Enhancing Cephalosporin C Biosynthesis through a 2A Peptide-Based Multigene Coexpression System.

Area of Science:

• Biotechnology
• Microbial Engineering
• Synthetic Biology

Background:

• Cephalosporin C (CPC) is a vital precursor for semisynthetic cephalosporins, industrially produced by Acremonium chrysogenum.
• Current metabolic engineering of A. chrysogenum is hindered by the lack of efficient multigene coexpression systems.

Purpose of the Study:

• To develop and apply a 2A peptide-based multigene coexpression system in A. chrysogenum.
• To evaluate the efficiency of various viral 2A peptides for coexpression.
• To enhance CPC production through targeted gene coexpression.

Main Methods:

• Systematic evaluation of ten virus-derived 2A peptides for self-cleavage efficiency in A. chrysogenum.
• Selection of the optimal P2A peptide for precalibrated, multigene expression.
• Coexpression of key CPC biosynthetic genes (cefEF, cefG) with a driver gene (ACRE_076110).

Main Results:

• P2A peptide demonstrated the highest self-cleavage efficiency (88.2%), enabling robust coexpression.
• Coexpression of cefEF and cefG with ACRE_076110 resulted in a 3.19-fold increase in CPC titer.
• Application in an industrial strain improved CPC yield by 22.2% (from 6.09 g/L to 7.45 g/L).

Conclusions:

• A functional 2A peptide-based multigene coexpression system was successfully established in A. chrysogenum.
• This system significantly enhances CPC production, offering a valuable tool for industrial fermentation.
• The developed system holds promise for advancing metabolic engineering strategies in A. chrysogenum.