title: Identification of genes that enhance cellulase protein production in yeast.
abstract: In order to enhance heterologous cellulase protein production in yeast, a plasmid harboring the endoglucanase gene from Clostridium thermocellum (Ctcel8A) was used to systematically transform a homozygous diploid yeast deletion strain collection. We identified 55 deletion strains that exhibited enhanced endoglucanase activity compared with that of the wild-type strain. Genes disrupted in these strains were classified into the categories of transcription, translation, phospholipid synthesis, endosome/vacuole function, ER/Golgi function, nitrogen starvation response, and cytoskeleton. The vps3Δ and vps16Δ strains, which have deletion in genes encoding components of the class C core vacuole/endosome tethering (CORVET) complex, also exhibited enhanced β-glucosidase activity when Ctcel8A was heterologously expressed. Moreover, multiple gene deletion strains were constructed by using the vps3Δ strain. Endoglucanase activity of the resulting rav1Δvps3Δ double deletion strain was exhibited higher than that of the rav1Δ or vps3Δ strains. Our genome-wide analyses using the yeast deletion strain collection identified useful genes that allow efficient expression of cellulase.
authors.affiliation: Department of Applied Molecular Bioscience, Yamaguchi University Graduate School of Medicine, Tokiwadai, Ube 755-8611, Japan. takao-k@yamaguchi-u.ac.jp
authors.forename: Takao
authors.initials: T
authors.lastname: Kitagawa
authors.forename: Katsunori
authors.initials: K
authors.lastname: Kohda
authors.forename: Kenro
authors.initials: K
authors.lastname: Tokuhiro
authors.forename: Hisashi
authors.initials: H
authors.lastname: Hoshida
authors.forename: Rinji
authors.initials: R
authors.lastname: Akada
authors.forename: Haruo
authors.initials: H
authors.lastname: Takahashi
authors.forename: Takao
authors.initials: T
authors.lastname: Imaeda
mesh_terms: D001709:Biotechnology / Q000379:methods*; D002480:Cellulase / Q000096:biosynthesis* / Q000737:chemistry* / Q000378:metabolism; D048013:Acetivibrio thermocellus / Q000737:chemistry*; D003599:Cytoskeleton / Q000378:metabolism; D011992:Endosomes; D000431:Ethanol / Q000737:chemistry; D005656:Fungal Proteins / Q000737:chemistry; D017353:Gene Deletion; D006720:Homozygote; D009584:Nitrogen / Q000737:chemistry; D010957:Plasmids / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D013696:Temperature; D014617:Vacuoles / Q000378:metabolism
chemical_list: D005656:Fungal Proteins; D000431:Ethanol; D002480:Cellulase; D009584:Nitrogen