1887

Abstract

Summary: Lactobacillus helveticus ITGL1 is able to hydrolyse many amino-acyl and dipeptidyl--nitroanilides. Analysis of heat inactivation kinetics, metal ion and protease inhibitor effects, and the subcellular location of aminopeptidase activities in both the parental strain and mutants deficient in lysyl--nitroanilide hydrolysis, led to the characterization of two cell-wall-associated aminopeptidases. APII and APIV. APII, which catalysed L-lysine -nitroanilide hydrolysis, was purified about 28-fold to homogeneity from cell-wall extracts of ITGL1 and characterized. The purified enzyme appeared to be monomeric, with a molecular mass of 97 kDa. Aminopeptidase activity was greatest at pH 6.5 and 50 C. APII was completely inhibited by bestatin, chelating agents such as EDTA or 1,10-phenanthroline and the divalent cations Zn and Cu. The activity of the EDTA-treated enzyme was restored by Co, Ca or Mn. Although APII was able to degrade several dipeptides and tripeptides with hydrophobic N-terminal amino acid (Leu, Ala), it was inactive on peptides containing Pro or Gly, and may thus contribute to the development of cheese flavour by processing bitter peptides.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-139-7-1441
1993-07-01
2021-05-17
Loading full text...

Full text loading...

/deliver/fulltext/micro/139/7/mic-139-7-1441.html?itemId=/content/journal/micro/10.1099/00221287-139-7-1441&mimeType=html&fmt=ahah

References

  1. Atlan D., Laloi P., Portalier R. 1989; Isolation and characterization of aminopeptidase-deficient Lactobacillus bulgaricus mutants.. Applied Environmental Microbiology 55:1717–1723
    [Google Scholar]
  2. Atlan D., Laloi P., Portalier R. 1990; X-prolyl-dipeptidyl aminopeptidase of Lactobacillus delbrueckii subsp. bulgaricus: characterization of the enzyme and isolation of deficient mutants.. Applied Environmental Microbiology 56:2174–2179
    [Google Scholar]
  3. Bartels H.J., Johnson M.E., Olson N.F. 1987; Accelerated ripening of Gouda cheese. II. Effect of freeze-shocked Lactobacillus helveticus on proteolysis and flavor development.. Milchwissenschaft 42:139–144
    [Google Scholar]
  4. Bradford M.M. 1976; A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding.. Analytical Biochemistry 72:248–254
    [Google Scholar]
  5. De Man J.C., Rogosa M., Sharpe E. 1960; A medium for the cultivation of lactobacilli.. Journal of Applied Bacteriology 23:130–135
    [Google Scholar]
  6. Ezzat N., El Soda M., Desmazeaud M., Ismail A. 1986; Peptide hydrolases from thermobacterium group of lactobacilli. III. Characterization of the intracellular exopeptidases.. Le Lait 66:445–451
    [Google Scholar]
  7. Hickey M.W., Hillier A.J., Jago G.R. 1983; Peptidase activities in lactobacilli.. Australian Journal of Dairy Technology 38:118–123
    [Google Scholar]
  8. Khalid N.M., Marth E.H. 1990a; Purification and partial characterization of a prolyl-dipeptidyl aminopeptidase from Lactobacillus helveticus CNRZ 32.. Applied Environmental Microbiology 55:381–388
    [Google Scholar]
  9. Khalid N.M., Marth E.H. 1990b; Partial purification and characterization of an aminopeptidase from Lactobacillus helveticus CNRZ 32.. Systematic and Applied Microbiology 13:311–319
    [Google Scholar]
  10. Kok J. 1990; Genetics of the proteolytic system of lactic acid bacteria.. FEMS Microbiology Reviews 87:15–42
    [Google Scholar]
  11. Kok J., Venema G. 1988; Genetics of proteinases of lactic acid bacteria.. Biochimie 70:475–488
    [Google Scholar]
  12. Laan H., Smid E.J., TAN P. S. T., Konings W.N. 1989; Enzymes involved in the degradation and utilization of casein in Lactococcus lactis. . Netherland Milk Dairy Journal 43:327–345
    [Google Scholar]
  13. Laemmli U. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4.. Nature, London 227:680–685
    [Google Scholar]
  14. Laloi P., Atlan D., BLANC B, Gilbert C., Portalier R. 1991; Cell-wall-associated proteinase of Lactobacillus delbrueckii subsp. bulgaricus CNRZ 397: differential extraction, purification and properties of the enzyme.. Applied Microbiology and Biotechnology 36:196–204
    [Google Scholar]
  15. Law B.A., Koshlstad J. 1983; Proteolytic system in lactic acid bacteria.. Antonie van Leeuwenhoek 49:225–245
    [Google Scholar]
  16. Miller J. 1972 Experiments in Molecular Genetics Cold Spring Harbor, NY:: Cold Spring Harbor Laboratory;
    [Google Scholar]
  17. Miyakawa H., Kobayashi S., Shimamura S., Tomita M. 1992; Purification and characterization of an aminopeptidase from Lactobacillus helveticus LHE-511.. Journal of Dairy Sciences 75:27–35
    [Google Scholar]
  18. Monnet V., Le BARS D., Gripon J.C. 1986; Specificity of a cell wall proteinase from Streptococcus lactis NCDO 763 towards bovine β-casein.. FEMS Microbiology Letters 36:127–131
    [Google Scholar]
  19. Monnet V., Bockelman W., Gripon J.C., Teuber M. 1989; Comparison of cell wall bound proteinases from Lactococcus lactis subsp. cremoris ACI and Lactococcus lactis subsp. lactis NCDO 763. II. Specificity towards bovine β-casein.. Applied Microbiology and Biotechnology 31:112–118
    [Google Scholar]
  20. Prost F., Chamba J.F. 1990; Rôle de l’activité aminopeptidasique des lactobacilles thermophiles de l’Emmental.. Reunion du Club des Bacteries Lactiques Jouy en Josas, France:
    [Google Scholar]
  21. Thomas T.D., Pritchard G.G. 1987; Proteolytic enzymes of dairy starter cultures.. FEMS Microbiology Reviews 46:245–268
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-139-7-1441
Loading
/content/journal/micro/10.1099/00221287-139-7-1441
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

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