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 journalArticle

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
Publication statusPublished - Sep 1997
Externally publishedYes

Fingerprint

Acetyl-CoA C-Acetyltransferase
Candida tropicalis
Alkanes
Candida
Yeast
Isoenzymes
Genes
Yeasts
Gene encoding
Saccharomyces cerevisiae Proteins
Molecular mass
Peroxisomes
Fungal Proteins
Recombinant Proteins
Chemical properties
Protein Sequence Analysis
Amino Acids
Glucose
Kinetics
Enzymes

Keywords

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

ASJC Scopus subject areas

  • Biochemistry

Cite this

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. / Kanayama, Naoki; Himeda, Yasukazu; Atomi, Haruyuki; Ueda, Mitsuyoshi; Tanaka, Atsuo.

In: Journal of Biochemistry, Vol. 122, No. 3, 09.1997, p. 616-621.

Research output: Contribution to journalArticle

@article{eab4fb3f13ab465397137270d86bb85d,
title = "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",
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.",
keywords = "Acetoacetyl-CoA thiolase, Candida tropicalis, Cytosol, Isozyme, Peroxisome",
author = "Naoki Kanayama and Yasukazu Himeda and Haruyuki Atomi and Mitsuyoshi Ueda and Atsuo Tanaka",
year = "1997",
month = "9",
language = "English",
volume = "122",
pages = "616--621",
journal = "Journal of Biochemistry",
issn = "0021-924X",
publisher = "Oxford University Press",
number = "3",

}

TY - JOUR

T1 - Expression of Acetoacetyl-CoA thiolase isozyme genes of n-alkane-assimilating yeast, candida tropicalis

T2 - Isozymes in two intracellular compartments are derived from the same genes

AU - Kanayama, Naoki

AU - Himeda, Yasukazu

AU - Atomi, Haruyuki

AU - Ueda, Mitsuyoshi

AU - Tanaka, Atsuo

PY - 1997/9

Y1 - 1997/9

N2 - 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.

AB - 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.

KW - Acetoacetyl-CoA thiolase

KW - Candida tropicalis

KW - Cytosol

KW - Isozyme

KW - Peroxisome

UR - http://www.scopus.com/inward/record.url?scp=0030838674&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0030838674&partnerID=8YFLogxK

M3 - Article

C2 - 9348092

AN - SCOPUS:0030838674

VL - 122

SP - 616

EP - 621

JO - Journal of Biochemistry

JF - Journal of Biochemistry

SN - 0021-924X

IS - 3

ER -