전체메뉴
검색
Article Search

JMB Journal of Microbiolog and Biotechnology

QR Code QR Code

Related articles in JMB

More Related Articles

Article

J. Microbiol. Biotechnol. 2004; 14(4): 777-782

Published online August 28, 2004

Copyright © The Korean Society for Microbiology and Biotechnology.

Overexpression of Arylsulfatase in E. coli and Its Application to Desulfatation of Agar

Jae Myung Lim , Yeon Hwa Jang 1, Hyeung Rak Kim 2, Young Tae Kim 3, Tae Jin Choi 3, Joong Kyun Kim 4 and Soo Wan Nam 5*

Department of Biotechnology and Bioengineering Pukyong National University Busan 608-737 Korea, 1Department of Biomaterial Control Dong-Eui University Busan 614-714 Korea, 2Division of Food Science and Biotechnology Pukyong National University Busan 608-737 Korea, 3Department of Microbiology Pukyong National University Busan 608-737 Korea, 4Department of Biotechnology and Bioengineering Dong-Eui University Busan 614-714 Korea, 5Department of Biotechnology and Bioengineering, Dong-Eui University, Busan 614-714, Korea

Abstract

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.

Keywords: Agar,arylsulfatase,desulfatation,E.coli,overexpression,Pseudoalteromonascarrageenovora