Expression of Acetoacetyl-CoA thiolase isozyme genes of n-alkane-assimilating yeast, candida tropicalis: Isozymes in two intracellular compartments are derived from the same genes

Naoki Kanayama, Yasukazu Himeda, Haruyuki Atomi, Mitsuyoshi Ueda, Atsuo Tanaka

Research output: Contribution to journalArticlepeer-review

9 Citations (Scopus)

Abstract

In the n-alkane-assimilating yeast Candida tropicalis, there are two isozymes of acetoacetyl-CoA thiolase, peroxisomal acetoacetyl-CoA thiolase (peroxisomal Thiolase I), and cytosolic acetoacetyl-CoA thiolase (cytosolic Thiolase I). We have previously isolated two genes (CT-T1A and CT-T1B) which encode Thiolase I. In order to compare the expressed products of Thiolase I isozyme-encoding genes in C. tropicalis, cytosolic Thiolase I was first purified from glucose-grown C. tropicalis in which the proliferation of peroxisomes and the expression of peroxisomal Thiolase I were repressed. Cytosolic Thiolase I was virtually identical to peroxisomal Thiolase I in molecular mass, kinetic and immuno-chemical properties, and primary structure at the N-terminus. Amino acid sequence analysis revealed that cytosolic Thiolase I was the mixture of products of two genes (CT-T1A and CT-T1B), as in the case of the peroxisomal enzyme. CT-T1A and CT-T1B were expressed independently in the yeast Saccharomyces cerevisiae and the recombinant proteins were purified. Recombinant Thiolase IA and IB exhibited practically identical enzymatic properties to cytosolic and peroxisomal Thiolase Is from C. tropicalis. These results revealed that cytosolic Thiolase I and peroxisomal Thiolase I were encoded not by different genes, but by the same genes (CT-T1A and CT-T1B) and are present as a mixture of products expressed by both genes, although their subcellular localizations are different.

Original languageEnglish
Pages (from-to)616-621
Number of pages6
JournalJournal of biochemistry
Volume122
Issue number3
DOIs
Publication statusPublished - Sep 1997
Externally publishedYes

Keywords

  • Acetoacetyl-CoA thiolase
  • Candida tropicalis
  • Cytosol
  • Isozyme
  • Peroxisome

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

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