Food Chemistry 428 (2023) 136709

Contents lists available at ScienceDirect

Food Chemistry
journal homepage: www.elsevier.com/locate/foodchem

Protein-polysaccharide nanoconjugates: Potential tools for delivery of

plant-derived nutraceuticals
Balwant S. Paliya a, 1, Vivek K. Sharma a, 1, Minaxi Sharma c, 1, Deepti Diwan d,
Quang D. Nguyen e, Tejraj M. Aminabhavi f, Gaurav Rajauria g, Brahma N. Singh a, b, *,
Vijai Kumar Gupta h, i, *
a
Herbal Nanobiotechnology Lab, Pharmacology Division, CSIR-National Botanical Research Institute, Lucknow 226001, India
b
Academy of Scientific and Innovation Research (AcSIR), Ghaziabad 201002, India
c
CARAH ASBL, Rue Paul Pastur, 11 – 7800 ATH, Belgium
d
Washington University School of Medicine, 4590 Children’s Place, Ste. 8200, Campus Box 8057, St. Louis MO63110, USA
e
Department of Bioengineering and Alcoholic Drink Technology, Institute of Food Science and Technology, Hungarian University of Agriculture and Life Sciences, H-1118
Budapest, Ḿ enesiút 45, Hungary
f
School of Advanced Sciences, KLE Technological University, Hubballi 580031, India
g
Department of Biological & Pharmaceutical Sciences, Munster Technological University, Tralee V92HD4V, Co. Kerry, Ireland
h
Biorefining and Advance Material Research Centre, SRUC, Barony Campus, Parkgate, Dumfries DG1 3NE, United Kingdom
i
Centre for Safe and Improved Food, SRUC, Kings buildings, West Mains Road, Edinburg EH9 3JG, United Kingdom

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

Keywords: Protein-polysaccharide nanoconjugates are covalently interactive networks that are currently the subject of
Proteins intense research owing to their emerging applications in the food nanotechnology field. Due to their biocom­
Polysaccharides patibility and biodegradability properties, they have played a significant role as wall materials for the formation
Nanoconjugates
of various nanostructures to encapsulate nutraceuticals. The food-grade protein-polysaccharide nanoconjugates
Delivery systems
Plant-derived nutraceuticals
would be employed to enhance the delivery and stability of nutraceuticals for their real use in the food industry.
The most common edible polysaccharides (cellulose, chitosan, pectin, starch, carrageenan, fucoidan, mannan,
glucomannan, and arabic gum) and proteins (silk fibroin, collagen, gelatin, soy protein, corn zein, and wheat
gluten) have been used as potential building blocks in nano-encapsulation systems because of their excellent
physicochemical properties. This article broadens the discussion of food-grade proteins and polysaccharides as
nano-encapsulation biomaterials and their fabrication methods, along with a review of the applications of
protein-polysaccharide nanoconjugates in the delivery of plant-derived nutraceuticals.

1. Introduction cardiovascular disease because they contains numerous nutraceutical

As an alternate approach to therapeutic agents, dietary nutraceut­
icals have emerged as different substitutes for conventional medication.
Dietary nutraceuticals are nutritional supplements that contain bioac­
tive ingredients found in food-origin products but at much higher con­
centrations and doses than active nutrients that we naturally absorb
through a regular, balanced diet (González-Sarrías et al., 2013). Several
epidemiological research studies and their meta-analysis concluded that
the regular consumption of fruits and vegetables prevents or reduces the
risk of chronic diseases such as type II diabetes, cancer, and
compounds such as polyphenols, flavonoids, anthocyanins, minerals,
vitamins, and other antioxidant compounds. These dietary nutraceutical
compounds are well known for their significant medicinal values and
health promoting effects (Hu et al., 2017). However, plant-based nu­
traceutical compounds showed low solubility and poor stability during
oral administration because of degradation via enzymes, gastrointes­
tinal (GI) tract-altered pH, and the presence of other bio-molecules. All
these hindrances affect the bioavailability of nutraceutical compounds
to target sites and cause low absorption and assimilation in the body.
Most of the dietary metabolites face a barrier during diffusion through

* Corresponding authors at: Herbal Nanobiotechnology Lab, Pharmacology Division, CSIR-National Botanical Research Institute, Lucknow 226001, India (B.N.
Singh). Biorefining and Advance Material Research Centre, SRUC, Barony Campus, Parkgate, Dumfries DG1 3NE, United Kingdom (V.K. Gupta).
E-mail addresses: bn.singh@nbri.res.in (B.N. Singh), vijai.gupta@sruc.ac.uk (V.K. Gupta).
1
These authors contributed equally.

