O 2-stable membrane-bound [NiFe]hydrogenase from a newly isolated Citrobacter sp. S-77

Shigenobu Eguchi, Ki Suk Yoon, Seiji Ogo

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Hydrogenases are of great interest due to their potential use in H 2-based technology. However, most hydrogenases are highly sensitive to O 2, which have been the major bottleneck in hydrogenase studies. Here we report an O 2-stable membrane-bound [NiFe]hydrogenase (MBH) purified from a newly isolated strain, S-77. According to the 16S rRNA gene sequence and phylogenetic analysis of the strain S-77, it belongs to the genus of Citrobacter. In vitro experiments using the cytoplasmic membrane of strain S-77 suggested that a cytochrome b acts as the physiological electron acceptor of the MBH. The purified MBH was composed of a dimer of heterodimers, consisting of two distinct subunits with the molecular weights of 58.5 and 38.5 kDa. The enzyme showed a specific activity for H 2-oxidation of 661U/mg, which is 35-fold greater than that for H 2-production of 18.7U/mg. Notably, the MBH showed a remarkable O 2-stability, maintaining almost 95% of its original activity even after incubation for 30 h in air at 4°C. These results suggest that the O 2-stable MBH may play an important role in the H 2-metabolic pathway under the aerobic conditions of Citrobacter sp. S-77. This is the first report of the purification and biochemical characterization of an O 2-stable MBH from the genus of Citrobacter.

Original languageEnglish
Pages (from-to)479-484
Number of pages6
JournalJournal of Bioscience and Bioengineering
Volume114
Issue number5
DOIs
Publication statusPublished - Nov 1 2012

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Citrobacter
Hydrogenase
Membranes
Dimers
Purification
Cytochromes b
Enzymes
Genes
Molecular weight
Metabolic Networks and Pathways
Proteins
rRNA Genes
Oxidation
Sequence Analysis
Electrons
Air
Molecular Weight
Cell Membrane
Technology
Experiments

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Applied Microbiology and Biotechnology
  • Bioengineering

Cite this

O 2-stable membrane-bound [NiFe]hydrogenase from a newly isolated Citrobacter sp. S-77. / Eguchi, Shigenobu; Yoon, Ki Suk; Ogo, Seiji.

In: Journal of Bioscience and Bioengineering, Vol. 114, No. 5, 01.11.2012, p. 479-484.

Research output: Contribution to journalArticle

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