Pseudomonas sp. DJ77 균주에서 Extradiol Dioxygenase를 암호화하는 phnQ 유전자의 클로닝과 대장균에서의 발현

Cloning of phnQ Gene Encoding Extradiol Dioxygenase from Pseudomonas sp. DJ77 and Its Expression in Escherichia coli

  • 발행 : 1997.03.01

초록

Pseudomonas sp. DJ77의 게놈 library로부터 phenanthrene 분해에 관련된 유전자를 포함하는 약 5-kb의 XhoI 절편을 pBLUESCRIPT SK(+)로 클로닝하였으며, 이 재조합 plasmid를 pUPX5라 명명하였다. 이 재조합 균주에 catechol과 2,3-dihydroxybiphenyl 용액을 분무하면 노란색의 meta-cleavage 화합물이 생성됨을 관찰 할 수 있었다. 그리고 효소 활성을 측정한 결과 catechol에서보다 2,3-dihydroxybiphenyl에 더 큰 활성을 나타냈다. 이 부분에 존재하는 2,3-dihydroxybiphenyl 1,2-dioxygenase 유전자의 위치를 결정하고, 이 유전자를 phnQ라 명명하였다.

We cloned the 5~kb Xlwl fragment containing gene responsible for degrad"tion of phenanthrene using pBLUES~ CRIPT SK( +) vector and E. coli XLI-Blue strain from the genomic library of Pseudomonas sp. 0177 and this recombinant plasmid was named pUPX5. The strain containing pUPX5 could produce a yellow meta-cleavage product using 2.3-dihydroxybiphenyl as a substrate. This strain have a higher activity toward 2,3-dihydroxybiphenyl than catechol. We sub cloned and localized the gene encoding 2.3-dihydroxybiphenyl-1.2-dioxygenase. which is designated as phn$\Omega$.

키워드

참고문헌

  1. Gene v.85 Nucleotide sequence and expression of the catechol 2,3-dioxygenase-encoding gene of phenol catabolizing Pseudomounas CF600 Bartklson, M.;V. Shingler
  2. Biochem. J. v.101 The meta-bolism of cresols by species of Pseudomonas Bayly, R.C.;S. Dagley;D. T. Gibson
  3. J. Biol. Chem. v.268 Purification and crystallization of 2,3-dihydroxybiphernyl 1,2-dioxygenase Eltis, L.D.;B. Hofmann;H.J. Hecht;H. Lunsdorf;K.N. Timmis
  4. Science v.270 Crystal structure of the biphenyl-cleaving extradiol dioxygenase from a PCB-degrading Pseu-domonad Han, S.G.;L.G. Eltis;K.N. Timmis;S.W. Muchmore;J.T. Bolin
  5. J. Bacteriol. v.172 Bacterial aromatic ring-cleavage enzymes are classified into two different gene families Harayama, S.;M. Rekik
  6. J. Bacteriol. v.100 Phenol and ben-zoate metabolism by Pseudomonas putida: regulation of tangential pathway Hegeman, G.D.;C.F. Feist
  7. FEMS Microbiol. Lett. v.118 Fuctional and struc-tural relationship of various extradiol aromatic ring-cleavage diosygenases of Pseudomonas origin Hirose, J.;N. Kimura;A. Suyama;A. Kobayashi;S. Hayashida;K. Furukawa
  8. Kor, J. Microbiol. v.24 Isolation of aromatic hydrocarbon-degrading bacteria and genetic characterization of their plasmid genes Kim, C.K.;J.W. Kim;Y.C. Kim;T.I. Mheen
  9. J. Bacteriol. v.177 Molecular and biochem-ical characterization of two meta-cleavage diosygenases involved in biphenyl and m-xylene degradation by Beijerinckia sp. strain B1. Kim, E.;G.J. Zylstra
  10. Kor. J. Microbiol. v.30 Molecular cloning of a gene cluster for phenanthrene degradation from Pseudomonas sp. strain DJ77 and its expression in Escherichia coli. Kim, Y.C.;K.S. Youn;M.S. Shin;H.S. Kim;M.S. Park;H.J. Park
  11. Kor. J. Microbiol. v.30 Nucleotide sequence of phnE gene encoding extradiol dioxygenase from Pseudomonas sp. strain DJ77 Kim, Y.C.;M.S. Shin;K.S. Youn;Y.S. Park;U. H. Kim
  12. J. Bacteriol. v.171 Cloning and sequencing of two tandem genes involved in degradation of 2,3-dihydroxybipheny1 to benzoic acid in the polychlorinated biphenyl-degrading soil bacterium Pseudomonas sp. strain KKS102 Kimbara, K.;T. Hashimoto;M. Fukuga;T. Koana;M. Takagi;M. Oishi;K. Yano
  13. Biochem. Biophys. Res. Comm. v.224 Structure of catechol 2,3-dioxygenase gene encoded in chromosomal DNA of Pseudomonas putida KF715 Lee, J.;J. Oh;K.R. Min;C.K. Kim;K.H. Min;K.S. Lee;Y.C. Kim;J.Y. Lim;Y. Kim
  14. Molecular cloning a laboratory manual Sambrook, J.;E.F. Fritsch;T. Maniatis
  15. Experiments with gene fusions Silhavy, T.J.;M.L. Berman;L.W. Enquist
  16. Biochemistry v.27 Cloning and nucleotide sequence of the 2,3-dihydroxybiphenyl dioxygenase gene from the PCB-degrading strain of Pseudomonas paucimobilis Q1 Taira, K.;N. Hayase;N. Arimura;S. Yamashita;T. Mi-yazaki;K. Furukawa
  17. J. Bacteriol. v.164 Toluene degradation by Pseudomonas putida F1 Zylstra, G.J.;D.T. Gilson