@article{wapshott-stehli_grunden_2021, title={In situ H2O2 generation methods in the context of enzyme biocatalysis}, volume={145}, ISSN={["1879-0909"]}, DOI={10.1016/j.enzmictec.2021.109744}, abstractNote={Hydrogen peroxide is a versatile oxidant that has use in medical and biotechnology industries. Many enzymes require this oxidant as a reaction mediator in order to undergo their oxygenation chemistries. While there is a reliable method for generating hydrogen peroxide via an anthraquinone cycle, there are several advantages for generating hydrogen in situ. As highlighted in this review, this is particularly beneficial in the case of biocatalysts that require hydrogen peroxide as a reaction mediator because the exogenous addition of hydrogen peroxide can damage their reactive heme centers and render them inactive. In addition, generation of hydrogen peroxide in situ does not dilute the reaction mixture and cause solution parameters to change. The environment would also benefit from a hydrogen peroxide synthesis cycle that does not rely on nonrenewable chemicals obtained from fossil fuels. Generation of hydrogen peroxide in situ for biocatalysis using enzymes, bioelectrocatalyis, photocatalysis, and cold temperature plasmas are addressed. Particular emphasis is given to reaction processes that support high total turnover numbers (TTNs) of the hydrogen peroxide-requiring enzymes. Discussion of innovations in the use of hydrogen peroxide-producing enzyme cascades for antimicrobial activity, wastewater effluent treatment, and biosensors are also included.}, journal={ENZYME AND MICROBIAL TECHNOLOGY}, author={Wapshott-Stehli, Hannah L. and Grunden, Amy M.}, year={2021}, month={Apr} }
 @article{somalinga_klemmer_arun_mathews_wapshott_grunden_2018, title={Cloning, Over-Expression, and Purification of Carbonic Anhydrase from an Extremophilic Bacterium: An Introduction to Advanced Molecular Biology}, volume={80}, ISSN={["1938-4211"]}, url={http://dx.doi.org/10.1525/abt.2018.80.1.29}, DOI={10.1525/abt.2018.80.1.29}, abstractNote={The photosynthetic bioreactor research program is a training platform appropriate for introducing advanced molecular biology techniques to undergraduate students and advanced high school biology students. For this advanced molecular biology training exercise, the enzyme carbonic anhydrase was cloned, over-expressed, purified, and functionally characterized. Carbonic anhydrases are industrially important enzymes with potential use in carbon sequestration and biofuel production. Alpha and beta carbonic anhydrases from Photobacterium profundum, a psychrophilic, halotolerant bacterium, were characterized in this study. Carbonic anhydrases that can withstand high salinity and are active at low temperatures can be transformed into oleaginous marine microalgae to enhance biofuel production. Our research program started with a three-day boot camp with lectures in relevant topics of molecular biology, microbiology, and research methods. After the boot camp, the lab phase of the project involved training students to perform polymerase chain reaction, DNA gel electrophoresis, DNA ligation, and bacterial transformation. In the final phase of the project, students were trained in recombinant protein over-expression and protein purification techniques. Here we report successful cloning and over-expression by high school students of two novel carbonic anhydrases from a psychrohalophile with application in biofuel production.}, number={1}, journal={AMERICAN BIOLOGY TEACHER}, publisher={University of California Press}, author={Somalinga, Vijayakumar and Klemmer, Hannah and Arun, Ashikha and Mathews, Stephanie and Wapshott, Hannah and Grunden, Amy M.}, year={2018}, month={Jan}, pages={29–34} }