1887

Abstract

The ß-galactosidase of DSM 20017 is encoded by two genes located on its chromosome. These genes designated and were cloned in NM 554 on an 8.65 kbp fragment inserted in vector pRB473. Deletion analysis of the originally cloned fragment revealed that both genes are required for the formation of a functional ß-galactosidase. and are transcribed as a single transcript of approximately 2.9 kbp starting 34 bp upstream of the translational start codon. The proteins derived from and share only 18-59% homology with other ß-galactosidases. The genes encoding the ß-galactosidase are scattered with multiple direct and inverted repeats of 9-12 bp. However, comparison with the plasmid-encoded ß-galactosidase revealed equal distribution of conserved amino acid residues and suggests that the genes have a common origin. Specific deletions or insertions resulting from the presence of the repeats were not observed. The ß-galactosidase was phenotypically expressed in NM 554 and LTH 1432. Its two genes can be used to replace antibiotic reporter genes to develop food-grade vectors and á-complementation systems for self-cloning in meat lactobacilli.

Loading

Article metrics loading...

/content/journal/micro/10.1099/13500872-141-12-3059
1995-12-01
2021-04-21
Loading full text...

Full text loading...

/deliver/fulltext/micro/141/12/mic-141-12-3059.html?itemId=/content/journal/micro/10.1099/13500872-141-12-3059&mimeType=html&fmt=ahah

