Schizosaccharomyces pombe minimum genome factory.

Area of Science:

• Biotechnology and Synthetic Biology
• Microbial Engineering
• Molecular Genetics

Background:

• Current fission yeast (Schizosaccharomyces pombe) protein production systems require optimization for cost-effectiveness and productivity.
• Many S. pombe genes are non-essential under nutrient-rich conditions and may hinder heterologous protein production by consuming energy.

Purpose of the Study:

• To engineer S. pombe strains optimized for heterologous protein production by deleting non-essential genes.
• To improve the economic viability and productivity of S. pombe as a host for diverse molecule synthesis.

Main Methods:

• Systematic deletion of non-essential genes using the Latour method to create large chromosomal deletions (approx. 500 kb).
• Transcriptome analysis of gene disruptants using microarrays to identify new promoters.
• Construction of protease-deficient S. pombe strains (single and multiple gene disruptions) to prevent product degradation.

Main Results:

• Established novel S. pombe strains with significant chromosomal deletions and auxotrophy for six nutrients, facilitating multi-plasmid co-expression.
• Developed efficient transformation methods for various S. pombe strains.
• Demonstrated increased production of human growth hormone in protease-deficient strains, indicating reduced protein degradation.

Conclusions:

• Engineering S. pombe by deleting non-essential and protease genes significantly enhances its utility as a host for heterologous protein production.
• The development of large-deletion and functional gene-deficient strains paves the way for highly effective and economical microbial cell factories.