based courses for undergraduate students; however, research project classes have been hardest hit. Unlike the lab-based lecture courses, where many of the lab instructors have resorted to virtual experiments and videos to substitute for actual lab experiments, senior year independent research courses, which are supposed to involve real lab-based research, are struggling to find an appropriate solution. A possible solution to cases where
“…While many published research-based curricula focus on general chemistry, organic chemistry, or biochemistry, comparatively few have been reported to enhance bioanalytical chemistry laboratories . Even fewer have extended to the analysis of proteins and the application of biological mass spectrometry, despite its increasingly prominent role in analytical chemistry .…”
Section: Introductionmentioning
confidence: 99%
“…8 While many published research-based curricula focus on general chemistry, organic chemistry, or biochemistry, comparatively few have been reported to enhance bioanalytical chemistry laboratories. 10 Even fewer have extended to the analysis of proteins and the application of biological mass spectrometry, despite its increasingly prominent role in analytical chemistry. 11 This is a significant oversight, as bioanalytical chemistry and biological mass spectrometry are both consistently used in the biomedical and pharmaceutical industries and are broadly applicable to students pursuing graduate school, professional school, or industry-related occupations following their undergraduate degree.…”
Course-based undergraduate research experiences (CUREs) integrate authentic research into undergraduate chemistry laboratories, introducing students to research while simultaneously reinforcing fundamental concepts. Despite their ubiquitous nature in bioanalytical research, few CUREs have been published applying the fundamental techniques of separations, spectroscopy, quantification, and mass spectrometry. To engage students in learning these increasingly essential bioanalytical techniques, we designed and implemented a semester-long project-based course centered around the purification, quantification, and identification of heterologously expressed proteins in five succinct and adaptable modules. Instructors can use these modules to form the foundation of a CURE specific to their research interests. The extensive commercial availability of plasmids for transformation combined with the modular approach to laboratory experiments enables convenient customization to accommodate diverse research goals. Instructors can tailor the modules to meet the curricular requirements and instrumentation capabilities of their institutions and can easily extend the research goals to incorporate more specialized analytical techniques, as needed. Through the implementation of the five modules, students apply the fundaments of acid−base chemistry, statistics, quantification strategies, spectrophotometry, separations, and mass spectrometry, thus covering the material required in most undergraduate introductory analytical courses. Instructors can then use these modules as a backbone to support student-led discovery-based investigations for the remainder of the course. Students demonstrate their understanding through the completion of a comprehensive, publication-style laboratory report as well as a poster presentation at a university-wide undergraduate research symposium. Since first offering this course in 2016, student evaluations have been exceedingly positive, with over 75% of students indicating that the course both increased their scientific skills as well as their confidence in their ability to succeed in further science courses. Furthermore, 22% of students reported they were "much more" or "extremely more" likely to enroll in a Ph.D. program in science, math, or engineering following the courses, emphasizing the impact that project-based laboratories can have on undergraduate chemists' career trajectories.
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