Hostname: page-component-78c5997874-ndw9j Total loading time: 0 Render date: 2024-11-06T07:44:52.672Z Has data issue: false hasContentIssue false
  • English
  • Français

Immunological and electrophoretic study of the proteolytic enzymes from various Lactococcus and Lactobacillus strains

Published online by Cambridge University Press:  01 June 2009

Masahiro Sasaki ,
Boukje W. Bosman  and
Paris S. T. Tan
Masahiro Sasaki
Affiliation:
Snow Brand European Research Laboratories BV, Zernikepark 6, NL-9747 AN Groningen, The Netherlands
Boukje W. Bosman
Affiliation:
Snow Brand European Research Laboratories BV, Zernikepark 6, NL-9747 AN Groningen, The Netherlands
Paris S. T. Tan*
Affiliation:
Snow Brand European Research Laboratories BV, Zernikepark 6, NL-9747 AN Groningen, The Netherlands
*
*For correspondence.
Get access Rights & Permissions [Opens in a new window]

Summary

Cell extracts of various lactobacilli and two Lactococcus strains were investigated for their immunoresponse with monoclonal and polyclonal antibodies raised against various proteolytic enzymes from Lc. lactis. Except for Lactobacillus casei SBT 2233, none of the lactobacilli proteins showed immunoresponse with the monoclonal antibodies. With polyclonal antibodies raised against aminopeptidases N and C and endopeptidase of Lc. lactis an immunoresponse was observed. However, the molecular masses of the reactive bands on the blot were considerably different from those of the corresponding lactococcal peptidases, except for the band that reacted with polyclonal antibodies against aminopeptidase C. The polyclonal antibodies raised against X-prolyl-dipeptidyl aminopeptidase and tripeptidase did not show any immunoreaction. As a control, all antibodies reacted with the lactococcal proteins on the blot, with molecular masses corresponding to those reported for the proteinases and peptidases. The results clearly showed that most of the proteolytic enzymes of lactobacilli were immunologically different from those of lactococci. The proteolytic enzymes in the cell-free extracts were separated by non-denaturing PAGE and visualized by zymogram staining. The electrophoretic pattern of the proteolytic enzymes of lactobacilli was different from that of Lc. lactis. Both experiments indicate that the enzymes of the proteolytic system of lactobacilli are different from those of lactococci.

Type
Original Articles
Information
Journal of Dairy Research , Volume 62 , Issue 4 , November 1995 , pp. 611 - 620
Copyright
Copyright © Proprietors of Journal of Dairy Research 1995

