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KMID : 1094720130180050897
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Biotechnology and Bioprocess Engineering
2013 Volume.18 No. 5 p.897 ~ p.902
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Heterologous expression and characterization of a recombinant thermophilic arylsulfatase from Thermotoga maritima
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Lee Dong-Geun
Shin Jae-Gyun
Jeon Myong-Je
Lee Sang-Hyeon
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Abstract
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Production of low sulfated agar or agarose from agar or agaropectins by enzymatic hydrolysis has advantages but a high melting temperature is needed. The arylsulfatase gene from thermophilic Thermotoga maritima was cloned and expressed in Escherichia coli W3110 with pCol-MICT as the vector. The gene was comprised of 1,782 bp and encoded a protein of 593 amino acids with a molecular weight of 65 kDa. The recombinant arylsulfatase was partially purified by heat treatment (70°C, 30 min) and characterized. The enzyme was prepared with a total protein content of 2.4 mg and a specific activity of 20.63 U/mg. Optimal temperature and pH of the enzyme were 80°C and 7.0, respectively, for hydrolysis of p-nitrophenyl sulfate and sulfate content of agar was diminished to 40% after a 12 h treatment at that condition. Enhanced electrophoretic movement of DNA was observed in enzymetreated agar gel compared to that in a non-treated agar gel. These results suggest that thermophilic arylsulfatase expressed in E. coli could be useful for producing a low sulfated agar and electrophoretic grade agarose.
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KEYWORD
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agar, agarose, arylsulfatase, thermophilic, Thermotoga maritima
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