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

  • Lim, Jae-Myung (Department of Biotechnology and Bioengineering, Pukyong National University) ;
  • Jang, Yeon-Hwa (Department of Biomaterial Control, Dong-Eui University) ;
  • Kim, Hyeung-Rak (Division of Food Science and Biotechnology, Pukyong National University) ;
  • Kim, Young-Tae (Department of Microbiology, Pukyong National University) ;
  • Choi, Tae-Jin (Department of Microbiology, Pukyong National University) ;
  • Kim, Joong-Kyun (Department of Biotechnology and Bioengineering, Pukyong National University) ;
  • Nam, Soo-Wan (Department of Biotechnology and Bioengineering, Dong-Eui University)
  • Published : 2004.08.01

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

References

  1. Carbohydr. Res. v.17 Marine polymers; A new procedure for the fractionation of agar Allan, G. G.;P. G. Johnson;Y. Lay;K. V. Sarkanen https://doi.org/10.1016/S0008-6215(00)81565-7
  2. J. Mol. Biol. v.90 The agarose double helix and its function in agarose gel structure Amott, S. https://doi.org/10.1016/0022-2836(74)90372-6
  3. Microbiology v.141 Arylsulphatase from Alteromonas carrageenovora Barbeyron, T.;P. Potin;C. Richard;O. Collin;B .Kloareg https://doi.org/10.1099/13500872-141-11-2897
  4. J. Mol. Biol. v.305 Crystal structure of an enzyme-substrate complex provides insight into the interaction between human arylsulfatase A and its substrates during catalysis Bulow, R. V.;B. Schmidt;T. Dierks;K. V. Figura;I. Uson https://doi.org/10.1006/jmbi.2000.4297
  5. J. Microbiol. Biotechnol. v.12 Rapid and simple method to prepare functional Pfu DNA polymerase expressed in Escherichia coli periplasm Chae, Y. K.;W. C. Jeon;K. S. Cho.
  6. Biochim. Biophys. Acta v.212 The isolation of arylsulphatse isoenzymes from Pseudomonas aeruginosa Delisle, G.;F. H. Milazzo https://doi.org/10.1016/0005-2744(70)90258-5
  7. Food Hydro. v.15 The influence of a mixed anionic system on the aggregation behavior of agarose Derbyshire, W;D. N. Hedges;P. J. lillford;I. T. Norton https://doi.org/10.1016/S0268-005X(00)00061-8
  8. J. Mar. Biotechnol. v.6 Arylsufatase in fish and shellfish of Veli Lake, South India Dhevendaran, K.;K. Maya
  9. Kor. J. Food Sci. Tech. v.31 Preparation of agarose from Gelidium amansii for gel electrophoresis using various purification methods and its resolution characteristics for DNA Do, J. R.;S. W. Oh
  10. Biochem. J. v.84 A note on the determination of the ester sulfate content of sulfated polysaccharides Dodgson, K. S.;R. G. Price
  11. Carbohydr. Res. v.16 The structure of agar. The fractionation of a complex mixture of polysaccharides Duckworth, M.;W. Yaphe https://doi.org/10.1016/S0008-6215(00)86113-3
  12. Eur. J. Biochem. v.221 An iducible arylsulfatase of Volvox carteri with properties suitable for a reporter-gene system: Purification, characterization and molecular cloning Hallmann, A.;M. Sumper https://doi.org/10.1111/j.1432-1033.1994.tb18723.x
  13. J. Bacteriol. v.139 Arylsulfatase in Salmonella typhymurium: Detection and influence of carbon source and tyramine on its synthesis Henderson, M. J.;F. H. Milazzo
  14. Dev. Biol. v.74 Arylsulfatase of sea-urchin sperm: Arylsulfatase as a lysine of sea-urchins Hoshi, M.;T. Moriya https://doi.org/10.1016/0012-1606(80)90436-4
