Trends in Food Science & Technology 104 (2020) 37–48

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Trends in Food Science & Technology

journal homepage: www.elsevier.com/locate/tifs

Prospecting the applications and discovery of peptide hydrogels in food

Luis M. De Leon Rodriguez a, Yacine Hemar b, *
a
School of Chemical Sciences, The University of Auckland, 23 Symonds St., Auckland, 1010, New Zealand
b
Department of Biotechnology and Food Engineering, Guangdong Technion Israel Institute of Technology, Shantou, 515063, Guangdong, China

A R T I C L E I N F O A B S T R A C T

Keywords: Background: The interest in peptide hydrogels of natural origin has dramatically increased given the potential
Hydrogels applications in several fields -e.g. biomedicine and nanotechnology. Interestingly, despite the current knowledge
Peptides on protein hydrolysates from food sources, which self-assemble and form gels, the extraction of single peptides
Food
that can form hydrogels from food products and/or their application in food and other areas remains poorly
Toxicity
Production
explored.
Scope and approach: This review provides a prospective analysis of the literature on the mechanism, production,
toxicity and potential applications of food derived peptide hydrogels.
Key findings and conclusions: Food products can be an important source of single peptides that form hydrogels,
and these can find applications in food science and other areas. However, research in this area is in its infancy
and its progress is limited due in part to the lack of 1) tools that will allow one to predict peptide fragments
within a food protein that can self-assemble and form gels and 2) efficient peptide purification protocols.
Therefore, more research will have to be directed on these areas in conjunction with optimization of recombi­
nant, and enzymatic/fermentation production protocols.

1. Introduction viable and cost-effective source of PHGs. Equally important is an un­

Research on peptides that self -assemble and form hydrogels have
attracted great attention given their inherent properties of biocompati­
bility, biodegradability, simple and reproducible preparation and the
availability of a vast variety of chemical functionalities and structures.
In the literature, one can find examples on the development of peptide
hydrogels (PHGs) in 1) biomedicine as drug controlled release agents,
scaffolds in tissue engineering and immunomodulation and antimicro­
bial agents (Hu et al., 2020; Liu, Zhang, Zhu, Liu, & Chen, 2019; Singh &
Peppas, 2014; Yadav, Chauhan, & Chauhan, 2020), 2) nanotechnology
as templates in the construction of three-dimensional nanoscale systems
(Rajagopal & Schneider, 2004), biosensing (Lian, Chen, Lu, & Yang,
2016) and semiconductors (Tao, Makam, Aizen, & Gazit, 2017) and 3)
organocatalysis (Huang, Yu, & Horng, 2020; Pelin et al., 2020).
Despite the numerous reports on PHGs their utilization in food
technology is basically unknown when considered as single ingredients.
Therefore, the objective of this review is to evaluate the potential use of
single peptides that can form hydrogels in foods. Important questions to
address are in relation to the source of PHGs and if this source warrants
their safe utilization in food products, or if some food products can be a
derstanding on the potential toxicity issues of PHGs and to identify
specific applications in food technology. There are several examples in
the literature in relation to protein hydrolysates from food sources that
aggregate and gel and on the use of self-assembled proteins in food
(Tomadoni, Capello, Valencia, & Gutiérrez, 2020). Therefore, in this
review we refer to relevant papers on this subject as the starting point for
the identification of single component PHGs. However, it is important to
remark that protein hydrolysates consist of a heterogenous mixture of
components which composition and nature can vary from batch to batch
because of variations in the source and/or on the enzymatic cleavage
process. This limitation highlights the need to identify the major
component(s) of protein hydrolysates that promote gelation (Lacou,
Léonil, & Gagnaire, 2016).
Peptides that can potentially form hydrogels derive from designed
synthetic sequences or peptide fragments borrowed from proteins found
in nature. In this review we will cover some of the main structural
characteristics found in these two groups of PHGs sources, their pro­
duction protocols with a focus on chemical and recombinant synthesis
and the most important types of PHGs.
Peptide hydrogels constitute one of the energy states that are

* Corresponding author.
E-mail address: y.hemar@auckland.ac.nz (Y. Hemar).

https://doi.org/10.1016/j.tifs.2020.07.025
Received 24 May 2020; Received in revised form 15 July 2020; Accepted 25 July 2020
Available online 2 August 2020
0924-2244/© 2020 Elsevier Ltd. All rights reserved.
L.M. De Leon Rodriguez and Y. Hemar
possible to the set of self-aggregating peptides (Ramos Sasselli, Halling,
Ulijn, & Tuttle, 2016; Wang, Liu, Xing, & Yan, 2016), all presenting
supramolecular structures similar to those of amyloid insoluble deposits
identified as important markers of a set of neurodegenerative diseases
(e.g. Alzheimer’s, Parkinson’s, Huntington’s and Prion diseases) and
protein aggregates related to type II diabetes, cancer and systemic
amyloidosis (Adamcik & Mezzenga, 2018). Hence, main concerns on
PHGs toxicity arise from this similarity and a thorough discussion will be
provided.
Finally, we report on the few examples of self-aggregating peptides
obtained from food products that are found in the literature and provide
a discussion of prospects in this area.
2. Peptide hydrogels
2.1. Peptide self-assembly
Peptide self-assembly is a process where individual molecules form
aggregates. These aggregates can be composed of unstructured, partially
structured or structured peptide chains, which are stabilized by non­
covalent forces including electrostatic interactions, solvophobic effects,
hydrogen bonds, van der Waals forces, and π– π stacking. These aggre­
gates can be further classified into amorphous and ordered depending on
the characteristics of the resultant structure at the nanoscale. Peptide
self-assembly is a thermodynamic and kinetic driven process where the
synergistic effect of the non-covalent interactions determines the state of
minimum energy of the formed nanostructures. Different metastable
states can be reached by modifying kinetic parameters such as temper­
ature, pH, counter-ions, concentration and solvent (Wang et al., 2016).
Therefore, depending on the conditions peptide ordered nanostructures
such as nanofibers, nanoribbons, nanobelts, micelles, nanospheres,
nanotubes, vesicles, planar membranes, etc. (Fichman et al., 2016) can
be obtained (Fig. 1). Peptide hydrogels correspond to one of these
trapped metastable states and generally consist of cross-linked peptide
Fig. 1. The energy landscape of peptide aggregation. The non-aggregated
peptides undergo folding and aggregation competing processes where the
molecules transit through various conformations directed by intermolecular
interactions. In a kinetically controlled pathway intermediate species can result
in various types of nano-aggregates- amorphous, fibres, spheres/particles, rib­
bons, tubes, including hydrogels (orange). These metastable states can further
move into the thermodynamically favoured state, a crystal (red). The relative
energy shown in the diagram do not represent absolute values. The figure in­
dicates that the different structures can possibly interconvert under specific
kinetic conditions. The figure was derived from Adamcik and Mezzenga (2018).
