• Journal of Microbiology and Biotechnology
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• v.14 no.4
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• pp.777-782
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• 2004

The arylsulfatase gene (astA, 984 bp ORF) from the P. carrageenovora genome was amplified by PCR and subcloned into the pET21a vector. When the constructed plasmid pAST-A1 (6.4 kb) was introduced into E. coli BL21(DE3), the transformant on the LB plate containing IPTG showed a hydrolyzing activity for 4-methylumbelliferyl sulfate and p-nitrophenyl sulfate. The highest arylsulfatase activity (2.1 unit/ml) was obtained at 10 mM IPTG. Most arylsulfatase activity was found in the cell lysate, whereas no significant activity was detected in the culture supernatant. The molecular weight of the recombinant enzyme was estimated to be 33.1 kDa by SDS-PAGE. After the reaction of agar with arylsulfatase for 12 h at $40^{\circ}C$, the gel strength of the agar increased by 2-fold, and 73% of the sulfate in the agar had been removed. This result suggests that arylsulfatase expressed in E. coli could be useful in the production of electrophoretic grade agarose.

• Microbiology and Biotechnology Letters
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• v.38 no.4
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• pp.428-433
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• 2010

Enzymatic hydrolysis of sulfate groups in agaropectin or agar simplifies the production process of high-quality or low sulfate-content agarose. This study was investigated that cell surface displayed arylsulfatase can be applied to desulfatation of agar for production of agarose. Sulfate content of agarose prepared by treatment of yeast surface-displayed arylsulfatase was decreased in a enzyme dosedependent manner. Especially, 35 unit/mL of yeast surface arylsulfatase attenuated sulfate content of agarose up to 0.2%. In the 0.6% agar(Junsei), 35 unit/mL enzyme treated at $40^{\circ}C$ for 3 h showed the lowest content of sulfate. Therefore, this result was determined to be the optimal condition to desulfatation of agar for production of agarose. In addition, the gel strength of yeast surface arylsulfatase treated agar and commercial agarose were compared. Agarose prepared by treatment of yeast surface arylsulfatase showed $559.8{\pm}0.12$ of gel strength, and it is a similar compared to the commercial agarose.