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Microbiology 152 (2006), 1857-1866; DOI  10.1099/mic.0.28702-0
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Microbiology 152 (2006), 1857-1866; DOI  10.1099/mic.0.28702-0
© 2006 Society for General Microbiology

Subcellular localization of glyoxylate cycle key enzymes involved in oxalate biosynthesis of wood-destroying basidiomycete Fomitopsis palustris grown on glucose

Shunsuke Sakai1,{dagger}, Tatsunori Nishide1,{ddagger}, Erman Munir2, Kei'ichi Baba1, Hiroshi Inui3, Yoshihisa Nakano3, Takefumi Hattori1 and Mikio Shimada1,§

1 Research Institute for Sustainable Humanosphere, Kyoto University, Uji, Kyoto 611-0011, Japan
2 University of North Sumatra, Jl. Bioteknologi No. 1 Kampus USU, Medan 20513, Indonesia
3 Department of Applied Biological Chemistry, University of Osaka Prefecture, Sakai, Osaka 599-8231, Japan

Correspondence
Takefumi Hattori
thattori{at}rish.kyoto-u.ac.jp

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.


Abbreviations: ACS, acetyl-CoA synthase; DAB, diaminobenzidine; FPICL1, Fomitopsis palustris isocitrate lyase 1; GLOX, glyoxylate; IDH, isocitrate dehydrogenase; MDH, malate dehydrogenase; MS, malate synthase; NTA, nitrilotriacetate; ODH, 2-oxoglutarate dehydrogenase; OXA, oxaloacetate acetylhydrolase; PNS, post-nuclear supernatant; PTS1, peroxisomal target signal 1; SDH, succinate dehydrogenase

{dagger}Present address: Department of Microbiology, Graduate School of Medicine, Kyoto University, Kyoto 606-8501, Japan.

{ddagger}Present address: Nisshin Foods Inc., Tokyo, Japan.

§Present address: Environmental and Biotechnological Frontier Engineering, Fukui University of Technology, Fukui, Japan.




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