1887

Abstract

species secrete large amounts of alkaline phosphatase (AP) enzymes that have not been characterized so far. An AP has been purified to homogeneity from cultures of IMRU 3570. The enzyme has a monomer size of 62 kDa and is processed in the culture to a 33 kDa protein as shown by immunoblotting. The enzyme was purified by ammonium sulfate precipitation, CM-Sephadex cationic exchange, chromatofocusing and HPLC Sphaerogel 3000SW filtration. The pure enzyme uses a variety of organic phosphorylated compounds as substrates. The N-terminal end of the mature protein was found to be RLREDPFTLGVASGDPHP. The gene has been cloned using as probe an oligomer based on the N-terminal sequence of the AP. encodes a protein of 62678 Da with low homology to the AP of . The gene was found to be homologous to three alkaline-phosphatase-encoding genes previously identified in the genome. On the basis of the optimal pH, substrate specificity and differences in amino acid sequence of motifs defining the active centre of APs, the AP uses a wide range of organic phosphate substrates and is different from the phosphatases of Gram-negative bacteria.

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2001-06-01
2020-09-29
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References

  1. Anba J., Bidaud M., Vasil M. L., Lazdunski A.. 1990; Nucleotide sequence of the Pseudomonas aeruginosa pho B gene, the regulatory gene for the phosphate regulon. J Bacteriol172:4685–4689
    [Google Scholar]
  2. Asturias J. A., Liras P., Martı́n J. F.. 1990; Phosphate control of pab S gene transcription during candicidin biosynthesis. Gene93:79–84[CrossRef]
    [Google Scholar]
  3. Au S., Roy K. L., Tigerstrom R. G.. 1991; Nucleotide sequence and characterization of the gene for secreted alkaline phosphatase from Lysobacter enzymogenes . J Bacteriol173:4551–4557
    [Google Scholar]
  4. Bibb M. J., Jones G. H., Joseph R., Buttner M. J., Ward J. M.. 1987; The agarase gene ( dag A) of Streptomyces coelicolor A3(2): affinity purification and characterization of the cloned gene product. J Gen Microbiol133:2089–2096
    [Google Scholar]
  5. Blanco J., Coque J. J. R., Velasco J., Martı́n J. F.. 1997; Cloning, expression in Streptomyces lividans and biochemical characterization of a endo-beta-1,4-xylanase of Thermomonospora alba ULJB1 with cellulose-binding. Appl Microbiol Biotechnol48:208–217[CrossRef]
    [Google Scholar]
  6. Chenust R. S., Bookstein C., Hulett F. M.. 1991; Separate promoters direct expression of pho AIII, a member of the Bacillus subtilis alkaline phosphatase multigene family, during phosphate starvation and sporulation. Mol Microbiol5:2181–2190[CrossRef]
    [Google Scholar]
  7. Cole S. T., Brosh R., Parkhill J.. 39 other authors 1998; Deciphering the biology of Mycobacterium tuberculosis from the complete genome sequence. Nature393:537–544[CrossRef]
    [Google Scholar]
  8. Daza A., Gil J. A., Vigal T., Martı́n J. F.. 1990; Cloning and characterization of a gene of Streptomyces griseus that increases production of extracellular enzymes in several Streptomyces . Mol Gen Genet222:384–392[CrossRef]
    [Google Scholar]
  9. Eder S., Shi L., Jensen K., Yamane K., Hulett F. M.. 1996; A Bacillus subtilis secreted phosphodiesterase/alkaline phosphatase is the product of a Pho regulon gene, phoD . Microbiology142:2041–2047[CrossRef]
    [Google Scholar]
  10. Fernández-Abalos J. M., Sánchez P., Coll P. M., Villanueva J. R., Santamarı́a R. I., Pérez P.. 1992; Cloning and nucleotide sequence of cel A1, an endo-beta-1,4-glucanase-encoding gene from Streptomyces halstedii JM8. J Bacteriol174:6368–6376
    [Google Scholar]
