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Identification of aryl-phospho-β-d-glucosidases in Bacillus subtilis

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Abstract

Four aryl-phospho-β-d-glucosidases were identified in Bacillus subtilis by using 4-methylumbelliferyl-phospho-β-d-glucopyranoside as a substrate. Two of these enzymes are the products of the bglA and bglH genes, previously suggested to encode aryl-phospho-β-d-glucosidases, while the other enzymes are encoded by the yckE and ydhP genes. Together, these four genes account for >99.9% of the glucosidase activity in B. subtilis on aryl-phospho-β-d-glucosides. yckE was expressed at a low and constant level during growth, sporulation, and spore germination, and was not induced by aryl-β-d-glucosides. ydhP was also not induced by aryl-β-d-glucosides. However, while ydhP was expressed at only a very low level in exponential-phase cells and germinating spores, this gene was expressed at a higher levels upon entry into the stationary phase of growth. Strains lacking yckE or ydhP exhibited no defects in growth, sporulation, or spore germination or in growth on aryl-β-d-glucosides. However, a strain lacking bglA, bglH and yckE grew poorly if at all on aryl-β-d-glucosides as the sole carbon source.

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Abbreviations

MU :

4-Methylumbelliferone

MUG :

4-Methylumbelliferyl-β-d-glucopyranoside

MUGal :

4-Methylumbelliferyl-β-d-galactopyranoside

MUG-P :

4-Methylumbelliferyl-β-d-glucopyranoside-6-phosphate

References

  • Anagnostopoulos C, Spizizen J (1961) Requirements for transformation in Bacillus subtilis. J Bacteriol 81:74–76

    Google Scholar 

  • Davis BJ (1964) Disc electrophoresis. II. Methods and application to human serum proteins. Ann NY Acad Sci 121:404–407

    CAS  Google Scholar 

  • Deutscher J, Galinier A, Martin-Verstraete I (2002) Carbohydrate uptake and metabolism. In: Hoch JA, Losick R, Sonenshein AL (eds) Bacillus subtilis and its closest relatives: from genes to cells. American Society for Microbiology, Washington, DC, pp 129–150

  • Ferrari FA, Nguyen A, Long D, Hoch JA (1983) Construction of an integrable plasmid for Bacillus subtilis. J Bacteriol 154:1513–1515

    CAS  PubMed  Google Scholar 

  • Ferrari E, Howard SM, Hoch JA (1985) Effect of sporulation mutations on subtilisin expression, assayed using a subtilisin-β-galactosidase gene fusion. In: Hoch JA, Setlow P (eds) Molecular biology of microbial differentiation. American Society for Microbiology, Washington, DC, pp 180–184

  • Guerot-Fleury AM, Shazand K, Frandsen N, Stragier P (1995) Antibiotic-resistance cassettes for Bacillus subtilis. Gene 167:335–336

    Article  CAS  PubMed  Google Scholar 

  • Krüger S, Hecker M (1995) Regulation of the putative bglPH operon for aryl-β-glucoside utilization in Bacillus subtilis. J Bacteriol 177:5590–5597

    PubMed  Google Scholar 

  • Krüger S, Hecker M (1996) Transcriptional analysis of bglPH expression in Bacillus subtilis: evidence for two distinct pathways mediating catabolite repression. J Bacteriol 178:2637–2644

    PubMed  Google Scholar 

  • Le Coq D, Lindner C, Krüger S, Steinmetz M, Stülke J (1995) New β-glucoside (bgl) genes in Bacillus subtilis: the bglP gene product has both transport and regulatory functions similar to those of BglF, its Escherichia coli homolog. J Bacteriol 177:1527-1535

    PubMed  Google Scholar 

  • LeDeaux JR, Grossman AD (1995) Isolation and characterization of kinC, a gene that encodes a sensor kinase homologous to the sporulation sensor kinases KinA and KinB in Bacillus subtilis. J Bacteriol 177:166–175

    CAS  PubMed  Google Scholar 

  • Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275

    CAS  Google Scholar 

  • Mason JM, Hackett RH, Setlow P (1988) Studies on the regulation of expression of genes coding for small, acid-soluble proteins of Bacillus subtilis spores using lacZ gene fusions. J Bacteriol 170:239–244