(For interpretation of the references to colour in this figure legend, the reader is
referred to the Web version of this article.)
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Trends in Food Science & Technology 104 (2020) 37–48
fiber/fibril networks entrapping large amounts of water.
The focus of this review is on single peptides that form hydrogels.
Moreover, through the text we will refer to other relevant nano­
aggregates as these occupy states that can potentially be reached during
the process of hydrogel formation as shown in Fig. 1. Hence, the im­
plications of obtaining these states should be kept in mind while
exploring applications of PHGs.
2.2. Peptide hydrogel formation
For peptides, one can visualize the formation of hydrogels through a
dynamic hierarchical assembly process comprising molecular self-
assembling, packing into nanostructures and cross-linking with entrap­
ment of water all driven by synergic combination of intermolecular non-
covalent interactions. This process results in a continuous three-
dimensional supramolecular network (Fig. 2a–d).
During the initial stages of the molecular self-assembling process
(typically called nucleation) peptides -folded or unfolded- have to un­
dergo a conformational change to adopt an aggregation prone confor­
mation (e.g. β-sheets) and thus self-assemble to form a minimally stable
assembly called the nucleus. The nucleus serves as template where other
peptides add to further elongate the system, thus transitioning into
protofibrils, fibrils and fibers (Aggeli et al., 2001) via a one-dimensional
nanostructure growth in the direction of the fibril axis, or nano-tapes via
a two-dimensional growth. A more detailed description of this process
was recently reported by Yuan et al., where the unfolded peptides
aggregate into solute rich nanoclusters via a liquid-liquid phase sepa­
ration stage and these nanoclusters acted as nucleation sites for the
formation of nanofibrils (Yuan et al., 2019). Peptide assembly continues
until the free peptide concentration falls below the critical fibrillar
concentration, resulting in fibrils/fibers nano-to micrometers long.
During the process fibrils/fibers interact with each other via entangle­
ment, branching or threading mechanisms (Hauptstein et al., 2018) and
with water to form a three-dimensional mesh.
The flow chart shown in Fig. 2e summarizes the mechanism of action
of hydrogel formation. Here it is important to highlight that hydrogel
formation is a kinetically controlled process (Fig. 1), hence it is highly
dependent not only on the peptide sequence but also in kinetic param­
eters such as concentration, pH, ionic strength and solvent. The effect of
these parameters on fibrillation has been thoroughly discussed in the
literature (Cao & Mezzenga, 2019; Wang et al., 2016). Additionally, one
can induce hydrogelation via the application of physical and chemical
strategies. Unfortunately, each peptide system has its unique energy
landscape (Fig. 1), hence, the set of conditions that will favour gelation
in a peptide are unique and not easily transferable to other systems.
2.3. Production of peptide hydrogels
Peptides are currently manufactured by chemical and microbial re­
combinant methodologies or a combination of both. Typically, the se­
lection of one or other process depends on the length of the peptide and
issues inherent to a specific sequence (e.g. aggregation).
Peptide production by chemical methods includes solution-phase
and solid-phase synthesis and is typically limited to peptides no longer
than 35 residues. Moreover, the chemical synthesis of longer peptides
can be achieved in liquid phase by the native chemical ligation method,
where one peptide fragment containing a C-terminal thioester reacts
with another fragment containing an N-terminal cysteine, resulting in
the formation of a new peptide bond (Agouridas et al., 2019).
The synthesis of peptides in solid phase (SPPS) constitutes the fastest
way to access relatively small amounts of material. The process consists
on building the peptide chain while attached to a solid polymeric sup­
port (Scheme 1) by using protected amino acids. For instance, in fluo­
renylmethoxycarbonyl (Fmoc)-SPPS the N-terminal amino group is
protected with the base labile Fmoc group while the functional groups of
the side chains (e.g. amino, thiol, carboxylic acid, hydroxy) are
L.M. De Leon Rodriguez and Y. Hemar
Fig. 2. Peptide hydrogel formation. a) tube representation of a random peptide conformation, b) top and c) side view of the peptide assembly into β-sheets, d) from
left to right ribbon, fibril/fibre formation, and cross-linking (Transmission Electron Microscopy image of actual peptide hydrogel where the white scale bar cor­
responds to 50 nm), e) Flow chart of the mechanism of action of hydrogel formation.
protected (PG) with acid labile moieties (e.g. tert-butoxycarbonyl (Boc),
trityl (Trt), tert-butyl (tBu), etc.). The SPPS is a cyclical process con­
sisting on the formation of amide bonds via coupling reagents in alter­
nation with the removal of Fmoc. The fact that the protected peptide
fragment is attached to a solid support reduces the purification steps to
simple solvent washings where the peptidyl resin is retained in a
filtration device and the reagents and side products are removed in the
liquid organic phase. Once the targeted peptide sequence is fully
assembled on the solid support, the peptide is cleaved from the resin and
the side-chain protecting groups removed by subjecting the resin to
strong acidic conditions (e.g. trifluoroacetic acid) in the presence of
compounds that scavenge reactive species (e.g. carbocations or radicals)
derived from the protecting groups. Finally, the peptide is purified by
reverse phase high performance liquid chromatography (HPLC) and
then lyophilized. However, despite of being a robust and well estab­
lished methodology, SPPS suffers of drawbacks, such as low yields,
usage of large amounts of toxic solvents (e.g. dimethylformamide, and
dichloromethane) and hazardous materials (e.g. benzotriazole derived
coupling reagents, etc.), and high purification costs (Isidro-Llobet et al.,
2019).
For the specific case of the chemical synthesis of peptide hydrogels
the above cited disadvantages of SPPS are further compounded by the
strong aggregating tendency of the peptide sequence, where in some
cases the synthesis cannot be completed. Thus, the SPPS of PHGs often
requires repeating amino acid couplings or Fmoc deprotection steps and
often results in mixtures of inseparable compounds containing peptide
fragments derived from incomplete residues couplings and/or
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Trends in Food Science & Technology 104 (2020) 37–48
diastereomeric peptides. Solution-phase peptide synthesis appears as an
alternative for the synthesis of aggregating peptides, but it is limited to
short sequences due to the need of isolation and characterization of
intermediates required at each step. With this perspective, one can
conclude that the application of PHGs derived from chemical synthesis
in food technology would be constrained to short peptide sequences
granted safety clearances are met.
Microbial recombinant peptide production appears as a robust, sus­
tainable and cost-effective alternative for long peptide sequences (>50
amino acids). This process employs the natural protein production ma­
chinery of cells—generally bacteria and yeast—to produce heterologous
peptides (Wegmüller & Schmid, 2014). An attractive aspect of recom­
binant peptide production is that safe organisms currently used in the
food industry (e.g. Saccharomyces cerevisiae) can be employed to
manufacture peptides, hence in principle minimizing safety concerns.
