A streamlined strain engineering workflow with genome-wide screening detects enhanced protein secretion in Komagataella phaffii.

ABSTRACT: Expression of secreted recombinant proteins burdens the protein secretion machinery, limiting production. Here, we describe an approach to improving protein production by the non-conventional yeast Komagataella phaffii comprised of genome-wide screening for effective gene disruptions, combining them in a single strain, and recovering growth reduction by adaptive evolution. For the screen, we designed a multiwell-formatted, streamlined workflow to high-throughput assay of secretion of a single-chain small antibody, which is cumbersome to detect but serves as a good model of proteins that are difficult to secrete. Using the consolidated screening system, we evaluated >19,000 mutant strains from a mutant library prepared by a modified random gene-disruption method, and identified six factors

SUBMITTER: Ito Y 

PROVIDER: S-EPMC9177720 | biostudies-literature | 2022 Jun

REPOSITORIES: biostudies-literature