Iterative evolution of Saccharomyces cerevisiae enhances recombinant protease secretion

Area of Science:

• Biotechnology
• Synthetic Biology
• Microbial Engineering

Background:

• Efficient secretion of heterologous proteases in Saccharomyces cerevisiae is hindered by host cytotoxicity and secretory stress.
• Existing yeast strains face limitations in producing high yields of complex recombinant enzymes.

Purpose of the Study:

• To develop an improved Saccharomyces cerevisiae chassis strain for enhanced heterologous protease production.
• To overcome bottlenecks in recombinant enzyme secretion and improve industrial biotechnology applications.

Main Methods:

• Iterative evolution of a yeast strain using ultraviolet (UV) and atmospheric and room temperature plasma (ARTP) mutagenesis.
• Development of a fluorescence-based screening pipeline for strain selection.
• Whole-genome sequencing to identify genetic adaptations in the evolved strain.

Main Results:

• The evolved strain, MA10Ah121, achieved nearly threefold higher protease production compared to the parental strain.
• MA10Ah121 demonstrated stable genetic performance across passages.
• Enhanced expression of two distinct collagenases (ColG and ColH) was achieved, indicating broad utility.

Conclusions:

• The developed yeast chassis (MA10Ah121) offers a robust and versatile platform for recombinant protease biosynthesis.
• Genomic adaptations, including chromosomal remodeling, underlie the strain's improved biosynthetic capacity and stress tolerance.
• This work advances the potential for using engineered yeast in industrial production of challenging recombinant enzymes.