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Modular metabolic flux control for kick-starting cascade catalysis through engineering customizable compartment
Qiang Ding1, Mengqi Ji1, Buhan Yao1
1School of Life Sciences, Anhui University, Hefei 230601, China; Key Laboratory of Human Microenvironment and Precision Medicine of Anhui Higher Education Institutes, Anhui University, Hefei 230601 Anhui, China; Anhui Key Laboratory of Modern Biomanufacturing, Hefei 230601 Anhui, China.
Researchers engineered a customizable microbial compartment to improve chemical biosynthesis. This compartmentalization enhanced the production of 2'-fucosyllactose (2'-FL) by co-localizing enzymes, boosting yields compared to free enzymes.
Area of Science:
- Synthetic Biology
- Metabolic Engineering
- Biocatalysis
Background:
- Microbial cell factories offer a sustainable route for producing high-value chemicals from renewable resources.
- Challenges in microbial biosynthesis include volatile precursors being consumed by pathway enzymes, hindering cascade catalysis.
- Intracellular compartmentalization is a strategy to spatially organize enzymes and improve metabolic efficiency.
Purpose of the Study:
- To develop a customizable intracellular compartment for pathway sequestration and spatially assembled cascade catalysis.
- To enhance the biosynthesis of 2 -fucosyllactose (2 -FL) using a compartmentalized microbial system.
- To demonstrate improved chemical titers and metabolic flux control through enzyme co-localization.
Main Methods:
- Designed a phase separation protein to form intracellular protein condensates, creating a customizable compartment in Escherichia coli.
- Utilized a short peptide interaction pair for modular assembly and recruitment of enzymes within the compartment.
- Expressed the 2 -FL salvage pathway heterogeneously within the engineered compartment as a proof-of-concept.
Main Results:
- Successfully constructed a customizable intracellular compartment capable of housing multiple enzymes.
- Demonstrated enhanced 2 -FL production, achieving a higher titer compared to wild-type and free enzyme systems.
- Showcased improved cascade catalysis and metabolic flux control due to enzyme co-localization within the compartment.
Conclusions:
- The developed customizable compartment effectively sequesters enzymatic pathways, enhancing cascade catalysis.
- This compartmentalization strategy significantly improves the production titer of target chemicals like 2 -FL.
- The findings provide a robust model for increasing titers and controlling metabolic flux in microbial cell factories through enzyme co-localization.