    CAS  PubMed  Google Scholar 

  • Miller J (1972) Assay of β-galactosidase. In: Experiments in molecular genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, pp 352–355

  • Nicholson WL, Setlow P (1990) Sporulation, germination and outgrowth. In: Harwood CR Cutting SM (eds) Molecular biological methods for Bacillus. John Wiley and Sons, Chichester, England, pp 391–450

  • Paidhungat M, Setlow B, Driks A, Setlow P (2000) Characterization of spores of Bacillus subtilis which lack dipicolinic acid. J Bacteriol 182:5505–5512

    Article  CAS  PubMed  Google Scholar 

  • Paidhungat M, Setlow P (2002) Spore germination and outgrowth. In: Hoch JA, Losick R, Sonenshein AL (eds) Bacillus subtilis and its closest relatives: from genes to cells. American Society for Microbiology, Washington, DC, pp 537–548

  • Rotman Y, Fields ML (1967) A modified reagent for dipicolinic acid analysis. Anal Biochem 22:168

    Google Scholar 

  • Saier MH Jr, Goodman SR, Maile RR, Moreno MS, Weyler W, Yang N, Paulsen IT (2002) Overview of transport in Bacillus subtilis. In: Hoch JA, Losick R, Sonenshein AL (eds) Bacillus subtilis and its closest relatives: from genes to cells. American Society for Microbiology, Washington, DC, pp 113–128

  • Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York

    Google Scholar 

  • Spizizen J (1958) Transformation of biochemically deficient strains of Bacillus subtilis by deoxyribonucleate. Proc Natl Acad Sci USA 44:1072–1078

    CAS  Google Scholar 

  • Stragier P, Bonamy C, Karmazyn-Campelli C (1988) Processing of a sporulation sigma factor in Bacillus subtilis: how morphological structure could control gene expression. Cell 52:697–704

    CAS  PubMed  Google Scholar 

  • Thompson J, Pikis A, Ruvinov SB, Henrissat B, Yamamoto H, Sekiguchi J (1998) The gene glvA of Bacillus subtilis 168 encodes a metal-requiring NAD(H)-dependent 6-α-phosphoglucosidase. Assignment to family 4 of the glycosylhydrolase superfamily. J Biol Chem 273:27347–27356

    Article  CAS  PubMed  Google Scholar 

  • Tobisch S, Glaser P, Krüger S, Hecker M (1997) Identification and characterization of a new β-glucoside utilization system in Bacillus subtilis. J Bacteriol 179:496–506

    CAS  PubMed  Google Scholar 

  • Wilson G, Fox CF (1974) The β-glucoside system of Eschericia coli. IV. Purification and properties of phospho-β-glucosidases A and B. J Biol Chem 249:5586–5598

    CAS  PubMed  Google Scholar 

  • Zhang, J, Aronson A (1994) A Bacillus subtilis bglA gene encoding phospho-β-glucosidase is inducible and closely linked to a NADH dehydrogenase-encoding gene. Gene 140:85–90

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

This work was supported by contracts from the 3M Corporation. We are grateful to A. Aronson for the gift of strain JZ30 and to Kasia Koziol-Dube for assistance with some experiments.

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Author notes

  1. Arturo Cabrera-Hernandez

    Present address: Center for Immunotherapy, University of Connecticut Health Center, Farmington, CT 06032, USA

Authors and Affiliations

  1. Department of Molecular, Microbial, and Structural Biology, University of Connecticut Health Center, Farmington, CT 06032, USA

    Barbara Setlow, Arturo Cabrera-Hernandez, Rosa Martha Cabrera-Martinez & Peter Setlow

  2. Departamento de Bioquimica, Instituto Politecnico Nacional, Escuela Nacional de Ciencas Biologicas, 11340, Mexico City, Mexico

    Rosa Martha Cabrera-Martinez

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  1. Barbara Setlow
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  2. Arturo Cabrera-Hernandez
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  4. Peter Setlow
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Correspondence to Peter Setlow.

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Setlow, B., Cabrera-Hernandez, A., Cabrera-Martinez, R.M. et al. Identification of aryl-phospho-β-d-glucosidases in Bacillus subtilis . Arch Microbiol 181, 60–67 (2004). https://doi.org/10.1007/s00203-003-0628-2

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