Enhancing myrcene biosynthesis in yeast through nuclear compartmentalization.
This research shows that engineering the yeast nucleus can enhance myrcene production, achieving a titer of 23.4 mg L-1. The findings may inform future strategies for microbial production of valuable compounds.
Where it sits
this study against the rest of the argireline corpusSummary and findings
This study measured myrcene production in engineered yeast, specifically Saccharomyces cerevisiae, by utilizing nuclear compartmentalization. The final titer achieved was 23.4 mg L-1 during flask-shaking fermentation. Two highly active myrcene synthases were identified from Snapdragon Oc15 and Picea abies.
Abstract
Myrcene is a high-value monoterpene extensively applied in the fragrance, flavor, and agricultural industries, yet its efficient microbial production remains challenging due to pathway competition and limited metabolic flux. Compartmentalization offers a unique strategy to spatially organize heterologous metabolic pathways in <i>Saccharomyces cerevisiae</i>, enabling improved pathway efficiency through physical separation from competing cytosolic metabolism. In this study, we engineered the <i>S. cerevisiae</i> nucleus as a synthetic metabolic compartment for myrcene biosynthesis. Screening of myrcene synthases identified two highly active enzymes from <i>Snapdragon Oc15</i> and <i>Picea abies</i> that function efficiently in <i>S. cerevisiae</i>. Myrcene production was detected only when myrcene synthase and the engineered GPP synthase mERG20p were co-localized to the nucleus, whereas cytosolic expression failed to yield detectable myrcene under their co-expression. Reconstruction of the complete mevalonate (MVA) pathway in the nucleus further increased myrcene titers. By identifying and optimizing rate-limiting steps, we substantially enhanced metabolic flux toward myrcene, achieving a final titer of 23.4 mg L<sup>-1</sup> in flask-shaking fermentation. This work demonstrates the feasibility of repurposing the yeast nucleus for myrcene efficient biosynthesis and provide a new strategy for further improving microbial production of myrcene.
Background
The study addresses the challenge of efficient microbial production of myrcene, a valuable monoterpene used in various industries. Previous efforts faced limitations due to pathway competition and metabolic flux constraints. This research explores the potential of nuclear compartmentalization in Saccharomyces cerevisiae to enhance myrcene biosynthesis.
Methods
The study involved engineering the nucleus of Saccharomyces cerevisiae to create a synthetic metabolic compartment for myrcene production. The population consisted of yeast cells, with specific myrcene synthases from Snapdragon Oc15 and Picea abies being screened for activity. The primary outcome measure was the titer of myrcene produced during flask-shaking fermentation.
Results
The primary endpoint showed a final titer of 23.4 mg L-1 of myrcene in flask-shaking fermentation. Myrcene production was only detected when the myrcene synthase and GPP synthase mERG20p were co-localized to the nucleus. The reconstruction of the complete mevalonate pathway in the nucleus further enhanced myrcene titers.
Interpretation
This study demonstrates a novel approach to improving myrcene production in yeast through nuclear compartmentalization. While the findings are statistically significant, the clinical relevance remains uncertain as the application of myrcene production in therapeutic contexts is not established. Limitations include the lack of human data and potential confounding factors related to the engineered yeast model.
Key findings
- Final titer of 23.4 mg L-1 in flask-shaking fermentation.
- Myrcene production detected only with nuclear co-localization of myrcene synthase and GPP synthase mERG20p.
- Reconstruction of the complete mevalonate pathway in the nucleus increased myrcene titers.
Limitations
- Not reported in abstract.
- Small-scale flask-shaking fermentation may not reflect larger-scale production.
- Lack of human data limits applicability to clinical settings.
- Potential confounding factors from engineered yeast model.