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. 1991 Aug;173(16):5159–5167. doi: 10.1128/jb.173.16.5159-5167.1991

Cloning, characterization, and expression in Escherichia coli of a gene encoding Listeria seeligeri catalase, a bacterial enzyme highly homologous to mammalian catalases.

A Haas 1, K Brehm 1, J Kreft 1, W Goebel 1
PMCID: PMC208208  PMID: 1860824

Abstract

A gene coding for catalase (hydrogen-peroxide:hydrogen-peroxide oxidoreductase; EC 1.11.1.6) of the gram-positive bacterium Listeria seeligeri was cloned from a plasmid library of EcoRI-digested chromosomal DNA, with Escherichia coli DH5 alpha as a host. The recombinant catalase was expressed in E. coli to an enzymatic activity approximately 50 times that of the combined E. coli catalases. The nucleotide sequence was determined, and the deduced amino acid sequence revealed 43.2% amino acid sequence identity between bovine liver catalase and L. seeligeri catalase. Most of the amino acid residues which are involved in catalytic activity, the formation of the active center accession channel, and heme binding in bovine liver catalase were also present in L. seeligeri catalase at the corresponding positions. The recombinant protein contained 488 amino acid residues and had a calculated molecular weight of 55,869. The predicted isoelectric point was 5.0. Enzymatic and genetic analyses showed that there is most probably a single catalase of this type in L. seeligeri. A perfect 21-bp inverted repeat, which was highly homologous to previously reported binding sequences of the Fur (ferric uptake regulon) protein of E. coli, was detected next to the putative promoter region of the L. seeligeri catalase gene.

