Dibenzothiophene desulfurizing enzymes from moderately thermophilic bacterium Bacillus subtilis WU-S2B: Purification, characterization and overexpression

Takashi Ohshiro, Yoshitaka Ishii, Toshiyuki Matsubara, Koichi Ueda, Yoshikazu Izumi, Kuniki Kino, Kotaro Kirimura

    Research output: Contribution to journalArticle

    26 Citations (Scopus)

    Abstract

    The moderately thermophilic bacterium Bacillus subtilis WU-S2B desulfurized dibenzothiophene (DBT) at 50°C through the selective cleavage of carbon-sulfur bonds. In this study, three enzymes involved in the microbial DBT desulfurization were purified and characterized. The first two enzymes, DBT monooxygenase (BdsC) and DBT sulfone monooxygenase (BdsA), were purified from the wild-type strain, and the last one, 2′-hydroxybiphenyl 2-sulfinic acid desulfinase (BdsB), was purified from the recombinant Escherichia coli overexpressing the gene, bdsB, with chaperonin genes, groEL/ES. The genes of BdsC and BdsA were also overexpressed. The molecular weights of BdsC and BdsA were determined to be 200 and 174 kDa, respectively, by gel filtration chromatography, suggesting that both enzymes had four identical subunits. BdsB had a monomeric structure of 40 kDa. The three enzymes were characterized and compared with the corresponding enzymes (DszC, DszA, and DszB) of mesophilic desulfurization bacteria. The specific activities of BdsC, BdsA, and BdsB were 84.2, 855, and 280 units/mg, respectively, and the latter two activities were higher than those of DszA and DszB. The heat stability and optimum temperature of BdsC, BdsA, and BdsB were higher than those of DszC, DszA, and DszB. Other enzymatic properties were investigated in detail.

    Original languageEnglish
    Pages (from-to)266-273
    Number of pages8
    JournalJournal of Bioscience and Bioengineering
    Volume100
    Issue number3
    DOIs
    Publication statusPublished - 2005

    Fingerprint

    Bacilli
    Desulfurization
    Bacillus subtilis
    Purification
    Bacteria
    Enzymes
    Genes
    Sulfinic Acids
    Chaperonins
    Chromatography
    Sulfur
    Escherichia coli
    Gel Chromatography
    Carbon
    Gels
    Hot Temperature
    Molecular Weight
    Molecular weight
    dibenzothiophene
    dibenzothiophene sulfone monooxygenase

    Keywords

    • Bacillus
    • Desulfinase
    • Desulfurization
    • Dibenzothiophene
    • Monooxygenase

    ASJC Scopus subject areas

    • Biotechnology
    • Bioengineering

    Cite this

    Dibenzothiophene desulfurizing enzymes from moderately thermophilic bacterium Bacillus subtilis WU-S2B : Purification, characterization and overexpression. / Ohshiro, Takashi; Ishii, Yoshitaka; Matsubara, Toshiyuki; Ueda, Koichi; Izumi, Yoshikazu; Kino, Kuniki; Kirimura, Kotaro.

    In: Journal of Bioscience and Bioengineering, Vol. 100, No. 3, 2005, p. 266-273.

    Research output: Contribution to journalArticle

    @article{50d649ba43324cd68c7624f71033e26b,
    title = "Dibenzothiophene desulfurizing enzymes from moderately thermophilic bacterium Bacillus subtilis WU-S2B: Purification, characterization and overexpression",
    abstract = "The moderately thermophilic bacterium Bacillus subtilis WU-S2B desulfurized dibenzothiophene (DBT) at 50°C through the selective cleavage of carbon-sulfur bonds. In this study, three enzymes involved in the microbial DBT desulfurization were purified and characterized. The first two enzymes, DBT monooxygenase (BdsC) and DBT sulfone monooxygenase (BdsA), were purified from the wild-type strain, and the last one, 2′-hydroxybiphenyl 2-sulfinic acid desulfinase (BdsB), was purified from the recombinant Escherichia coli overexpressing the gene, bdsB, with chaperonin genes, groEL/ES. The genes of BdsC and BdsA were also overexpressed. The molecular weights of BdsC and BdsA were determined to be 200 and 174 kDa, respectively, by gel filtration chromatography, suggesting that both enzymes had four identical subunits. BdsB had a monomeric structure of 40 kDa. The three enzymes were characterized and compared with the corresponding enzymes (DszC, DszA, and DszB) of mesophilic desulfurization bacteria. The specific activities of BdsC, BdsA, and BdsB were 84.2, 855, and 280 units/mg, respectively, and the latter two activities were higher than those of DszA and DszB. The heat stability and optimum temperature of BdsC, BdsA, and BdsB were higher than those of DszC, DszA, and DszB. Other enzymatic properties were investigated in detail.",
    keywords = "Bacillus, Desulfinase, Desulfurization, Dibenzothiophene, Monooxygenase",
    author = "Takashi Ohshiro and Yoshitaka Ishii and Toshiyuki Matsubara and Koichi Ueda and Yoshikazu Izumi and Kuniki Kino and Kotaro Kirimura",
    year = "2005",
    doi = "10.1263/jbb.100.266",
    language = "English",
    volume = "100",
    pages = "266--273",
    journal = "Journal of Bioscience and Bioengineering",
    issn = "1389-1723",
    publisher = "Elsevier",
    number = "3",

