Plant-Derived Proteins and Peptides as Potential Immunomodulators.

The immune response of humans may be modulated by certain biopeptides. The present study aimed to determine the immunomodulatory potential of plant-derived food proteins and hydrolysates obtained from these proteins via monocatalytic in silico hydrolysis (using ficin, stem bromelainm or pepsin (pH > 2)). The scope of this study included determinations of the profiles of select bioactivities of proteins before and after hydrolysis and computations of the frequency of occurrence of selected bioactive fragments in proteins (parameter A), frequency/relative frequency of the release of biopeptides (parameters AE, W) and the theoretical degree of hydrolysis (DHt), by means of the resources and programs available in the BIOPEP-UWM database. The immunomodulating (ImmD)/immunostimulating (ImmS) peptides deposited in the database were characterized as well (ProtParam tool). Among the analyzed proteins of cereals and legumes, the best precursors of ImmD immunopeptides (YG, YGG, GLF, TPRK) turned out to be rice and garden pea proteins, whereas the best precursors of ImmS peptides appeared to be buckwheat (GVM, GFL, EAE) and broad bean (LLY, EAE) proteins. The highest number of YG sequences was released by stem bromelain upon the simulated hydrolysis of rice proteins (AE = 0.0010-0.0820, W = 0.1994-1.0000, DHt = 45-82%). However, antibacterial peptides (IAK) were released by ficin only from rice, oat, and garden pea proteins (DHt = 41-46%). Biopeptides (YG, IAK) identified in protein hydrolysates are potential immunomodulators, nutraceuticals, and components of functional food that may modulate the activity of the human immune system. Stem bromelain and ficin are also active components that are primed to release peptide immunomodulators from plant-derived food proteins.

Characteristics of Potential Protein Nutraceuticals of Plant Origin with Antioxidant Activity.

This study used selected plant proteins and the tools available in the BIOPEP-UWM database to profile proteins and release antioxidant nutraceuticals from their primary structures. The frequency of the occurrence of fragments with antioxidant activity in a protein sequence (the A parameter) was determined. A simulated monocatalytic proteolysis was carried out using ficin or stem bromelain or pepsin (pH > 2), and the theoretical degree of hydrolysis (DHt) and the frequency (including relative frequency) of the release of fragments with a particular antioxidant activity by a selected enzyme (the AE and W parameters, respectively). Both barley hordoindolines and the protein group of "actins and other rice proteins" were characterised by the best antioxidant potential. On the other hand, among the main analysed cereal protein groups or species, the best nutraceutical sources included kafirins, rice glutelins and α-gliadins. Potentially the most nutraceutical molecules were released by pepsin (HL, VY, PHQ and PWQ biopeptides) from gliadins, but the most analysed proteins were hydrolysed (66% on average) and the DHt for ficin and bromelain amounted to 27% and 31%, respectively. However, based on the calculated AE mean values, it can be concluded that nutraceuticals were more frequently released from rice protein structures (IY and VY biopeptides), and less frequently released from barley and other cereal protein species, which may be of significance in the context of designing nutraceutical food.

Isolation of Oat (Avena sativa L.) Total Proteins and Their Prolamin Fractions for 2D Electrophoresis.

Appropriate sample preparation is essential to obtaining good results of two-dimensional gel electrophoresis (2-DE). For various reasons (particularly phenolic compounds, proteolytic enzymes, and cell-wall mucilages) the extraction of proteins from plant material, among them oat proteins, is difficult. During isolation all soluble substances that may interfere with the analysis (especially isoelectric focusing) are removed, and proteins of interest are separated from the remains. However, the applied procedure of isolation cannot be too extensive, because additional stages cause loss of the proteins.In this chapter, we describe a simple procedure for the isolation of oat total proteins and their prolamin fractions prior to 2-DE, without necessity of considerable purification. It can be used for oat protein fractionation, measurement of oat protein concentration, and their 2-DE analysis, with particular reference to prolamin fractions. The presented routine includes modified methods of plant seed proteins extraction and sequential Osborne extraction, based on oat protein solubility differences.