Transposon Mutagenesis in Purple Sulfur Photosynthetic Bacteria: Identification of hypF, Encoding a Protein Capable of Processing [NiFe] Hydrogenases in α, β, and γ Subdivisions of the Proteobacteria

Barna Fodor, G. Rákhely, Ákos T. Kovács, K. Kovács

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37 Citations (Scopus)

Abstract

A random transposon-based mutagenesis system was optimized for the purple sulfur phototrophic bacterium Thiocapsa roseopersicina BBS. Screening for hydrogenase-deficient phenotypes resulted in the isolation of six independent mutants in a mini-Tn5 library. One of the mutations was in a gene showing high amino acid sequence similarity to HypF proteins in other organisms. Inactivation of hydrogen uptake activity in the hypF-deficient mutant resulted in a dramatic increase in the hydrogen evolution capacity of T. roseopersicina under nitrogen-fixing conditions. This mutant is therefore a promising candidate for use in practical biohydrogen-producing systems. The reconstructed hypF gene was able to complement the hypF-deficient mutant of T. roseopersicina BBS. Heterologous complementation experiments, using hypF mutant strains of T. roseopersicina, Escherichia coli, and Ralstonia eutropha and various hypF genes, were performed. They were successful in all of the cases tested, although for E. coli, the regulatory region of the foreign gene had to be replaced in order to achieve partial complementation. RT-PCR data suggested that HypF has no effect on the transcriptional regulation of the structural genes of hydrogenases in this organism.

Original languageEnglish
Pages (from-to)2476-2483
Number of pages8
JournalApplied and Environmental Microbiology
Volume67
Issue number6
DOIs
Publication statusPublished - Jun 2001

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Chromatiaceae
Thiocapsa roseopersicina
Proteobacteria
photosynthetic bacteria
mutagenesis
Mutagenesis
transposons
sulfur
mutants
bacterium
protein
gene
Hydrogenase
Genes
Proteins
proteins
hydrogen
Hydrogen
Cupriavidus necator
biohydrogen

ASJC Scopus subject areas

  • Biotechnology
  • Environmental Science(all)
  • Microbiology

Cite this

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title = "Transposon Mutagenesis in Purple Sulfur Photosynthetic Bacteria: Identification of hypF, Encoding a Protein Capable of Processing [NiFe] Hydrogenases in α, β, and γ Subdivisions of the Proteobacteria",
abstract = "A random transposon-based mutagenesis system was optimized for the purple sulfur phototrophic bacterium Thiocapsa roseopersicina BBS. Screening for hydrogenase-deficient phenotypes resulted in the isolation of six independent mutants in a mini-Tn5 library. One of the mutations was in a gene showing high amino acid sequence similarity to HypF proteins in other organisms. Inactivation of hydrogen uptake activity in the hypF-deficient mutant resulted in a dramatic increase in the hydrogen evolution capacity of T. roseopersicina under nitrogen-fixing conditions. This mutant is therefore a promising candidate for use in practical biohydrogen-producing systems. The reconstructed hypF gene was able to complement the hypF-deficient mutant of T. roseopersicina BBS. Heterologous complementation experiments, using hypF mutant strains of T. roseopersicina, Escherichia coli, and Ralstonia eutropha and various hypF genes, were performed. They were successful in all of the cases tested, although for E. coli, the regulatory region of the foreign gene had to be replaced in order to achieve partial complementation. RT-PCR data suggested that HypF has no effect on the transcriptional regulation of the structural genes of hydrogenases in this organism.",
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AU - Rákhely, G.

AU - Kovács, Ákos T.

AU - Kovács, K.

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