References

  1. Birnboim H.C., Doly J. 1979; A rapid alkaline extraction procedure for screening recombinant plasmid DNA.. Nucleic Acids Res 7:1513–1523
    [Google Scholar]
  2. Biswas I., Gruss S., Ehrlich S.D., Maguin E. 1993; High-efficiency gene inactivation and replacement system for grampositive bacteria.. J Bacteriol 175:3628–3635
    [Google Scholar]
  3. Boizet B., Villeval D., Slos P., Novel M., Novel G., Mercenier A. 1988; Isolation and structural analysis of the phospho-β-galactosidase gene from Streptococcus lactis Z268.. Gene 62:249–261
    [Google Scholar]
  4. Bringel F., Hubert J.-C. 1990; Optimized transformation by electroporation of Lactobacillus plantarum strains with plasmid vectors.. Appl Microbiol Biotechnol 33:664–670
    [Google Scholar]
  5. Bullock W.O., Fernandez J.M., Short J.M. 1987; XLl-Blue: a high efficiency plasmid transforming recK Escherichia coli strain with β-galactosidase selection.. Biotechniques 5:376–378
    [Google Scholar]
  6. Burchhardt G., Bahl H. 1991; Cloning and analysis of the β-galactosidase-encoding gene from Clostridium thermosulfurogenesEMI.. Gene 106:13–19
    [Google Scholar]
  7. Buvinger W.E., Riley M. 1985; Nucleotide sequence of Klebsiellapneumoniae lac genes.. J Bacteriol 163:850–857
    [Google Scholar]
  8. Chassy B.M., Thompson J. 1983; Regulation of lactose- phosphoenolpyruvate-dependent phosphotransferase system and β-d-phosphogalactosidegalactohydrolase activities in Lactobacillus casei.. J Bacteriol 154:1195–1203
    [Google Scholar]
  9. David S., Stevens H., van Riel M., Simons G., de Vos W.M. 1992; Leuconostoclactis β-galactosidase is encoded by two overlapping genes.. J Bacteriol 174:4475–4481
    [Google Scholar]
  10. Dretzen G., Bellard M., Sassone-Corsi P., Chambon P. 1981; Recovery of DNA fragments from agarose gels using DEAE paper.. Anal Biochem 170:38–44
    [Google Scholar]
  11. Farrow J.A.E. 1980; Lactose hydrolysing enzymes in Streptococcus lactis and Streptococcus cremoris and also in some other species of streptococci.. J Appl Bacteriol 49:493–503
    [Google Scholar]
  12. Flickinger J.L., Porter E.V., Chassy B.M. 1986; Molecular cloning of a plasmid-encoded β-galactosidase from Lactobacillus casei.. Abstr Annu Meet Am Soc Microbiol 86:156
    [Google Scholar]
  13. Germond J.E., Lapierre L., Delley M., Mollet B. 1993; Novel IS element (ISL3) located next to the Lactobacillus bulgaricuslacZgene.. FEMS Microbiol Rep 12:A13
    [Google Scholar]
  14. Haima P., van Sinderen D., Schotting H., Bron S., Venema G. 1990; Development of a β-galactosidase α-complementation system for molecular cloning in Bacillus subtilis.. Gene 86:63–69
    [Google Scholar]
  15. Hammes W.P., Bantleon A., Min S. 1990; Lactic acid bacteria in meat fermentation.. FEMS Microbiol Rev 87:165–174
    [Google Scholar]
  16. Hanahan F.C.H. 1985; Techniques for transformation of E. coli.. In DNA Cloning 1 pp. 109–135 Edited by Glover D.M. Oxford & Washington, DC: IRL Press;
    [Google Scholar]
  17. Hancock K.R., Rockman E., Young C.A., Pearce L., Maddox I.S., Scott D.B. 1991; Expression and nucleotide sequence of the Clostridium acetobutylicum β-galactosidase gene cloned in Escherichia coli.. J Bacteriol 173:3084–3095
    [Google Scholar]
  18. Hashiba H., Takiguchi R., Jyoho K., Aoyama K. 1992; Establishment of a host-vector system in Lactobacillus helveticus with β-galactosidase activity as a selection marker.. Biosci Biotechnol Biochem 56:190–194
    [Google Scholar]
  19. Hawley D.K., McClure W.R. 1983; Compilation and analysis of Escherichia coli promoter DNA sequences.. Nucleic Acids Res 11:2237–2255
    [Google Scholar]
  20. Hengstenberg W., Reiche B., Eisermann R., Fischer R., KeBler U., Tarrach A., de Vos W.M., Kalbitzer H.-R., Glaser S. 1989; Structure and function of proteins involved in sugar transport by the PTS of Gram-positive bacteria.. FEMS Microbiol Rev 63:35–42
    [Google Scholar]
  21. Herman R.E., McKay L.L. 1986; Cloning and expression of the β-d-galactosidasegene from Streptococcus thermophilus in Escherichia coli.. Appl Erniron Microbiol 52:45–50
    [Google Scholar]
  22. Herrchen M., Legler G. 1984; Identification of an essential carboxylate group at the active site of lacZ β-galactosidase from Escherichia coli.. Eur J Biochem 138:527–531
    [Google Scholar]
  23. Hirata H., Fukazawa T., Negoro S., Okada H. 1986; Structure of a jS-galactosidase gene of Bacillus stearothermophilus.. J Bacteriol 166:722–727
    [Google Scholar]
  24. Kalnins A., Otto K., Rüther U., Müller-Hill B. 1983; Sequence of the lacZ gene in E. coli.. EMBO J 2:593–597
