Identification and functional analysis of the genes encoding dibenzothiophene-desulfurizing enzymes from thermophilic bacteria

Kotaro Kirimura, Koji Harada, Hidekazu Iwasawa, Takeomi Tanaka, Yuichiro Iwasaki, Toshiki Furuya, Yoshitaka Ishii, Kuniki Kino

    Research output: Contribution to journalArticle

    34 Citations (Scopus)

    Abstract

    Thermophilic bacteria Bacillus subtilis WU-S2B and Mycobacterium phlei WU-F1 desulfurize dibenzothiophene (DBT) and alkylated DBTs through specific cleavage of the carbon-sulfur bonds over a temperature range up to 52°C. In order to identify and functionally analyze the DBT-desulfurization genes, the gene cluster containing bdsA, bdsB, and bdsC was cloned from B. subtilis WU-S2B. The nucleotide and amino acid sequences of bdsABC show homologies to those of the other known DBT-desulfurization genes and enzymes; e.g. a nucleotide sequence homo logy of 61.0% to dszABC of the mesophilic bacterium Rhodococcus sp. IGTS8 and 57.8% to tdsABC of the thermophilic bacterium Paenibacillus sp. A11-2. Deletion and subcloning analysis of bdsABC revealed that the gene products of bdsC, bdsA and bdsB oxidized DBT to DBT sulfone (DBTO2), converted DBTO2 to 2′-hydroxybiphenyl-2-sulfinate (HBPSi), and desulfurized HBPSi to 2-hydroxybiphenyl (2-HBP), respectively. Resting cells of a recombinant Escherichia coli JM109 harboring bdsABC converted DBT to 2-HBP over a temperature range of 30-52°C, indicating that the gene products of bdsABC were functional in the recombinant. The activities of DBT degradation at 50°C and DBT desulfurization (2-HBP production) at 40°C in resting cells of the recombinant were approximately five times and twice, respectively, as high as those in B. subtilis WU-S2B. The recombinant E. coli cells also degraded alkylated DBTs, such as 2,8-dimethylDBT and 4,6-dimethylDBT. The nucleotide sequences of B. subtilis WU-S2B bdsABC and the corresponding genes from M. phlei WU-F1 were found to be completely identical to each other although the strains are genetically different.

    Original languageEnglish
    Pages (from-to)703-713
    Number of pages11
    JournalApplied Microbiology and Biotechnology
    Volume65
    Issue number6
    DOIs
    Publication statusPublished - 2004 Nov

    Fingerprint

    Functional analysis
    Gene encoding
    Desulfurization
    Bacteria
    Enzymes
    Genes
    Bacillus subtilis
    Nucleotides
    Mycobacterium phlei
    Escherichia coli
    Paenibacillus
    Rhodococcus
    Bacilli
    Temperature
    Multigene Family
    dibenzothiophene
    Amino acids
    Sulfur
    Amino Acid Sequence
    Carbon

    ASJC Scopus subject areas

    • Biotechnology
    • Microbiology
    • Bioengineering
    • Microbiology (medical)

    Cite this

    Identification and functional analysis of the genes encoding dibenzothiophene-desulfurizing enzymes from thermophilic bacteria. / Kirimura, Kotaro; Harada, Koji; Iwasawa, Hidekazu; Tanaka, Takeomi; Iwasaki, Yuichiro; Furuya, Toshiki; Ishii, Yoshitaka; Kino, Kuniki.

    In: Applied Microbiology and Biotechnology, Vol. 65, No. 6, 11.2004, p. 703-713.

    Research output: Contribution to journalArticle

    Kirimura, Kotaro ; Harada, Koji ; Iwasawa, Hidekazu ; Tanaka, Takeomi ; Iwasaki, Yuichiro ; Furuya, Toshiki ; Ishii, Yoshitaka ; Kino, Kuniki. / Identification and functional analysis of the genes encoding dibenzothiophene-desulfurizing enzymes from thermophilic bacteria. In: Applied Microbiology and Biotechnology. 2004 ; Vol. 65, No. 6. pp. 703-713.
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    abstract = "Thermophilic bacteria Bacillus subtilis WU-S2B and Mycobacterium phlei WU-F1 desulfurize dibenzothiophene (DBT) and alkylated DBTs through specific cleavage of the carbon-sulfur bonds over a temperature range up to 52°C. In order to identify and functionally analyze the DBT-desulfurization genes, the gene cluster containing bdsA, bdsB, and bdsC was cloned from B. subtilis WU-S2B. The nucleotide and amino acid sequences of bdsABC show homologies to those of the other known DBT-desulfurization genes and enzymes; e.g. a nucleotide sequence homo logy of 61.0{\%} to dszABC of the mesophilic bacterium Rhodococcus sp. IGTS8 and 57.8{\%} to tdsABC of the thermophilic bacterium Paenibacillus sp. A11-2. Deletion and subcloning analysis of bdsABC revealed that the gene products of bdsC, bdsA and bdsB oxidized DBT to DBT sulfone (DBTO2), converted DBTO2 to 2′-hydroxybiphenyl-2-sulfinate (HBPSi), and desulfurized HBPSi to 2-hydroxybiphenyl (2-HBP), respectively. Resting cells of a recombinant Escherichia coli JM109 harboring bdsABC converted DBT to 2-HBP over a temperature range of 30-52°C, indicating that the gene products of bdsABC were functional in the recombinant. The activities of DBT degradation at 50°C and DBT desulfurization (2-HBP production) at 40°C in resting cells of the recombinant were approximately five times and twice, respectively, as high as those in B. subtilis WU-S2B. The recombinant E. coli cells also degraded alkylated DBTs, such as 2,8-dimethylDBT and 4,6-dimethylDBT. The nucleotide sequences of B. subtilis WU-S2B bdsABC and the corresponding genes from M. phlei WU-F1 were found to be completely identical to each other although the strains are genetically different.",
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    AU - Tanaka, Takeomi

    AU - Iwasaki, Yuichiro

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