1887

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Volume 143, Issue 5

Cloning of a protopectinase gene of and its expression in Free

Abstract

A protopectinase (PPase)-encoding gene, from was cloned by colony hybridization using two oligonucleotide probes synthesized from the N-terminal amino acid sequences of native PPase SE1 and one peptide from a lysyl endopeptidase digest. Nucleotide sequencing revealed that contains an ORF encoding a 367 amino acid protein. Mature PPase SE3 is composed of 340 amino acids and the N-terminus of the ORF appeared to correspond to a signal peptide and a propeptide processed by a KEX2-like proteinase. The deduced amino acid sequence of was 65.4, 56.7, 58.1, 61.8 and 48.9% homologous to the polygalacturonases of and respectively. One domain, which might interact with polygalacturonic acid, is highly conserved not only in fungal polygalacturonases but also in bacterial and plant polygalacturonases. was expressed in but three forms (the mature form, a glycosylated form and an uncharacterized processed form) of PPase SE3 were present among the products.

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Keyword(s): gene cloning , protopectinase , Saccharomyces cerevisiae , Trichosporon penicillatum and yeast polygalacturonase
© Society Society for General Microbiology 1997
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1997-05-01
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References

  1. Atkinson R. G, Gardner R. C. 1993; A polygalacturonase gene from kiwifruit (Actinidia deliciosa). Plant Physiol 103:669–670
     [Google Scholar]
  2. Becker D. M., Guarente L. 1991; High-efficiency transformation of yeast by electroporation. Methods Enzymol 194:182–187
     [Google Scholar]
  3. Brennan S. O., Peach R. J., Bathurst I.C. 1990; Specificity of yeast KEX2 protease for variant human proalbumins is identical to the in vivo specificity of the hepatic proalbumin convertase. J Biol Chem 265:21494–21497
     [Google Scholar]
  4. Bussink H. J. D., Buxton F. P., Visser. J. 1991; Expression and sequence comparison of the Aspergillus niger and Aspergillus tubigensis genes encoding polygalacturonase II. Curr Genet 19:467–474
     [Google Scholar]
  5. Calvin N. M., Hanawalt P. C. 1988; High-efficiency transformation of bacterial cells by electroporation. J Bacteriol 170:2797–2801
     [Google Scholar]
  6. Caprari C., Richter A., Bergmann C., Lo Cicero S., Salvi G., Cervone F., De Lorenzo G. 1993; Cloning and characterization of a gene encoding the endopolygalacturonase of Fusarium moniliforme. Mycol Res 97:497–505
     [Google Scholar]
  7. Gurr S. J., Unkles S. E., Bergmann C., Kinghorn J. R. 1987; The structure and organization of nuclear genes of filamentous fungi. In Gene Structure in Eukaryotic Microbes,. Edited by J. R. Kinghorn. Oxford : IRL Press. pp:93–139
     [Google Scholar]
  8. Iguchi K., Kishida M., Sakai T. 1996; Purification and characterization of three extracellular protopectinases with polygalacturonase activities from Trichosporon penicillatum. Biosci Biotechnol Biochem 60:603–607
     [Google Scholar]
  9. Julius D., Brake A., Blair L., Kunisawa R., Thorner J. 1984; Isolation of the putative structural gene for lysine-argininecleaving endopeptidase required for processing of yeast prepro-α-factor,. Cell 37:1075–1089
     [Google Scholar]
  10. Kitamoto N., Kimura T., Kito Y., Ohmiya K., Tsukagoshi N. 1993; Structural features of a polygalacturonase gene cloned from Aspergillus oryzae KBN616. FEMS Microbiol Lett 111:37–42
     [Google Scholar]
  11. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
     [Google Scholar]
  12. Niogret M. F., Dubald M., Mandaron P., Mache R. 1991; Characterization of pollen polygalacturonase encoded by several cDNA clones in maize. Plant Mol Biol 17:1155–1164
     [Google Scholar]
  13. Rose M. D., Broach J. R. 1991; Cloning genes by complementation in yeast. Methods Enzymol 194:195–230
     [Google Scholar]
  14. Saarilahti H. I., Heino P., Pakkanen R., Kalkkinen N., Palva I., Palva E.T. 1990; Structural analysis of the pehA gene and characterization of its protein product, endopolygalacturonase, of Erwinia carotovora subspecies carotovora. Mol Microbiol 4:1037–1044
     [Google Scholar]
  15. Sakai T., Okushima M. 1978; Purification and crystallization of a protopectin-solubilizing enzyme from Trichosporon penicillatum. Agric Biol Chem 46:667–676
     [Google Scholar]
  16. Sakai T., Okushima M., Yoshitake S. 1984; Purification, crystallization and some properties of endopolygalacturonase from Kluyveromyces fragilis. Agric Biol Chem 48:1951–1961
     [Google Scholar]
  17. Sakai T., Sawada M., Katsuragi T., Tonomura K. 1989; Possible role of cell wall mannan in the secretion of the pectinsolubilizing enzyme of Trichosporon penicillatum SNO-3. Agric Biol Chem 53:9–18
     [Google Scholar]
  18. Sakai T., Sakamoto T., Hallaert J., Vandamme E. J. 1993; Pectin, qectinase, and protopectinase : production, properties, and applications. Adv Appl Microbiol 39:213–294
     [Google Scholar]
  19. Sambrook J., Fritsch E. F., Maniatis T. 1992; Molecular Cloning: a Laboratory Manual, 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.
     [Google Scholar]
  20. ScottCraig J.S., Panaccione D. G., Cervone F., Walton J. D. 1990; Molecular Cloning: a Laboratory Manual, 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.
     [Google Scholar]
  21. Wach A., Pick H., Philippsen P. 1994; Procedures for isolating yeast DNA for different purposes. In Molecular Genetics of Yeast : a Practical Approach,. Edited by J. R. Johnston. Oxford: IRL Press. pp:1–16
     [Google Scholar]
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Cloning of a protopectinase gene of Trichosporon penicillatum and its expression in Saccharomyces cerevisiae
Microbiology 143, 1657 (1997); https://doi.org/10.1099/00221287-143-5-1657
/content/journal/micro/10.1099/00221287-143-5-1657
/content/journal/micro/10.1099/00221287-143-5-1657
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