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Purification and characterization of a post-proline dipeptidyl aminopeptidase from Streptococcus cremoris AM2

Published online by Cambridge University Press:  01 June 2009

Mary Booth ,
Ide Ni Fhaoláin ,
P. Vincent Jennings  and
Gerard O'Cuinn
Mary Booth
Affiliation:
Department of Biochemistry, University College, Galway and Department of Life Sciences, Regional Technical College, Galway, Ireland
Ide Ni Fhaoláin
Affiliation:
Department of Life Sciences, Regional Technical College, Galway, Ireland
P. Vincent Jennings
Affiliation:
Department of Life Sciences, Regional Technical College, Galway, Ireland
Gerard O'Cuinn
Affiliation:
Department of Life Sciences, Regional Technical College, Galway, Ireland
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Summary

The present study describes the purification of a post-proline dipeptidyl aminopeptidase from the cytoplasm of Streptococcus cremoris AM2. On the basis of its elution from a calibrated Sephadex G200 column, the enzyme had a molecular weight of 117000 and exhibited a broad pH optimum activity between 6·0 and 9·0. The activity was most comprehensively inhibited by phenylmethylsulphonylfluoride and more modestly inhibited by 1,10-phenanthroline and 8-hydroxyquinoline but not by EDTA. A range of peptides containing either proline or alanine as the penultimate amino acid residue could act as substrates. The presence of proline on the carboxy side of the scissile bond prevented hydrolysis. However the enzyme could release Pro-Pro from Pro-Pro-Gly-Phe-Ser-Pro. The significance of this substrate specificity is considered in the context of removal of either single proline residues or prolylproline sequences from oligopeptides during cheese ripening.

Type
Original Articles
Information
Journal of Dairy Research , Volume 57 , Issue 1 , February 1990 , pp. 89 - 99
Copyright
Copyright © Proprietors of Journal of Dairy Research 1990

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References

REFERENCES

Biede, S. L. & Hammond, E. G. 1979 Swiss cheese flavor; 1 Chemical analysis. Journal of Dairy Science 62 227–237CrossRefGoogle Scholar
Booth, M., Donnelly, W. J., Nifhaolain, I., Jennings, P. V. & O'Cuinn, G. 1990 Proline-specific peptidases of Streptococcus cremoris AM2. Journal of Dairy Research 57 79–88CrossRefGoogle Scholar
Booth, M., Donnelly, W. J., Nifhaolain, I., Jennings, P. V. & O'Cuinn, G. 1989 Preparation of subcellular fractions from strains of Streptococcus cremoris. Biochemical Society Transactions 17 385–386CrossRefGoogle Scholar
Browne, P. & O'Cuinn, G. 1983 An evaluation of the role of a pyroglutamyl peptidase, a post-proline cleaving enzyme and a post-proline dipeptidyl aminopeptidase, each purified from the soluble fraction of guinea-pig brain, in the degradation of thyroliberin in vitro. European Journal of Biochemistry 137 75–87CrossRefGoogle Scholar
Casey, M. G. & Meyer, J. 1985 Presence of X-prolyl-dipeptidyl peptidase in lactic acid bacteria. Journal of Dairy Science 68 3212–3215CrossRefGoogle ScholarPubMed
Fukasawa, K. M. & Harada, M. 1981 Purification and properties of dipeptidyl peptidase IV from Streptococcus mitis ATCC 9811 Archives of Biochemistry and Biophysics 210 230–237CrossRefGoogle ScholarPubMed
García-Alvárez, N., Bordallo, C., Gascon, S. & Suarez-Renduele, P. 1985 Purification and characterisation of a thermosensitive X-prolyl dipeptidyl aminopeptidase (dipeptidyl aminopeptidase yscV) from Saccharomyces cerevisiae. Biochimica et Biophysica. Acta 832 119–125Google Scholar
Heymann, E. & Mentlein, R. 1986 Complementary action of dipeptidyl peptidase IV and minopeptidase M in the digestion of β-casein. Journal of Dairy Research 53 229–236CrossRefGoogle ScholarPubMed
Hickey, M. W., Hillier, A. J. & Jago, G. R. 1983 Peptidase activities in lactobacilli. Australian Journal of Dairy Technology 38 118–123Google Scholar
Johnson, W. C. & Lindsay, A. J. 1939 An improved universal buffer. Analyst 64 490–492CrossRefGoogle Scholar
Kato, T., Nagatsu, T., Kimura, T. & Sakakibara, S. 1978 Fluorescence assay of X-prolyl dipeptidyl aminopeptidase activity with a new fluorogenic substrate. Biochemical Medicine 19 351–359CrossRefGoogle ScholarPubMed
Meyer, J. & Jordi, R. 1987 Purification and characterisation of X-prolyl dipeptidyl aminopeptidase from Lactobacillus lactis and from Streptococcus thermophilus. Journal of Dairy Science 70 738–745CrossRefGoogle ScholarPubMed
Mills, O. E. & Thomas, T. D. 1981 Nitrogen sources for growth of lactic Streptococci in milk. New Zealand Journal of Dairy Science and Technology 16 43–55Google Scholar
Mou, L., Sullivan, J. J. & Jago, G. R. 1975 Peptidase activities in Group N streptococci. Journal nf Dairy Research 42 147–155CrossRefGoogle Scholar
O'Connor, B. & O'Cuinn, G. 1986 Post-proline dipeptidyl aminopeptidase from synaptosomal membranes of guinea-pig brain: a possible role for this activity in the hydrolysis of His-ProNH2, arising from the action of synaptosomal membrane pyroglutamate aminopeptidase on thyroliberin. European Journal of Biochemistry 154 329–335CrossRefGoogle Scholar
O'Keeffe, R. B., Fox, P. F. & Daly, C. 1976 Contribution of rennet and starter proteases to proteolysis in Cheddar cheese. Journal of Dairy Research 43 97–107CrossRefGoogle Scholar
O'Keefe, A. M., Fox, P. F. & Daly, C. 1978 Proteolysis in cheddar cheese: role of coagulant and starter bacteria. Journal of Dairy Research 45 465–477CrossRefGoogle Scholar
Ruettimann, K. W. & Ladisch, M. R. 1987 Casein micelles, structure, properties and enzymic coagulation. Enzyme Microbiology and Technology 9 578–589CrossRefGoogle Scholar
Sullivan, J. J. & Jago, G. R. 1972 The structure of bitter peptides and their formation from casein. Australian Journal of Dairy Technology 27 98–104Google Scholar
Visser, S., Exterkate, F. A., Slangen, C. J. & DeVeer, G. J. C. M. 1986 Comparative study of action of cell wall proteinases from various strains of Streptococcus cremoris on bovine αs1-, β- and κ-casein. Applied and Environmental Microbiology 52 1162–1166CrossRefGoogle ScholarPubMed
Walter, R., Simmons, W. H. & Yoshimoto, T. 1980 Proline specific endo- and exopeptidases. Molecular and Cellular Biochemistry 30 111–127CrossRefGoogle ScholarPubMed
Yoshimoto, T. & Tsuru, D. 1982 Proline-specifie dipeptidyl aminopeptidase from Flavobacterium meningosepticum. Journal of Biochemistry 91 1899–1906CrossRefGoogle ScholarPubMed