Moreover, unlike chemical synthesis recombinant production is limited
to peptides containing natural amino acids. The utilization of recombi­
nant methods to prepare self-assembling peptides was reviewed by Kyle
et al. (Kyle, Aggeli, Ingham, & McPherson, 2009), where the progress on
the recombinant production of collagen, elastin and designed
self-assembling peptides was discussed. An important drawback on the
use of recombinant processes to generate PHGs is the low recoveries of
final product, which is attributed to either enzymatic cleavage and/or
inefficient purification. A remarkable example of the successful appli­
cation of recombinant technologies on the preparation of PHGs is the
case of QQRFEWEFEQQ (P11-4) (Riley, Aggeli, Koopmans, & McPher­
son, 2009). This peptide was recovered in relatively high amounts (90
L.M. De Leon Rodriguez and Y. Hemar
Scheme 1. General overview of the chemical synthesis of peptides in solid
phase using the Fmoc-tBu strategy.
mg/L bacteria culture from 2.5 g protein/L bacteria culture) when
produced in an optimized Escherichia coli strain with a vector containing
a trimer peptide repeat fused to a protein which accumulates in recov­
erable inclusion bodies (a strategy commonly used to elude protease
cleavage). The peptide was recovered from the inclusion bodies after a
detergent lysis step and the peptide released from the protein with a
cyanogen bromide cleavage (Riley et al., 2009). Despite this successful
production of P11-4 reports on the preparation of PHGs by recombinant
methods are scarce (Sonmez, Nagy, & Schneider, 2015; Zerfaß, Brauk­
mann, Nietzsche, Hobe, & Paulsen, 2015).
2.4. Types of peptide hydrogels
In general, PHGs can be classified based on their origin into designed
and natural derived peptides. Designed peptides are man-made and
follow specific rules in relation to the nature and position of the amino
acids that conform a peptide sequence. The goal is to achieve specific
secondary structures to drive assembly. The most common type of
designed peptides exploits amphiphilicity to induce peptide assembly
and hydrogelation. The discovery of peptides of natural origin that self-
assemble and that could potentially form gels is typically accomplished
by scrutinizing known protein structures to uncover sub-sequences
which attain specific intra- or inter-molecular self-assembling second­
ary structures (Valéry, Pandey, & Gerrard, 2013). These peptide frag­
ments are then synthesized and tested for hydrogelation. If hydrogel
formation is not attained, then further sequence/chemical modifications
follow.
Peptide hydrogels can be further classified based on the secondary-
structure attained by the assembling peptides, that is α-helices,
β-sheets, β-hairpins and coiled coils (Mart, Osborne, Stevens, & Ulijn,
2006). Among the secondary structures, β-sheets are the most widely
studied, hence these will be discussed further (Rodriguez, Hemar, Cor­
nish, & Brimble, 2016).
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Trends in Food Science & Technology 104 (2020) 37–48
The main feature of the β-sheet structure is an adjacent parallel or
anti-parallel peptide strand arrangement driven by hydrogen bonding.
Strands stacking is also observed in the nanostructure of β-sheet PHGs
and is typically attributed to hydrophobic interactions.
Hydrogel forming β-sheet peptides can be further grouped based on
the nature and sequence of amino acids. The most important group
consists of sequences containing alternating hydrophobic and hydro­
philic amino acids, an arrangement resulting in a facial amphiphilic
character, with hydrophobic side chains groups facing one side and
hydrophilic groups facing the opposite side relative to the peptide
backbone (see (RADA)4 in Fig. 3). Inside this group, one can cite that of
ionic-complementary peptides (Chen, 2005), comprising alternating
negatively and positively charged amino acids in the hydrophilic face.
This ordering of residues of opposite charges drives aggregation via
electrostatic interactions.
Another group of β-sheet PHGs present a head-to-tail amphiphilicity
and includes two types: one group consists of peptides composed of a
sub-sequence of hydrophobic residues attached to a shorter sub-
sequence of hydrophilic amino acids (see LIVAGD in Fig. 3), and
another group involves peptides modified with hydrophobic acyl chain
(s) (see C16-KTTKS in Fig. 3). Aggregation in these peptides is controlled
either by the hydrophobic or the hydrophilic portions, depending on
their length and β-sheet formation propensity of the fragments (Hamley,
2011).
Finally, another group of β-sheet PHGs covers those peptides that do
not completely adhere to an alternating hydrophobic/hydrophilic
sequence arrangement. Some examples include the P11 family of pep­
tides (Aggeli et al., 2003) (see P11-4 in Fig. 3), peptide sequences derived
from proteins found in nature and peptides containing the diphenyla­
lanine fragment (see Fmoc-FF in Fig. 3).
2.5. Peptide hydrogels current uses/applications
The applications of PHGs in biomedicine, biosensing and biocatalysis
has been well documented in the literature. Therefore, we refer the
interested reader to recent reviews indicated in the introduction section.
Herein, the main focus of the discussion will be about the correlation
between the properties of PHGs and the application. Moreover, we also
provide a brief overview on synthetic peptides that could be potentially
used in food products.
The utility of a PHG is determined by its atomic, nano-, and supra­
molecular structure. Thus, for example the properties of the functional
groups of the amino acids present in PHGs and their spatial arrangement
within the nanostructure determine the potential bioactivity of the
system. For instance, a combination of certain amino acids might confer
a PHG with cell attachment and proliferation properties or the capability
to retain drugs or proteins within the fibrillar mesh (Ishida, Watanabe,
Oshikawa, Ajioka, & Muraoka, 2019). These characteristics are exploi­
ted in cell culture, tissue engineering and drug delivery applications (J.
Li & Mooney, 2016; Uzunalli & Guler, 2020). Details of the nano- and
supramolecular structure of PHGs (e.g. the fibril/fiber cross-section and
length, inter- and stacking strand distances) can be determined by im­
aging techniques such as transmission electron microscopy (TEM),
cryogenic-TEM, atomic force microscopy (AFM), and by X-ray diffrac­
tion or small angle scattering methods (XRD or SAXS).
An important property of PHGs is its mechanical strength as this is a
parameter that allows to determine the material physical limitations,
hence its compatibility with a desired application.
The mechanical properties of PHGs are determined by rheological
characterization. Typical rheology experiments measure the storage
modulus (G′ ) (a measure of a material stiffness) and the loss modulus
(G′′ ) (a measure of the material liquid-like flow properties), as a function
of time, oscillatory frequency at constant strain, and oscillatory strain at
constant frequency. The generated data allows one to determine not
only if a material forms a gel (G’ > G′′ ) and how stiff it is, but it also
provides insights that allow to understand the mechanism of gelation.
L.M. De Leon Rodriguez and Y. Hemar
Fig. 3. Examples of β-sheet peptide hydrogels. In general, the hydrophilic residues are represented in blue and the hydrophobic in red. Other molecular components
and residues outside an alternating hydrophobic/hydrophilic sequence are coloured black. (For interpretation of the references to colour in this figure legend, the
reader is referred to the Web version of this article.)