https://doi.org/10.1016/j.foodchem.2023.136709
Received 18 December 2022; Received in revised form 20 June 2023; Accepted 21 June 2023
Available online 28 June 2023
0308-8146/© 2023 Elsevier Ltd. All rights reserved.
B.S. Paliya et al.
the plasma membrane lipid bilayer, which profoundly limits the appli­
cations of plant-derived nutraceuticals. To sidestep the following prob­
lems, dietary nutraceutical compounds should be protected by
encapsulating or entrapping them with nanocarriers to enhance their
stability during GI tract transfer at the target site (Gao et al., 2021).Fig. 1
With the rapid advancement of material science and chemistry over
the last few decades, the use of nanomaterials has found its way into
sectors such as pharmacy, clinical research, and treatments. Synthetic
polymers-based nanomaterials showed considerable clinical success,
and then studies were focused on searching for natural sources to
fabricate biopolymer-based nanomaterials to enhance the stability and
bioavailability of bioactive ingredients because they were relatively
safe. Several biopolymers have currently been modified and utilized for
various applications in food industry (Ahmad et al., 2022).
Polysaccharides are the most abundantly polymer and are ubiqui­
tously present in almost every plants, animals and microbes. These
natural substances have a wide range of characteristic properties that
are required for sustaining living systems. Along with bio-molecules,
DNA, RNA, proteins, and polysaccharides are also essential for build­
ing various biological components required in a living system, such as
signaling molecules, antigens for cell recognition, cell–cell communi­
cations, and molecular components. They impart a crucial role in cell
structure, adhesion, the immune system, and therapeutic prevention
(Song et al., 2022). Recently, polysaccharides have gained much
attention as promising biomaterials due to their properties such as being
renewable, biodegradable, and environmentally biocompatible. More
importantly, their vast availability from natural sources makes them
useful biomaterials as wall materials. Polysaccharides are hydrophilic,
non-toxic, and easily digestible by different enzymes present in the
body. These properties make polysaccharides a perfect material for the
fabrication of nanoparticles and are used as carriers to deliver many
drugs and therapeutic metabolites. In terms of medical applications,
polysaccharides and nanoparticles showed high drug loading efficiency,
drug stability, easy diffusibility, and targeted delivery (Nourbakhsh
et al., 2020). Therefore, researchers constantly develop polysaccharide-
based nanomaterials for various biomedical applications. This concept
also promotes the idea of fabricating polysaccharide-based nanocarriers
that are used to protect nutraceuticals and other bioactive food
metabolites.
Similarly, protein polymers are also abundantly found in nature as
macromolecules that are derived from plants, animals, algae, bacteria,
and fungi. They are biocompatible, easily processed, and biodegradable
in the environment without leaving toxic waste (Song et al., 2022).
However, proteins are highly susceptible to enzymatic degradation and
denaturation when administered into the body. Protein administrations
showed low bioavailability and a high biological passage barrier and
Fig. 1. Electrostatic polymer crosslinking.
2
Food Chemistry 428 (2023) 136709
therefore required repeated consumption for better results. Researchers
are trying to depict reliable methods to improve in vivo protein stability
so it can be successfully utilized for immunological, therapeutic and
nutraceutical purposes (Liu et al., 2022). Proteins and polymer-based
nanoparticles served as potential agents for a variety of food and drug
delivery systems due to their unique structural architecture. Protein-
based nanostructures showed enhanced stability of nutraceuticals both
in vitro and in vivo due to the entrapment of proteins in nanoparticles
(Song et al., 2022; Sun et al., 2021).
However, several important characteristics of polymer should be
considered, such as its immunogenicity inside the body (it does not
activate the immune response), biocompatibility, and safety. A promi­
nent polymer should contain characteristic physical and chemical
properties suitable for nanocomposite synthesis and their morphological
alteration. As a result, the synthesized nanocomposite is biodegradable
in vivo conditions, such as polymer’s physical properties such as size,
molecular weight, and nanostructure formation, after nutraceutical
metabolites are encapsulated, determine polymer degradation kinetics
inside the body (Aranaz et al., 2021). Different crosslinked biopolymers
have also been investigated for the encapsulation of nutraceutical sub­
stances in addition to delivery methods or carriers. For nutraceuticals to
perform as intended, suitable encapsulating materials are crucial. Un­
doubtedly, the distribution of nutraceuticals in food applications can
only be accomplished with edible ingredients. For the encapsulation of
nutraceuticals, food-grade biopolymer-based systems, such as proteins
and polysaccharides, have chosen.
The current review discusses the synthesis and application of
protein-polysaccharide nanoconjugates as dietary nutraceutical carriers
for the safe and efficient delivery of bioactive nutraceuticals to target
specific sites inside the body for better assimilation. Firstly, a variety of
suitable natural polysaccharides and proteins were discussed, together
with their availability in nature, their advantages, and the problems
associated with their application as nanocarriers for nutraceutical
compounds. The following section covered dietary nutraceuticals
derived from food and plant-based metabolites that are currently used as
dietary supplements, as well as limitations encountered during oral
administration of these nutraceutical supplements, such as stability,
degradation, and low solubility. Further, the review summarized
research studies on protein-polysaccharide-based nanoconjugates for
enhancing stability and bioavailability of plant-derived dietary nutra­
ceuticals along with a review of the possible research gap, challenges
and future perspectives.
2. Plant-derived nutraceutical compounds and their
applications in healthcare
Nutraceuticals refer to a food-derived nutritional component that
provides medical or health benefits, including the prevention and
treatment of disease. In other terms, nutraceuticals are products isolated
or purified from foods that are generally sold in medicinal forms not
usually associated with food. The term “nutraceutical” was coined in
1989 by Dr. Stephen D. Feliz, who is also considered the father of
nutraceuticals. So, the word nutraceuticals are derived from nutrition
and pharmaceuticals. In the past few decades, changes in the method of
agriculture have resulted in the use of harmful chemicals, pesticides, and
the artificial ripening of fruits and vegetables, significantly depleting the
nutritional values of foods. The inadequate supply of the daily dose of
nutrition such as vitamins, minerals, proteins, carbohydrates, fats, and
other nutrients will ultimately manifest itself in the form of various
ailments in the long run. To achieve better nutrition, nutraceuticals play
a prominent role in bridging the gap.
Plant-derived nutraceuticals demonstrated the numerous health
benefits with no or minimum side-effects. The varieties of functional
foods such as fortified foods, fiber, plant extracts, amino acids, vitamins,
minerals, biologically active phytochemicals, and herbal products all
come under the single umbrella of nutraceutical category. Plant-derived
B.S. Paliya et al. Food Chemistry 428 (2023) 136709

Table 1
Plant-based nutraceutical compounds and their therapeutic effects.
Compound Source Therapeutic effect References

