Biofuel CellsPurifying engineered enzymes

to generate electricity

Tila HidalgoAlief ISDHastings High School

Zhilei Chen, PhDArtie McFerrin Department

Department of Chemical Engineering

Biofuel Cells

Biofuel cells have great potential for real world application.

The Research Objective

•Current Issues•Not a lot of power generated•Poor connections •Loss of enzyme from the fuel cell

To improve power and

longevity of the fuel

cell.

My Classroom Project:

Bacterial Transformation and Protein Purification.◦A project for my AP Biology students.

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College Board Objectives Addressed

In addition to lab skills based around the inquiry model of investigation that is part of the AP Biology curriculum the following college board objectives will be covered: ◦ Essential knowledge 4.A.1: The subcomponents of biological molecules and their sequence determine

the properties of that molecule.◦ Essential knowledge 2.B.2: Growth and dynamic homeostasis are maintained by the constant

movement of molecules across membranes.◦ Essential knowledge 3.A.1: DNA, and in some cases RNA, is the primary source of heritable information. ◦ Essential knowledge 3.C.1: Changes in genotype can result in changes in phenotype.

Phenotypes are determined through protein activities. To foster student understanding of this concept, instructors can choose an illustrative example such as:

◦ Enzymatic reactions ◦ Transport by proteins ◦ Synthesis ◦ Degradation

Genetic engineering techniques can manipulate the heritable information of DNA and, in special cases, RNA.

To foster student understanding of this concept, instructors can choose an illustrative example such as: ◦ Electrophoresis ◦ Plasmid-based transformation ◦ Restriction enzyme analysis of DNA ◦ Polymerase Chain Reaction (PCR)

Protein Purification Objective:

Students in groups will research, develop, implement and present

a protocol and results of a protein purification using protein

from transformed bacteria.

The project will culminate in group presentation where the process will be evaluated for a percent yield, purity, and cost

effectiveness.

Day 1

Pre-test

Teacher lecture introducing the project

and presenting my summer E3 experience and engineering as a

field of study.

Homework assignment: Identify units we have

studied throughout the year that connect to my summer

research. What ways are your life better because of what

engineers do?

Day 2-3

Students will perform the bacterial transformation. I plan to use the Biorad pGlo transformation.

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Day 4-6

Research days. I want students to do a

literature review of protein purification. They will use this

to write a presentation answering the questions:

•Why is protein purification important? •What are ways that it is done? •What are limitations to these protocols? •What properties of the pGlo protein could be used to purify it? •What is your proposed protocol to purify pGlo?•The protocol will include a cost projection, specific steps to follow to purify the protein, and a method to calculate a percent yield.

Day 7-8- “Lab Meeting”

Students will present their findings, and after will develop a final protocol to purify the pGlo.

•The collective answer should be that the cells need to by lysed, and using the hydrophobic properties of the pGlo protein it can be purified on a column that address that property. •Methods of cell lysis can vary from group to group as well as specific reagents to wash and elute the protein, but they should all have common chemical properties.

Day 9-11- Protein Purification.www.bio-rad.com/webroot/web/software/.../pGLO_GFP_031208.pp

Day 9-11

Students will perform their experiments.

They will then analyze their results using electrophoresis, and calculate a percent yield.

www.bio-rad.com/webroot/web/software/.../pGLO_GFP_031208.pp

Day 12-13

Final Presentations

PurityQuantityFinal CostPercent yield Error analysis

Post Test

Rubric Rough Draft

Requirement Points

Methods Clearly Stated 20

Quantative and Qualative Data was included. (picture of the gel and yield)

20

Cost of protocol/g of product 10

Percent yield calculated 10

Error analysis (where can your project be improved and why? What would your next step be?)

20

Citation, organization, general presentation

10

Acknowledgements

TAMU E3 program National Science Foundation Nuclear Power Institute Dr. Chen and Dr. Guan Chris Skinner