Enzymatic generation of peptides from potato proteins by selected proteases and characterization of their structural properties

Abstract

The use of low grade starting material for the generation of peptides with bioactivity properties is of interest. The proteins from the potato starch industry byproduct is a promising source, as several health benefits may be associated with their hydrolysates. The efficiency of selected proteases (Novo Pro-D, Alcalase, Flavourzyme, and Papain), exhibiting different substrate specificities and cleavage action modes, to hydrolyze potato protein isolate (patatin and protease inhibitors) was investigated. Novo Pro-D resulted in the lowest degree of hydrolysis, whereas Alcalase, Flavourzyme, and Papain exhibited a high catalytic efficiency for the hydrolysis of potato proteins. The degree of hydrolysis behaved in a concentration dependent manner. However, the end-product profile (peptides and free amino acids) was dependent not only on the protease specificity, its cleavage action mode (endo/exo) and the availability of the intermediate substrates but also on the contribution of the protease inhibitors to the reaction kinetics through their inhibitory effects. Indeed, the dependence of the exo-activity on the catalytic efficiency of the endo-action of protease was shown to be significant. Papain generated more unique peptide sequences with homology assessment matching several potato proteins when compared with Flavourzyme. This can be attributed to the high exo-peptidase activity of Flavourzyme resulting in the generation of shorter peptides which were difficult to match. Flavourzyme produced more peptides originated from patatin fraction, whereas Papain resulted in the release of more peptides corresponding to the protease inhibitor fractions. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:420-429, 2016.

Enzymatic hydrolysis of black cricket (Gryllus assimilis) proteins positively affects their antioxidant properties

“…This fact may be related to the synergistic action of proteases with different mechanisms of action and specificities on protein hydrolysis. Alcalase TM 2.4L is a B. licheniformis serine protease that contains subtilisin endoproteinase activity and high specificity for cleavage of aromatic amino acids (Pimentel, Alves, Harnedy, FitzGerald, & Oliveira, 2019; Waglay & Karboune, 2016). Neutrase TM 0.8L is an endoprotease that has high affinity for hydrophobic amino acids (Aguilar et al., 2019), and can promote (as well as Alcalase TM 2.4L) an increase in the number of target sites available for the Flavourzyme TM 500L action, which justifies the synergistic effect of the ternary combination of the applied proteases (Nchienzia, Morawicki, & Gadang, 2010).…”

Processing, Valorization and Application of Bio-Waste Derived Compounds from Potato, Tomato, Olive and Cereals: A Review

“…During processing of potatoes 5-12 m 3 of potato fruit juice accumulates per ton [119,120]. This leads to more than two million tons of potato fruit juice in the European Union per year [121].…”

“…Waglay and Karboune [120] examined the proteases Flavourzyme, Alcalase, Papain and Novo Pro-D for hydrolysis of proteins isolated from potato fruit juice and figured out that Alcalase, Flavourzyme and Papain possessed high catalytic activities whereas Novo Pro-D showed a low degree of hydrolysis. Further differences were measured with regard to the end products: Papain generated unique peptides that can be related to protease inhibitor fractions, whereas Flavourzyme mainly produced peptides which arise from protease-inhibitors.…”

Application of the Plackett-Burman design to determine the main factors affecting the anti-oxidative activity of goat's milk casein hydrolyzed by Alcalase and papain [pdf]

“…However, if the content of papain is too high, the substrate is excessively digested. If the polypeptide with antioxidant activity is digested into a short peptide, it loses its antioxidant activity (Waglay and Karboune, 2016).…”