Pseudomonas sp. W7로부터 Agarase 유전자의 Cloning 및 Escherichia coli에서의 발현

  • 하정철 (부산수산대학교 생물공학과) ;
  • 김구택 (부산수산대학교 생물공학과) ;
  • 김성구 (부산수산대학교 생물공학과) ;
  • 유주현 (연세대학교 식품생물공학과) ;
  • 공인수 (부산수산대학교 생물공학과)
  • Published : 1995.12.01

Abstract

A marine bacterium which produces extracelluar agarase was isolated from sea water. Isolated strain was identified as Pseudomonas sp. by the morphological and biochemical properties (1). HindIII restriction fragment of 3.2 kb from Pseudomonas genomic DNA was cloned into pUC19 to obtain recombinant plasmid pJA1 which enables E. coli JM83 to produce agarase. Most of agarase produced in E. coli was secreted into the culture medium. The enzyme (pJA1) showed the highest agarase activity during the stationary phase (20 hrs) of E. coli. The optimum temperature and pH were 40$\circ$C and 7.8, respectively. Restriction gene map anlaysis revealed that it has different restriction pattern with three kind of agarase gene reported.

Keywords

References

  1. 한국산업미생물학회 춘계학술발표대회 Program Purification of Extracellular Agarase from Marine Bacterium 하순득;공인수;공재열
  2. J. Food Science and Technol. v.14 Hayashi,K.;Nonaka,K.
  3. 日本農藝化學會 講演要旨集 Property and production of neoagarooligosaccharides 洞野敏明;德永隆久;日高秀昌;三好熙三;北川應進;平賀哲男;片所 功
  4. Food Chemical v.4 洞野敏明
  5. Carbohydr. Res. v.66 Enzymic cleavage of the α-linkages in agarose, to yield agarooligosaccharides Young,K.S.;S.S.Bhattacharjee;W.Yaphe
  6. J. Biochem. v.113 The action of a bacterial agarase on agarose, porphyran and alkali treated porphyran Duckworth,M.;J.R.Turvey
  7. J. Biochem. v.113 The specificity of agarase from a Cytophaga species Duckworth,M.;J.R.Turvey
  8. Eur. J. Biochem. v.135 β-AgaraseⅠand Ⅱ from Pseudomonas atlantica: purification and some properties Morrice,L.M.;M.W.Melean;F.B.Williamson;W.F.Long
  9. Eur. J. Biochem. v.187 Purification and characterization of a novel β-agarase from Vibrio sp. AP-2 Aoki,T.;T.Araki;M.Kitamikado
  10. Mol. Gen. Genet. v.209 The agarase gene (dagA) of Streptomyces coelicolor A3(2): nucleotide sequence and transcriptional analysis Buttner,M.J.;I.M.Fernley;M.J.Bibb
  11. J. Mol. Biol. v.3 A procedure for the isolation of deoxyribonucleic acid from microorganisms Marmur,J.
  12. A laboratory manual 2nd ed. Sambrook,J.;E.F.Fritsch;Maniatis
  13. Nucleic Acids Res. v.9 Rapid and efficient cosmid cloning Ish-Horowicz,D.;J.F.Burke
  14. Anal. Biochem. v.114 A rapid boiling method for the preparation of bacterial plasmids Holmes,D.S.;Quigley,M.
  15. Molecular cloning: a laboratory manual Maniatis,T.;E.F.Fritsch;J.Sambrook
  16. J. Mol. Biol. v.53 The calcium dependent bacteriophage DNA infection Mandel,M.;A.Higa
  17. J. Bacteriol. v.42 Gran,H.H.
  18. J. Biol. Chem. v.153 Notes on sugar determination Somogyi,M.
  19. Mol. Gen. Genet. v.86 Cloning and expression in Escherichia coli Cornelis,P.;C.Digneffe;K.Willinot
  20. Appl. Environ. Microbiol. v.59 no.5 Purification and characterization of a new agarase from a marine bacterium, Vibrio sp. strain JT0107 Sugano,Y.;T.Matsumoto