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Applied and Environmental Microbiology, June 2001, p. 2476-2483, Vol. 67, No. 6
Institute of Biophysics, Biological Research
Center, Hungarian Academy of Sciences, and Department of
Biotechnology, University of Szeged, H-6726 Szeged, Hungary
Received 16 November 2000/Accepted 28 January 2001
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.
0099-2240/01/$04.00+0 DOI: 10.1128/AEM.67.6.2476-2483.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
Transposon Mutagenesis in Purple Sulfur Photosynthetic Bacteria:
Identification of hypF, Encoding a Protein Capable
of Processing [NiFe] Hydrogenases in
, 
, and 
Subdivisions of the Proteobacteria
*
Corresponding author. Mailing address: Department
of Biotechnology, University of Szeged, Temesvári krt. 62, H-6726 Szeged, Hungary. Phone: 36 62 544 351. Fax: 36 62 544 352. E-mail: kornel{at}nucleus.szbk.u-szeged.hu.
Applied and Environmental Microbiology, June 2001, p. 2476-2483, Vol. 67, No. 6
0099-2240/01/$04.00+0 DOI: 10.1128/AEM.67.6.2476-2483.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
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