Curcumin Turmeric Anticancer, antiinflammatory, antioxidant, antimicrobial, antimutagenic, cardiovascular, Diabetes (Hu et al., 2017)
Resveratrol Grape skin and Seeds Anti-inflammatory, cardioprotective, anti-carcinogenic, neuroprotective and vasorelaxant (Wu et al., 2017)
Quercetin Apples, grapes and onion Anti-Quorum sensing, antioxidant (Farrag et al., 2018)
Rutin Buckwheat and citrus fruit Antibacterial, antiallergic, antitumor, vasoactive, anti-inflammatory, hypolipidemic, antihypertensive (Ganeshpurkar &
and antiplatelet Saluja, 2017)
Genistein Soy-based foods Anticancer (Thangavel et al.,
2019)
Catechin and Tea, apples, chocolate, Antioxidative and lowering hypertension, inflammation, and hyperlipidemia (Liu et al., 2020)
EGCG and green tea
β-Carotene Carrots, pumpkins, maize, Cardioprotective and anticancer (Jaswir et al., 2011)
and tomato
Astaxanthin Phaffiayeast, shrimp, crab, Anticancer, antioxidant anti-diabetes, CVD, and neurodegenerative disorders (Olaizola, 2007)
and salmon
Myricetin Grapes, walnut, and Anti-diabetic, anticancer, immunomodulatory, analgesic and antihypertensive (Taheri et al., 2020)
berries
Chrysin Honey, mushrooms and Lowers neurotoxicity and oxidative stress, antioxidant, anti-inflammatory, andanticancer (Naz et al., 2019)
propolis
Lycopene Tomato, grapefruit, and Anticancer (prostate cancer), cardiovascular diseases, decreased risk of chronic diseases & antioxidant (Dhiman & Bhalla,
apricot 2019)
Lutein Spinach, lettuce, and eggs Antioxidant and prevents cataracts, Inhibits diabetic retinal vascular disease, and eye detachment, bone (Li et al., 2020)
remodeling activities, hepatoprotective, anti-inflammatory & anti-arthritis
Kaempferol Tea, strawberries,and Anticancer & antioxidant (Chen & Chen, 2013)
tomatoes
Saponin Soybeans, legumes, and Anti-inflammatory, hypocholesterolemic, and antifungal (Marrelli et al., 2016)
maize
Naringenin Citrus fruit Antioxidant and effective in lowering visceral adiposity and plasma lipids, anti-inflammatory, (Jaswinder et al.,
antiestrogenic, anticancer 1996)
Apigenin Celery and parsley Anti-toxicant, anti-inflammatory, and anticancer, amnesia and alzheimer’s disease, diabetes, insomnia (Ali et al., 2019)
and depression
Chlorogenic Green coffee and apples Effective against cancer, diabetes, antioxidant, hepatoprotective, hypoglycaemic (Marques & Farah,
acid 2009)
Gingerol Ginger Anti-cancer, antioxidant, anti-inflammatory, antiemetic, antiulcer, and antiplatelet, anti-diabetic and (Kausar et al., 2022)
lipid-lowering activities
Cinnamic acid Cinnamon Anticancer diabetes and neurological disorders (Ruwizhi &
Aderibigbe, 2020)

bioactive nutraceutical compounds include polyphenols, flavanones,
anthocyanins, proanthocyanidins, flavanones, ellagitannins, ellagic
acid, flavan-3-ols, and procyanidins (Hu et al., 2017). Nutraceuticals are
of two kinds: traditional and non-traditional. Traditional nutraceuticals
are simply natural whole foods, whereas non-traditional nutraceuticals
are fortified nutraceuticals and recombinant nutraceuticals, which are
made artificially with the help of biotechnology. Nutraceuticals are
further classified into three major categories: (a) the first one is based on
the source from which the nutraceuticals are extracted form plants,
animals, and microbes; (b) based on their chemical nature; (c) based on
its mechanism of action such as anti-cancer anti-inflammatory proper­
ties antioxidants and osteogenetic properties lowering the low-density
lipoprotein.
Now a day’s varieties of nutraceuticals are available in the market.
However, only those products are in demand that contains nutraceut­
icals belonging to antioxidants, omega-3 fatty acids, and probiotics. The
most widely used nutraceutical with antioxidant properties comes from
plant polyphenolic compounds, vitamins, carotenoids, and minerals,
especially those of plant origin. Phytosterols and anthocyanins are
gaining popularity as potential nutraceutical supplements due to their
numerous biological activities. Polyphenolic nutraceuticals confer their
health benefits through mechanisms that include direct scavenging of
reactive oxygen species (ROS), enhancing the activity of detoxifying
enzymes against carcinogens, promoting cancerous cell apoptosis, and
maintaining DNA repair (Shahidi, 2012). Different types of nutraceuti­
cal compounds derived from natural sources along with their healthcare
benefits are summarized in Table 1.
3. Limitations associated with plant-derived nutraceutical
compounds
Plant-derived nutraceuticals, in addition to a healthy diet, provides
significant health benefits. They demonstrated the efficient clinical po­
tential of preventing and treating many chronic diseases and harmful
disorders. However, adequate in vivo assimilation and the potential
benefit of these nutraceuticals were not achieved due to low solubility in
lipid or aqueous phase. For example, curcumin is often referred to as a
“wonder phytocompound” because of its versatile therapeutic effects at
multitarget sites, including cancer cells. However, due to its poor solu­
bility (aqueous solubility is 11 ng/ml), physical instability (degradation
in water at different physiological pH levels), and low bioavailability at
the target site, curcumin could not exhibit its real therapeutic effects
under in-vivo conditions (Metzler et al., 2013). Phyto-nutraceuticals
showed complex formation in the GI tract with other compounds,
restricted permeability across mucus-secreting epithelium cells, and
optimal release from the food matrix. Moreover, carotenoids were
entrapped inside the cellular architecture of the food matrix, but it did
not release properly in the GI tract (Rimbach et al., 2008).
Similarly, the low solubility of plant polyphenols is one of the
physiological parameters for low bioavailability. Plant polyphenols are
subjected to self-oxidation in the alkaline pH of the GI tract. When food
passes through the GI tract, the acidic pH (2.0 to 4.0) offers the degra­
dation of the food matrix via several protein-degrading enzymes, and at
this pH, polyphenols are stable. However, at different alkaline pH levels
(8.0 to 9.0) of the small intestine, mostly polyphenols destabilize and
degrade. For example, EGCG gets degraded and oxidized very fast in the
intestinal environment of high pH. Anthocyanins showed weak stability,
which results in low absorption in blood serum. Curcumin exhibited
high stability in alkaline pH 8.0 to 9.0, but degrades at pH below 7.4. In

