Subcellular localization of glyoxylate cycle key enzymes involved in oxalate biosynthesis of wood-destroying basidiomycete Fomitopsis palustris grown on glucose.
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.
Sakai
Shunsuke
S
Research Institute for Sustainable Humanosphere, Kyoto University, Uji, Kyoto 611-0011, Japan.
Nishide
Tatsunori
T
Research Institute for Sustainable Humanosphere, Kyoto University, Uji, Kyoto 611-0011, Japan.
Munir
Erman
E
University of North Sumatra, Jl. Bioteknologi No. 1 Kampus USU, Medan 20513, Indonesia.
Baba
Kei'ichi
K
Research Institute for Sustainable Humanosphere, Kyoto University, Uji, Kyoto 611-0011, Japan.
Inui
Hiroshi
H
Department of Applied Biological Chemistry, University of Osaka Prefecture, Sakai, Osaka 599-8231, Japan.
Nakano
Yoshihisa
Y
Department of Applied Biological Chemistry, University of Osaka Prefecture, Sakai, Osaka 599-8231, Japan.
Hattori
Takefumi
T
Research Institute for Sustainable Humanosphere, Kyoto University, Uji, Kyoto 611-0011, Japan.
Shimada
Mikio
M
Research Institute for Sustainable Humanosphere, Kyoto University, Uji, Kyoto 611-0011, Japan.
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
D003470:Culture Media; D006038:Glyoxylates; D010070:Oxalates; D008292:Malate Synthase; D007522:Isocitrate Lyase; D005947:Glucose