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Selected References

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  1. BEERS R. F., Jr, SIZER I. W. A spectrophotometric method for measuring the breakdown of hydrogen peroxide by catalase. J Biol Chem. 1952 Mar;195(1):133–140. [PubMed] [Google Scholar]
  2. Bachmann B. J. Linkage map of Escherichia coli K-12, edition 8. Microbiol Rev. 1990 Jun;54(2):130–197. doi: 10.1128/mr.54.2.130-197.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bethards L. A., Skadsen R. W., Scandalios J. G. Isolation and characterization of a cDNA clone for the Cat2 gene in maize and its homology with other catalases. Proc Natl Acad Sci U S A. 1987 Oct;84(19):6830–6834. doi: 10.1073/pnas.84.19.6830. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bortolussi R., Vandenbroucke-Grauls C. M., van Asbeck B. S., Verhoef J. Relationship of bacterial growth phase to killing of Listeria monocytogenes by oxidative agents generated by neutrophils and enzyme systems. Infect Immun. 1987 Dec;55(12):3197–3203. doi: 10.1128/iai.55.12.3197-3203.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Boyd J., Oza M. N., Murphy J. R. Molecular cloning and DNA sequence analysis of a diphtheria tox iron-dependent regulatory element (dtxR) from Corynebacterium diphtheriae. Proc Natl Acad Sci U S A. 1990 Aug;87(15):5968–5972. doi: 10.1073/pnas.87.15.5968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. CLAYTON R. K. Purified catalase from Rhodopseudomonas spheroides. Biochim Biophys Acta. 1959 Nov;36:40–47. doi: 10.1016/0006-3002(59)90067-8. [DOI] [PubMed] [Google Scholar]
  7. Chakraborty T., Goebel W. Recent developments in the study of virulence in Listeria monocytogenes. Curr Top Microbiol Immunol. 1988;138:41–58. [PubMed] [Google Scholar]
  8. Claiborne A., Fridovich I. Purification of the o-dianisidine peroxidase from Escherichia coli B. Physicochemical characterization and analysis of its dual catalatic and peroxidatic activities. J Biol Chem. 1979 May 25;254(10):4245–4252. [PubMed] [Google Scholar]
  9. Claiborne A., Malinowski D. P., Fridovich I. Purification and characterization of hydroperoxidase II of Escherichia coli B. J Biol Chem. 1979 Nov 25;254(22):11664–11668. [PubMed] [Google Scholar]
  10. Dallmier A. W., Martin S. E. Catalase and superoxide dismutase activities after heat injury of Listeria monocytogenes. Appl Environ Microbiol. 1988 Feb;54(2):581–582. doi: 10.1128/aem.54.2.581-582.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Dallmier A. W., Martin S. E. Catalase, superoxide dismutase, and hemolysin activities and heat susceptibility of Listeria monocytogenes after growth in media containing sodium chloride. Appl Environ Microbiol. 1990 Sep;56(9):2807–2810. doi: 10.1128/aem.56.9.2807-2810.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Fita I., Rossmann M. G. The active center of catalase. J Mol Biol. 1985 Sep 5;185(1):21–37. doi: 10.1016/0022-2836(85)90180-9. [DOI] [PubMed] [Google Scholar]
  14. Flamm R. K., Hinrichs D. J., Thomashow M. F. Introduction of pAM beta 1 into Listeria monocytogenes by conjugation and homology between native L. monocytogenes plasmids. Infect Immun. 1984 Apr;44(1):157–161. doi: 10.1128/iai.44.1.157-161.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Fridovich I. Biological effects of the superoxide radical. Arch Biochem Biophys. 1986 May 15;247(1):1–11. doi: 10.1016/0003-9861(86)90526-6. [DOI] [PubMed] [Google Scholar]
  16. Goldberg I., Hochman A. Purification and characterization of a novel type of catalase from the bacterium Klebsiella pneumoniae. Biochim Biophys Acta. 1989 May 31;991(2):330–336. doi: 10.1016/0304-4165(89)90124-4. [DOI] [PubMed] [Google Scholar]
  17. Heimberger A., Eisenstark A. Compartmentalization of catalases in Escherichia coli. Biochem Biophys Res Commun. 1988 Jul 15;154(1):392–397. doi: 10.1016/0006-291x(88)90698-5. [DOI] [PubMed] [Google Scholar]
  18. Henikoff S. Unidirectional digestion with exonuclease III in DNA sequence analysis. Methods Enzymol. 1987;155:156–165. doi: 10.1016/0076-6879(87)55014-5. [DOI] [PubMed] [Google Scholar]
  19. Horwitz M. S., Loeb L. A. Structure-function relationships in Escherichia coli promoter DNA. Prog Nucleic Acid Res Mol Biol. 1990;38:137–164. doi: 10.1016/s0079-6603(08)60710-2. [DOI] [PubMed] [Google Scholar]
  20. Köhler S., Leimeister-Wächter M., Chakraborty T., Lottspeich F., Goebel W. The gene coding for protein p60 of Listeria monocytogenes and its use as a specific probe for Listeria monocytogenes. Infect Immun. 1990 Jun;58(6):1943–1950. doi: 10.1128/iai.58.6.1943-1950.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  22. Leblond-Francillard M., Gaillard J. L., Berche P. Loss of catalase activity in Tn1545-induced mutants does not reduce growth of Listeria monocytogenes in vivo. Infect Immun. 1989 Aug;57(8):2569–2573. doi: 10.1128/iai.57.8.2569-2573.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Loewen P. C., Switala J. Multiple catalases in Bacillus subtilis. J Bacteriol. 1987 Aug;169(8):3601–3607. doi: 10.1128/jb.169.8.3601-3607.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Loewen P. C., Triggs B. L. Genetic mapping of katF, a locus that with katE affects the synthesis of a second catalase species in Escherichia coli. J Bacteriol. 1984 Nov;160(2):668–675. doi: 10.1128/jb.160.2.668-675.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Loewen P. C., Triggs B. L., Klassen G. R., Weiner J. H. Identification and physical characterization of a Col E1 hybrid plasmid containing a catalase gene of Escherichia coli. Can J Biochem Cell Biol. 1983 Dec;61(12):1315–1321. doi: 10.1139/o83-168. [DOI] [PubMed] [Google Scholar]