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

REFERENCES

Arora, G. & Lee, B. H. 1992 Purification and characterization of aminopeptidase from Lactobacillus casei ssp. casei LLG. Journal of Dairy Science 75 700–710CrossRefGoogle Scholar
Bockelmann, W., Schulz, Y. & Teuber, M. 1992 Purification and characterization of an aminopeptidase from Lactobacillus delbrueckii subsp. bulgaricus. International Dairy Journal 2 95–107CrossRefGoogle Scholar
Bosman, B. W., Tan, P. S. T. & Konings, W. N. 1990 Purification and characterization of a tripeptidase from Lactococcus lactis subsp. cremoris Wg2. Applied and Environmental Microbiology 56 1839–1843CrossRefGoogle ScholarPubMed
Holck, A. & Næs, H. 1991 Cloning, sequencing and expression of the gene encoding the cell-envelope-associated proteinase from Lactobacillus paracasei subsp. paracasei NCDO 151. Journal of General Microbiology 138 1353–1364CrossRefGoogle Scholar
Khalid, N. M. & Marth, E. H. 1990 Partial purification and characterization of an aminopeptidase from Lactobacillus helveticus CNRZ 32. Systemic and Applied Microbiology 13 311–319CrossRefGoogle Scholar
Klein, J. R., Klein, U., Schad, M. & Plapp, R. 1993 Cloning, DNA sequence analysis and partial characterization of pepN, a lysyl aminopeptidase from Lactobacillus delbrückii ssp. lactis DSM7290. European Journal of Biochemistry 217, 105–114CrossRefGoogle ScholarPubMed
Kok, J. & De Vos, W. M. 1994 The proteolytic system of lactic acid bacteria. In Genetics and Biotechnology of Lactic Acid Bacteria, pp. 169–210 (Eds Gasson, M. J. and Vos, W. M. de). Glasgow: Blackie Academic and ProfessionalCrossRefGoogle Scholar
Kok, J., Leenhouts, K. J., Haandrikman, A. J., Ledeboer, A. M. & Venema, G. 1988 Nucleotide sequence of the cell wall proteinase gene of Streptococcus cremoris Wg2. Applied and Environmental Microbiology 54 231–238CrossRefGoogle ScholarPubMed
Laan, H., Smid, E. J., De Leij, L., Schwander, E. & Konings, W. N. 1988 Monoclonal antibodies to the cell-wall-associated proteinase of Lactococcus lactis subsp. cremoris Wg2. Applied and Environmental Microbiology 54 2250–2256CrossRefGoogle Scholar
Law, B. A. 1984 Flavour development in cheeses. In Advances in the Microbiology and Biochemistry of Cheese and Fermented Milk, pp. 187–208 (Eds Davies, F. L. and Law, B. A.). London: Elsevier Applied ScienceGoogle Scholar
Mayo, B., Kok, J., Venema, K., Bockelmann, W., Teuber, M., Reinke, H. & Venema, G. 1991 Molecular cloning and sequence analysis of the X-prolyl-dipeptidyl-aminopeptidase gene from Lactococcus lactis subsp. cremoris. Applied and Environmental Microbiology 57 38–44CrossRefGoogle ScholarPubMed
Meyer-Barton, E. C., Klein, J. R., Imam, M. & Plapp, R. 1993 Cloning and sequence analysis of the X- prolyl-dipeptidyl-aminopeptidase gene (pepX) from Lactobacillus delbrückii ssp. lactis DSM7290. Applied Microbiology and Biotechnology 40 82–89CrossRefGoogle ScholarPubMed
Miyakawa, H., Kobayashi, S., Shimamura, S. & Tomita, M. 1992 Purification and characterization of an aminopeptidase from Lactobacillus helveticus LHE-511. Journal of Dairy Science 75 27–35CrossRefGoogle Scholar
Nardi, M., Chopin, M. -C., Chopin, A., Cals, M. M. & Gripon, J. -C. 1991 Cloning and DNA-sequence analysis of an X-prolyl-dipeptidyl-aminopeptidase gene from Lactococcus laclis subsp. lactis NCDO 763. Applied and Environmental Microbiology 57 45–50CrossRefGoogle ScholarPubMed
Neviani, E., Boquien, C. -Y., Monnet, V., Phan Than, L. & Gripon, J. -C. 1989 Purification and characterization of an aminopeptidase from Lactococcus laclis subsp. cremoris AM2. Applied and Environmental Microbiology 55 2308–2314CrossRefGoogle ScholarPubMed
Pritchard, G. G. & Coolbear, T. 1993 The physiology and biochemistry of the proteolytic system in lactic acid bacteria. FEMS Microbiology Reviews 12 179–206CrossRefGoogle ScholarPubMed
Sarath, G. 1989 Proteinase assay methods. In Proteolytic Enzymes: A practical approach, pp. 25–54 (Eds Beyond, R. J. and Bond, J. S.). Oxford: IRL PressGoogle Scholar
Sasaki, M., Bosman, B. W. & Tan, P. S. T. 1995 Comparison of proteolytic activities in various lactobacilli. Journal of Dairy Research 62 601–610CrossRefGoogle ScholarPubMed
Tan, P. S. T., Chapot-Chartier, M.P., Pos, K. M., Rousseau, M., Boquien, C. -Y., Gripon, J. -C. & Konings, W. N. 1992 a Localization of peptidases in lactococci. Applied and Environmental Microbiology 58 285–290CrossRefGoogle ScholarPubMed
Tan, P. S. T. & Konings, W. N. 1990 Purification and characterization of an aminopeptidase from Lactococcus lactis subsp. cremoris Wg2. Applied and Environmental Microbiology 56 526–532CrossRefGoogle ScholarPubMed
Tan, P. S. T., Poolman, B. & Konings, W. N. 1993 Review article. The proteolytic enzymes of Lactococcus laclis. Journal of Dairy Research 60 269–286CrossRefGoogle Scholar
Tan, P. S. T., Pos, K. M. & Konings, W. N. 1991 Purification and characterization of an endopeptidase from Lactococcus lactis subsp. cremoris Wg2. Applied and Environmental Microbiology 57 3593–3599CrossRefGoogle ScholarPubMed
Tan, P. S. T., Van Alen-Boerrigter, I. J., Poolman, B., Siezen, R. J., De Vos, W. N. & Konings, W. N. 1992 b Characterization of the Lactococcus lactis pepN gene encoding an aminopeptidase homologous to mammalian aminopeptidase N. FEBS Letters 306 9–16CrossRefGoogle ScholarPubMed
Visser, S. 1993 Proteolytic enzymes and their relation to cheese ripening and flavour. An overview. Journal of Dairy Science 76 329–350CrossRefGoogle Scholar
Van Boven, A., Tan, P. S. T. & Konings, W. N. 1988 Purification and characterization of a dipeptidase from Streptococcus cremoris Wg2. Applied and Environmental Microbiology 54 43–49CrossRefGoogle ScholarPubMed
Zevaco, C., Monnet, V. & Gripon, J. -C. 1990 Intracellular X-prolyl dipeptidyl peptidase from Lactococcus lactis ssp. lactis: purification and properties. Journal of Applied Bacteriology 68 357–366CrossRefGoogle Scholar