3
B.S. Paliya et al.
contrast, trans-resveratrol showed stability at an acidic pH, whereas it
was unstable at a pH above 6.8 (Amri et al., 2012; Hu et al., 2017).
Poor bioavailability of nutraceuticals is also attributed to the phos­
pholipid bilayer and mucus secretion of epithelial cells. Lipid-soluble
nutraceuticals transport excess more easily across cell membranes
compared to aqueous-soluble ingredients. According to research, the
chemical composition of nutraceuticals has a significant impact on the
absorption rate of nutrient-active ingredients (Actis-Goretta et al.,
2013).The presence of pro-oxidants in biological systems causes varia­
tions in the bioactive properties of nutraceutical compounds via
different hydrolysis and redox reactions. The study revealed that ca­
rotenoids and omega-3 fatty acids were subjected to oxidation in the GI
tract environment due to their nature as pro-oxidants. Protein and
peptide-based nutraceuticals metabolize through breakdown and enzy­
matic action in the acidic milieu of the stomach and GI tract. However,
some of the nutraceuticals get oxidized by enzymes and hamper overall
nutraceuticals’ performance. For instance, CYP450 enzymes oxidized
plant-derived nutraceutical bioactive compounds such as genistein and
nobiletin (Metzler et al., 2013).
4. Food-grade nanocarriers
Plant polyphenols have been shown to have positive health effects,
but their low bioavailability continues to restrict their use as prospective
nutraceuticals in the food business. After reaching the stomach lumen, a
relatively little amount of plant polyphenols travels through the
bloodstream for digestion and metabolism. Thus, it is essential to
improve the bioavailability of plant-derived nutraceuticals in order to
encourage their metabolism and absorption and enhance their health
benefits. According to reports, encapsulated delivery systems improve
bioavailability, boost bioactivity, and protect drugs from harmful
environmental factors (Prateeksha et al., 2022).
Despite the fact that a variety of wall materials can be used for
encapsulation, certain characteristics, including affinity, film-forming
capacity, degradability, intestinal resistance, and viscosity, should be
optimized before the choice of delivery materials (Song et al., 2022).
Gums, proteins, and polysaccharides can all be used to make edible
delivery materials. Because of this, it is important to clarify edible
nanoencapsulation materials, particularly for proteins and poly­
saccharides, in order to increase the bioavailability of dietary phyto­
chemicals as nutraceuticals and ensure their successful utilisation in the
food sector.
4.1. Polysaccharides as nanocarriers for nutraceutical delivery
Polysaccharides-based nanodelivery systems have enormous appli­
cations as an additional functional food supplement with nutraceutical
value. The nanoformulation of polysaccharides and processed food
preparations contain a variety of bioactive ingredients that directly in­
fluence health benefits, including clinical symptoms and disease pre­
vention. Because polysaccharides have health benefits, nanocarriers can
also be used as energy ingredients and dietary agents in food resources.
Currently, processed foods’ nutrition and energy contents are main­
tained by using various research techniques by retaining the nutraceu­
tical content using encapsulation technologies during food processing.
Therefore, future research should focus on incorporating polysaccharide
nanomaterials with food-origin ingredients to provide more stability to
nutraceuticals. Moreover, different tastes, colors, and textures of food
can easily be achieved using polysaccharide-based nanocarriers (Tang
et al., 2022). This section discusses various aspects of the natural origin
of polysaccharides and their role as nutraceutical carriers in the food
industry (Table 2).
4.1.1. Cellulose
Cellulose is a high-molecular-weight homopolymer that is a renew­
able resource derived from plants, algae stocks, and hulls. Naturally, it is
4
Food Chemistry 428 (2023) 136709
present in plant cell walls in the form of nested bundles of microfibrils.
Disrupting their natural structure by chemical and mechanical methods
imparts cellulose nanofibrils. From plants, two kinds of nanocellulose
are obtained including nanofibrils (crystalline and amorphous structure)
and nanocrystals (crystalline structure). Whereas microfibrils obtained
from bacteria are referred to as bacterial nanocellulose. The size and
shape of all types of nanocellulose varies depending on their source.
Regarding health benefits, nanofibrils of cellulose showed a positive
bioactive effect against irritable bowel disease, inflammatory bowel
syndrome, neurodegenerative diseases, and immunoregulation (Patil
et al., 2022). Nanocellulose gains its research demand due to its char­
acteristic properties such as mechanical strength, biodegradability, and
biocompatibility. They can easily be adsorbed on oil–water and air–­
water interfaces and are considered perfect agents for foams or emulsion
stabilizers. Nanocellulose acts as fillers or oil part replacers in the for­
mation of nanoemulsions. Due to self-assembly, a gel-like network forms
when interacting with an aqueous solvent. Nanocellulose applications
can be employed by exploiting its surface modification through chemi­
cal functionalization or by using food ingredients in combination such as
biopolymers and biosurfactants. Among several methods, sulfuric acid
hydrolysis technique is the most frequently used method for nanocrystal
production. Acid hydrolysis causes the generation of sulphate half-ester
groups at the nanocellulose surface, due to which negative charge
repulsion forces impart colloidal stability in an aqueous solution (Chen
et al., 2009). For nanofiber production, the mechanical shear method
was used to break up plant cell walls, which fabricates nano-sized fibers
that were less crystalline and longer in size (Kontturi et al., 2016).
The isolated nanocellulose is amphiphilic, and this physicochemical
property facilitates both emulsion and foam stabilization through oil–­
water and water–air interface adsorption. In an aqueous solution,
nanocellulose can form a network, which means it can be utilized as an
agent for food texture modification. The stabilization of emulsion using
colloidal particles is called Pickering emulsion. Nanocellulose is more
hydrophilic in nature as compared to hydrophobic, thereby showing
better oil-in-water adsorption and emulsion stabilization. Food in­
gredients entrapped in emulsion get enhanced stability in nanocellulose
and stabilized processed food emulsion for a longer period. It was re­
ported that the optimized amount of nanocellulose in the emulsion
provided six months of stability without phase separation or settlement.
Pickering emulsions by combining cellulose nanocrystals, water-soluble
cellulose derivatives, and tannic acid (TA) were prepared (Hu et al.,
2016). TA mixed with the emulsion showed considerable stability to
dehydration and rehydration due to the formation of a protective TA-
nanocellulose complex around oil droplets. Several natural food in­
gredients show similar stability in nanocellulose emulsion, which is
considered advantageous for nutraceutical stability in some foods. Soy
protein isolate (SPI) and cellulose nanocrystals (CNC) were used to
construct a novel nanocomposite as carriers for curcumin delivery
(Wang et al., 2020). The nanocomposite’s physical stability depends on
the CNC and SPI mass ratios. The SPI-CNC complex is small in size
(197.7 nm) with a low polydispersity index (0.14) when the SPI-CNC
mass ratio is kept at 6:1. These nanocarriers are relatively stable at
different pH levels from 3 to 9 and various temperatures from 30 to
90 ◦ C. Particles are stable at salt concentrations up to 40 mmol/L and
show a high encapsulation efficiency of 88.3%. The nanocomplex is
effectively digested in the GI tract, and the released carrier components
are sustained metabolically. These polysaccharide-based nanocarriers
were effective for delivering non-polar nutraceuticals.
Several studies have been done on stabilizing nutraceutical essential
oils, namely cinnamaldehyde, eugenol and limonene using encapsula­
tion approach in cellulose nanocrystals and microfibrillated cellulose.
these essential oil-encapsulating nanoparticles exhibited a strong anti­
bacterial activity (Mikulcová et al., 2016). Authors also concluded that
the antibacterial effect was dependent primarily on direct interaction
between emulsion droplets and pathogens, while the impact of free, non-
encapsulated oil was only marginal. The other study reported the best
 B.S. Paliya et al.
Table 2
Natural polysaccharides used for nanoparticles synthesis.
Polysaccharides origin sources Molecular structure References

