Cleavage and Purification of Tagless L-DOPA 2,3 dioxygenase from Streptomyces hygroscopicus jingganensis (JING) for crystallization

Or-El Ankori ’25

Faculty Mentor: Dr. Colabroy

What is the broader impact of your summer research? That is, why should the world care about your work?

The Colabroy lab is interested in an enzyme class called extradiol dioxgenases which cleave catecholic rings, a chemical structure found abundantly in nature within a structural part of trees called lignin. This cleaving reaction then generates a product that re-cyclizes into a compound that can be used as a precursor for antibiotics, feedstocks, and tumor agents. The work out lab does centers around probing this reaction through the usage of enzymes from different bacterial species and modified substrates (made by our collaborators) so that we can ultimately use this knowledge to understand the kinetic activity of this reaction and channel it into productive upcycling. Waiting for grade

The “little why” of my project centered around an end goal of crystallizing pShjingDDO, the L-DOPA 2,3-Dioxygenase from Streptomyces hygroscopicus jingganensis. Since our lab has a crystallographer collaborator, my role was to optimize a cheaper way to cleave and purify the histidine tag off of our enzyme of interest so that it remains soluble in solution for crystallization. 

What has summer research taught you about yourself and the field that you researched and/or studied?

This summer, I learned more about what it feels like to work outside of a college environment, in the closest experience I’ve yet had to a 9-5. It was interesting for me to adjust to a new work-life balance in which I had more free time (unlike school, which is all-encompassing) and imagine that this is what life could look like in the future. Additionally, it was both awesome and trying to work (and live with) the same people throughout the summer; not everything was breezy all the time, but I was glad to have had the opportunity to work on my communication skills and we are all still very good friends! I loved working in lab this summer because while I got to sample biochemical techniques in past courses (during lab), this experience allowed me to be immersed in a wet-lab experience. My past research experience has not been linear, and I’m excited to continue my summer work into the semester!

What advice would you give to students considering a future research opportunity similar to the one you participated in?

When you live alone (especially over the summer, when the dining hall is not open and no one is telling you what to do) there is a lot of self regulation to do! It’s a good opportunity to work on your self-discipline and see what adult life feels like a little more.