Harnessing a P450 fatty acid decarboxylase from

Area of Science:

• Biotechnology
• Synthetic Biology
• Enzyme Engineering

Background:

• Alkenes are crucial industrial chemicals.
• Microbial biosynthesis offers a sustainable production route.
• Fatty acid decarboxylases are key enzymes for alkene synthesis.

Purpose of the Study:

• To develop a direct microbial biosynthesis pathway for terminal alkenes from fermentable sugars.
• To characterize the Macrococcus caseolyticus P450 fatty acid decarboxylase (OleTMC).
• To engineer and optimize an E. coli strain for enhanced alkene production.

Main Methods:

• Characterization of OleTMC activity on various fatty acids.
• Engineering of a de novo alkene biosynthesis pathway in E. coli.
• Introduction of a redox system (CamA-CamB) to overcome electron transfer limitations.
• In silico protein modeling to analyze enzyme-substrate interactions.

Main Results:

• The engineered E. coli strain (EcNN101) produced odd-chain terminal alkenes from glucose.
• Optimized strains achieved a production of 17.78 mg/L alkenes.
• Incorporation of a redox system increased alkene production by approximately 2.8-fold.
• In silico modeling provided insights into OleTMC's substrate specificity.

Conclusions:

• Direct microbial biosynthesis of terminal alkenes from sugars is feasible using engineered pathways.
• OleTMC is a promising enzyme for alkene production.
• Redox system engineering significantly enhances the efficiency of microbial alkene biosynthesis.