    }

    TY - JOUR

    T1 - Dibenzothiophene desulfurizing enzymes from moderately thermophilic bacterium Bacillus subtilis WU-S2B

    T2 - Purification, characterization and overexpression

    AU - Ohshiro, Takashi

    AU - Ishii, Yoshitaka

    AU - Matsubara, Toshiyuki

    AU - Ueda, Koichi

    AU - Izumi, Yoshikazu

    AU - Kino, Kuniki

    AU - Kirimura, Kotaro

    PY - 2005

    Y1 - 2005

    N2 - The moderately thermophilic bacterium Bacillus subtilis WU-S2B desulfurized dibenzothiophene (DBT) at 50°C through the selective cleavage of carbon-sulfur bonds. In this study, three enzymes involved in the microbial DBT desulfurization were purified and characterized. The first two enzymes, DBT monooxygenase (BdsC) and DBT sulfone monooxygenase (BdsA), were purified from the wild-type strain, and the last one, 2′-hydroxybiphenyl 2-sulfinic acid desulfinase (BdsB), was purified from the recombinant Escherichia coli overexpressing the gene, bdsB, with chaperonin genes, groEL/ES. The genes of BdsC and BdsA were also overexpressed. The molecular weights of BdsC and BdsA were determined to be 200 and 174 kDa, respectively, by gel filtration chromatography, suggesting that both enzymes had four identical subunits. BdsB had a monomeric structure of 40 kDa. The three enzymes were characterized and compared with the corresponding enzymes (DszC, DszA, and DszB) of mesophilic desulfurization bacteria. The specific activities of BdsC, BdsA, and BdsB were 84.2, 855, and 280 units/mg, respectively, and the latter two activities were higher than those of DszA and DszB. The heat stability and optimum temperature of BdsC, BdsA, and BdsB were higher than those of DszC, DszA, and DszB. Other enzymatic properties were investigated in detail.

    AB - The moderately thermophilic bacterium Bacillus subtilis WU-S2B desulfurized dibenzothiophene (DBT) at 50°C through the selective cleavage of carbon-sulfur bonds. In this study, three enzymes involved in the microbial DBT desulfurization were purified and characterized. The first two enzymes, DBT monooxygenase (BdsC) and DBT sulfone monooxygenase (BdsA), were purified from the wild-type strain, and the last one, 2′-hydroxybiphenyl 2-sulfinic acid desulfinase (BdsB), was purified from the recombinant Escherichia coli overexpressing the gene, bdsB, with chaperonin genes, groEL/ES. The genes of BdsC and BdsA were also overexpressed. The molecular weights of BdsC and BdsA were determined to be 200 and 174 kDa, respectively, by gel filtration chromatography, suggesting that both enzymes had four identical subunits. BdsB had a monomeric structure of 40 kDa. The three enzymes were characterized and compared with the corresponding enzymes (DszC, DszA, and DszB) of mesophilic desulfurization bacteria. The specific activities of BdsC, BdsA, and BdsB were 84.2, 855, and 280 units/mg, respectively, and the latter two activities were higher than those of DszA and DszB. The heat stability and optimum temperature of BdsC, BdsA, and BdsB were higher than those of DszC, DszA, and DszB. Other enzymatic properties were investigated in detail.

    KW - Bacillus

    KW - Desulfinase

    KW - Desulfurization

    KW - Dibenzothiophene

    KW - Monooxygenase

    UR - http://www.scopus.com/inward/record.url?scp=28744452205&partnerID=8YFLogxK

    UR - http://www.scopus.com/inward/citedby.url?scp=28744452205&partnerID=8YFLogxK

    U2 - 10.1263/jbb.100.266

    DO - 10.1263/jbb.100.266

    M3 - Article

    C2 - 16243275

    AN - SCOPUS:28744452205

    VL - 100

    SP - 266

    EP - 273

    JO - Journal of Bioscience and Bioengineering

    JF - Journal of Bioscience and Bioengineering

    SN - 1389-1723

    IS - 3

    ER -