Gene conversion-associated successive engineering of modular polyketide synthases.

Area of Science:

• Biochemistry
• Synthetic Biology
• Metabolic Engineering

Background:

• Modular polyketide synthases (PKSs) are crucial for secondary metabolite production.
• Rational engineering of PKSs often faces challenges with element incompatibility, leading to reduced productivity.
• The cinnamomycin biosynthetic gene cluster (cmm BGC) serves as a model system.

Purpose of the Study:

• To develop a PKS engineering strategy that mimics evolutionary gene conversion.
• To enable successive engineering of modular PKS for de novo compound generation.
• To investigate the role of the KS domain in extender unit selection.

Main Methods:

• Simulating gene conversion events within the cmm BGC.
• Reprogramming the modular PKS architecture.
• Analyzing the function of the intra-module KS domain.

Main Results:

• Successfully generated a macrolide with predicted structural features.
• Demonstrated the feasibility of successive PKS engineering using a gene conversion-inspired approach.
• Identified the KS domain's role in proofreading extender units during polyketide synthesis.

Conclusions:

• The gene conversion-associated strategy offers a promising route for modular PKS engineering.
• This method can overcome limitations in de novo PKS construction.
• Further exploration of this approach could unlock new avenues for natural product synthesis.