Week 1 HW: Final Project Ideas

1. What would you like to biomanufacture and why? Micronutrients package: iron compounds (heme/ferrous iron), vitamin A, iodine (iodide), zinc compounds (zinc glycinate, zinc gluconate, zinc citrate, etc.), and folate.

Micronutrient deficiency is a major health challenge in Africa that disproportionately affects women of reproductive age, infants, and children under five. See https://link.springer.com/article/10.1186/s43054-026-00603-7

Over half of all children in Eastern and Southern Africa are deficient in essential vitamins and minerals (UNICEF). 321+ million people are deficient in Iodine. ⅔+ of women of reproductive age in Africa are micronutrient deficient. A healthy diet is also too expensive to afford, even for non-poor people (2023 UN Economic Commission for Africa, see https://openknowledge.fao.org/server/api/core/bitstreams/052b1978-0f1f-4db9-8a1d-49bd5b4a19ce/content)

2. What living system would you use?

Survey of current literature on biomanufacturing of compounds in my package:

1. Ferrous iron compound: E.coli can reduce insoluble Fe3+ into soluble Fe2+ https://pubmed.ncbi.nlm.nih.gov/18394146/ Potential oxidative/metabolic stress issue
2. Vitamin A: Oxidative cleavage of plant-derived provitamin A carotenoids by the enzyme beta-carotene 15,15’-dioxygenase (CMO) E.coli and yeast have been engineered to convert simple sugars into vitamin A (https://www.sciencedirect.com/science/article/pii/S2405805X24001157)
3. Iodide: dehalogenating bacteria break down anthropogenic organic iodine to release free iodide: https://pubs.acs.org/esthag/article-abstract/57/6/2625/1726040/Microbial-Contributions-to-Iodide-Enrichment-in?redirectedFrom=fulltext E.coli included but a closer survey needed (because I just skimmed this)
4. Zinc gluconate: Glucose oxidation – hydrolysis – Zinc chelation (Gluconobacter accumulates gluconic acid in growth medium; Pseudomonas, Bacillus, and Gluconacetobacter secrete gluconic acid to dissolve mineralized zinc in soil)
5. Folate: synthesized de novo in bacteria (including E. coli, I think) https://pdb101.rcsb.org/global-health/antimicrobial-resistance/drugs/antibiotics/folate-synthesis/folate-synthesis

Likely chassis: E.coli (for zinc gluconate, I guess chemical synthesis and supplements also work if we don’t want to bother with Bacillus and co-culture, which is sure to affect the cultivation conditions and may both drag down efficiency and increase cost) – potentially do directed evolution to select strains more robust under metabolic/oxidative stress

3. How would you use automation?

Automated cultivation conditions control: depends on how I want the end product to be delivered (partially inspired by iGEM Japan United, so maybe I’ll integrate a chromosome degradation plasmid and then bake these bacteria into cookies :D), but anyways, automated medium pH monitoring and control would be one use of automation.

Automated cookie bakers maybe?

4. How much would you need to make, and how long would it take?

[1] How much? Over half of children in southern and eastern Africa: ~140 million Over ⅔ of women of reproductive age: ~407 million So let’s make 700 million servings. It’s impractical to produce and deliver servings meant for daily intake (too much packaging), so maybe one intake per month. Each serving includes 240 to 540 mg of iron (more for women due to menstruation), 2.7 to 8.1 mg of iodine (8.1 for breastfeeding or pregnant women), vitamin A tbd on more medical research, 240 to 360 mg of zinc, and 4500 to 12000 mcg of folate. Different packages for different ages and genders (and lactation/pregnancy).

[2] How long? Hard to ascertain the precise rate of synthesis :( Extraction also takes time.

5. What don’t you know yet?

• Precise rates of synthesis
• Precise extraction & purification protocols
• The method of delivery
• How much vitamin A is needed per person per month

Final Project