https://pages.htgaa.org/2026b/paulina-liceaga/index.htmlAbout me Recent biotech engineering graduate 🌿🩷 interested in learning more about the future of biomanufacturing ! Contact info Homework, Labs, and Projects See the nav bar on the left to look through my work for How To Biomanufacture Almost Anything (Fall 2026)!Hugoenhttps://pages.htgaa.org/2026b/paulina-liceaga/week-01-hw-project-ideas/index.htmlMon, 01 Jan 0001 00:00:00 +0000https://pages.htgaa.org/2026b/paulina-liceaga/week-01-hw-project-ideas/index.htmlProject Idea: Cell-Free Manufacturing of chemotherapeutic Etoposide Precursor (1) What you would like to biomanufacture and why? The goal is to biomanufacture podophyllotoxin, a natural precursor used in the production of the chemotherapeutic drug etoposide, which is used primarily to treat testicular cancer and small-cell lung cancer. Podophyllotoxin is traditionally obtained from mayapple plants, particularly Podophyllum hexandrum (Himalayan mayapple), which contains higher concentrations of podophyllotoxin than the more common American mayapple, Podophyllum peltatum. The reliance on these plants for extraction creates both a supply challenge and a conservation concern, particularly for P. hexandrum, which has experienced significant pressure from harvesting. There has already been work toward producing podophyllotoxin in alternative biological systems, including engineered tobacco plants. However, I am interested in exploring whether a more controllable biomanufacturing platform, such as a microbial chassis or cell-free system, could eventually provide a more flexible and scalable route to production. My aim is to both: contribute to a more reliable supply of an important pharmaceutical precursor, potentially making its downstream production more accessible and less dependent on plant harvesting, while also reducing pressure on the native plant populations from which podophyllotoxin is traditionally obtained.https://pages.htgaa.org/2026b/paulina-liceaga/week-02-hw-dna-design/index.htmlMon, 01 Jan 0001 00:00:00 +0000https://pages.htgaa.org/2026b/paulina-liceaga/week-02-hw-dna-design/index.htmlPart 0: Set up in Kernel The location of my files in Kernel is in the HTBAA26_Genspace repository, file name Paulina Liceaga Part 1: Define your expression target The target molecule is a single enzyme found in the Himalayan Mayapple plant, OMT3 or Pluviatolide O-methyltransferase, involved in the biosynthesis of etoposide, a chemotherapeutic compound of the topoisomerase inhibitor family (PubMed:26359402). The accession code for this protein in UniProt is A0A0N9HMN6 and its genetic sequence is as follows:https://pages.htgaa.org/2026b/paulina-liceaga/week-03-hw-cell-free/index.htmlMon, 01 Jan 0001 00:00:00 +0000https://pages.htgaa.org/2026b/paulina-liceaga/week-03-hw-cell-free/index.htmlA. COMPARE CELL-FREE REACTION COMPOSITIONS A1. Why does the concentration of potassium glutamate and magnesium glutamate differ between compositions? Which is more sensitive? The concentrations differ because each formulation was optimized for its own conditions. The lysate strain and preparation method change how much Mg²⁺ and K⁺ the extract already carries over, and the energy system matters too. Jewett & Swartz (2004) had to lower Mg²⁺ when switching from PEP to pyruvate, because PEP binds Mg²⁺ more tightly (11.3 mM total in Cytomim vs about 23 mM in PANOx-SP). Olsen et al. (2026) likewise re-tuned Mg²⁺ and phosphate for each new lysate batch. The two ions also do different jobs. K⁺ is the main cation that sets ionic strength, so it is needed at roughly 80-362 mM across formulations. Mg²⁺ is a cofactor for ribosomes and polymerases, needed at about 8-10 mM.https://pages.htgaa.org/2026b/paulina-liceaga/final-project/index.htmlMon, 01 Jan 0001 00:00:00 +0000https://pages.htgaa.org/2026b/paulina-liceaga/final-project/index.html