ISSN:
1432-072X
Keywords:
Candida tropicalis
;
Saccharomyces cerevisiae
;
Peroxisomes
;
Isocitrate lyase
;
GAL7 promoter
;
High level expression
Source:
Springer Online Journal Archives 1860-2000
Topics:
Biology
Notes:
Abstract 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.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/BF00245389
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