Additionally, by combining rheology with other characterization tech­
niques (e.g. TEM) one can further determine physical parameters such as
the network mesh size, which corresponds to the average distance be­
tween consecutive cross-links -an important parameter to consider for
drug delivery applications.
The PHGs discussed in the previous section present G′ values ranging
from 7 to 28,000 Pa, when measured under similar conditions (~1 wt %
peptide concentration at pH 7 and 25 ◦ C) (Rodriguez et al., 2016). With
these values in mind, one can have an idea on the potential applications
of PHGs. For instance, a soft PHG (low G′ ) might not be useful as support
in bone regeneration applications since the elasticity of bone tissue is
high (>30 kPa). However, soft materials could serve as media for bone
cell culture, since a G′ of 50–100 Pa is the estimated minimum range
required to maintain mammalian cells in suspension. Also, stiff PHGs
can find applications in the development of new nanomaterials. Addi­
tionally, G′ reference values of food products can give us an idea of the
applicability of PHGs in food science, for instance a set yoghurt made
with skim milk has a G’ of ~100 Pa (Zuo, Hemar, Hewitt, & Saunders,
2008), while that of a cooked meat batter can exceed 100 kPa (Glorieux,
Steen, De Brabanter, Foubert, & Fraeye, 2018).
Here, it is important to highlight that the mechanical properties of a
PHG not only depend on the rheology measurement conditions but also
on physicochemical factors such as peptide concentration, pH, temper­
ature, nature and concentration of salt, etc.
It is also important to mention that while the number of PHGs that
have entered clinical trials for biomedical applications is scarce
(Aswathy, Narendrakumar, & Manjubala, 2020), the number of com­
panies that supply PHGs has been increasing -Puramatrix (Corning),
HydroMatrix (Sigma), Biogelx, PGD-HydroGels (PeptiGelDesign) and
Manchester Biogel, to cite a few. This highlights the increase interest in
the field of PHGs.
Reports on the uses of single peptides that form hydrogels derived
from food products is limited (vide infra). However, one can find reports
on synthetic self-assembling peptides that were designed as emulsifiers
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Trends in Food Science & Technology 104 (2020) 37–48
(Wibowo, Wang, Shao, Middelberg, & Zhao, 2017) (Hui, Wibowo, &
Zhao, 2016; Mondal et al., 2017) and in some cases a potential appli­
cation in food was stated. Among the most promising peptide candidates
for food applications one can mention five tripeptides (KYW, KFF, KYF,
FFD and DFF), which were shown to have temperature dependent
emulsion-stabilizing capabilities at a 40 mM concentration in water in
presence of rapeseed oil (a food-regulated oil) (Scott, McKnight, Tuttle,
& Ulijn, 2016). Interestingly, it was shown that the peptides that formed
the more stable emulsions (KYW, KFF and KYF) self-assembled into a
fibrillar structure and were capable of forming gels, while the peptides
that formed the less stable emulsions (FFD and DFF) assembled into
bilayer-like structures. However, the emulsifying ability of these short
peptides is typically affected by some physicochemical conditions (e.g.
low temperature, presence of salt, etc.). Therefore, the chemical deriv­
atization of short peptides with either polyaromatic chemical groups (e.
g. fluorenylmethylcarbonyl (Fmoc) (e.g. Fmoc-FF in Fig. 3) or long acyl
chains (e.g. C16-KTTKS in Fig. 3) has been applied as a strategy to
generate more robust peptide based emulsifying agents (Bai et al.,
2014). However, the attachment of polyaromatic groups raises toxicity
concerns which hampers further applications in food products.
Contrarily, acylated peptides are considered less toxic. The C13-KR
peptide is a recent example of acylated peptide-based emulsifier (Lv
et al., 2019). This peptide showed good emulsifying properties under
acidic conditions and had better stability to high temperature and high
salt concentration than Tween 80 and whey protein isolate, which are
widely used traditional emulsifiers.
2.6. Toxicity
Safety concerns of peptide hydrogels are primarily attributed to the
structural link with disease-related amyloids. However, to assert on the
toxicity of PHGs as additives in food products, one should first consider
the stability and permeability of peptides to the different barriers of the
gastrointestinal (GI) tract and thus evaluate the potential for peptides to
L.M. De Leon Rodriguez and Y. Hemar
reach the circulatory system.
Peptides are stable in the mouth and oesophagus given the short
retention times and the insignificant level of peptidases. Medium and
large, but short peptides, are unstable in the stomach given the presence
of high concentrations of acid and pepsin present in the gastric fluid. The
small intestine contains numerous proteases and peptidases; thus, it
constitutes the major barrier for peptide integrity. Yet, very small pep­
tides (2–3 amino acids long) are stable in the small intestine. Finally, the
myriad of microbes present in the large intestine can either consume
peptides via fermentation or cleave them via secreted proteases (Smart,
Gaisford, & Basit, 2014). If peptides survive their journey through the
digestive system, their absorption from the intestinal lumen into the
systemic circulation can still be constrained. The plausible pathways for
the absorption of peptides from the intestinal lumen are: paracellular
diffusion, passive diffusion through enterocytes, endocytosis and
transport via carrier systems. In general, it is known that peptide ab­
sorption through these pathways is limited to small molecules (<600
Da) with an apparent selectivity for neutral and hydrophobic peptide
sequences. Thus, a consensus found in the literature is that peptide oral
bioavailability at physiologically relevant concentrations is limited to
di- and tripeptides (Miner-Williams, Stevens, & Moughan, 2014; Wang,
Xie, & Li, 2019).
In the literature, there are only a few reports on the oral bioavail­
ability of peptides in humans. In a recent review, Qingbiao et al. (Xu,
Hong, Wu, & Yan, 2019) compiled information on the oral bioavail­
ability of a series of peptides, mostly di- and tripeptides, and that
reached blood concentrations in the low μg/mL range. Interestingly,
there is a report where three relatively long peptide fragments derived
from κ-casein (106–169) and αs1-casein (1–23 and 1–21) were found in
serum of human adult subjects after oral administration of yoghurt and
cow milk. For the case of the κ-casein (106–169), the authors detected
the peptide in serum up to 8 h after oral intake of milk or yoghurt and
found a maximum average concentration of ~3 μg/mL in subjects who
ingested yoghurt.
One can further bring to the reader’s attention two peptide drugs
(<1.5 KDa), which are orally administered Desmopressin and Cyclo­
sporin A. These peptides are cyclic (via a disulphide bond for Desmo­
pressin) and contain non-natural amino acids within their sequence
(Fig. 4), features that protects them against digestive enzymes. Despite
these somehow structural similarities the bioavailability of Desmo­
pressin is <1% and that of Cyclosporin A is >80% (Smart et al., 2014).
