A Self-Assembling Protein Hydrogel Technology for Enzyme Incorporation onto Electrodes in Biofuel Cells

1. A Self-Assembling Protein Hydrogel Technology for Enzyme Incorporation onto Electrodes in Biofuel Cells Chris Skinner El Campo ISD Zhilei Chen, PhD Artie McFerrin Department of Chemical Engineering Texas A&M University

2. Protein Engineering • DNA for the desired protein is identified, and cut out with restriction enzymes. • Plasmid DNA is cut with the same restriction enzymes. • The pieces are put together with an enzyme called ligase to create a new plasmid.

3. Transformation and Purification • Newly engineered plasmid is then put into E. coli through a process called transformation. • Transformed E. coli is then grown and induced to make the newly engineered enzyme. • Once harvested, the proteins must be purified, or isolated • During the engineering of the protein, it was "tagged" to make isolation easier • The solution containing the protein is poured through a filter system

4. Purification (continued) • The protein is eluted from the filter using a buffer that has a higher affinity for the molecule • The collected protein solution undergoes electrophoresis to demonstrate its purity

5. Fuel Cell Construction Mix enzyme with Multi-walled carbon nanotubes (MWNT’s) •Fix enzyme to electrodes •Argarose + MWNT + enzyme

6. STAAR/EOC OBJECTIVES • P.1.A demonstrate safe practices during laboratory and field investigations • P.1.B demonstrate an understanding of the use and conservation of resources and the proper disposal or recycling of materials • P.2.A know the definition of science and understand that it has limitations, as specified in chapter 112.39, subsection (b)(2) of 19 TAC • P.2.B know that scientific hypotheses are tentative and testable statements that must be capable of being supported or not supported by observational evidence. Hypotheses of durable explanatory power which have been tested over a wide variety of conditions are incorporated into theories

7. STAAR/EOC OBJECTIVES (cont.) • P.2.C know that scientific theories are based on natural and physical phenomena and are capable of being tested by multiple independent researchers. Unlike hypotheses, scientific theories are well‐established and highly‐reliable explanations, but may be subject to change as new areas of science and new technologies are developed • P.2.D distinguish between scientific hypotheses and scientific theories • P.2.E design and implement investigative procedures, including making observations, asking well‐defined questions, formulating testable hypotheses, identifying variables, selecting appropriate equipment and technology, and evaluating numerical answers for reasonableness • P.2.H make measurements with accuracy and precision and record data using scientific notation and International System (SI) units

8. STAAR/EOC OBJECTIVES (cont.) • P.2.I identify and quantify causes and effects of uncertainties in measured data • P.2.J organize and evaluate data and make inferences from data, including the use of tables, charts, and graphs • P.2.K communicate valid conclusions supported by the data through various methods such as lab reports, labeled drawings, graphic • organizers, journals, summaries, oral reports, and technology‐based reports • P.2.L express and manipulate relationships among physical variables quantitatively, including the use of graphs, charts, and equations • P.3.A in all fields of science, analyze, evaluate, and critique scientific explanations by using empirical evidence, logical reasoning, and experimental and observational testing, including examining all sides of scientific evidence of those scientific explanations, so as to encourage critical thinking by the student

9. STAAR/EOC OBJECTIVES (cont.) • P.3.B communicate and apply scientific information extracted from various sources such as current events, news reports, published journal articles, and marketing materials • P.3.E research and describe the connections between physics and future careers • P.5.D identify examples of electric and magnetic forces in everyday life • P.5.E characterize materials as conductors or insulators based on their electrical properties • P.5.F design, construct, and calculate in terms of current through, potential difference across, resistance of, and power used by electric circuit elements connected in both series and parallel combinations.

10. Enzymes as an Alternative Fuel Source • Day 1 • Bacterial Reproduction • Guided Activty: Pglo • DNA replication • Genetic Engineering • Snail fuel video Greenlaw, Mackinley, “DOD: Cyborg Snails Make Good Batteries, Online Video Clip, YouTube, March 14, 2012, June 25, 2012, http://www.youtube.com/watch?v=-RVaXjZ8MlQ

11. Enzymes as an Alternative Fuel Source • REDOX Reactions • Reversible reactions • Liberation of electrons • Electrical Circuit • Anode • Cathode • Connecting wires (MWNT) • Pre-test • Pre-lab worksheet

12. Enzymes as an Alternative Fuel Source • Day 2 • Construction of control fuel cell • Glucose oxidase (Anode) • Laccase (Cathode) • Buffer • Construction of working fuel cell • Glucose oxidase (Anode) • Laccase (Cathode) • Buffer • MWNT’s

13. Engineering Design • Day 3 Engineering Design • 1. Asking Questions vrs. Defining Problems2. Developing and Using Models3. Planning & Carrying Out Investigations4. Analyzing and Interpreting Data5. Using Mathematics and Computational Thinking6. Constructing Explanations and Designing Solutions7. Engaging in Argument From Evidence8. Obtaining, Evaluating and Communicating InformationSource: NAS, A Framework for K-12 Science Education

14. Engineering Design

15. Engineering Design • Define the Problem – In many cases, done for you by a client • Brainstorm • Research and Generate Ideas – gather background information on different aspects of the problem • Identify Criteria, Constraints & Performance Specifications – define what the system must do (compare to Design Specs) • Explore Ideas and Invention • Analysis & Selection – at this stage you have structured the problem, and can now apply sophisticated analysis techniques to examine the performance of the design. You have some potentially viable designs (use categories Cost, Safety, Performance, Reliability to Rank each) • Develop Detailed Design • Model or Prototyping and Testing • Test & Evaluate • Refine • Production • Communicate Results

16. Assessment • Presentation

17. Acknowledgements • TAMU E3 program • National Science Foundation • Nuclear Power Institute • Dr. Zhilei Chen • Dr. Dongli Guan • Tila Hidalgo • Dr. Cheryl Page • Matthew Pariyothorn