1887

Abstract

The structural gene () coding for the extracellular lipase of PAO1 has been cloned on plasmid pSW118. Nucleotide sequence analysis revealed a gene of 936 bp. codes for a proenzyme of 311 amino acids including a leader sequence of 26 amino acids. The mature protein was predicted to have a of 30134, an isoelectric point of 5.6, and a consensus sequence (IGHSHGG) typical of lipases. Furthermore it is highly homologous (>60%) to other lipases from various pseudomonads. The gene failed to hybridize detectably with genomic DNA from other species except , even under relaxed stringency. Located 220 bp downstream of the gene, is an open reading frame (ORF2, ) which encodes a hydrophilic protein (283 amino acids; 33587) that shows some homology to the gene product of . In complementation tests of lipase-defective mutants, was shown to be necessary for expression of active extracellular lipase in PAO1.

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1992-07-01
2021-10-19
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References

  1. Andreoli P. M., Cox M. M. J., Farin F., Wohlfarth S. 1989 Molecular cloning and expression of genes encoding lipolytic enzymes European Patent no. 0 334 462 A1
    [Google Scholar]
  2. Aoyama S., Yoshida N., Inouye S. 1988; Cloning, sequencing and expression of the lipase gene from Pseudomonas fragi IFO-12049 in E. coli. FEBS Letters 242 36 40
    [Google Scholar]
  3. Bagdasarian M., Lurz R., Rückert B., Franklin F. C. H., Bagdasarian M. M., Frey J., Timmis K. N. 1981; Specific-purpose plasmid cloning vectors. II. Broad host range, high copy number, RSF1010-derived vectors, and a host-vector system for gene cloning in Pseudomonas. Gene 16 237 247
    [Google Scholar]
  4. Bally M., Ball G., Badere A., Lazdunski A. 1991; Protein secretion in Pseudomonas aeruginosa: The xcpA gene encodes an integral inner membrane protein homologous to Klebsiella pneumoniae secretion function protein Pu1O. Journal of Bacteriology 173 479 486
    [Google Scholar]
  5. Brady L., Brzozowski A. M., Derewenda Z. S., Dodson E., Dodson G., Tolley S., Turkenburg J. P., Christiansen L., Huge-Jensen B., Norskov L., Thim L., Menge U. 1990; A serine protease triad forms the catalytic centre of a triacylglycerol lipase. Nature, London 343 767 770
    [Google Scholar]
  6. Chen S. T., Jordan E. M., Wilson R. B., Draper R. K., Clowes R. C. 1987; Transcription and expression of the exotoxin A gene of Pseudomonas aeruginosa. Journal of General Microbiology 133 3081 3091
    [Google Scholar]
  7. Chung G. H., Lee Y. P., Yoo O. J., Rhee J. S. 1991; Overexpression of a thermostable lipase gene from Pseudomonas fluorescens in Escherichia coli. Applied Microbiology and Biotechnology 35 237 241
    [Google Scholar]
  8. Cox M. M. J., Andreoli P. M., Gerritse G., Quax W. J. 1991; Molecular cloning, characterization and expression of a lipase gene from Pseudomonas species. In Book of Abstracts of Third International Symposium on Pseudomonad Biology and Biotechnology in Trieste p. 153
    [Google Scholar]
  9. Deininger P. L. 1983; Random subcloning of sonicated DNA: Application to shotgun DNA sequence analysis. Analytical Biochemistry 129 216 223
    [Google Scholar]
  10. Deretic V., Gill J. F., Chakrabarty A. M. 1987; Pseudomonas aeruginosa infection in cystic fibrosis: nucleotide sequence and transcriptional regulation of the algD gene. Nucleic Acids Research 15 4567 4581
    [Google Scholar]
  11. Feller G., Thiry M., Gerday C. 1990; Sequence of a lipase gene from the antarctic psychrotroph Moraxella TA144. Nucleic Acids Research 18 6431
    [Google Scholar]
  12. Feller G., Thiry M., Gerday C. 1991; Nucleotide sequence of the lipase gene lip3 from the antarctic psychrotroph Moraxella TA144. Biochimica et Biophysica Acta 1088 323 324
    [Google Scholar]