Chitosan Animals & Shells of marine crustaceans, crabs, lobsters, shrimps, fishes and insect (Divya & Jisha, 2018)
Fungi scales

Cellulose Plants Wheat, bamboo, cotton, jute, potato, sugar beet pulp, tuber vegetables, (Patil et al., 2022)
hemp, barley

Pectin Plants Cell wall and intracellular layer of plant cells, fruits such as apples, lemons (Dos Santos et al., 2015)
oranges
5

Starch Plants Rice, grains, potato, fruits, wheat, maize (Liu et al., 2020)

Chondroitin Animals Animal cartilage (Garnjanagoonchorn et al., 2007)

Sulfate

Food Chemistry 428 (2023) 136709

(continued on next page)
 B.S. Paliya et al.
Table 2 (continued )
Polysaccharides origin sources Molecular structure References

Heparin Animals Mucosal tissue of animal (van der Meer et al., 2017)

Fucoidan Plants & Algae Brown seaweeds (Fucusvesiculosus&Undariapinnatifida) (Dos Santos, Grenha, & biology,
2015)
6

Carrageenan Plants & Algae Red seaweed (Irish moss) (Dos Santos & Grenha, 2015)

Food Chemistry 428 (2023) 136709

(continued on next page)
 B.S. Paliya et al.
Table 2 (continued )
Polysaccharides origin sources Molecular structure References

Mannan Plants Yeast, algae, ivory nuts & coffee beans (Razan, 2021)

Arabic gum Plants Acacia species (Acacia seyal and Acacia senegal) (Jaafar, 2019)

Glucomannan Plants Tubers of amorphophallus konjac (Behera & Ray, 2017)

7

Tragacanth gum Plants Astragalus sp. (Taghavizadeh et al., 2021)

Food Chemistry 428 (2023) 136709

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B.S. Paliya et al. Food Chemistry 428 (2023) 136709

antimicrobial activity in cellulose nanocrystal-stabilized cinnamalde­

hyde essential oil (Saini et al., 2017). Vitamins A and E were used to
immobilize on cellulose acetate nanofibers and electron-dense cellulose
acetate nanofibers (Taepaiboon et al., 2007). The synthesized nanofibers
demonstrated 45% and 83% vitamin A and E loading capacity, respec­

(Cheng et al., 2011)

tively with sustained release of vitamins. This is a great research
establishment showing the use of polysaccharide nanoemulsions as
nutraceutical carriers. Gallic acid an antioxidant molecule was encap­
References

sulated via fabricating functionalized cellulose nanocrystals (CNC)

(Criado et al., 2016). The synthesis process includes a redox reaction
between hydrogen peroxide and ascorbic acid and the subsequent
mixing of gallic acid. The functionalized nanocrystals exhibited a strong
antioxidant activity of 1.9–20 mM Trolox eq./mg CNC.

