Purification of Pyruvate Dehydrogenase Complex from an Extreme Thermophile, Bacillus Caldolyticus, and Its Thermal Stability

Yasuaki Hiromasa, Yoichi Aso, Shoji Yamashita

Research output: Contribution to journalArticle

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Abstract

Pyruvate dehydrogenase multienzyme complex was purified from Bacillus caldolyticus. The complex was composed of four polypeptides with molecular masses of 39.8, 41.7, 53.7, and 57.5kDa estimated by SDS-PAGE and they were presumed to be pyruvate decarboxylase (E1, dimeric), lipoate acetyltransferase (E2), and lipoamide dehydrogenase (E3) on the analogy of those from Bacillus stearothermophilus. E1 and E3 were stable at pH 5.7-10.2 and 4.5-11.3, respectively. Halves of E1 and E3 activity were abolished by incubation for 30min at 65°C and 85°C, respectively. Loss of overall activity was principally due to inactivation of E1. Structural changes in the complex incubated at high temperature were studied by fluorescence spectroscopy. The results suggested that the thermal denaturation of the complex proceeded through at least two different steps: inactivations of E1 and E3, and the former process is accompanied by a reduction of the complex size.

Original languageEnglish
Pages (from-to)1062-1066
Number of pages5
JournalBioscience, Biotechnology, and Biochemistry
Volume57
Issue number7
DOIs
Publication statusPublished - Jan 1 1993
Externally publishedYes

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Dihydrolipoyllysine-Residue Acetyltransferase
Pyruvate Decarboxylase
Multienzyme Complexes
Dihydrolipoamide Dehydrogenase
Pyruvate Dehydrogenase Complex
Geobacillus stearothermophilus
Fluorescence Spectrometry
Bacilli
Bacillus
Purification
Polyacrylamide Gel Electrophoresis
Thermodynamic stability
Hot Temperature
Peptides
Denaturation
Temperature
Fluorescence spectroscopy
Molecular mass
Pyruvic Acid
Oxidoreductases

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Analytical Chemistry
  • Biochemistry
  • Applied Microbiology and Biotechnology
  • Molecular Biology
  • Organic Chemistry

Cite this

Purification of Pyruvate Dehydrogenase Complex from an Extreme Thermophile, Bacillus Caldolyticus, and Its Thermal Stability. / Hiromasa, Yasuaki; Aso, Yoichi; Yamashita, Shoji.

In: Bioscience, Biotechnology, and Biochemistry, Vol. 57, No. 7, 01.01.1993, p. 1062-1066.

Research output: Contribution to journalArticle

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