  13. Filloux A., Bally M., Ball G., Akrim M., Tommassen J., Lazdunski A. 1990; Protein secretion in Gram-negative bacteria: transport across the outer membrane involves common mechanisms in different bacteria. EMBO Journal 13 4323 4329
    [Google Scholar]
  14. Frenken L. G. J., Bos J. W., Visser C., Verrips C. T. 1991; Isolation and characterization of the Pseudomonas glumae lipase gene and a second gene essential for lipase secretion. In Book of Abstracts of Third International Symposium on Pseudomonad Biology and Biotechnology in Trieste p. 153
    [Google Scholar]
  15. Gambello M. J., Iglewski B. H. 1991; Cloning and characterization of the Pseudomonas aeruginosa lasR gene, transcriptional activator of elastase expression. Journal of Bacteriology 173 3000 3009
    [Google Scholar]
  16. Gilbert E. J., Cornish A., Jones C. W. 1991; Purification and properties of extracellular lipase from Pseudomonas aeruginosa EF2. Journal of General Microbiology 137 2223 2229
    [Google Scholar]
  17. Götz F. 1991; Staphylococcal lipases and phospholipases. In Lipases: Structure, Mechanism and Genetic Engineering (GBF monograph no. 16) pp. 285 292 Edited by Alberghina L., Schmid R. D., Verger R. Weinheim: VCH;
    [Google Scholar]
  18. Götz F., Popp F., Korn E., Schleifer K. H. 1985; Complete nucleotide sequence of the lipase gene from Staphylococcus hyicus cloned in Staphylococcus carnosus. Nucleic Acids Research 13 5895 5906
    [Google Scholar]
  19. Gray G. L., Smith D. H., Baldridge J. S., Harkins H. N., Vasil M. L., Chen E. Y., Heyneker H. L. 1984; Cloning, nucleotide sequence, and expression in Escherichia coli of the exotoxin A structural gene of Pseudomonas aeruginosa. Proceedings of the National Academy of Sciences of the United States of America 81 2645 2649
    [Google Scholar]
  20. Henikoff S. 1984; Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing. Gene 28 351 359
    [Google Scholar]
  21. Holloway B. W., Krishnapillai V., Morgan A. F. 1979; Chromosomal genetics of Pseudomonas. Microbiological Reviews 43 73 102
    [Google Scholar]
  22. Ihara F., Kageyama Y., Hirata M., Nihira T., Yamada Y. 1991; Purification, characterization, and molecular cloning of lactonizing lipase from Pseudomonas species. Journal of Biological Chemistry 266 18135 18140
    [Google Scholar]
  23. Jaeger K.-E., Wohlfarth S., Winkler U. K. 1991; Extracellular lipase of Pseudomonas aeruginosa. In Lipases: Structure, Mechanism and Genetic Engineering (GBF monograph no. 16) pp. 381 384 Edited by Alberghina L., Schmid R. D., Verger R. Weinheim: VCH;
    [Google Scholar]
  24. Jaeger K.-E., Adrian F.-J., Meyer H. E., Hancock R. E. W., Winkler U. K. 1992; Extracellular lipase from Pseudomonas aeruginosa is an amphiphilic protein. Biochima et Biophysica Acta in the Press
    [Google Scholar]
  25. Jorgensen S., Skov K. W., Diderichsen B. 1991; Cloning, sequence, and expression of a lipase gene from Pseudomonas cepacia: Lipase production in heterologous hosts requires two Pseudomonas genes. Journal of Bacteriology 173 559 567
    [Google Scholar]
  26. Kaudelka H., Wohlfarth S., Winkler U. 1992; Molecular characterization of two lipase-defective mutants of Pseudomonas aeruginosa. BioEngineering 8 (Abstracts of Gemeinsame Frühjahrsta-gung der VAAM und DHGM) 79
    [Google Scholar]
  27. Kugimiya W., Otani Y., Hashimoto Y., Takagi Y. 1986; Molecular cloning and nucleotide sequence of the lipase gene from Pseudomonas fragi. Biochemical and Biophysical Research Communications 141 185 190
    [Google Scholar]
  28. Kyte J., Doolittle R. F. 1982; A simple method for displaying the hydropathic character of a protein. Journal of Molecular Biology 157 105 132
    [Google Scholar]
  29. Lee C. Y., Iandolo J. J. 1986; Lysogenic conversion of staphylococcal lipase is caused by insertion of the bacteriophage L54a genome into the lipase structural gene. Journal of Bacteriology 166 385 391
    [Google Scholar]
  30. Marmur J. 1961; A procedure for the isolation of deoxyribonucleic acid from micro-organisms. Journal of Molecular Biology 3 208 218