4.1.2. Chitosan
Chitosan is a natural, biodegradable, and biocompatible biopolymer
obtained through the deacetylation of chitin. Physicochemically, it is
whitish, inelastic, hard, cationic, and strongly basic. Structurally, chi­
tosan is a repeating unit of two types of alternating-unit glucosamine
units with some portion of N-acetyl glucosamine, linked via β-(1–4)-
glycosidic linkage. Chitosan finds its versatile application in the research
fields of agriculture, biomedical industries, food industries, genetic en­
gineering, pollution remedies, and water treatment (Divya & Jisha,
2018). Its mucoadhesive property allows attachment to different mucus
surfaces, and the surface positive charge facilitates drug loading. Both
properties synergistically facilitate the sustained release of drugs and
nutraceuticals at the target site. Its positive charge promoted the inter­
action among various negatively charged polymers and molecules’
functional groups, leading to the whole nanoassembly with targeting
Molecular structure

properties. Chitosan is inexpensive, easily moldable to various shaped

scaffolds, and non-toxic; thus, chitosan nanoparticles are widely used as
carrier molecules, and their mucoadhesive properties allow the delivery
of chitosan nanoparticles via oral administration, intravaginal, intra­
nasal, intraocular, intratracheal, and intrapulmonary routes. The poly­
mer is used as dietary fibre in many countries, including the USA, to
reduce fat absorption. Its polymeric nature allows encapsulation and the
formation of the nanocomposite of several nutraceutical ingredients to
provide stability and target delivery. The most noticeable feature of
chitosan is its non-immunogenic response, which does not cause severe
inflammation of the immune system. Chitosan nanoparticles are strong
antibacterial agents due to their surface positive charge, which provides
a contamination-free coating for food ingredients (Ahmad et al., 2022).
Chitosan is thought to help with the intestinal absorption of many
bioactive nutraceutical compounds, particularly flavonoids. Catechin
and epigallocatechin, flavonoids present in green tea, are susceptible to
degradation in the intestinal environment, which hinders adequate ab­
sorption of both plant metabolites. Chitosan encapsulation significantly
improved the stability and intestinal absorption of both flavonoids.
Curcumin has vast therapeutic properties, but due to its low solubility
Aureobasidium pullulans

and fast metabolism, its bioavailability after oral administration is low.

The mucoadhesive chitosan-coated curcumin nanoparticles were
developed to have a prolonged retention time while adhering to the
colon mucosal surface, leading to sustained release of curcumin with
sources

enhanced bioavailability and an improved anti-cancer effect against

colorectal cancer cells. A high-pressure, time-recycle homogenization
for alginate-chitosan nanosphere preparation and size optimization was
studied (Hu et al., 2017). The synthesized alginate-chitosan encapsu­
lated epigallocatechin with 80.1% efficiency and exhibited antioxidant
Fungus

activity in 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging of

origin

8.8% and 69.3% for 2.6 and 6.9 pH, respectively. An improved green tea
Table 2 (continued )

