About This Project
We have designed a novel protein scaffold onto which we can attach the enzymes from any pathway, creating an assembly line of chemical reactions that lead to a reaction rate increase. We are engineering this system to produce the plant hormone inodole-3-acetic acid (IAA) as a proof of concept. Once we successfully produce IAA at higher rates than free enzymes in solution, we will use our highly modular scaffold with other enzymes from a range of pathways significant to research and industry
Ask the ScientistsJoin The Discussion
What is the context of this research?
Enzymes are biological catalysts which speed up chemical reactions, and are often part of sequential pathways. The speed of these chemical reactions is essential to the viability and efﬁciency of processes across many industries including pharmaceuticals, waste management and food production. However, many of these reactions are slowed down by signiﬁcant distances between enzyme catalysts in solution, which can then also lead to unwanted reactions involving intermediate substrates in multi-step reactions.
What is the significance of this project?
We are aiming to speed up enzymatic reactions so that we can increase levels of productivity. This research may have real world implications because speeding up biochemical reactions could lower the cost and time of production for many everyday items, such as antibiotics, laundry detergent, and fermented food and beverages.
Furthermore, our research may lead to the production of a scaffold which could act as an important research tool, due to the modularity of our protein scaffold. This is because any enzyme with a Spy/Snoop tag will covalently bond to the corresponding catcher on our scaffold.
Additionally, our protein scaffold has the potential to be used in bioremediation to degrade environmental pollutants such as plastics and petrochemicals, making them non-toxic.
What are the goals of the project?
We have decided to focus on creating the plant hormone indole-3-acetic acid as a proof of concept for our project. We hypothesize that when using the self-assembling prefolding protein scaffold to create indole-3-acetic acid, we can significantly speed up the rate of reaction. If the data supports our hypothesis this will be a proof of concept of this protein scaffold increasing rate of reaction. Theoretically then we could use this system to increase the speed of many other enzymatic reactions.
These reagents and consumables are of integral importance to our project, particularly because much of the funding is required to carry out confirmation testing. Funding our project ensures research into our system can continue.
Genetic sequencing with the Ramaciotti Centre for Genomics is used to confirm the plasmids being transformed into our E.coli cells.Gel electrophoresis reagents are essential to the running of SDS-Page gels and Western Blots to confirm the proteins are being expressed by our E.coli cells. This includes MES buffer, GelRed, agarose, DNA ladder, loading dye, and GelCode Blue. Once these gels confirm the creation of these proteins, protein purification reagents such as SCX and SAX resins are used to separate and purify the proteins required for the scaffold from the other cellular components. Laboratory consumables refers to pipette tips, petri dishes, and other disposable equipment required for the project.
Achieving these research milestones is essential to our project. The proof of concept is to be expected to be completed between midway and the end of September. The validation of our scaffold will be an iterative process throughout the project with experiments such as fluorescent protein markers to confirm the orthogonal nature of our Spy/Snoop tags and catchers. Time has been left before the iGEM wiki freeze to account for delays in progress.
Aug 24, 2018
Aug 26, 2018
Grow up DH5a and T7 Express E. coli and make competent
Aug 30, 2018
Protein expression alphaPFD-s and betaPFD-s + Assembly of Scaffold
Aug 30, 2018
Protein expression, purification of IaaH and IaaM
Aug 30, 2018
Protein expression, fluorescent proteins
Meet the Team
Assemblase is an interdisciplinary team, made up of eleven undergraduate students with backgrounds in the medical sciences, cell and molecular biology, engineering, education, and law. They look forward to being able to be experimentally creative within the 'synthetic biology space', particularly in the context of an exciting student-led research opportunity. The team are all enthusiastic about scientiﬁc research and excited about our project!
Nick is currently an undergraduate at UNSW studying Science, majoring in Genetics in his final 8 months of study. He loves science fiction and the hope it brings people which meets his passion for real science believing bioinformatics and synthetic biology are the future. He aspires to help people through scientific progress and to set the wheels in motion for social change through his career.
Linda is currently in her third year of studying a Bachelor’s degree in Science and Education, majoring in Biotechnology. She is interested in how bacteria are able to communicate with one another through the magic of quorum sensing, which is an area within science that she is hoping to explore more of in the future. Apart from science, eating and napping, Linda also really loves cats!
Megan is currently three years into an Advanced Science/Law degree at the University of New South Wales, majoring in Cell and Molecular Biology. She ended up doing Science because she wants to change the way that science is regulated, and spend more time involving researchers in government policy which directly affects them.
Emily is a third year Science/Law student majoring in Microbiology at the University of New South Wales. Growing up in a small coastal town inspired her interest in marine microbiology and the good wife. Synthetic biology's potential in harnessing complex biological mechanisms from nature to design unique systems is something she hopes to pursue further.
Bec is passionate about microbes, robots and wine. Although many view this as strange, some see the broad potential for innovation that comes with the intersection of these interests. From synthetic biology and biohacking to smart homes and viticulture, she looks forward to applying the skills from her Advanced Science and Engineering dual degree to develop the new technologies of tomorrow.
Tobias is majoring in molecular biology and neurobiology. He is interested in further exploring the molecular aspects of the function of the brain to help find understand the basis of neurodegenerative diseases. He hopes, in the future, to see synthetic biology play a key role in solving problems of the brain.
Alex is a 3rd year Advanced Science student majoring in Biotechnology and Genetics. He is most passionate about technologies that aim to benefit the planet. Currently he is learning how to integrate computer science, maths, and biology, to create informative models of complex systems. In future he would like to explore the growing field of mushroom biotechnology!
Tyler is in his third year of Advanced Science, studying Biotechnology and Genetics. Inspired by the likes of Craig Venter, he wants to create life forms and biological robots to further society and bring us into the future.
Brian is in his 4th year of his Advanced Science/Arts degree, studying Microbiology and Music. He hopes to help expand the synthetic biologist's toolkit and advance our understanding of the world that we live in. He is fascinated by synthetic biology as it is driven by innovation - who knows where it will take us next?
Patricia is a third year Advanced Science student, majoring in Genetics and Biotechnology. Patricia has a keen interest in computer science, and would like to be able to combine her love for science and computing to help solve biological problems. Outside of science, she is passionate about travel, dogs and entrepreneurship.
Ahmad Zeeshan siddiqui
Zeeshan is a final year Molecular and Cell Biology student at UNSW. He is excited by the potential of recombinant DNA technologies as well as believing that bioinformatics is the future of medicine. In his spare time he enjoys football, playing the guitar and getting involved with the biotech society on campus.
Nothing posted yet.
To achieve the protein scaffold, we using the self-assembling prefoldin protein due to it's stability in solutions and ease of use. We are utilizing the Spy and Snoop Tag/Catcher systems to covalently attach the enzymes to our scaffold.
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