# Enabling Plant Natural Product Biosynthesis by Debugging Heterologous Protein Expression in Yeast

> **NIH NIH R35** · UNIVERSITY OF DELAWARE · 2023 · $395,836

## Abstract

PROJECT SUMMARY
The Blenner Research Group at Clemson University is broadly focused on using genetic engineering and
synthetic biology to improve microbial systems for applications in human health, sustainability, and national
defense. Our research is aimed at developing the capabilities to accurately and precisely control the properties
of oleaginous yeast – Yarrowia lipolytica. This yeast has naturally evolved the capability to produce a significant
amount of lipid. As such, they are useful for producing biochemicals derived from lipids, such as omega-3 fatty
acids and fatty alcohols. As oleaginous yeast are considered non-conventional and in many cases non-model
systems, there were few genetic engineering tools available to manipulate these systems. The Blenner Research
Group has invested significant time and energy building finely-tuned promoters, inducible promoters, temporal
controlled promoters, and a suite of CRISPR-Cas9 technologies for rapid genome editing. Significant progress
in lipid and oleochemical production was due to Y. lipolytica's innate high flux capacity through intermediates
and cofactors needed for lipid biosynthesis. Our broad goal over the next five years are to increase the diversity
of products that are made in oleaginous yeast. There are over 200,000 known natural products structures and
natural products make up the majority of therapeutic compounds. Many bioactive natural products are derived
from plants; however, isolation from plants is uncertain, can be extremely costly, and in some cases, agriculture
cannot provide enough material. Several classes of plant natural products, including terpenoids, flavonoids, and
alkaloids, are biosynthesized using fundamental building blocks from high flux pathways in Y. lipolytica. As such,
we hypothesize oleaginous yeast are ideally suited for high titer natural product biosynthesis. This proposal
focuses on addressing a common problem across all natural product heterologous pathways – difficulty
expressing foreign proteins. We seek to identify known and novel responses to common mode of heterologous
protein expression failure. Once specific genes associated with each modes of failure are identified, these can
serve as indicators to identify a new gene's mode of failure. These can also be used to help identify solutions to
improve expression based on the mode of failure. The resulting improvements can be stacked into platform
strains and used to more easily screen natural product pathway libraries. We will focus on flavonoids due to their
diverse chemistry and known therapeutic effects; however, we expect these platform strains to be useful for
making other natural products as well.

## Key facts

- **NIH application ID:** 10691233
- **Project number:** 5R35GM133803-06
- **Recipient organization:** UNIVERSITY OF DELAWARE
- **Principal Investigator:** Mark Alan Blenner
- **Activity code:** R35 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2023
- **Award amount:** $395,836
- **Award type:** 5
- **Project period:** 2019-08-01 → 2025-05-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10691233

## Citation

> US National Institutes of Health, RePORTER application 10691233, Enabling Plant Natural Product Biosynthesis by Debugging Heterologous Protein Expression in Yeast (5R35GM133803-06). Retrieved via AI Analytics 2026-05-21 from https://api.ai-analytics.org/grant/nih/10691233. Licensed CC0.

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