Integrated human practice 

Switching our project focus 

       We initially wanted to mitigate climate change by creating a co-culture between an anaerobic, acetogenic bacteria E. limosum and E.coli. Upon discussing our project with Dr. Kuk- Jeong Chin, an anaerobic bacteria specialist at Georgia State University, and the Toulouse INSA-UPS iGEM team, we realized the difficulties in growing anaerobic bacteria, especially in a Biosafety level 1 lab like ours. Therefore, we explored other alternatives to how to mitigate the effects of climate change. 

Social concern 

       Combustion of fossil fuels has contributed heavily to current global warming, leaving multiple vulnerable populations susceptible to diseases, natural disasters due to weather changes, and more that is directly caused by global warming. Algal biofuels have been explored as an alternative to foss il fuels, but algal biofuels cost more to produce in comparison with fuels made from other sources (Rafa et al, 2021). The cheaper cost of fossil fuels is one of various reasons why it is sold more comp ared to algal biofuels. Researchers have attributed the high costs of algal biofuels to production met hods. According to research, the equipment to grow the algae as well as other factors contribute to the high cost of algal biofuel production (Rafa et al, 2021).  Scientists are aware and understand algal biofuels are more environmentally friendly compared to fossil fuels, but it was also important to fin d strategies to cut down the cost. We particularly learned this upon making a connection with  a co ntact at the National Renewable Energy Laboratory, a lab funded by the Department of Energy that researches more sustainable methods for generating energy. Amongst the many research methods this company focuses on, we were interested in their strategies to cut down on costs on algal biof uel development.

       However, we thought it was critical to determine whether or not the people that do not work in the algal biofuel industry nor are on our  IGEM team also have an interest in mitigating climate change through alternative fuels. To do this, we surveyed the youth community ranging from age 16 to 20 and a few adults between the ages of 26 and 30. We decided to distribute surveys mainly to young people since they are instrumental in shaping the future and will have to endure the severe effects of climate change. We received feedback specifically from high school students as well as additional college students. This survey is ongoing but the individuals that have participated in the survey so far were willing to switch from fossil fuels to algal biofuels if it were to mitigate climate change. A  few of this population (ages 16-17)  expressed how they did not know much about biofuels. We decided to complete an educational outreach event to clarify questions participants had about biofuels, especially to students who didn’t know much about biofuels, and included interactive activities as described under Educational Outreach. After seeing the students express how the diagrams and other strategies we used further helped them understand biofuels and climate change, we plan to complete more educational outreach sessions to answer questions students may have about biofuels, especially for students that were survey participants but did not attend the last educational outreach event we hosted.

       Most participants in the survey expressed that they would be willing to pay more for biofuels. However, the range of how much more  participants were willing to buy the algal biofuel was limited to between $1 and $20; this is based on only participants that expressed an exact price. Then, a few participants either did not state how much higher that they wanted to pay for algal biofuels or indicated they wanted to pay lower for biofuels due to concerns with safety. Although the price of algal biofuels is expected to be higher than fossil fuels, this survey shows that there is a limit to how much higher people are willing to pay for algal biofuels, providing more evidence how important it is to make algal biofuels  more cost-effective to be sold on a large scale.

First Set of Survey Results

      1. A pre-survey was distributed before the educational outreach event (attendees of the educational outreach events). In total, 5 people filled out this survey.

      2. A post-survey was distributed after the educational outreach event to high school students (attendees of the educational outreach events). 2 out of the 5 people that filled out the pre-survey filled out the post-survey.

Second Set of Survey Results

      These results are compiled from students attending Georgia State University that did not attend the educational outreach event but still wanted to participate in the survey. Four parti cipants were given this survey and also were able to refer to the slides presented at the educational outre ach event while completing the survey.

Connecting with scientists

      Scientists have confirmed that E.coli from wastewater stimulated algae growth (Higgins et al, 2014). Upon discovering this research, we thought of an idea to create a co-culture with both E.coli and Chlorella minutissima. The plan would be to genetically modify E.coli with the IAM pathway for indole-3-acetic acid (IAA) production, so the IAA hormone can stimulate the growth of microalgae. Then, the next step would be to genetically modify the microalgae with genes encoding for enzymes that enhance the production of lipids. Lipids extracted from microalgae are then manufactured into biofuels. Our goal is that these modifications and the co-culture will result in a much more sustainable and cost-effective approach for algal biofuel development.

Dr. Gilbert 

      To further  investigate how to create a co-culture with both E.coli and microalgae, we sch eduled a meeting with Eric Knoshaug, who is a scientist at the National Laboratory for Renewable energy (NREL). His research is focused on converting algal biomass to biofuels and chemicals of interest. His research entails various methods including genetic engineering and lipid extraction. At NREL, they have partners that take the algae and grow them outdoors in hopes to try to turn the project into a large-scale process. 

      Key points that Eric Knoshaug highlighted are listed below. We decided it would be great to integrate methods from this list that work well in our current lab space to further improve the research project design.

      1. Place the microalgae in flasks in a lighted incubator since the driving force of lipid production for microalgae is based on how much light the microalgae can capture. However, a test tube as opposed to a flask is much more ideal.

      2. The microalgae need to remain mixing for maximum growth since the algae can settle, causing the cells on top to get all the light as opposed to the cells on the bottom do not get all the light. One method to ensure mixing is occurring is placing the flasks containing the algae on a shaking platform. Another consideration to ensure the algae remains mixing is connecting an aquarium pump to the flasks, so carbon dioxide is brought to the flasks but also creates mixing since it keeps the algae floating.

      3. Use an algae growth media.

      4. The first step is to grow one tube of the microalgae in the right concentration of antibiotic to test how antibiotics will affect the microalgal growth.

      5. The next experiment should be growing microalgae to see how fast the microalgae grow without the co-culture. The reason for this is to establish the boundaries of what optical densities to grow the microalgae at.

      a. Start growing the algal cultures at an optical density at 0.1 and see how fast the microalgae gets to an optical density of 1 or an upper density where self-shading occurs, creating growth curves. To ensure self-shading doesn’t occur, the research team could come in daily to take outhalf the culture every morning to keep the algal culture dilute.

      b. Checking OD readings every 2 or 3 hours would create an ideal growth curve.

      6. Typically, the bacteria should be added to the space occupied with the microalgae when the microalgae are at an optical density at 0.3 or 0.4 or we can stick the E.coli right when we inoculate our tubes.

      7. The negative controls we can consider would include the algae media, algae, and no E.coli

      8. The positive control could include algae media, algae, and E.coli It would be important to maintain an OD of 0.1 for algae and an OD of 0.1 for E.coli after the co-culture is created. 

      When asked if our research project would be a more sustainable alternative to market to fuel industries, Dr. Knoshaug stated, “ We are actually working with plant hormones ourselves to help the microalgae growth, so you guys are on the right track; if the co-culture you have chosen proves to be of value, absolutely!”. 

Eric Knoshaug 
Georgia state igem team meeting with Eric Knoshaug 

Future Direction

      Along with surveying community members to ensure we meet community needs through our current research design, our aim for continuing human practice is to continue meeting with professionals to discuss how to improve the efficiency of the research project in hopes of finding a way to make algae biofuels more cost-effective and sustainable for worldwide commercial success.