Purification and Characterization of a Carboxypeptidase Y from Kluyveromyces fragilis JSB95

A carboxypeptidase Y was purified to homogeneity from the crude cell extracts of Kluyveromyces fragilis JSB95 grown in yeast, peptone, and dextrose broth using an FPLC® system equipped with ion-exchange and gel filtration columns. The enzyme was purified 216-fold over the crude extract; recovery was...

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Published in:Journal of dairy science 1998-03, Vol.81 (3), p.647-654
Main Authors: Transfiguracion, Julia C., Lee, Byong H., Park, Seung Y., Van der Voort, Fred R.
Format: Article
Language:English
Subjects:
Biological and medical sciences
bitterness removal
carboxypeptidase Y
cheese ripening
Food industries
Food microbiology
Fundamental and applied biological sciences. Psychology
Kluyveromyces fragilis
Milk and cheese industries. Ice creams
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creator Transfiguracion, Julia C.
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description A carboxypeptidase Y was purified to homogeneity from the crude cell extracts of Kluyveromyces fragilis JSB95 grown in yeast, peptone, and dextrose broth using an FPLC® system equipped with ion-exchange and gel filtration columns. The enzyme was purified 216-fold over the crude extract; recovery was 18%. The estimated molecular mass was 56 kDa and consisted of two subunits. The maximum activity of the purified enzyme was obtained at pH 6.0 and 25°C and was strongly inhibited by diisopropylphosphofluoridate and phenylmethylsulfonylfluoride as well as by some metal ions. The Michaelis constant and maximum velocity values for n-benzoyl-L-tyrosine-p-nitroanilide substrate were 5.1mM and 22.58 μM/min per mg of protein; for Cbz-Phe-Ala, values were 2.98mM and 22.58 μM per min/mg of protein. Carboxypeptidase Y showed hydrolytic activity against some hydrophobic peptides of the tryptic digests of αs1- and β-caseins.
doi_str_mv 10.3168/jds.S0022-0302(98)75619-X
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source Elsevier ScienceDirect Journals; Elektronische Zeitschriftenbibliothek - Freely accessible e-journals
subjects Biological and medical sciences
bitterness removal
carboxypeptidase Y
cheese ripening
Food industries
Food microbiology
Fundamental and applied biological sciences. Psychology
Kluyveromyces fragilis
Milk and cheese industries. Ice creams
title Purification and Characterization of a Carboxypeptidase Y from Kluyveromyces fragilis JSB95
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