Team:Baltimore BioCrew/Gold

Human Practices: Gold

Implementing our Investigated Project

Our team has identified the potential collateral damage of having an exponentially growing population of genetically modified E. coli. We realized that we did want our colonies to widely grow in order to achieve their maximum plastic degradation threshold, but are unsure of how their introduction to an aquatic ecosystem might affect its environment, aquatic organisms, and the people that live off these lifeforms. In an attempt to evade such detrimental repercussions, as a team, we’ve decided to concoct a kill switch of sorts- one that acts upon programmed changes in temperature so that it can accommodate the seasonal changes of Baltimore.

During the summer, when tourism is at an all time high, is when the E. coli will be most active. This is to combat the increases in pollution that follow in an increase in human activity. During the winter, when people are mainly indoors is when it can be inferred that pollution in the Inner Harbor will be at its annual low because of the deficit of human activity. Since there is a relatively low need for our plastic degrading organisms during those winter times, a kill switch can be activated to control the population. By having a more wholesome controlling of the E. coli cells we, as a community, can better monitor them and prevent them from seriously harming our harbor if plans go awry.

Ex: We implement our E. coli cells into the Inner Harbor 1.5 months before winter conditions that would cause our E. coli to self-destruct. We study the aquatic conditions to observe the changes that our G.M.O. may be responsible for.

Say, we notice that there our E. coli cells are making the local crab population sick and its population has decreased by 10%- we now know that our G.M.O’s are incompatible with the Harbor so we won’t reintroduce them after the winter. If we do decide to, they will be further modified to try to solve the crab problem.

A Functional Proof of Concept for the Baltimore BioCrew's BioBrick Device

Based upon our rigorous research efforts, our proof of concept is based upon the meticulous construction of our BioBricks, one of which being the BioBrick for the lipase gene (BBa_K2187001). This BioBrick serves to replace MHETASE, a distinguishing enzyme which gives Ideonella sakaiensis it’s renowned plastic-degrading properties. The other enzyme that fulfills and progresses this property is called PETASE, which we substituted with the Chlorogenate Esterase gene (BBa_K2187001).

With the 2 known plastic-degrading enzymes of Ideonella sakaiensis being substituted with our BioBricks, which serve identical functions, our team believes that this is enough to prove that our genetic modification will work.

We know that Lipase and Chlorogenate Esterase will serve to replace the 2 genes that are unable to physically acquire, so with this is mind, we used the predetermined Prefix/Suffix, Promoter, RBS, and Terminator to build our BioBricks off of. Once we had the forward and reverse primers included into our gene sequence, we had the complete set of nucleotides with all fulfilled requirements; with this, we used these BioBricks and transformed them into E. coli cells through a plasmid. The antibiotic resistance segregated our transformed cells, and with this, our E. coli cells should be capable of expressing the said enzymes that instigate the decomposition of PET and MHET plastics.

From transforming our E. coli cells to express our MHETASE and PETASE substitutes, we are hopeful that our results are in accordance with our hypothesis.

The BioCrew's Visionary Aspiration to Accommodate the "Real World"

The issue that we, the Baltimore BioCrew, investigated was the significant pollution of plastic in our community Inner Harbor. Using synthetic biology, we wanted to realistically rid such waste through the implementation of a genetically modified E.coli colony that degrades PET plastics using two enzymes from Ideonella Sakaiensis, a newly discovered Japanese bacterium.

We realize that under real-world conditions, placing genetically engineered E. coli into a reputable body of water valued by the large population of Maryland is not a very flattering proposal.

Taking the negative stigma placed around E. coli into consideration, along with the potential risk of marine ecosystems' safety and the possibility of this bacteria evolving, the Baltimore BioCrew wanted to establish a project design that would make our proposal seem less like a stunt and more like a scientific solution.

To demonstrate our whole system of using E. coli K12 as our organism “model”, the team deemed that it was advantageous in the sense that it was stable, inexpensive, quick to procreate.

Since we are currently running test trials on our E. coli, we are confident that if it behaves as we hypothesize it to, the many potential uses for our G.M.O. would be systematic. We could install vacuums onto sailboats that collect plastics to further degrade them in some sort of hypothetical compartment; we could form a new type of disposal bin, much like recycling, to collect the plastic and leave it in the E.coli solution to disassemble itself; we could even have large industrial plastic collection tanks that accumulate plastics from junk yards and dissolve them there.

With this being said, in addition to the incorporation of the hydrolytic enzymes within our bacteria, the bacteria could be able to use the very PET plastics as a carbon source, allowing it to procreate with very minimal measures.

Rather than perpetuating the painting of a picture where E. coli derives a detrimental connotation associated with it, further ideas involve giving it the ability to convert the carbon it consumes into energy that could power technology ranging from light bulbs to hydroelectric wheels.

Our ideas may seem ambitious, but E. coli's ability to reproduce quickly could make our genetically engineered bacteria much more than a large energy supply- it could be the global solution to a problem we’ve been facing for decades.