  11. Furukawa K., Hasunuma K., Shinohara Y.. 1987; Characterization of Pi-repressible enzymes secreted in culture media by Neurospora crassa wild-type cells and null-type mutants. J Bacteriol169:4790–4795
    [Google Scholar]
  12. Garcı́a-González M. D., Martı́n J. F., Vigal T., Liras P.. 1991; Characterization, expression in Streptomyces lividans , and processing of the amylase of Streptomyces griseus IMRU 3570: two different amylases are derived from the same gene by an intracellular processing mechanism. J Bacteriol173:2451–2458
    [Google Scholar]
  13. Goldman S., Hecht K., Eisenberg H., Mevarech M.. 1990; Extracellular Ca2+-dependent inducible alkaline phosphatase from the extremely halophilic archaebacterium Haloarcula marismortui . J Bacteriol172:7065–7070
    [Google Scholar]
  14. Golovan S., Wang G., Zhang J., Forsberg C. W.. 2000; Characterization and overproduction of the Escherichia coli app A encoded bifunctional enzyme that exhibits both phytase and acid phosphatase activities. Can J Microbiol46:59–71[CrossRef]
    [Google Scholar]
  15. Ha N. C., Kim Y. O., Oh T. K., Oh B. H.. 1999; Preliminary X-ray crystallographic analysis of a novel phytase from a Bacillus amyloliquefaciens strain. Acta Crystallogr D Biol Crystallogr55:691–693[CrossRef]
    [Google Scholar]
  16. Ha N. C., Oh B. C., Shin S., Kim H. J., Oh T. K., Kim Y. O., Choi K. Y., Oh B. H.. 2000; Crystal structures of a novel, thermostable phytase in partially and fully calcium-loaded states. Nat Struct Biol7:147–153[CrossRef]
    [Google Scholar]
  17. van Hartingsveldt W., van Zeijl C. M., Harteveld G. M.. 7 other authors 1993; Cloning, characterization and overexpression of the phytase-encoding gene ( phyA ) of Aspergillus niger . Gene127:87–94[CrossRef]
    [Google Scholar]
  18. von Heijne G.. 1986; A new method for predicting signal sequence cleavage sites. Nucleic Acids Res14:4683–4690[CrossRef]
    [Google Scholar]
  19. Hopwood D. A., Bibb M. J., Chater J. K., Kieser T., Bruton C. J., Kieser H. M., Smith C. P., Ward J. M., Schrempf H.. 1985; Genetic Manipulation in Streptomyces: a Laboratory Manual Norwich, UK: John Innes Foundation;
    [Google Scholar]
  20. Hulett F. M., Kim E. E., Bookstein C., Kapp N. V., Edwards C. W., Wyckoff H. W.. 1991; Bacillus subtilis alkaline phosphatases III and IV. Cloning, sequencing and comparisons of deduced amino acid sequence with Escherichia coli alkaline phosphatase three-dimensional structure. J Biol Chem266:1077–1084
    [Google Scholar]
  21. Kantrowitz E. R.. 1994; Structure and function of Escherichia coli alkaline phosphatase. In Phosphate in Microorganisms, Cellular and Molecular Biology pp319–328 Edited by Torriani-Gorini A.. Yagil E., Silver S.. Washington, DC: American Society for Microbiology;
    [Google Scholar]
  22. Kerovuo J., Lauraeus M., Nurminen P., Kalkkinen N., Apajalahti J.. 1998; Isolation, characterization, molecular gene cloning, and sequencing of a novel phytase from Bacillus subtilis . Appl Environ Microbiol64:2079–2085
    [Google Scholar]
  23. Laemmli U. K.. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature227:680–685[CrossRef]
    [Google Scholar]
  24. Lee T., Makino K., Shinagawa H., Amemura M., Nakata A.. 1989; Phosphate regulon in members of the family Enterobacteriaceae : comparison of the pho B- pho R operons of E. coli , Shigella dysenteriae and Klebsiella pneumoniae . J Bacteriol171:6593–6599
    [Google Scholar]
  25. Leloir L. F., Cardini C. E.. 1957; Characterization of phosphorous compounds by acid lability. Methods Enzymol3:840–859
    [Google Scholar]
  26. Lim D., Golovan S., Forsberg C. W., Jia Z.. 2000; Crystal structures of Escherichia coli phytase and its complex with phytate. Nat Struct Biol7:108–113[CrossRef]
    [Google Scholar]