  26. MARGOLIASH E., NOVOGRODSKY A., SCHEJTER A. Irreversible reaction of 3-amino-1:2:4-triazole and related inhibitors with the protein of catalase. Biochem J. 1960 Feb;74:339–348. doi: 10.1042/bj0740339. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Mainou-Fowler T., MacGowan A. P., Postlethwaite R. Virulence of Listeria spp.: course of infection in resistant and susceptible mice. J Med Microbiol. 1988 Oct;27(2):131–140. doi: 10.1099/00222615-27-2-131. [DOI] [PubMed] [Google Scholar]
  28. Mead D. A., Szczesna-Skorupa E., Kemper B. Single-stranded DNA 'blue' T7 promoter plasmids: a versatile tandem promoter system for cloning and protein engineering. Protein Eng. 1986 Oct-Nov;1(1):67–74. doi: 10.1093/protein/1.1.67. [DOI] [PubMed] [Google Scholar]
  29. Melik-Adamyan W. R., Barynin V. V., Vagin A. A., Borisov V. V., Vainshtein B. K., Fita I., Murthy M. R., Rossmann M. G. Comparison of beef liver and Penicillium vitale catalases. J Mol Biol. 1986 Mar 5;188(1):63–72. doi: 10.1016/0022-2836(86)90480-8. [DOI] [PubMed] [Google Scholar]
  30. Mengaud J., Vicente M. F., Chenevert J., Pereira J. M., Geoffroy C., Gicquel-Sanzey B., Baquero F., Perez-Diaz J. C., Cossart P. Expression in Escherichia coli and sequence analysis of the listeriolysin O determinant of Listeria monocytogenes. Infect Immun. 1988 Apr;56(4):766–772. doi: 10.1128/iai.56.4.766-772.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Mulvey M. R., Sorby P. A., Triggs-Raine B. L., Loewen P. C. Cloning and physical characterization of katE and katF required for catalase HPII expression in Escherichia coli. Gene. 1988 Dec 20;73(2):337–345. doi: 10.1016/0378-1119(88)90498-2. [DOI] [PubMed] [Google Scholar]
  32. Murray W. W., Rachubinski R. A. The nucleotide sequence of complementary DNA and the deduced amino acid sequence of peroxisomal catalase of the yeast Candida tropicalis pK233. Gene. 1987;61(3):401–413. doi: 10.1016/0378-1119(87)90202-2. [DOI] [PubMed] [Google Scholar]
  33. Murthy M. R., Reid T. J., 3rd, Sicignano A., Tanaka N., Rossmann M. G. Structure of beef liver catalase. J Mol Biol. 1981 Oct 25;152(2):465–499. doi: 10.1016/0022-2836(81)90254-0. [DOI] [PubMed] [Google Scholar]
  34. Rocourt J., Alonso J. M., Seeliger H. P. Virulence comparée des cinq groupes génomiques de Listeria monocytogenes (sensu lato). Ann Microbiol (Paris) 1983 May-Jun;134A(3):359–364. [PubMed] [Google Scholar]
  35. Sakajo S., Nakamura K., Asahi T. Molecular cloning and nucleotide sequence of full-length cDNA for sweet potato catalase mRNA. Eur J Biochem. 1987 Jun 1;165(2):437–442. doi: 10.1111/j.1432-1033.1987.tb11457.x. [DOI] [PubMed] [Google Scholar]
  36. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Schellhorn H. E., Hassan H. M. Transcriptional regulation of katE in Escherichia coli K-12. J Bacteriol. 1988 Sep;170(9):4286–4292. doi: 10.1128/jb.170.9.4286-4292.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Tai S. P., Holmes R. K. Iron regulation of the cloned diphtheria toxin promoter in Escherichia coli. Infect Immun. 1988 Sep;56(9):2430–2436. doi: 10.1128/iai.56.9.2430-2436.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Tartaglia L. A., Storz G., Ames B. N. Identification and molecular analysis of oxyR-regulated promoters important for the bacterial adaptation to oxidative stress. J Mol Biol. 1989 Dec 20;210(4):709–719. doi: 10.1016/0022-2836(89)90104-6. [DOI] [PubMed] [Google Scholar]
  40. Triggs-Raine B. L., Doble B. W., Mulvey M. R., Sorby P. A., Loewen P. C. Nucleotide sequence of katG, encoding catalase HPI of Escherichia coli. J Bacteriol. 1988 Sep;170(9):4415–4419. doi: 10.1128/jb.170.9.4415-4419.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Triggs-Raine B. L., Loewen P. C. Physical characterization of katG, encoding catalase HPI of Escherichia coli. Gene. 1987;52(2-3):121–128. doi: 10.1016/0378-1119(87)90038-2. [DOI] [PubMed] [Google Scholar]
  42. Wayne L. G., Diaz G. A. A double staining method for differentiating between two classes of mycobacterial catalase in polyacrylamide electrophoresis gels. Anal Biochem. 1986 Aug 15;157(1):89–92. doi: 10.1016/0003-2697(86)90200-9. [DOI] [PubMed] [Google Scholar]
  43. Welch D. F., Sword C. P., Brehm S., Dusanic D. Relationship between superoxide dismutase and pathogenic mechanisms of Listeria monocytogenes. Infect Immun. 1979 Mar;23(3):863–872. doi: 10.1128/iai.23.3.863-872.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Woodbury W., Spencer A. K., Stahman M. A. An improved procedure using ferricyanide for detecting catalase isozymes. Anal Biochem. 1971 Nov;44(1):301–305. doi: 10.1016/0003-2697(71)90375-7. [DOI] [PubMed] [Google Scholar]
  45. Yamada Y., Saito H., Tomioka H., Jidoi J. Susceptibility of micro-organisms to active oxygen species: sensitivity to the xanthine-oxidase-mediated antimicrobial system. J Gen Microbiol. 1987 Aug;133(8):2007–2014. doi: 10.1099/00221287-133-8-2007. [DOI] [PubMed] [Google Scholar]
  46. von Ossowski I., Mulvey M. R., Leco P. A., Borys A., Loewen P. C. Nucleotide sequence of Escherichia coli katE, which encodes catalase HPII. J Bacteriol. 1991 Jan;173(2):514–520. doi: 10.1128/jb.173.2.514-520.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]

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