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Identification of the first archaeal arylsulfatase from Pyrococcus furiosus and its application to desulfatation of agar

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Abstract

A gene encoding a putative arylsulfatase from the hyperthermophilic archaeon Pyrococcus furiosus was identified, cloned, and expressed as a fusion protein with a Sce VMA intein and chitin binding domain (CBD) residue. The gene (PF1345) from P. furiosus encoding a 35 kDa protein showed some similarity (17 ∼ 19%) with other arylsulfatases from the bacteria. The recombinant fusion arylsulfatase was overexpressed in E. coli and partially purified. Its molecular mass was estimated to be 90 kDa by SDS-PAGE. The optimal temperature and pH for arylsulfatase activity were found to be 45°C and 9.5, respectively. Various divalent cations (Ca2+, Mg2+, Co2+, Cu2+, Zn2+, and Mn2+) slightly activated the arylsulfatase activity in a narrow range of concentrations (below 0.5 mM), whereas Zn2+ concentrations above 2.0 mM significantly inhibited the activity. After the reaction of agar with recombinant fusion arylsulfatase for 12 h at 50°C, 75% of the sulfate in the agar was removed, and the DNA migration was greatly enhanced. Therefore, the arylsulfatase in this study could be applicable for the production of electrophoretic grade agarose by removing sulfate groups in agar.

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Authors and Affiliations

  1. Department of Biomaterial Control (BK21 program), Dong-Eui University, Busan, 614-714, Korea

    Keong-Tsul Jung, Soo-Wan Nam, Byung-Woo Kim & Sung-Jong Jeon

  2. Department of Biotechnology & Bioengineering, Dong-Eui University, Busan, 614-714, Korea

    Soo-Wan Nam & Sung-Jong Jeon

  3. Department of Life Science and Biotechnology, Dong-Eui University, Busan, 614-714, Korea

    Byung-Woo Kim

  4. Blue-Bio Industry RIC, Dong-Eui University, Busan, 614-714, Korea

    Soo-Wan Nam, Byung-Woo Kim & Sung-Jong Jeon

  5. Division of Life Sciences, Korea Polar Research Institute, Incheon, 406-840, Korea

    Han-Woo Kim

  6. Division of Electron Microscopic Research, Korea Basic Science Institute (KBSI), Daejeon, 305-333, Korea

    Dong-Ju You

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  1. Keong-Tsul Jung
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Correspondence to Sung-Jong Jeon.

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Jung, KT., Kim, HW., You, DJ. et al. Identification of the first archaeal arylsulfatase from Pyrococcus furiosus and its application to desulfatation of agar. Biotechnol Bioproc E 17, 1140–1146 (2012). https://doi.org/10.1007/s12257-012-0228-6

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  • DOI: https://doi.org/10.1007/s12257-012-0228-6

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