Peptidases from Maclura Pomifera for Preparation of Food Protein Hydrolysates : Purification by Single-Step Chromatography and Characterization of Pomiferin I

Our objective was to isolate peptidases from the latex of Maclura pomifera fruits and use them to hydrolyze food proteins, as well as to purify and characterize the main peptidase. Two partially purified proteolytic extracts were prepared by ethanol (EE) and acetone (AE) precipitation from an aqueou...

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Detalles Bibliográficos
Autores principales: Reyes Jara, Andrea Milagros, Corrons, María Alicia, Salese, Lucía, Liggieri, Constanza Silvina, Bruno, Mariela Anahí
Formato: Articulo
Lenguaje:Inglés
Publicado: 2020
Materias:
Ciencias Exactas
Biología
Plant peptidase
Maclura pomifera
Food protein hydrolysate
Purification
Acceso en línea:http://sedici.unlp.edu.ar/handle/10915/131902
Aporte de:
SEDICI (UNLP) de Universidad Nacional de La Plata
id I19-R120-10915-131902
record_format dspace
institution Universidad Nacional de La Plata
institution_str I-19
repository_str R-120
collection SEDICI (UNLP)
language Inglés
topic Ciencias Exactas
Biología
Plant peptidase
Maclura pomifera
Food protein hydrolysate
Purification
spellingShingle Ciencias Exactas
Biología
Plant peptidase
Maclura pomifera
Food protein hydrolysate
Purification
Reyes Jara, Andrea Milagros
Corrons, María Alicia
Salese, Lucía
Liggieri, Constanza Silvina
Bruno, Mariela Anahí
Peptidases from Maclura Pomifera for Preparation of Food Protein Hydrolysates : Purification by Single-Step Chromatography and Characterization of Pomiferin I
topic_facet Ciencias Exactas
Biología
Plant peptidase
Maclura pomifera
Food protein hydrolysate
Purification
description Our objective was to isolate peptidases from the latex of Maclura pomifera fruits and use them to hydrolyze food proteins, as well as to purify and characterize the main peptidase. Two partially purified proteolytic extracts were prepared by ethanol (EE) and acetone (AE) precipitation from an aqueous suspension of exuded fruit latex. EE was used to hydrolyze food proteins with a ratio of 0.19 caseinolytic units (Ucas) per mg of substrate. Different values of hydrolysis degree were observed for hydrolysates of egg white, soy protein isolate, and casein at 180 min (9.3%, 31.1%, and 29.1%, respectively). AE was employed to purify a peptidase which exhibited an isoelectric point (pI) of 8.70 and whose abundance in AE was 28.3%. This enzyme was purified to homogeneity using a single-step procedure by cation-exchange chromatography, achieving an 8.1-fold purification and a yield of 16.7%. The peptidase was named pomiferin I and showed a molecular mass of 63,177.77 Da. Kinetic constants (KM 0.84 mM, Vmax 27.50 uM s−1, kcat 72.37 s−1, and kcat/KM 86.15 mM−1 s−1) were determined employing N-α-carbobenzoxy-L-alanyl-p-nitrophenyl ester as substrate. Analysis by PMF showed only partial homology of pomiferin I with a serine peptidase from a species of the same family.
format Articulo
Articulo
author Reyes Jara, Andrea Milagros
Corrons, María Alicia
Salese, Lucía
Liggieri, Constanza Silvina
Bruno, Mariela Anahí
author_facet Reyes Jara, Andrea Milagros
Corrons, María Alicia
Salese, Lucía
Liggieri, Constanza Silvina
Bruno, Mariela Anahí
author_sort Reyes Jara, Andrea Milagros
title Peptidases from Maclura Pomifera for Preparation of Food Protein Hydrolysates : Purification by Single-Step Chromatography and Characterization of Pomiferin I
title_short Peptidases from Maclura Pomifera for Preparation of Food Protein Hydrolysates : Purification by Single-Step Chromatography and Characterization of Pomiferin I
title_full Peptidases from Maclura Pomifera for Preparation of Food Protein Hydrolysates : Purification by Single-Step Chromatography and Characterization of Pomiferin I
title_fullStr Peptidases from Maclura Pomifera for Preparation of Food Protein Hydrolysates : Purification by Single-Step Chromatography and Characterization of Pomiferin I
title_full_unstemmed Peptidases from Maclura Pomifera for Preparation of Food Protein Hydrolysates : Purification by Single-Step Chromatography and Characterization of Pomiferin I
title_sort peptidases from maclura pomifera for preparation of food protein hydrolysates : purification by single-step chromatography and characterization of pomiferin i
publishDate 2020
url http://sedici.unlp.edu.ar/handle/10915/131902
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