    [Google Scholar]
  25. Knauf H.J., Vogel R.F., Hammes W.P. 1992; Cloning, sequence and phenotypic expression of katA, which encodes the catalase of Lactobacillus sake LTH 677.. Appl Enoiron Microbiol 58:832–839
    [Google Scholar]
  26. Leong-Morgenthaler P., Zwahlen M.C., Hottinger H. 1991; Lactose metabolism in Lactobacillus bulgaricus: analysis of the primary structure and expression of the genes involved. J Bacteriol 173:1951–1957
    [Google Scholar]
  27. McKay L., Miller A. III Sandine W.E., Elliker P.R. 1970; Mechanisms of lactose utilization by lactic acid streptococci: enzymatic and genetic analyses.. J Bacteriol 102:804–809
    [Google Scholar]
  28. de Man J.C., Rogosa M., Sharpe M.E. 1960; A medium for the cultivation of lactobacilli.. J Appl Bacteriol 23:130–135
    [Google Scholar]
  29. Mayo B., González B., Area P., Suárez J.E. 1994; Cloning and expression of the plasmid encoded β-D-galactosidase gene from a Lactobacillus plantarumstrain of dairy origin.. FEMS Microbiol Lett 122:145–151
    [Google Scholar]
  30. Mollet B., Delley M. 1990; Spontaneous deletion formation within the /Lgalactosidase gene of Lactobacillus bulgaricus.. J Bacteriol 172:5670–5676
    [Google Scholar]
  31. Obst M., Hehn R., Vogel R.F., Hammes W.P. 1992; Lactose metabolism in Lactobacillus curvatus and Lactobacillus sake.. FEMS Microbiol Lett 97:209–214
    [Google Scholar]
  32. Oshima A., Tsuji A., Nagao Y., Sakuraba H., Suzuki Y. 1988; Cloning, sequencing, and expression of cDNA for human β-galactosidase.. Biochem Biophys Res Commun 157:238–244
    [Google Scholar]
  33. Romano A.H., Brino G., Peterkofsky A., Reizer J. 1987; Regulation of β-galactoside transport and accumulation in heterofermentative lactic acid bacteria.. J Bacteriol 169:5589–5596
    [Google Scholar]
  34. van Rooijen R.J. 1993 Characterisation of the Lactococcuslactis lactose genes and regulation of their expression. PhD thesis, NIZO, The Netherlands
    [Google Scholar]
  35. 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]
  36. Sanger F., Nicklen S., Coulson A.R. 1977; DNA sequencing with chain-terminating inhibitors.. Proc Natl Acad Sci USA 745463–5467
    [Google Scholar]
  37. Schmidt B.F., Adams R.M., Requadt C., Power S., Meinzer S.E. 1989; Expression and nucleotide sequence of the Lactobacillus bulgaricus β-galactosidase gene cloned in Escherichia coli.. J Bacteriol 171:625–635
    [Google Scholar]
  38. Schroeder C.J., Robert C., Lenzen G., McKay L.L., Mercenier A. 1991; Analysis of the lacZ sequences from two Streptococcus thermophilus strains: comparison with the Escherichia coli and Lactobacillus bulgaricus β-galactosidase sequences.. J Gen Microbiol 137:369–380
    [Google Scholar]
  39. Shimizu-Kadota M. 1988; Cloning and expression of the phospho-β-galactosidase genes on the lactose plasmid and the chromosome of Lactobacillus casei C257 in Escherichia coli.. Biochemistry 70:523–529
    [Google Scholar]
  40. Tabor S., Richardson C.C. 1989; Selective inactivation of the exonuclease activity of bacteriophage T7 DNA polymerase by in vitro mutagenesis.. J Biol Chem 264:6447–6458
    [Google Scholar]
  41. Tichaczek P.S., Vogel R.F., Hammes W.P. 1993; Cloning and sequencing of cur A encoding curvacin A, the bacteriocin produced by Lactobacillus curvatus LTH 1174.. Arch Microbiol 160:279–283
    [Google Scholar]
  42. Tichaczek P.S., Vogel R.F., Hammes W.P. 1994; Cloning and sequencing of sakP encoding sakacin P, the bacteriocin produced by Lactobacillus sake LTH 673.. Microbiology 140:361–367
    [Google Scholar]
  43. de Vos W.M., Simons G. 1988; Molecular cloning of lactose genes in dairy lactic streptococci: the phospho-β-galactosidase and β-galactosidase genes and their expression products.. Biochimie 70:461–473
    [Google Scholar]
  44. de Vos W.M., Boerrigter I., van Rooijen R.J., Reiche B., Hengstenberg W. 1990; Characterization of the lactose-specific enzymes of the phosphotransferase system in Lactococcuslactis.. J Biol Chem 265:22554–22560
    [Google Scholar]
  45. Wetmur J.G. 1991; DNA probes: applications of the principles of nucleic acid hybridization.. Crit Rev Biochem Mol Biol 26:227–259
    [Google Scholar]
  46. Zabin I. 1982; β-galactosidase a-complementation.A model of protein-protein interaction.. Mol Cell Biochem 49:87–96
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/13500872-141-12-3059
Loading
/content/journal/micro/10.1099/13500872-141-12-3059
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error