  15. FEMS Microbiol. Rev. v.24 Riding the sulfur cycle - metabolism of sulfonates and sulfate esters in Gram-negative bacteria Kertesz, M. A.
  16. J. Microbiol. Biotechnol. v.12 Overproduction of Bacillus mcerans cyclodextrin glucanotransferase in E. coli by coexpression of GroEL/ES chaperone Kwon, M. J.;S. L. Park;S. K. Kim;S. W. Nam
  17. J. Microbiol. biotechnol. v.13 High-level expression in escherichia coli of alkaline phosphatase from Thermus caldophilus GK24 and purification of the recombinant enzyme Lee, J. H.;Y. D. Cho;J. J. Choi;Y. J. Lee;H. S. Hoe;H. K. Kim;S. T. Kwon
  18. J. Microbiol. Biotechnol. v.13 Expression and characterization of uropathogenic Escherichia coli adhesin protein linked to cholera toxin A2B subunits in Escherichia coli TB1 Lee, Y. H.;D. K. Ryu;B. O. Kim;S. N. Pyo
  19. Biochem. Eng. J. v.12 Efficient secretory overexpression of Bacillus subtilis pectate lyase in Escherichia coli and single-step purification Matsumoto, T.;D. Katsura;A. Kondo;H. Fukuda https://doi.org/10.1016/S1369-703X(02)00075-X
  20. Carbohydr. Polym. v.49 Isolation and characterization of soluble sulfated polysaccharide from the red seaweed Gracilaria cornea Melo, M. R. S.;J. P. A. Feitosa;A. L. P. Freitas;R. C. M. de Paula https://doi.org/10.1016/S0144-8617(02)00006-1
  21. J. Biol. Chem. v.273 Arylsulfatase from Klebsiella pneumoniae carries a formylglycine generated from a serine Miech, C.;T. Dierks;T. Selmer;K. von Figura;B. Schmidt https://doi.org/10.1074/jbc.273.9.4835
  22. Comp. Biochem. Physiol. v.41 Lysosomal enzymes in aquatic species II. Distribution and particle properties of thermally acclimated muscle lysosomes of rainbow trout Salmo gairdeneri Milanesi, A. A.;J. W. C. Bind
  23. Appl. Environ. Microbiol. v.39 Formation and purification of Serratia marcescens arylsulfatase Murooka, Y.;M. H. Yim;T. Harada
  24. J. Biol. Chem. v.145 Regulation of derepressed synthesis of arylsulfatase by tyramine oxidase in Salmonella typhimurium Murooka, Y.;T. Harada
  25. Trends Biotechnol. v.15 Biotechnology and the red seaweed polysaccharide industry: Status, needs and prospects Renn, D. https://doi.org/10.1016/S0167-7799(96)10069-X
  26. Biochim. Biophys. Acta v.86 A method of preparing agarose Russell, B.;T. H. Mead;A. Polson https://doi.org/10.1016/0304-4165(64)90171-0
  27. Eur. J. Biochem. v.238 The evolutionary conservation of a novel protein modification, the conversion of cysteine to serinesemialdehyde in arylsulfatase from Volvox carteri Selmer, T.;A. Hallmann;B. Schmidt;M. Sumper;K. von Figura https://doi.org/10.1111/j.1432-1033.1996.0341z.x
  28. Fisheries Sci. v.58 Purification and characteriation of arylsulfatase produced from Sphingomonas sp. nov. Seo, H. J.;J. H. Kim;D. S. Byun;H. R. Kim
  29. Geochim. Cosmochim. Acta v.58 Sulfonates: A new class of organic sulfur compounds in marine sediments Vairavamurthy, M. A.;W. Zhou;T. Eglinton;B. Manowitz https://doi.org/10.1016/0016-7037(94)90200-3
  30. Appl. Environ. Microbiol. v.65 Low-molecular-weight sulfonates, a major substrate for sulfate reducers in marine microbial mats Visscher, P. T.;R. F. Gritzer;E. R. Leadbetter
  31. The Handbook of Environmental Chemistry v.1A Zehnder, A. J. B.;S. H. Zinder;O. Hutzinger(ed.)