High level expression of isocitrate lyase gene of n-alkane-utilizing yeast Candida tropicalis in Sacchromyces cerevisiae
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The genomic DNA of peroxisomal isocitrate lyase (ICL) isolated from an n-alkane-assimilating yeast, Candida tropicalis, was truncated to utilize the original open reading frame under the control of the GAL7 promoter and was expressed in Saccharomyces cerevisiae. The recombinant ICL was synthesized as a functionally active enzyme with a specific activity similar to the enzyme purified from C. tropicalis, and was accounted for approximately 30% of the total extractable proteins in the yeast cells. This recombinant enzyme was easily purified to homogeneity. N-Terminal amino acid sequence, molecular masses of native form and subunit, amino acid composition, peptide maps, and kinetic parameters of the recombinant ICL were essentially the same as those of ICL purified from C. tropicalis. From these facts, S. cerevisiae was suggested to be an excellent microorganism to highly express the genes encoding peroxisomal proteins of C. tropicalis.
Key wordsCandida tropicalis Saccharomyces cerevisiae Peroxisomes Isocitrate lyase GAL7 promoter High level expression
sodium dodecylsulfate-polyacrylamide gel electrophoresis
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- 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: 275–279Google Scholar
- Okada H, Ueda M, Sugaya T, Atomi H, Mozaffar S, Hishida T, Teranishi Y, Okazaki K, Takechi T, Kamiryo T, Tanaka A (1987) Catalase gene of the yeast Candida tropicalis. Sequence analysis and comparison with peroxisomal and cytosolic catalases from other sources. Eur J Biochem 170: 105–110CrossRefGoogle Scholar
- 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: 127–134Google Scholar