Abstract
Malate synthase, one of the key enzymes in the glyoxylate cycle, was purified from peroxisomes of alkane-grown yeast, Candida tropicalis. The enzyme was mainly localized in the matrix of peroxisomes, judging from subcellular fractionation followed by exposure of the organelles to hypotonic conditions. The molecular mass of this peroxisomal malate synthase was determined to be 250,000 daltons by gel filtration on a Sepharose 6B column as well as by ultracentrifugation. On sodium dodecylsulfate/polyacrylamide slab-gel electrophoresis, the molecular mass of the subunit of the enzyme was demonstrated to be 61,000 daltons. These results revealed that the native form of this enzyme was homo-tetrameric. Peroxisomal malate synthase showed the optimal activity pH at 8.0 and absolutely required Mg2+ for enzymatic activity. The K m values for Mg2+, acetyl-CoA and glyoxylate were 4.7 mM, 80 μM and 1.0 mM, respectively.
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Beevers H (1979) Microbodies in higher plants. Ann Rev Plant Physiol 30:159–193
Bowden L, Lord JM (1978) Purification and comparative properites of microsomal and glyoxysomal malate synthase from castor bean endosperm. Plant Physiol 61:259–265
Dixon GH, Kornberg HL, Lund P (1960) Purification and properties of malate synthetase. Biochim Biophys Acta 41:217–233
Durchschlag H, Biedermann G, Eggerer H (1981) Large-scale purification and some properties of malate synthase from baker's yeast. Eur J Biochem 114:255–262
Falmagne P, Wiame JM (1973) Purification et caractérisation partielle des deux malate synthases d'Escherichia coli. Eur J Biochem 37:415–424
Fukui S, Tanaka A (1979) Yeast peroxisomes. Trends Biochem Sci 4:246–249
Goodman DBP, Davis WL, Jones RG (1980) Glyoxylate cycle in toad urinary bladder: Possible stimulation by aldosterone. Proc Natl Acad Sci USA 77:1521–1525
Jones CT (1980) Is there a glyoxylate cycle in the liver of the fetal guinea pig? Biochem Biophys Res Commun 95:894–856
Köller W, Kindl H (1977) Glyoxylate cycle enzymes of the glyoxysomal membrane from cucumber cotyledons. Arch Biochem Biophys 181:236–248
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275
Miernyk JA, Trelease RN (1981) Malate synthase from Gossypium hirsutum. Phytochem 20:2657–2663
Nabeshima S, Tanaka A, Fukui S (1977) Effect of carbon sources on the level of glyoxylate cycle enzymes in n-alkane-utilizable yeasts. Agric Biol Chem 41:275–279
Osumi M, Miwa N, Teranishi Y, Tanaka A, Fukui S (1974) Ultrastructure of Candida yeasts grown on n-alkanes. Appearance of microbodies and its relationship to high catalase activity. Arch Microbiol 99:181–201
Osumi M, Fukuzumi F, Teranishi Y, Tanaka A, Fukui S (1975) Development of microbodies in Candida tropicalis during incubation in a n-alkane medium. Arch Microbiol 103:1–11
Sundaram TK, Chell RM, Wilkinson AE (1980) Monomeric malate synthase from a thermophilic Bacillus. Arch Biochem Biophys 199:515–525
Tanaka A, Osumi M, Fukui S (1982) Peroxisomes of alkane-grown yeast: Fundamental and practical aspects. Ann NY Acad Sci 386:183–199
Uchida M, Ueda M, Matsuki T, Okada H, Tanaka A, Fukui S (1986) Properties of isocitrate lyase from an alkane-utilizable yeast, Candida tropicalis. Agric Biol Chem 50:127–134
Ueda M, Tanaka A, Fukui S (1982) Peroxisomal and mitochondrial carnitine acetyltransferases in alkane-grown yeast Candida tropicalis. Eur J Biochem 124:205–210
Ueda M, Okada H, Tanaka A, Osumi M, Fukui S (1983) Induction and subcellular localization of enzymes participating in propionate metabolism in Candida tropicalis. Arch Microbiol 136:169–176
Ueda M, Tanaka A, Fukui S (1984) Characterization of peroxisomal and mitochondrial carnitine acetyltransferases purified from alkane-grown Candida tropicalis. Eur J Biochem 138:445–449
Ueda M, Yamanoi K, Morikawa T, Okada H, Tanaka A (1985) Peroxisomal localization of enzymes related to fatty acid β-oxidation in an n-alkane-grown yeast, Candida tropicalis. Agric Biol Chem 49:1821–1828
Woodcock E, Merrett MJ (1978) Purification and immunochemical characterization of malate synthase from Euglena gracilis. Biochem J 173:95–101
Zwart KB, Veenhuis M, Plat G, Harder W (1983) Characterization of glyoxysomes in yeasts and their transformation into peroxisomes in response to changes in environmental conditions. Arch Microbiol 136:28–38
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Okada, H., Ueda, M. & Tanaka, A. Purification of peroxisomal malate synthase from alkane-grown Candida tropicalis and some properties of the purified enzyme. Arch. Microbiol. 144, 137–141 (1986). https://doi.org/10.1007/BF00414723
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DOI: https://doi.org/10.1007/BF00414723