High level expression of isocitrate lyase gene of n-alkane-utilizing yeast Candida tropicalis in Saccharomyces cerevisiae.
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 micro-organism to highly express the genes encoding peroxisomal proteins of C. tropicalis.
Oda
K
K
Department of Industrial Chemistry, Faculty of Engineering, Kyoto University, Japan.
Atomi
H
H
Ueda
M
M
Kondo
J
J
Teranishi
Y
Y
Tanaka
A
A
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D002175:Candida / Q000201:enzymology* / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D004790:Enzyme Induction; D015966:Gene Expression Regulation, Fungal; D007522:Isocitrate Lyase / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D010446:Peptide Fragments / Q000737:chemistry; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics*
D000596:Amino Acids; D010446:Peptide Fragments; D011994:Recombinant Proteins; D007522:Isocitrate Lyase