Random mutagenesis and transcriptomics-guided rational engineering in Zygosaccharomyces rouxii for elevating...

Area of Science:

• Biotechnology
• Metabolic Engineering
• Industrial Microbiology

Background:

• D-arabitol is a valuable compound with diverse industrial applications.
• Current biomanufacturing of D-arabitol is limited by suboptimal yeast chassis performance and poorly understood synthesis pathways.
• Enhancing D-arabitol production is crucial for meeting industrial demand.

Purpose of the Study:

• To improve the D-arabitol production capabilities of Zygosaccharomyces rouxii.
• To overcome limitations in yeast chassis performance for enhanced biomanufacturing.
• To develop a robust and efficient process for high-titer D-arabitol production.

Main Methods:

• Atmospheric and room temperature plasma mutagenesis was employed to generate a mutant strain (Z. rouxii M075).
• Transcriptome-guided metabolic engineering focused on co-expressing key pathway enzymes to create strain ZR-M3.
• Optimized fed-batch fermentation in a 5 L bioreactor was performed for large-scale production.

Main Results:

• The mutant strain Z. rouxii M075 achieved a D-arabitol titer of 42.0 g/L, a 50% increase over the wild type.
• Strain ZR-M3, developed through metabolic engineering, reached 48.9 g/L of D-arabitol after 96 hours.
• Optimized fed-batch fermentation of ZR-M3 resulted in a D-arabitol titer of 152.8 g/L at 192 hours, with a productivity of 0.8 g/L/h.

Conclusions:

• The developed Z. rouxii strains show significantly enhanced D-arabitol production capabilities.
• Metabolic engineering strategies, guided by transcriptomics, are effective in improving biomanufacturing performance.
• This research offers a promising foundation for efficient industrial-scale D-arabitol bioproduction.