1887

Abstract

The gene coding for the proline iminopeptidase (Pip) of pv. was cloned in an strain using a selective medium containing the dipeptide -Ala--Leu as the sole source of -leucine. Nucleotide sequencing of this gene revealed a 939 bp open reading frame encoding a 312 amino acid protein (35 126 Da). The deduced amino acid sequence showed 47% identity with the Pip from A fusion gene was overexpressed in under the control of the promoter. The Pip of hydrolysed -prolyl--nitroanilide with the highest efficiency, but was also able to hydrolyse -alanyl--nitroanilide and -alanyl--nitroanilide. The molecular mass of Pip was found to be 35 kDa by SDS-PAGE and 120 kDa by gel filtration, suggesting that the active enzyme is a multimer.

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1996-10-01
2021-05-13
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References

  1. Albertson N.H., Koomey M. 1993; Molecular cloning and characterization of a proline iminopeptidase gene from Neisseria gonorrhoeae . Mol Microbiol 9:1203–1211
    [Google Scholar]
  2. Allaker R.P., Young K.A., Hardie J.M. 1994; Production of hydrolytic enzymes by oral isolates of Eikenella corrodens . FEMS Microbiol Lett 123:69–74
    [Google Scholar]
  3. Atlan D., Gilbert C., Blanc B., Portalier R. 1994; Cloning, sequencing and characterization of the pepIP gene encoding a proline iminopeptidase from Lactobacillus delbrueckii subsp. bulgaricus CNRZ 397. Microbiology 140:527–535
    [Google Scholar]
  4. Arraj J.A., Marinus M.-G. 1983; Phenotypic reversal in dam mutants of Escherichia coli K-12 by recombinant plasmid containing the dam + gene. J Bacteriol 153:562–565
    [Google Scholar]
  5. Bradford M.M. 1976; A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254
    [Google Scholar]
  6. Chesnney R.H., Scott J.R., Vapnek O. 1979; Integration of the plasmid prophages PI and P7 into the chromosome of Escherichia coli . J Mol Biol 130:161–173
    [Google Scholar]
  7. Díaz E., Timmis K.N. 1995; Identification of functional residues in a 2-hydroxymuconic semialdehyde hydrolase. J Biol Chem 270:1–9
    [Google Scholar]
  8. Dow J.M., Fan M.J., Newman M.-A., Daniels M.J. 1993; Differential expression of conserved protease genes in crucifer- attacking pathovars of Xanthomonas campestris . Appl Environ Microbiol 59:3996–4003
    [Google Scholar]
  9. Ehrenfreund P., Mollay C., Kreil G. 1992; Purification and properties of an iminopeptidase from culture media of Streptomyces plicatus . Biochem Biophys Res Commun 184:1250–1255
    [Google Scholar]
  10. García J.L., Buesa J.M. 1986; An improved method to clone penicillin acylase genes: cloning and expression in Escherichia coli of penicillin G acylase from Kluyvera citrophila . J Biotechnol 3:187–195
    [Google Scholar]
  11. Gilbert G, Atlan D., Blanc B., Portalier R. 1994; Proline iminopeptidase from Lactobacillus delbrueckii subsp. bulgaricus CNRZ 397: purification and characterization. Microbiology 140:537–542
    [Google Scholar]
  12. Hawley D. K., McClure W. R. 1983; Compilation and analysis of Escherichia coli promoter DNA sequences. Nucleic Acids Res 11:2237–2255
    [Google Scholar]
  13. Hayase N., Taira K. 1990; Pseudomonas putida KF715 bphABCD operon encoding biphenyl and polychlorinated biphenyl degradation: cloning, analysis and expression in soil bacteria. J Bacteriol 172:1160–1164
    [Google Scholar]
  14. Hessing J.G.M. 1983 The primary structure of atropinesterase from Pseudomas putida. PhD thesis University of Leiden;