This difference is attributed to the very high lipophilicity of Cyclosporin
A, a property that allows the peptide to partition across the lipid
membranes of intestinal cells. Therefore, one can conclude that the
chances to find in the circulatory system orally consumed PHGs, which
are non-cyclic and hydrophilic, is low. However, highly hydrophobic
Fig. 4. Chemical structures of Desmopressin and Cyclosporin A. The residues highlighted in green correspond to D-amino acids, in blue refer to N-methylated residues
and in pink to non-proteogenic units. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)
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Trends in Food Science & Technology 104 (2020) 37–48
peptides and peptides containing at least two cysteines (which could
generate a cyclic product) can potentially pass the GI tract and be
absorbed.
If PHGs could reach concentrations in blood in the low μg/mL range,
then as a second step to evaluate their toxicity one should investigate the
mechanism of hydrogelation to determine if PHGs could self-assemble at
these concentrations. As indicated above hydrogelation involves a
nucleation step where unfolded peptides undergo a conformational
change to adopt a self-assembly prone structure (e.g. β-sheets). Hence, if
PHGs self-assemble at low concentrations then they could potentially act
as nucleation points for the assembly of other peptides or proteins found
in vivo. This option is important since it has been recognized that protein
amyloid formation driven by aggregation is a generic feature of all
proteins. In fact, a genomic survey indicated that almost all proteins
(>98.7%) contain at least one self-complementary short sequence
capable of forming amyloid fibrils (Goldschmidt, Teng, Riek, & Eisen­
berg, 2010).
Literature reports indicate that self-assembly of PHGs can occur at
concentrations ranging from 1000 μg/mL down to 8 μg/mL (for the case
of some acylated peptides) (Castelletto, Gouveia, Connon, & Hamley,
2013), the latter value is well below the critical gel concentration (5–30
mg/mL) (Rodriguez et al., 2016). This allow us to conclude that except
for some acylated peptides, most PHGs—man-made or naturally deri­
ved—are safe to use in food.
To the best of our knowledge there is no report on toxicity of peptide
hydrogels present in food products in humans. However, there are some
antecedents which associate the ingestion of protein amyloid fibrils with
reactive amyloidosis in mice (Cui, Kawano, Hoshii, Liu, & Ishihara,
2008). Cao and Mezzenga (2019) proposed three potential toxicity
pathways of protein amyloids. We believe that these pathways can also
apply to peptide hydrogels, hence they are included here.
1) Cross-seeding effect. The basic idea is that PHGs could potentially act
as templates for amyloid precursor proteins to aggregate more
rapidly. For instance, it has been shown that fibrillar seeds of various
protein sources including milk protein (casein) promote Aβ40 and
Aβ42 (related to Alzheimer’s disease) association (Ono et al., 2014;
Yau & Tycko, 2018). A recent review on amyloid cross-seeding has
been recently reported (Chaudhuri, Prajapati, Anand, Dubey, & Kar,
2019). Jean et al. (Jean, Foley, & Vaux, 2017) reported that
non-natural hydrogel formation could also physically change both
the intracellular and extracellular environments of cells, disrupting
processes, from motility to molecular transport, and overall cell
survival.
2) Nanotoxicity. This idea builds on the notion that structures of
nanometer dimensions may alter the physical, chemical, and
L.M. De Leon Rodriguez and Y. Hemar
biological properties of adjacent environments. However, for PHGs,
where intermolecular interactions play a major role in the assembly,
this potential toxicity is to be regarded very low against the
competing cross-seeding effect.
3) Toxicity by inducing microbiota imbalances. PHGs could promote
formation of biofilms, which in turn could alter the balance of mi­
croorganisms in humans.
2.7. Additional issues related to peptide self-assembling
An undesired consequence of peptide aggregation is precipitation
(Fig. 1) as this can lead not only to toxicity problems as indicated above
but also to unpleasant organoleptic perception of the food product. For
instance, acidic sport drinks supplemented with enzymatic soy protein
hydroxylate present peptide sedimentation upon prolonged chilled
storage, thus prompting to consumer complaints. The chemical char­
acterization of the sediment revealed the presence of several peptide
fractions derived from a limited region of the β-conglycinin α subunit
(310–362) (Nakamori et al., 2014). These peptides were richer in hy­
drophobic residues than the peptides found in solution and comprised
fragments with either acidic or basic isoelectric points. These results led
to a tentative sedimentation mechanism in which a core sediment is
initially formed by isoelectric precipitation, followed by the attachment
of hydrophobic peptides which in turn accelerate aggregation and
precipitation.
With this example one can envision possible situations where the
addition of PHGs to a complex food sample could lead to precipitate
formation.
3. Food sources of peptide hydrogels
In general, one can claim that food proteins are non-toxic, nutri­
tionally rich, easily digestible, and inexpensive, hence, an initial hy­
pothesis is that food proteins constitute an attractive source of PHGs.
Food proteins can be broadly classified into animal, plant and fungi
according to their origin. Examples of each type of proteins are: 1) milk
proteins, egg proteins, blood proteins, meat proteins, insect proteins,
etc. for proteins of animal origin, 2) cereals proteins (wheat, corn,
barley, oats, rice), legumes and pulses proteins (peas, soybeans, lupins,
lentils), tubers proteins (potato), oil seeds proteins (rapeseed, cotton­
seed, peanut, sunflower, hemp seed), pseudocereals proteins (amaranth,
chia), edible seeds proteins (quinoa, buckwheat), algae, etc. for proteins
of plant origin, and 3) proteins of fungi origin.
A total of 23 food proteins have been identified to assemble into
amyloid fibrils (Cao & Mezzenga, 2019), where proteins of animal
sources have received most scientific attention. A large proportion of
reports on physicochemical conditions (e.g. temperature, salt concen­
tration, enzyme, etc.) that promote fibril formation and gelation are
found for milk proteins. However, the characterization of individual
peptides—subproducts of hydrolysis—that self-assemble and form
hydrogels is scarce.
In terms of the properties (the emulsifying, foaming, etc.) of self-
assembling materials relevant in food science, one can state that the
benefits of peptides relative to proteins lie: 1) in their homogeneity and
easy accessibility as they do not require prior partial denaturation and 2)
in their capability to diffuse more rapidly to the interface through their
enhanced solubility and, hence, to adsorb more rapidly at the air/water
or lipid/water interface (Lacou et al., 2016).
3.1. Peptides from enzymatic protein hydrolysis
Peptides can be initially obtained as a complex mixture from the
enzymatic hydrolysis of proteins found in food (Lacou et al., 2016). The
sequence of the peptide fragments or hydrolysate composition is
determined by the bond cleavage specificity of the enzyme and the
physicochemical conditions used during hydrolysis (e.g. amount of
43
Trends in Food Science & Technology 104 (2020) 37–48
enzyme, hydrolysis time, pH, temperature, ionic strength). There are
several commercial enzymes with well stablished residue bond-cleavage
specificity (e.g. trypsin, papain, pronase, pepsin, bromelain, alcalase and
chymotrypsin) and there are in silico methods (Peptide Cutter in
https://web.expasy.org/peptide_cutter/) that can predict the type and
number of peptides that will theoretically result from the treatment of a
protein with a specific enzyme (Song et al., 2018). However, a protein
enzymatic cleavage is greatly influenced by the conditions of the hy­
drolysis depending on how such conditions affect the tertiary structure
of the protein. Thus, for example a fully denatured protein will result in a
different hydrolysate protein profile when compared to a protein that
maintains certain structure, hence limiting enzyme accessibility to
exposed protein fragments only. To purify individual peptides the pro­
tein hydrolysate is commonly subjected to a series of separation steps
(fraction separation based on charge, size and hydrophobicity and
chromatography) and the purified single peptides sequences are iden­
tified, typically from tandem mass spectrometry.