polyphenol-derived epigallocatechin gallate (EGCG) potential was

examined against human melanoma cells using nanotechnology (Sid­
Polysaccharides

diqui et al., 2014). The team developed chitosan nanoparticles encap­

sulated EGCG for an oral delivery system. The nano-EGCG demonstrated
Pullulan

an eight-fold dose advantage over native EGCG. Moreover, nano-EGCG

treated melanoma cell line showed significant apoptosis induction and

8
B.S. Paliya et al.
cell cycle arrest. Also, nano-EGCG exhibited growth inhibition in tumor
xenografts.
Chitosan nanoparticles containing lycopene were created using the
ionic gelation method along with various concentrations of chitosan
(100–200 mg) and sodium tripolyphosphate (Dhiman & Bhalla, 2019).
The nanocomposite showed 89.4% lycopene entrapment efficiency and
83.5% release efficacy. These results suggested that lycopene-loaded
chitosan nanoparticles exhibited enhanced lycopene accumulation and
can be an effective agent for target delivery of lycopene. Resveratrol, a
polyphenolic compound majorly found in grape skin and peanuts, was
loaded with chitosan nanoparticles via the ionic cross-linking method
(Wu et al., 2017). Resveratrol nanoparticles exhibited an average par­
ticles size range of 172 to 217 nm, and the particles are stable at UV
storage. Resveratrol-loaded nanoparticles demonstrated low toxicity to
normal hepatocyte L02 cells and showed promising antiproliferative
activity against liver carcinoma SMMC 7721 cells, indicating synthe­
sized nanoparticles’ efficacy for chemotherapy use. Quercetin is another
dietary flavonoid commonly present in fruits, grains, vegetables, and
plant leaves. The ionic gelation method was used to develop quercetin-
loaded chitosan nanoparticles (Zhang et al., 2008). A study against
hepatocancer indicated that quercetin-loaded chitosan nanoparticles are
more effective for hepatoprotection. A chitosan nanocarrier for nar­
ingenin flavonoid has used and found that nanoparticles are highly
effective to reduce the viability of breast cancer T47D cells (Winarti
et al., 2015). Naringenin-loaded chitosan nanoparticles were developed
for bone development therapy (Malathy & Iyer, 2018).
4.1.3. Pectin
Gellan gum/pectin blend nanoparticles were prepared using the
ionotropic gelation method for the encapsulation of resveratrol and
tested for nanoparticle permeability and sustained release pattern in the
Caco-2 cell model and mucus-secreting tripe co-culture model (Prezotti
et al., 2020). They reported that in a low pH acidic medium, particles
can load high concentrations of the drug (greater than80%) and remain
attached to polymeric nanoparticles for 2 h, and in a pH 6.8 medium.
These particles exhibited sustain release of resveratrol up to 85% in 30 h.
Pectin nanoparticles were also developed using food-grade material
from cassava roots to coat the dietary nutraceutical, β-carotene (Coelho
et al., 2020). The pectin nanoparticles were synthesized with the aim of
GI delivery of lipophilic dietary nutraceuticals to improve bioavail­
ability. The particles are of 21.3 nm mean size, mucoadhesive, and
showed improved stability up to 90 days at 4 ◦ C, bioavailability, and
antioxidant activity.
Hyperoside (HYP) is a 3-O-galactoside of quercetin present in plants
and has excellent antioxidant, antibacterial, antiinflammatory, anti­
tumor, anticoagulant, and antidiabetic activities. Hyperoside, on the
other hand, is only sparingly soluble in water and has a low bioavail­
ability (Li et al., 2022). A HYP and zein-tea polyphenol-loaded pectin
nanocomplex was prepared (Wang et al., 2021). The freeze-dried
nanocomplex showed improved water-redispersibility and high stabil­
ity at pH 2.0–8.0 and 0–500 mM ionic strength conditions. Moreover,
nanocomplexes also displayed enhanced antioxidant potential as
compared to free HYP and time-lapse release of nutraceutical metabo­
lites in GI conditions.
4.1.4. Starch
Starch is a natural biopolymer present as plant carbohydrate storage.
Starch-based nanomaterials, as biodegradable, non-toxic, biocompat­
ible, have a potential application for the regulated release and protec­
tion of different nutraceuticals due to their role as capping and
stabilizing agents. Its low solubility in cold water limits its versatile use.
Starch nanoparticles exhibited some advantages over other poly­
saccharides such as enzymatic hydrolysis, pH sensitivity, and mucoad­
hesive properties. They could be employed as nanofillers to enhance the
mechanical and barrier qualities of biocomposites, according to recent
studies. It is being looked at how they may be used in the food business,
9
Food Chemistry 428 (2023) 136709
which is constantly looking for new and inventive ways to create
effective and sustainable systems. As a result, an exponentially growing
number of studies have lately been devoted to creating biocomposites by
combining starch nanoparticles with other biopolymeric matrices.
Additionally, the use of starch nanoparticles in the nanoencapsulation of
many nutraceuticals has improved their resistance to simulated diges­
tive conditions, cellular uptake, and the delivery of active ingredients in
cancer cell lines.
Owing to its ease of metabolism and degradation, it has a wide range
of applications in food modification. EGCG-loaded starch nanoparticles
were fabricated with a loading efficiency of 84.4% (Liu et al., 2020). The
synthesized nanoparticles demonstrated sustained release of nutra­
ceuticals in an activated GI environment. Other independent research
groups synthesized rutin-loaded starch nanoparticles following the
ultrasonication method to improve the bioavailability of rutin. The re­
searchers discovered increased bioavailability in stimulated gastric
conditions as well as significant antioxidant activity (Remanan & Zhu,
2021). Spherical starch nanoparticles were synthesized and loaded with
quercetin using nanoprecipitation with 0.1 M hydrochloric acid as a
non-solvent. They reported considerable effects on quercetin release
kinetics and antioxidant activity (Farrag et al., 2018).
4.1.5. Carrageenan
Carrageenan polysaccharide has been extensively utilized for com­
mercial applications in food, pharmaceutical, and cosmetic industries.
Carrageenan is sulfated higher molecular weight polysaccharide ob­
tained from different species of red sea weeds belonging to Rhodophycea
family. The key sources of carrageenan are Eucheuma, Chondrus, Hyp­
nea, Solieria, Cripus, Sarconema, and Iridaea. Galactose and anhy­
drogalactose units are joined together by glycosidic bonds to form
carrageenan (Aga et al., 2021). It creates a curled structure because it is
a flexible anionic polysaccharide. Sulfate and hydroxyl groups found in
carrageenans can be altered through oxidation, methylation, sulfation,
and acetylation to enhance their physicochemical characteristics. In the
food industry, carrageenan is used in food industry as a stabilizing,
gelling, thickening, and emulsifying agent. It enhances the texture of
cottage cheese, puddings, and dairy desserts. For making sausages,
patties, and low-fat hamburgers, it is used as binders and stabilizers.
Carrageenan also demonstrates exceptional benefits when compared to
other encapsulating materials. Carrageen, for instance, demonstrates
greater stability than pectin. Thus, carrageenan could be an excellent
nanocarrier for nutraceuticals because of its superior properties, such as
non-toxicity, biocompatibility, biodegradability, and low immunoge­