    [Google Scholar]
  15. Hiraoka B., Harada M. 1991; Use of l-prolyl-leucylglycinamide (MIF-1) for the high-performance liquid chromatographic determination of proline iminopeptidase activity in rat liver. J Chromatogr 563:142–146
    [Google Scholar]
  16. Horn J.M., Harayama S., Timmis K.N. 1991; DNA sequence determination of the TOL plasmid (pWWO) xyllGFJ genes of Pseudomonas putida-. implications for the evolution of aromatic catabolism. Mol Microbiol 5:2459–2474
    [Google Scholar]
  17. Kato K., Kawahara K., Takahashi T., Kakinum A. 1980a; sPurification of α-amino acid ester hydrolase from Xanthomonas citri . Agric Biol Chem 44:1069–1074
    [Google Scholar]
  18. Kato K., Kawahara K., Takahashi T., Kakinum A. 1980b; Substrate specificity of α-amino acid ester hydrolase from Xanthomonas citri . Agric Biol Chem 44:1075–1081
    [Google Scholar]
  19. Kitazono A., Yoshimoto T., Tsuru D. 1992; Cloning, sequencing and high expression of the proline iminopeptidase gene from Bacillus coagulans. . J Bacteriol 174:7919–7925
    [Google Scholar]
  20. Kitazono A., Kitano A., Tsuru D., Yoshimoto T. 1994a; Isolation and characterization of the prolyl aminopeptidase gene (pap) from Aeromonas sobria: comparison with the Bacillus coagulans enzyme. J Biochem 116:818–825
    [Google Scholar]
  21. Kitazono A., Ito K., Yoshimoto T. 1994b; Prolyl aminopeptidase is not a sulfhydryl enzyme: identification of the active serine residue by site-directed mutagenesis. J Biochem 116:943–945
    [Google Scholar]
  22. Klein J.R., Schmidt U., Plapp R. 1994; Cloning, heterologous expression, and sequencing of a novel proline iminopeptidase gene, pepl, from Lactobacillus delbrueckii subsp.lactis DSM 7290. Microbiology 140:1133–1139
    [Google Scholar]
  23. Laemmli U.K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
    [Google Scholar]
  24. MacCarthy J.E.G., Brimacombe R. 1994; Prokaryotic translation : the interactive pathway leading to initiation. Trends Genet 10:402–407
    [Google Scholar]
  25. Menn F.M., Gerben J.Z., Gibson D.T. 1991; Location and sequence of the todF gene encoding 2-hydroxy-6-oxohepa-2,4- dienoate hydrolase in Pseudomonas putida . Gene 104:91–94
    [Google Scholar]
  26. Ninomiya K., Kawatani K., Tanaka S., Kawata S., Makisumi S. 1982; Purification and properties of a proline iminopeptidase from apricot seeds. J Biochem 92:413–421
    [Google Scholar]
  27. Sambrook J., Fritsch E.J., Maniatis T. 1989 Molecular Cloning: a Laboratory Manual Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  28. Sanger F., Nicklen S., Coulson A.R. 1977; DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467
    [Google Scholar]
  29. Sarid S., Berger A., Katchalski E. 1959; Proline iminopeptidase. J Biol Chem 234:1740–1744
    [Google Scholar]
  30. Szwajcer-Dey E., Rasmussen J., Meldal M., Breddman K. 1992; Proline-specific endopeptidases from microbial sources: isolation of an enzyme from Xanthomonas sp. J Bacteriol 174:2454–2459
    [Google Scholar]
  31. Verniere C., Provost O., Civerolo E.L., Gambin O., Jacquemoud Collet J.P., Luisette J. 1993; Evaluation of the biolog substrate utilization system to identify and assess metabolic variation among strains of Xanthomonas campestris pv. citri. Appl Environ Microbiol 59:243–249
    [Google Scholar]
  32. Yoshimoto T., Saeki T., Tsuru D. 1983; Proline iminopeptidase from Bacillus megaterium: purification and characterization. J Biochem 93:469–477
    [Google Scholar]
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