title: Subcellular localization of glyoxylate cycle key enzymes involved in oxalate biosynthesis of wood-destroying basidiomycete Fomitopsis palustris grown on glucose.
abstract: This study investigated the subcellular localization of key enzymes of the glyoxylate cycle, i.e. isocitrate lyase (ICL; EC 4.1.3.1) and malate synthase (EC 2.3.3.9), that function constitutively in coordination with oxalate biosynthesis of glucose-grown Fomitopsis palustris. The ICL purified previously from F. palustris is termed FPICL1. Subcellular fractionation analysis of the cell homogenate by the sucrose density-gradient method showed that both key enzymes were present in peroxisomes, whereas acetyl-CoA synthase (EC 6.2.1.1) and oxalate-producing oxaloacetate acetylhydrolase (EC 3.7.1.1) were cytosolic. The peroxisomal localization of FPICL1 was further confirmed by electron microscopic and immunocytochemical analysis with anti-FPICL1 antibody. In addition, the peroxisomal target signal, composed of SKL at the C terminus of the cDNA encoding FPICL1, was found, which also suggests that FPICL1 is peroxisomal. Accordingly, it is postulated that transportation of succinate from peroxisomes to mitochondria, and vice versa, for the transportation of isocitrate or citrate, occurs in glucose-grown F. palustris for the constitutive metabolic coordination of the TCA and glyoxylate cycles with oxalate biosynthesis.
authors.affiliation: Research Institute for Sustainable Humanosphere, Kyoto University, Uji, Kyoto 611-0011, Japan.
authors.forename: Shunsuke
authors.initials: S
authors.lastname: Sakai
authors.affiliation: Research Institute for Sustainable Humanosphere, Kyoto University, Uji, Kyoto 611-0011, Japan.
authors.forename: Tatsunori
authors.initials: T
authors.lastname: Nishide
authors.affiliation: University of North Sumatra, Jl. Bioteknologi No. 1 Kampus USU, Medan 20513, Indonesia.
authors.forename: Erman
authors.initials: E
authors.lastname: Munir
authors.affiliation: Research Institute for Sustainable Humanosphere, Kyoto University, Uji, Kyoto 611-0011, Japan.
authors.forename: Kei'ichi
authors.initials: K
authors.lastname: Baba
authors.affiliation: Department of Applied Biological Chemistry, University of Osaka Prefecture, Sakai, Osaka 599-8231, Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Inui
authors.affiliation: Department of Applied Biological Chemistry, University of Osaka Prefecture, Sakai, Osaka 599-8231, Japan.
authors.forename: Yoshihisa
authors.initials: Y
authors.lastname: Nakano
authors.affiliation: Research Institute for Sustainable Humanosphere, Kyoto University, Uji, Kyoto 611-0011, Japan.
authors.forename: Takefumi
authors.initials: T
authors.lastname: Hattori
authors.affiliation: Research Institute for Sustainable Humanosphere, Kyoto University, Uji, Kyoto 611-0011, Japan.
authors.forename: Mikio
authors.initials: M
authors.lastname: Shimada
mesh_terms: D000595:Amino Acid Sequence; D001487:Basidiomycota / Q000201:enzymology* / Q000254:growth & development; D003470:Culture Media; D003600:Cytosol / Q000201:enzymology; D005947:Glucose / Q000378:metabolism*; D006038:Glyoxylates / Q000378:metabolism*; D007522:Isocitrate Lyase / Q000235:genetics / Q000378:metabolism; D008292:Malate Synthase / Q000378:metabolism; D008969:Molecular Sequence Data; D010070:Oxalates / Q000378:metabolism*; D020675:Peroxisomes / Q000201:enzymology; D017422:Sequence Analysis, DNA; D013347:Subcellular Fractions / Q000201:enzymology*; D014934:Wood
chemical_list: D003470:Culture Media; D006038:Glyoxylates; D010070:Oxalates; D008292:Malate Synthase; D007522:Isocitrate Lyase; D005947:Glucose