3.2. Peptides from fermentation
The production of peptides via fermentation comprises the exposure
of food proteins to microorganisms, such as yeasts, fungi, or bacteria.
These microorganisms use their own enzymes to hydrolyse the proteins
into shorter peptides.
Both processes (enzymatic hydrolysis and fermentation) have been
widely applied for the identification of bioactive peptides from food
protein digests (Aluko, 2018) but reports on the practical utilization for
the purification and identification of peptide hydrogels are scarce as we
will discuss later.
At this point one can try to address the question if some food prod­
ucts can be a viable and cost-effective source of PHGs? An accurate
answer can be given if one could compare the cost of producing a given
peptide by chemical methods versus the cost of obtaining it from a food
source.
Typically, the cost of producing a peptide by chemical means de­
pends on the number of amino acids in the sequence and the degree of
purity required for the final product. Thus, for instance, the synthesis of
1 g of an eight-residue peptide with a >98% final purity will cost ~ USD
1420 (according to pricing from https://www.biobasic.com/peptides
-pricing/).
Now, let us assume that one wants to obtain an octapeptide fragment
via enzymatic hydrolysis from αs1-casein, which is the most abundant
protein found in milk (9–15 g/L). In an ideal scenario—assuming com­
plete enzymatic cleavage of the casein fraction—one will end up with a
protein hydrolysate containing ~4% peptide mass (considering an
octapeptide with a MW of 933 Da divided by the MW of as1-casein)
relative to the initial amount of as1-casein (i.e. ~ 470 mg/L of milk). The
amount of peptide in this ideal situation appoints food products as an
attractive source of PHGs. However, this calculation does not account
for the losses of product incurred during the purification of the peptide.
One must remember that the protein hydrolysate consists of a complex
mixture of peptides, hence the purification of the desired octapeptide
will not be a simple task. In fact, in a recent review article by Chakrabarti
et al. (Chakrabarti, Guha, & Majumder, 2018), the authors identified the
lack of appropriate and scalable production methods as one of the fac­
tors that have delayed the commercial application of bioactive peptides
derived from food. Hence, one would expect a similar issue for PHGs. A
caveat to consider with PHGs is their self-assembly capacity, since it is
this property which could be sometimes advantageous when trying to
purify these peptides from a complex mixture. For instance, recently
Suwal et al. (Suwal et al., 2019) reported that the purification of an
eight-residue peptide derived from β-lactoglobulin can be easily per­
formed by consecutive aqueous washings/centrifugation of a protein
hydrolysate where the peptide flocculated at concentrations above 2
mg/mL due to self-aggregation. Thus, the authors obtained the peptide
with a 91% purity. Unfortunately, data on recovery yields of PHGs from
L.M. De Leon Rodriguez and Y. Hemar
protein hydrolysates is lacking, which complicates the completion of our
cost analysis, but it is presumably low. However, given the possibility of
a simple purification of PHGs from foods appoints these as a potential
source of PHGs.
The identification of PHGs in food proteins is further complicated in
part due to the lack of predictive tools that will allow one to screen for
peptide sequences within proteins that could potentially form gels. To
the best of our knowledge there are only two reports where attempts to
correlate the sequence of di- and tri-peptide sequences with gel prop­
erties were made (Frederix et al., 2015; F.; Li et al., 2019). Thus, a focus
on this area should be prioritized by the scientific community.
3.3. Gelatine based peptide hydrogels
Gelatine is one of the most widely used hydrogel forming materials
derived from food. Gelatine is typically obtained from the thermal acid
or alkaline denaturization-hydrolysis of collagen I from bovine and
porcine sources. This process leads to the release and partial hydrolysis
of the protein chains conforming the triple helix of collagen (a collagen I
monomer consists of left-handed polyproline II helices; two α1 chains
and one α2 chain intertwined into a right-handed triple-helix, composed
of ~1050 amino acids). Although the preparation of gelatines from
different sources have been reported in the literature (Mariod & Fadul,
2013), type A (obtained from acid hydrolysis of pig skin) and type B
(derived from basic hydrolysis of beef hides or bones) gelatines are the
most commonly used.
Gelatine hydrogel formation is thermally induced by heating. When
cooling below the gelation transition temperature (Tg ~ 30 ◦ C) the
gelatine molecules transition from a random coil conformation to an off
register partial triple helix structure, which further forms a physically
cross-linked network of gelatin chains stabilized by hydrogen bonding.
Physically linked gelatine hydrogels and aggregates are widely used in
food products (e.g. desserts, candies, jellied meat, ice creams, bakery
goods, etc.) as clarification agents, stabilizers, thickeners, emulsifiers,
texturizers, and protective materials; and in pharmaceutical applica­
tions as foaming, emulsifying and wetting agents (Rashid et al., 2019).
Physically cross-linked gelatine hydrogels exhibit poor mechanical
and enzymatic stability, which makes them unsuitable for biomedical
applications. Hence, in order to improve stability, gelatine hydrogels are
produced by chemical cross-linking protocols in which the gelatine tri­
ple helix units are connected via strong covalent bonds. These bonds are
either introduced by chemically modifying the gelatine peptides prior
gelation, introducing additional reactive molecules during gelation or
by the action of enzymes. The chemically cross-linked gelatines are
finding application as drug delivery agents, in tissue engineering and
wound dressings among others (Jaipan, Nguyen, & Narayan, 2017).
3.4. Peptide hydrogels from milk
While milk from different mammalian species (cow, sheep, goat,
mare, etc.) is consumed, here the focus will be on bovine milk which is
the most commercially available. Normal bovine milk is a complex
system that contains proteins (30–35.8 g/L), fat (37–50 g/L), lactose
(47–50 g/L), ash (6.8–7.4 g/L) and water (O’Regan, Ennis, & Mulvihill,
2009).
There are two main groups of milk proteins which are categorized
according to their solubility at pH 4.6. The non-soluble fraction of
bovine milk, the caseins, which comprise about 80% of the total nitro­
gen in milk, consist of four nonglobular phosphoproteins, αs1- (~40%),
αs2- (~10%), β- (~35%), and κ-caseins (~15%), that associate into mi­
celles of about 200 nm in diameter. The caseins micelles are stabilized
by strong electrostatic interactions with calcium phosphate bridges and
weaker noncovalent intermolecular interactions between the casein
proteins. The soluble whey fraction contains ~60% of β-lactoglobulin
(β-lg), ~20% α-lactalbumin (α-la), ~3% bovine serum albumin (BSA),
and ~10% of Immunoglobulin (Ig) (O’Regan et al., 2009).
