Analysis of genes involved in nitrate reduction in Clostridium perfringens

Katsuyo Fujinaga, Yuki Taniguchi, Yezhou Sun, Seiichi Katayama, Junzaburo Minami, Osamu Matsushita, Akinobu Okabe

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

We have conducted the genetic analysis of fermentative nitrate reduction in Clostridium perfringens, a strict anaerobic bacterium. Nitrate reductase (NarA) was purified from the cytoplasmic fraction of the organism. Using a degenerate primer designed from its N-terminal amino acid sequence, a 9.5 kb fragment containing seven ORFs was cloned. The molecular mass and N-terminal amino acid sequence predicted from the nucleotide sequence of ORF 4 coincided with those determined for the purified NarA, indicating that ORF 4 corresponds to a narA gene. ORFs 5 and 6 encode a 15.4 kDa ferredoxin-like protein containing four iron-sulfur clusters and a 45 kDa protein homologous to NADH oxidase, respectively. Analyses involving primer extension and Northern blotting revealed that these three ORFs are transcribed as a polycistronic message. The ORF 5- and ORF 6-encoded proteins were shown by immunablotting to be synthesized by cells grown in the presence of nitrate. Thus, these two proteins are likely to function as electron-transfer components in nitrate reduction in C. perfringens. The 9.5 kb fragment and a downstream region of 6.1 kb do not contain any genes involved in nitrate uptake or nitrite reduction. Instead, all 5 ORFs downstream of ORF 6 are homologous to genes reported for molybdopterin biosynthesis, unlike the genomic organization already determined for the respiratory and assimilatory nitrate-reduction systems. The evolutionary relationships between these two nitrate-reduction systems and the fermentative one based on the results of comparative genetic analysis are discussed.

Original languageEnglish
Pages (from-to)3377-3387
Number of pages11
JournalMicrobiology
Volume145
Issue number12
Publication statusPublished - Dec 1999
Externally publishedYes

Fingerprint

Clostridium perfringens
Nitrates
Open Reading Frames
Genes
Nitrate Reductase
Amino Acid Sequence
Proteins
Ferredoxins
Anaerobic Bacteria
Nitrites
Sulfur
Northern Blotting
Iron
Electrons

Keywords

  • Clostridium perfringens
  • Ferredoxin-like protein
  • Molybdopterin guanine dinucleotide
  • NADH oxidase
  • Nitrate reductase

ASJC Scopus subject areas

  • Microbiology

Cite this

Fujinaga, K., Taniguchi, Y., Sun, Y., Katayama, S., Minami, J., Matsushita, O., & Okabe, A. (1999). Analysis of genes involved in nitrate reduction in Clostridium perfringens. Microbiology, 145(12), 3377-3387.

Analysis of genes involved in nitrate reduction in Clostridium perfringens. / Fujinaga, Katsuyo; Taniguchi, Yuki; Sun, Yezhou; Katayama, Seiichi; Minami, Junzaburo; Matsushita, Osamu; Okabe, Akinobu.

In: Microbiology, Vol. 145, No. 12, 12.1999, p. 3377-3387.

Research output: Contribution to journalArticle

Fujinaga, K, Taniguchi, Y, Sun, Y, Katayama, S, Minami, J, Matsushita, O & Okabe, A 1999, 'Analysis of genes involved in nitrate reduction in Clostridium perfringens', Microbiology, vol. 145, no. 12, pp. 3377-3387.
Fujinaga K, Taniguchi Y, Sun Y, Katayama S, Minami J, Matsushita O et al. Analysis of genes involved in nitrate reduction in Clostridium perfringens. Microbiology. 1999 Dec;145(12):3377-3387.
Fujinaga, Katsuyo ; Taniguchi, Yuki ; Sun, Yezhou ; Katayama, Seiichi ; Minami, Junzaburo ; Matsushita, Osamu ; Okabe, Akinobu. / Analysis of genes involved in nitrate reduction in Clostridium perfringens. In: Microbiology. 1999 ; Vol. 145, No. 12. pp. 3377-3387.
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