Customized optimization of metabolic pathways by combinatorial transcriptional engineering

Area of Science:

• Metabolic Engineering
• Synthetic Biology
• Systems Biology

Background:

• Balancing metabolic flux in engineered heterologous pathways is crucial for high yield and productivity.
• Existing methods struggle to tailor pathways for specific host organisms and metabolic backgrounds.

Purpose of the Study:

• To develop a simple, efficient, and programmable method for rapid tuning of gene expression in heterologous metabolic pathways.
• To demonstrate the efficacy of the COMPACTER approach in optimizing xylose and cellobiose utilizing pathways in yeast.

Main Methods:

• A library of mutant pathways was created using de novo assembly of promoter mutants with varying strengths.
• High-throughput screening and selection were employed to identify optimal pathway variants.
• The COMPACTER method was applied to engineer xylose and cellobiose utilization pathways in yeast.

Main Results:

• COMPACTER enabled the generation of a highly efficient xylose utilizing pathway and the most efficient cellobiose utilizing pathway reported in literature.
• The engineered pathways achieved near-highest and highest efficiencies, respectively, in both laboratory and industrial yeast strains.
• The optimized pathways exhibited host-specificity, highlighting the method's ability to tailor pathways.

Conclusions:

• COMPACTER is a powerful and programmable approach for optimizing metabolic pathways by combinatorial transcriptional engineering.
• The method allows for rapid tuning of gene expression and pathway flux, leading to improved yield and productivity.
• COMPACTER facilitates the creation of tailored metabolic pathways for specific host organisms, overcoming limitations of existing methods.