title: Genome-wide identification of the targets for genetic manipulation to improve L-lactate production by Saccharomyces cerevisiae by using a single-gene deletion strain collection.
abstract: To identify genome-wide targets for gene manipulation for increasing L-lactate production in recombinant Saccharomyces cerevisiae strains, we transformed all available single-gene deletion strains of S. cerevisiae with a plasmid carrying the human L-lactate dehydrogenase gene, and examined L-lactate production in the obtained transformants. The thresholds of increased or decreased L-lactate production were determined based on L-lactate production by the standard strain in repetitive experiments. L-lactate production data for 4802 deletion strains were obtained, and deletion strains with increased or decreased L-lactate production were identified. Functional category analysis of genes whose deletion increased L-lactate production revealed that ribosome biogenesis-related genes were overrepresented. Most deletion strains for genes related to ribosome biogenesis exhibited increased L-lactate production in 200-ml batch cultures. We deleted the genes related to ribosome biogenesis in a recombinant strain of S. cerevisiae with a genetic background different from that of the above deletion strains, and examined the effect of target gene deletion on L-lactate production. We observed that deletion of genes related to ribosome biogenesis leads to increased L-lactate production by recombinant S. cerevisiae strains, and the single-gene deletion strain collection could be utilized in identifying target genes for improving L-lactate production in S. cerevisiae recombinant strains.
authors.affiliation: Department of Bioinformatic Engineering, Graduate School of Information Science and Technology, Osaka University, 1-5 Yamadaoka, Suita, Osaka 565-0871, Japan. Electronic address: hirasawa@ist.osaka-u.ac.jp.
authors.forename: Takashi
authors.initials: T
authors.lastname: Hirasawa
authors.forename: Masakado
authors.initials: M
authors.lastname: Takekuni
authors.forename: Katsunori
authors.initials: K
authors.lastname: Yoshikawa
authors.forename: Aki
authors.initials: A
authors.lastname: Ookubo
authors.forename: Chikara
authors.initials: C
authors.lastname: Furusawa
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Shimizu
mesh_terms: D017353:Gene Deletion; D015966:Gene Expression Regulation, Fungal; D018390:Gene Targeting; D005800:Genes, Fungal; D005818:Genetic Engineering; D016681:Genome, Fungal; D006801:Humans; D007770:L-Lactate Dehydrogenase / Q000235:genetics* / Q000378:metabolism; D019344:Lactic Acid / Q000096:biosynthesis*; D010957:Plasmids; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D015203:Reproducibility of Results; D012270:Ribosomes / Q000235:genetics* / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000145:classification / Q000235:genetics* / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism; D014170:Transformation, Genetic
chemical_list: D011994:Recombinant Proteins; D029701:Saccharomyces cerevisiae Proteins; D019344:Lactic Acid; D007770:L-Lactate Dehydrogenase
keywords: Ribosome; Saccharomyces cerevisiae; Single-gene deletion strain collection; l-Lactate