  27. Liras P., Martı́n J. F., Villanueva J. R.. 1977; Sequential expression of macromolecule biosynthesis and candicidin formation in Streptomyces griseus . J Gen Microbiol102:269–277[CrossRef]
    [Google Scholar]
  28. Martı́n J. F.. 1989; Molecular mechanisms for the control by phosphate of the biosynthesis of antibiotics and other secondary metabolites. In Regulation of Secondary Metabolism in Actinomycetes pp213–237 Edited by Shapiro S.. Boca Raton, FL: CRC Press;
    [Google Scholar]
  29. Martı́n J. F., McDaniel L. E.. 1975; Kinetics of biosynthesis of polyene macrolide antibiotics in batch cultures: cell maturation time. Biotechnol Bioeng17:925–938[CrossRef]
    [Google Scholar]
  30. Martı́n J. F., Naharro G., Liras P., Villanueva J. R.. 1979; Isolation of mutants deregulated in phosphate control of candicidin biosynthesis. J Antibiot32:600–606[CrossRef]
    [Google Scholar]
  31. Martı́n J. F., Marcos A. T., Martı́n A., Asturias J. A., Liras P.. 1994; Phosphate control of antibiotic biosynthesis at the transcriptional level. In Phosphate in Microorganisms: Cellular and Molecular Biology pp140–147 Edited by Torriani-Gorini A.. Yagil E., Silver S.. Washington, DC: American Society for Microbiology;
    [Google Scholar]
  32. Mitchell D. B., Vogel K., Weimann B. J., Pasamontes L., van Loon A. P.. 1997; The phytase subfamily of histidine acid phosphatases: isolation of genes for two novel phytases from the fungi Aspergillus terreus and Myceliophthora thermophila . Microbiology143:245–252[CrossRef]
    [Google Scholar]
  33. Moos M., Yen Nguyen N., Liu T.-Y.. 1988; Reproducible high yield sequencing of proteins electrophoretically separated and transfered to an inert support. J Biol Chem263:6005–6008
    [Google Scholar]
  34. Park T., Lee J. H., Kim H. K., Hoe H. S., Kwon S. T.. 1999; Nucleotide sequence of the gene for alkaline phosphatase of Thermus caldophilus GK24 and characteristics of the deduced primary structure of the enzyme. FEMS Microbiol Lett180:133–139[CrossRef]
    [Google Scholar]
  35. Piddington C. S., Houston C. S., Paloheimo M., Cantrell M., Miettinen-Oinonen A., Nevalainen H., Rambosek J.. 1993; The cloning and sequencing of the genes encoding phytase ( phy ) and pH 2·5-optimum acid phosphatase ( aph ) from Aspergillus niger var. awamori . Gene133:55–62[CrossRef]
    [Google Scholar]
  36. Pond J. L., Eddy C. K., Mackenzie K. F., Conway T., Borecky D. J., Ingram L. O.. 1989; Cloning, sequencing and characterization of the principal acid phosphatase, the pho C product from Zymomonas mobilis . J Bacteriol171:767–774
    [Google Scholar]
  37. Powar V. K., Jagannathan V.. 1982; Purification and properties of phytate-specific phosphatase from Bacillus subtilis . J Bacteriol151:1102–1108
    [Google Scholar]
  38. Sambrook J., Fritsch E. F., Maniatis T.. 1989; Molecular Cloning: a Laboratory Manual , 2nd edn. Cold Spring Harbor, NY: Cold Springer Harbor Laboratory;
    [Google Scholar]
  39. Schlochtermeier A., Walter S., Schroder J., Moorman M., Schrempf H.. 1992; The gene encoding the cellulase (Avicelase) Cel1 from Streptomyces reticuli and analysis of protein domains. Mol Microbiol6:3611–3621[CrossRef]
    [Google Scholar]
  40. Towbin H., Staehelin T., Gordon J.. 1992; Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Biotechnology24:145–149
    [Google Scholar]
  41. Urabe H., Lenzini M. W., Mukaide M., Dusart J., Nakano M. M., Ghuysen J. M., Ogawara H.. 1990; β-Lactamase expression in Streptomyces cacaoi . J Bacteriol172:6427–6434
    [Google Scholar]
  42. Vigal T., Gil J. A., Daza A., Martı́n J. F., Garcı́a-González M. D.. 1991; Cloning, characterization and expression of an α-amylase gene from Streptomyces griseus IMRU 3570. Mol Gen Genet225:278–288[CrossRef]
    [Google Scholar]
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