    [Google Scholar]
  31. Nakanishi Y., Kurono Y., Koide Y., Beppu T. 1989; Recombinant DNA, bacterium of the genus pseudomonas containing it, and process for preparing lipase by using it. European Patent No. 0 331 376 A2
    [Google Scholar]
  32. Nakanishi Y., Watanabe H., Washizu K., Narahashi Y., Kurono Y. 1991; Cloning, sequencing and regulation of the lipase gene from Pseudomonas sp. M-12-33. In Lipases: Structure, Mechanism and Genetic Engineering (GBF monograph no. 16) pp. 263 266 Edited by Alberghina L., Schmid R. D., Verger R. Weinheim: VCH;
    [Google Scholar]
  33. Nishioka T., Chihara-Shiomi M., Yoshikawa K., Inagaki M., Yamamoto Y., Hiratake J., Baba N., Oda J. 1991; Lipase from Pseudomonas sp.: Reactions, cloning, and amino acid sequence analysis. In Lipases: Structure, Mechanism and Genetic Engineering (GBF monograph no. 16) pp. 253 262 Edited by Alberghina L., Schmid R. D., Verger R. Weinheim: VCH;
    [Google Scholar]
  34. Ohkawa I., Shiga S., Kageyama M. 1979; An esterase on the outer membrane of Pseudomonas aeruginosa for the hydrolysis of long chain acyl esters. Journal of Biochemistry 86 643 656
    [Google Scholar]
  35. O’Hoy K., Krishnapillai V. 1987; Recalibration of the Pseudomonas aeruginosa PAO chromosome map in time units using high-frequency-of-recombination donors. Genetics 115 611 618
    [Google Scholar]
  36. Palmeros B., Tamayo E. M., Ramirez G., Najera R., Soberon-Chaves G. 1991; Cloning and characterization of a Pseudomonas aeruginosa lipase. In Book of Abstracts of Third International Symposium on Pseudomonad Biology and Biotechnology in Trieste p. 167
    [Google Scholar]
  37. Sambrook J., Fritsch E. F., Maniatis T. 1989 Molecular Cloning: a Laboratory Manual , 2nd Edn.. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  38. Sanger F., Nicklen S., Coulson A. R. 1977; DNA sequencing with chain terminating inhibitors. Proceedings of the National Academy of Sciences of the United States of America 74 5463 5467
    [Google Scholar]
  39. Schrag J. D., Li Y., Wu S., Cygler M. 1991; Ser-His-Glu triad forms the catalytic site of the lipase from Geotrichum candidum. Nature, London 351 761 764
    [Google Scholar]
  40. Sillero A., Ribeiro J. M. 1989; Isoelectric points of proteins: Theoretical determination. Analytical Biochemistry 179 319 325
    [Google Scholar]
  41. Staden R. 1980; A new computer method for the storage and manipulation of DNA gel reading data. Nucleic Acids Research 8 3673 3694
    [Google Scholar]
  42. Stuer W., Jaeger K. E., Winkler U. K. 1986; Purification of extracellular lipase from Pseudomonas aeruginosa. Journal of Bacteriology 168 1070 1074
    [Google Scholar]
  43. Vasil M. L., Chamberlain C., Grant C. C. R. 1986; Molecular studies of Pseudomonas exotoxin A gene. Infection and Immunity 52 538 548
    [Google Scholar]
  44. Watson M. E. E. 1984; Compilation of published signal sequences. Nucleic Acids Research 13 5145 5164
    [Google Scholar]
  45. West S. E. H., Iglewski B. H. 1988; Codon usage in Pseudomonas aeruginosa. Nucleic Acids Research 16 9323 9335
    [Google Scholar]
  46. Winkler U., Scholle H., Bohne L. 1975; Mutants of Serratia marcescens lacking cyclic nucleotide phosphodiesterase activity and requiring cyclic 3′,5′-AMP for the utilization of various carbohydrates. Archives of Microbiology 104 189 196
    [Google Scholar]
  47. Winkler F. K., D’Arcy A., Hunziker W. 1990; Structure of human pancreatic lipase. Nature, London 343 771 774
    [Google Scholar]
  48. Wohlfarth S., Winkler U. K. 1988; Chromosomal mapping and cloning of the lipase gene of Pseudomonas aeruginosa. Journal of General Microbiology 134 433 440
    [Google Scholar]
  49. Yanisch-Perron C., Vieira J., Messing J. 1985; Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mpl8 and pUC19 vectors. Gene 33 103 119
    [Google Scholar]
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