44
Trends in Food Science & Technology 104 (2020) 37–48
Peptides/protein mixtures derived from milk products have been
generated by enzymatic hydrolysis and fermentation. However, the
identification of a main component causative of hydrogelation is rare
(Madadlou & Abbaspourrad, 2018). Next, we report the few cases where
specific PHGs were identified from milk proteins.
3.4.1. Peptide hydrogels from milk hydrolysates
Several aggregating peptides were identified from bovine whey
protein hydrolysed with Bacillus licheniformis protease (a glutamyl
endopeptidase) (Creusot & Gruppen, 2007). The sequence of the major
aggregating component corresponded to β-lg AB [f1–45] (Fig. 5). This
peptide was identified together with several overlapping sub-sequences
which also corresponded to aggregating peptides, such as β-lg AB
[f12–45] and β-lg AB [f1–33]. The authors attributed the aggregating
properties of these peptides in part to hydrophobic interactions
conferred in large proportion to stretch β-lg AB [f20–34]. The aggre­
gating capability of β-lg AB [f1–45] gives the peptide stability to further
hydrolysis (as indicated by the presence of three cleavage positions
within the sequence namely Asp11, Asp33, and Glu44). Other aggre­
gating peptides found after hydrolysis were β-lg AB [f90–108]-S-S-α-la
[f50–113], α-la [f12–49]-S-S-α-la [f50–113], β-lg AB [f90–108]-S-S-β-lg
AB [f90–108], β-lg A [f90–157], and β-lg AB [f135–157/158] (Fig. 5).
In another work, Pouliot et al. (2009), isolated the peptide β-lg AB
[f1–8] (Fig. 5) from a peptide mixture obtained during the concentration
by reverse osmosis of a tryptic hydrosylate of whey protein. The peptide
β-lg AB [f1–8] was identified as the entity responsible to form aggre­
gates. Aggregates of β-lg AB [f1–8] were obtained at basic pH (8–10) and
were related to a β-sheet structure. Furthermore, the authors proposed
that β-lg AB [f1–8] is the major hydrophobic domain involved in the
aggregation process of β-lg AB [f1–45].
The β-lg AB [f1–8] peptide consists of eight amino acids with
sequence (LIVTQTMK) in which the first seven are either hydrophobic or
neutral and the last one is hydrophilic. In principle, this primary struc­
ture provides a head to tail amphiphilic nature to this peptide, which
explains the peptide capacity to self-assemble and form supramolecular
hydrogels (Guy, Tremblay, Voyer, Gauthier, & Pouliot, 2011). Further­
more, a study of synthetic analogues of β-lg AB [f1–8] showed that the
hydrophobic character of the peptide is the predominant factor driving
peptide self-assembly. For instance, substitution of Lys with Ala not only
did not affect peptide assembly or hydrogel formation but favoured their
formation over a wide pH range (2.0–10.0). This was attributed to a
decrease in the repulsive electrostatic interactions introduced by the
charged Lys. A similar behaviour was observed for the N-acetylated
derivative of β-lg AB [f1–8] and for an analogue with a reverse sequence
(KMTQTVIL). This again was attributed to an increase in the peptide’s
hydrophobic character and reduction of electrostatic repulsive in­
teractions of the N-terminal amino group (Guy & Voyer, 2012).
The effect of several physicochemical factors on the hydrogelation of
peptide β-Ig f1-8 were studied by Guy et al. (Guy et al., 2011). In general,
it was found that β-Ig f1-8 transitioned from random coils to a β-sheet
secondary structure in a pH range of 9–11 and formed hydrogels at
concentrations higher than 2.5 mg/mL within this pH range. Hydrogel
formation was not relevantly affected by changes in ionic strength or
temperature.
Recently, it was demonstrated that the octapeptide β-Ig f1-8 is
capable of stabilizing bovine micellar caseins by promoting protein
dissociation and further encapsulation into peptide gels, which derives
into stable supramolecular structures via strong hydrophobic in­
teractions (Suwal et al., 2019).
Importantly, there is a report on the cytotoxic effect of β-Ig f1-8,
which was attributed to the nanostructures formed by the peptide.
These observations warrant the need of further studies to assess the safe
utilization of PHGs derived from food sources (Jacquot, Gauthier,
Drouin, & Boutin, 2010).
L.M. De Leon Rodriguez and Y. Hemar Trends in Food Science & Technology 104 (2020) 37–48

Fig. 5. Peptide fragments of bovine β-Ig A (A) and α-Ia B (B) identified to form aggregates (Creusot & Gruppen, 2007).

3.4.2. Peptide hydrogels from milk fermentation Table 1 shows six aggregating peptides that were identified after
A mixture of peptides was extracted from the liquid phase of a curd, sequential hydrolysis of soy glycinin with chymotrypsin and trypsin (soy
which was the product of the fermentation with S. thermophilus, L. casei, glycinin has a strong tendency to aggregate upon hydrolysis with
and B. bifidum in a 7:1:2 ratio. Importantly, the peptide mixture pre­ chymotrypsin) (Kuipers & Gruppen, 2008). An important feature of
sented antimicrobial properties and formed hydrogels. However, the these peptides is that they are highly hydrophobic, which is highlighted
authors of this work did not determine which peptides in the mixture
had antimicrobial or gel formation capabilities (Manna, Ghosh, &
Mandal, 2019). Table 1
Peptide sequences identified from plant protein aggregates.
