Metabolic pathway assembly using docking domains from type I cis-AT polyketide synthases

Area of Science:

• Metabolic Engineering
• Synthetic Biology
• Biocatalysis

Background:

• Engineered metabolic pathways often lack natural organization, leading to flux imbalances and reduced biocatalytic efficiency.
• Modular polyketide synthases (PKSs) utilize an assembly line mechanism for polyketide synthesis.

Purpose of the Study:

• To develop a strategy for assembling cascade enzymes using docking domains from type I cis-AT PKS.
• To enhance biocatalytic efficiency and increase target production in engineered microbial systems.

Main Methods:

• The mimic PKS enzyme assembly line (mPKSeal) strategy was developed, recruiting cascade enzymes tagged with docking domains.
• This strategy was applied to the astaxanthin biosynthetic pathway in engineered Escherichia coli.
• The effectiveness of docking pairs from the same or different cis-AT PKS classes was evaluated.

Main Results:

• Astaxanthin production was increased by 2.4-fold through mPKSeal-mediated multienzyme assembly.
• Docking pairs from various cis-AT PKS classes proved effective for enzyme assembly.
• The study demonstrated enhanced biocatalytic efficiency and increased target production.

Conclusions:

• The mPKSeal strategy offers a viable approach to address cascade catalytic efficiency challenges in metabolic engineering.
• This method highlights the potential of engineered enzyme assembly for improving microbial cell factory performance.
• The findings pave the way for more efficient biocatalytic production of valuable compounds.