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Purification and characterization of arylsulfatase from Sphingomonas sp. AS6330

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Abstract

Arylsulfatase was purified from Sphingomonas sp. AS6330 through ionic exchange, hydrophobic- and gel-chromatographies. The purity increased 12,800-fold with approximately 19.1% yield against cell homogenate. The enzyme was a monomeric protein with apparent molecular weight of 62 kDa as determined by sodium dodecylsulfate-polyacrylamide gel electrophoresis, and 41 kDa as determined by gel filtration. The enzyme had optimum reaction conditions for hydrolysis of sulfate ester bonds in agar and p-nitrophenyl sulfate (NPS) at pH 7.0 and 45°C, with a specific activity of 3.93 and 97.2 U, respectively. The enzyme showed higher activity towards agar than other sulfated marine polysaccharides such as porphyran, fucoidan and carrageenan. The K m and V max of the enzyme for hydrolysis of NPS were 54.9 μM and 113 mM/min, respectively. With reaction of 200 g agar with 100 U arylsulfatase for 8 h at 45°C, gel strength increased 2.44-fold, and 97.7% of the sulfate in the agar was hydrolyzed.

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Acknowledgement

This work was supported by Korea Research Foundation Grant (KRF-2001-015-HP0001).

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Authors and Affiliations

  1. Division of Food Science and Biotechnology, Pukyong National University, 608–737, Busan , South Korea

    D.-S. Byun, J.-S. Choi & H.-R. Kim

  2. Department of Biological Science, Pusan National University, 609–735, Busan , South Korea

    J.-H. Kim & W.-C. Choi

  3. Department of Food Science, Louisiana State University, Baton Rouge, LA 70803, USA

    J. S. Godber

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  1. J.-H. Kim
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  2. D.-S. Byun
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  3. J. S. Godber
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  4. J.-S. Choi
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  5. W.-C. Choi
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  6. H.-R. Kim
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Correspondence to H.-R. Kim.

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Kim, JH., Byun, DS., Godber, J.S. et al. Purification and characterization of arylsulfatase from Sphingomonas sp. AS6330. Appl Microbiol Biotechnol 63, 553–559 (2004). https://doi.org/10.1007/s00253-003-1463-8

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  • DOI: https://doi.org/10.1007/s00253-003-1463-8

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