Isolation and characterization of genes responsible for naphthalene degradation from thermophilic naphthalene degrader, Geobacillus sp. JF8

Daisuke Miyazawa, Le Thi Ha Thanh, Akio Tani, Masaki Shintani, Nguyen Hoang Loc, Takashi Hatta, Kazuhide Kimbara

Research output: Contribution to journalArticle

Abstract

Geobacillus sp. JF8 is a thermophilic biphenyl and naphthalene degrader. To identify the naphthalene degradation genes, cis-naphthalene dihydrodiol dehydrogenase was purified from naphthalene-grown cells, and its N-terminal amino acid sequence was determined. Using a DNA probe encoding the N-terminal region of the dehydrogenase, a 10-kb DNA fragment was isolated. Upstream of nahB, a gene for dehydrogenase, there were two open reading frames which were designated as nahAc and nahAd, respectively. The products of nahAc and nahAd were predicted to be alpha and beta subunit of ring-hydroxylating dioxygenases, respectively. Phylogenetic analysis of amino acid sequences of NahB indicated that it did not belong to the cis-dihydrodiol dehydrogenase group that includes those of classical naphthalene degradation pathways. Downstream of nahB, four open reading frames were found, and their products were predicted as meta-cleavage product hydrolase, monooxygenase, dehydrogenase, and gentisate 1,2-dioxygenase, respectively. A reverse transcriptase-PCR analysis showed that transcription of nahAcAd was induced by naphthalene. These findings indicate that we successfully identified genes involved in the upper pathway of naphthalene degradation from a thermophilic bacterium.

Original languageEnglish
Article number44
JournalMicroorganisms
Volume8
Issue number1
DOIs
Publication statusPublished - Jan 2020

Fingerprint

Geobacillus
Genes
Oxidoreductases
gentisate 1,2-dioxygenase
Open Reading Frames
Dioxygenases
Protein Sequence Analysis
DNA Probes
Hydrolases
Mixed Function Oxygenases
Reverse Transcriptase Polymerase Chain Reaction
naphthalene
Amino Acid Sequence
Bacteria
DNA

Keywords

  • Geobacillus
  • Naphthalene
  • Thermophile

ASJC Scopus subject areas

  • Microbiology
  • Virology
  • Microbiology (medical)

Cite this

Isolation and characterization of genes responsible for naphthalene degradation from thermophilic naphthalene degrader, Geobacillus sp. JF8. / Miyazawa, Daisuke; Thanh, Le Thi Ha; Tani, Akio; Shintani, Masaki; Loc, Nguyen Hoang; Hatta, Takashi; Kimbara, Kazuhide.

In: Microorganisms, Vol. 8, No. 1, 44, 01.2020.

Research output: Contribution to journalArticle

Miyazawa, Daisuke ; Thanh, Le Thi Ha ; Tani, Akio ; Shintani, Masaki ; Loc, Nguyen Hoang ; Hatta, Takashi ; Kimbara, Kazuhide. / Isolation and characterization of genes responsible for naphthalene degradation from thermophilic naphthalene degrader, Geobacillus sp. JF8. In: Microorganisms. 2020 ; Vol. 8, No. 1.
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