Biochemical characterization of two glutamate dehydrogenases with different cofactor specificities from a hyperthermophilic archaeon Pyrobaculum calidifontis

Taisuke Wakamatsu, Chisato Higashi, Taketo Ohmori, Katsumi Doi, Toshihisa Ohshima

研究成果: Contribution to journalArticle査読

4 被引用数 (Scopus)

抄録

Two putative glutamate dehydrogenase (GDH) genes (pcal_1031 and pcal_1606) were found in a sulfur-dependent hyperthermophilic archaeon, Pyrobaculum calidifontis. The two genes were then expressed in Escherichia coli, and both of the recombinant gene products showed GDH activity. The two enzymes were then purified to homogeneity and characterized in detail. Although both purified GDHs had a hexameric structure and neither exhibited allosteric regulation, they showed different coenzyme specificities: one was specific for NAD+, the other for NADP+ and different heat activation mechanisms. In addition, there was little difference in the kinetic constants, optimal temperature, thermal stability, optimal pH and pH stability between the two enzymes. The overall sequence identity between the two proteins was very high (81 %), but was not high in the region recognizing the 2′ position of the adenine ribose moiety, which is responsible for coenzyme specificity. This is the first report on the identification of two GDHs with different coenzyme specificities from a single hyperthermophilic archaeon and the definition of their basic in vitro properties.

本文言語英語
ページ(範囲)379-389
ページ数11
ジャーナルExtremophiles
17
3
DOI
出版ステータス出版済み - 5 2013

All Science Journal Classification (ASJC) codes

  • 微生物学
  • 分子医療

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