Peptidases from Maclura Pomifera for Preparation of Food Protein Hydrolysates: Purification by Single-Step Chromatograph
- PDF / 902,898 Bytes
- 18 Pages / 439.37 x 666.142 pts Page_size
- 58 Downloads / 197 Views
Peptidases from Maclura Pomifera for Preparation of Food Protein Hydrolysates: Purification by Single-Step Chromatography and Characterization of Pomiferin I Andrea Milagros Reyes Jara 1,2 & María Alicia Corrons 3 & Lucía Salese 1,3 & Constanza Silvina Liggieri 3,4 & Mariela Anahí Bruno 1,3 Received: 18 March 2020 / Accepted: 29 September 2020/ # Springer Science+Business Media, LLC, part of Springer Nature 2020
Abstract
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-αcarbobenzoxy-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. Keywords Plant peptidase . Maclura pomifera . Food protein hydrolysate . Purification
Introduction Enzymes are biocatalysts employed in numerous industries—such as food, cosmetics, and wastewater treatment [1, 2]. The use of enzymes in industrial processes presents advantages with respect to traditional chemical reactions as neither high temperatures nor organic solvents
* Mariela Anahí Bruno [email protected] Extended author information available on the last page of the article
Applied Biochemistry and Biotechnology
or extreme pH values are required. Moreover, reaction specificity and product purity increase while environmental impact is reduced [3]. Proteases—EC 3.4—are hydrolases that cut peptide bonds and stand out in the global market due to their multiple applications in biotechnology including meat, milk and soy processing, organic synthesis, and leather treatment, among others [3, 4]. These enzymes can be extracted from different sources as animal, plant, fungal, and bacterial [5]. Proteolysis produces changes in chemical, physical, biological, and immunological properties of protein substrates. For example, hydrolysis of food proteins can achieve improvement of nutritional and functional properties, retardation of deterioration, prevention of undesired interactions, removal of toxic or inhibitory factors, or change of fl
Data Loading...
