Purification of peroxisomal malate synthase from alkane-grownCandida tropicalisand some properties of the purified enzyme
✍ Scribed by Hirofumi Okada; Mitsuyoshi Ueda; Atsuo Tanaka
- Publisher
- Springer
- Year
- 1986
- Tongue
- English
- Weight
- 792 KB
- Volume
- 144
- Category
- Article
- ISSN
- 0302-8933
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✦ Synopsis
Malate synthase, one of the key enzymes in the glyoxylate cycle, was purified from peroxisomes of alkanegrown 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 Mg z+ for enzymatic activity. The Km values for Mg 2 +, acetyl-CoA and glyoxylate were 4.7 raM, 80 gM and 1.0 raM, respectively.