3.5. Peptide hydrogels from hen egg white Protein origin Nanostructure Peptides sequence
(Reference)
Recently, it was shown that the peptide mixture resulting from the Soy (Kuipers Precipitate NAMFVPHYNLNANSIIYALNGR,
thermal acid hydrolysis of hen egg white lysozyme forms injectable & Gruppen, RPSYTNGPQEIYIQQGNGIFGMIFPGCPSTY,
hydrogels in PBS buffer in presence of MgCl2. Hydrogelation of the 2008) NAMFVPHYNLNANSIIYALNGR,
NGIYSPHWNLNANSVIY,
hydrolysate was attributed to fragments 49–101 and 53–101 which
NGIYSPHWNLNANSVIYVTR,
contain the disulfide bridges by Cys64–Cys80 and Cys76–Cys94, NGSHLPSYLPYPQMIIVVQGK
respectively (Yang, Li, Yao, Yu, & Ma, 2019). Canola (Hong Spheres (DQPLVII, FVRYHE, GFAYVVQ, GLEETI, GLYLP,
et al., 2017) GLYLPSFF, GLYLPTFF, MVLPK, MVLPQ,
3.6. Peptide hydrogels from plant proteins PRGPPQSPQDN, PRPFYLAGN, SALRALP,
SHWIYN, SHWIYNTGDQPLVE, SLGVPPQLGNA,
SPKISYVVQGMGI, SVLRGLPLEVI,
There are several proteins derived from plants (i.e. potato, rice, SYTLPILQYIRL, TGDQPLLVII)a
wheat, soy, pea, peanut, faba bean, kidney bean, red bean, mung bean (GPQQRPPLLQQ, PFQKTMPGPS,
and cottonseed) that are known to form aggregates and/or gels under PQGPQQRPPLLQ, PQGPQQRPPLLQQ,
PQQRPPLLQQ, SPQQRPPLLQQ, TLPILQYIRL)b
specific conditions (Cao & Mezzenga, 2019; Josefsson et al., 2019;
Soy ( Fibrils GVAWWMYNNEDTPVVAVSIIDTNSLEc,
Josefsson et al., 2020; Langton et al., 2020; Mackintosh et al., 2009; Josefsson SILSGFTLEFLEHAFSVc,
Munialo, Martin, van der Linden, & de Jongh, 2014; Nicolai & Chasse­ et al., 2019) LDIFLSIVDMNEGALLLPHFd,
nieux, 2019; Ridgley & Barone, 2013; Ridgley, Ebanks, & Barone, 2011; ADFLLFVLSGRAILTLVNNDe,
Tang & Wang, 2010; Tang, Zhang, Wen, & Huang, 2010; Zhao, Liu, LSSTQAQQSYLQGFSHNILETSFHSEFEe
Potato ( Fibrils GGGIKGIIPAIILEFLEGQLQEVDf,
Zhao, Ren, & Yang, 2011; Zhou, Zhang, Yang, Wang, & Qian, 2014). One Josefsson PALLSLSVATRLAQEDf,
can also find examples of medical applications of nanostructured plant et al., 2020) PAFASIRSLNYKQLLLLSLGTGTNSEFf,
proteins (Reddy & Yang, 2011). However, to the best of our knowledge APCANGIFRYNSg
single peptide sequences derived from plant proteins causative of Supscripted letters: a = peptides derived from cruciferin, b = peptides derived
hydrogelation have not been identified. Instead, one can find reports on from napin, c = peptides derived from glycinin G1, d = peptides derived from
peptide mixtures which assemble into nanostructures. Next, we provide β-conglycinin (α-chain), e = peptides derived from β-conglycinin (β-chain) f =
a summary of those cases where peptides of plant origin have been peptides derived from patatin, g = peptide derived from a serine protease in­
identified as potential components of aggregates. hibitor. Bold letters indicate sites of amino acid variations.

45
L.M. De Leon Rodriguez and Y. Hemar
as the main factor contributing to their poor solubility which further
induces the formation of insoluble aggregates.
Two well-defined nanoparticles were produced in the presence of
Ca2+ and peptides derived from the alkali hydrolysis of a cruciferin
extract - one of the major constituent proteins found in Canola meal
(Hong, Akbari, & Wu, 2017). Further characterization showed that the
nanoparticles consist of 26 peptides (the peptide composition of one of
the nanoparticles is shown in Table 1), which primarily adopted β-sheet
and turn conformations, and that particle formation was mainly driven
by electrostatic and hydrophobic interactions.
Five peptides derived from the hydrolysis of nanofibrils formed by
soy proteins were identified (Table 1) (Josefsson et al., 2019). The
peptide ability to form fibrils was further assessed by subjecting the
synthetically made peptides to fibrillation conditions (pH 2, 50 ◦ C with
and without agitation). All peptides, except ADFLLFVLSGRAILTLVNND
formed short fibrils which were associated with some degree of β-sheet
structure. The authors of this work concluded the existence of several
fibrillation pathways for the mixture of soy protein isolate.
More recently, four peptide fragments derived from two different
proteins were identified as the major components of the nanofibrils
obtained from a potato protein isolate (Table 1) (Josefsson et al., 2020).
Importantly, the authors of this work did not determine whether fibrils
contained only one type of peptide or a combination.
3.7. Potential usage of hydrogels of single peptides in food
Most of the potential applications of PHGs in food science can be
extrapolated from those reported in the literature for self-assembled
food protein hydrolysates. However, the relatively high production
costs of PHGs limits their current use to premium food technologies.
Among these, one can highlight the use of PHGs as: 1) carriers for nu­
trients and drugs, packaging materials, antioxidants, and antimicrobials
(Cao & Mezzenga, 2019); 2) carriers for bioactive peptides obtained
from animal by-products (Abaee, Mohammadian, & Jafari, 2017); 3)
disrupters of protein structure for reduction of the allergenicity of some
food proteins (Bechaux, Gatellier, Le Page, Drillet, & Sante-Lhoutellier,
2019); 4) materials for microbial compartmentalization (Johnston et al.,
2020); and as microbial biosensors (Tomadoni et al., 2020).
Moreover, once the production limitations of PHGs are solved then
these compounds can be applied as: 1) thickening and gelling in­
gredients; 2) additives for meat products, in order to lower the cooking
loss compared to a control meat, to increase water-binding capacity and
to induce specific texture properties (Lacou et al., 2016); 3) building
blocks” for design and development of food-grade micro- and nanonet­
work structures (Ramos et al., 2017), hence acting as texturizing agents
or fat replacers or nutraceutical nanocarriers; 4) substitutes of milk
proteins in yoghurts; and 5) source of protein-based nutrients.
Furthermore, PHGs can readily be applied in those cases requiring
relatively small amount of material such as emulsifiers and protein
stabilizers (Suwal et al., 2019).
4. Conclusions
This literature review allows one to conclude that food products can
be an important source of single component PHGs and that these can
find important applications not only in food science but they can also be
part to the repertoire of PHGs scrutinized for applications in biomedi­
cine, nanotechnology and other areas. However, research in this area is
in its infancy and its progress is limited due in part to the lack of 1) tools
that will allow one to predict peptide fragments within a food protein
that can self-assemble and form gels and 2) efficient peptide purification
protocols. Therefore, more research will have to be directed on these
areas in conjunction with optimization of enzymatic/fermentation
production protocols. Finally, one should consider that when adding
single component PHGs to a heterogeneous complex food sample the
aggregation capacity of the peptides may be either enhanced or
46
Trends in Food Science & Technology 104 (2020) 37–48
impaired given the presence of other molecules (polysaccharides, lipids
and other proteins). This warrants further studies that will allow one to
understand the extent to which PHGs can be an important part of the
final functional properties, including the sensory properties of food
products and their mechanism of action.
Author contributions
L.M. De Leon-Rodriguez conducted the review and wrote this
manuscript. Y. Hemar designed, revised, and corrected this manuscript.
Funding
This research did not receive any specific grant rom funding agencies
in the public, commercial, or not-for-profit organizations.
Declaration of competing interest
Nothing declared.
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