Harnessing Metalloprotease Wss1 to Enhance Methanol Utilization

Area of Science:

• Biotechnology
• Synthetic Biology
• Microbial Engineering

Background:

• Methylotrophic bacteria are key for one-carbon (C1) feedstock assimilation in biomanufacturing.
• Poor tolerance to C1 compounds like formaldehyde limits their industrial application.
• Formaldehyde causes DNA-protein cross-linking (DPC), inhibiting growth and C1 assimilation.

Purpose of the Study:

• To enhance formaldehyde tolerance and C1 assimilation in Escherichia coli.
• To investigate the role of DNA-protein cross-linking (DPC) in limiting C1 utilization.
• To engineer a robust E. coli strain for efficient methanol bioconversion.

Main Methods:

• Overexpression of the metalloproteinase SpWss1 from Schizosaccharomyces pombe in E. coli.
• Assessing formaldehyde tolerance and DPC alleviation in engineered strains.
• Quantifying methanol assimilation and production of valuable chemicals.

Main Results:

• Fine-overexpression of SpWss1 alleviated DPC damage and enhanced formaldehyde tolerance.
• Engineered E. coli showed a 10-fold increase in methanol assimilation (142 mM vs 14 mM).
• The modified strain consumed up to 309 mM methanol and enhanced production of triacetic acid lactone and fatty acids.

Conclusions:

• Addressing DNA-protein cross-linking is crucial for optimizing C1 assimilation in microbial systems.
• SpWss1 engineering offers a promising strategy for developing high-performance C1-utilizing E. coli.
• This work provides insights for sustainable chemistry and industrial biotechnology applications.