nicity (Aga et al., 2021).
The LYS-CRG nanocomplex was developed as a novel carrier to
improve the stability and in vitro sustain the release of curcumin
nutraceuticals (Sun et al., 2021). The particles demonstrated excellent
encapsulation and loading efficiency, measured at 96.2% and 2.31%,
respectively. The encapsulated curcumin showed increased retention
time, thermal stability, and photostability. The incorporation of
carrageenan-derived bionanocomposites for food packaging offered a
significant increase in food quality, stability at a wide range of tem­
peratures, and inhibition of contaminating microbial growth. Food
packaging with carrageenan-based nanocomposite reduces the risk of
environmental contamination due to the high biodegradable index of
carrageenan (Aga et al., 2021). As compared to distribution methods
without carrageenan, the delivery methods containing carrageenan
have a number of important advantages. For instance, carrageenan can
boost loading effectiveness, increase release amount, and improve
bioactive ingredient retention rate (Dong et al., 2021). Additionally,
carrageenan protects other components of the delivery system by
lowering their susceptibility to enzymes. This improves the stability of
the loaded compounds in GI conditions and allows for effective delivery
of particular bioactive components (Dong et al., 2021). Carrageenan can
help with the distribution of bioactive compounds in many ways, but
there are still a lot of issues that need to be resolved and new obstacles to
B.S. Paliya et al.
overcome before it can be effectively used in the food sector.
4.1.6. Fucoidan
Due to their advantageous effects on human health, the use of nu­
traceutical substances is currently expanding. They have limited sta­
bility throughout food production and human metabolism, low
bioavailability, and low cellular absorption due to their sensitivity,
which limits their bioactivity. In response, effective nanotechnology tool
for the delivery and protection of bioactive chemicals is nano­
encapsulation. In this respect, fucoidan, a natural anionic sulfated
polymer found in algae species and brown seaweed, has a promising use
in bioactive metabolite delivery due to its biodegradability and eco­
nomic cost. Fucoidan is chiefly composed of L-fucose and sulphate ester
groups. However, other sugar units such as D-galactose, D-mannose, D-
xylose, L-rhamnose, and uronic acid are also found in fucoidan isolated
from different algal sources (Dos Santos & Grenha, 2015). Fucoidan’s
therapeutic effects include anti-oxidant, anti-inflammatory, anti-tumor,
anti-viral, anti-thrombotic, and immunoregulatory activity.
Fucoidan and chitosan-based nanoparticles loaded with quercetin
were designed as nutraceutical functional foods. The NPs were synthe­
sized using the polyelectrolyte method with different fucoidan/chitosan
weight ratios (Barbosa et al., 2019). The team reported that when
keeping the weight ratio at 1:1, nanoparticles showed stability with
increasing pH or in a stimulated gastric environment. Also, quercetin
encapsulation with fucoidan or chitosan prevents its degradation during
oral applications and enhances its bioavailability. In another study,
EGCG was loaded with fucoidan and noncomplex hyaluronan with
polyethylene glycol-gelatine. The nanoassembly targeted macrophage
activation and pro-inflammatory signal production. Subsequently, one
independent research group improved resveratrol solubility, chemical
stability, and bioavailability by fabricating a zein-fucoidan nano­
assembly using a ratio of 2:1, loaded them with resveratrol. The syn­
thesized nanoassembly demonstrated sustained release under in vitro
digestion (Liu et al., 2022).
4.1.7. Mannan and glucomannan
Mannan is a mannose-based polysaccharide polymer linked by a
β-1,4-glycosidic bond. It is widely distributed in plants, fungi, bacteria,
and yeast. It is present in nature in association with different com­
pounds, such as glucomannan. Mannan is non-toxic and digestible,
making it a suitable polymer for nanocarrier development. Mannan
medicinal activity includes immune boosting and pathogen defence (Liu
et al., 2018). Glucomannan is a water-soluble polymer that is considered
a dietary fiber. It is a part of the hemicellulose found in some plant
species’ cell walls. Most of the time, glucomannan is a straight-chain
polymer with some little branching. The component sugars have a
1.6:1 ratio of β-(1 → 4)-linked D-mannose and D-glucose. Through β-(1
→ 6)-glucosyl linkages, there is around 8% branching. Food additive
glucomannan is used as a thickening and emulsifier.
A novel nanogel was developed, comprising konjac glucomannan-
chitosan and the crosslink agent 1-ethyl-3-(3-dimethylaminopropyl)/
N-hydroxysuccinimide. The fabricated nanogel was used to encapsulate
curcumin and investigated for stability and bioavailability (Wu et al.,
2021). The author has also reported that konjac glucomannan-chitosan
nanogel encapsulation imparts high encapsulation efficiency and sta­
bility to curcumin in the GI environment. Moreover, the encapsulated
curcumin showed sustained release in simulated GI conditions.
4.1.8. Arabic gum
Arabic gum is a branching molecule that is mostly made of a highly
branched polysaccharide with a little amount of a protein-
polysaccharide complex. Arabic gum, a polysaccharide derived from
exudates of Acacia senegal and Acacia seyal trees, is formed viscous
solutions, emulsions, and gels. It has been considered essential wall
materials for a variety of nanoencapsulated food ingredients due to their
excellent physicochemical properties, including flexibility,
10
Food Chemistry 428 (2023) 136709
biocompatibility, biodegradability, and reactive sites for molecular in­
teractions. It is made up of D-galactose, D-glucuronic acid, L-rhamnose,
and L-arabinose in a highly branched configuration. Arabic gum is
particularly unique compared to other gums due to its high protein
content and unique functional qualities. Arabic gum is stable in acidic
environments (pH 4.5–5.0) with Newtonian flow and soluble in cold
water (Tan et al., 2016). Gum solution has a lesser viscosity than other
gums, and only reaches a viscous state at 30–50% concentration. The
ability of arabic gum to act as an emulsifier for flavours and oils is a
crucial functional aspect. Arabic gum is considered a Grossly Recognized
As a Safe excipient. It is widely utilized in food industry as a thickening,
foam stabilizing, coating, and emulsifying agent, especially in the food
industry. Therapeutically, arabic gum has been reported for the treat­
ment of diabetes mellitus, oxidative stress, and hypertension (Ali et al.,
2009).
The storage and delivery of curcumin based on oil-in-water emul­
sions by combining chitosan and arabic gum has developed. The pre­
pared nanostructures displayed a retention level greater than 85% over
time, as well as an encapsulation efficiency and loading efficiency of
90% and 3.8%, respectively. Lastly, the created nanostructure was able
to slow down curcumin’s GI medium liberation process, increasing
bioavailability (Tan et al., 2016). Arabic gum nanoparticles are used to
load gallic acid to increase its bioavailability. The author reported that
arabic gum-encapsulated gallic acid nanoparticles are highly selective
for targeted cancer lines and show remarkably low toxicity toward
normal cell lines. These results suggested the potential use of Arabic gum
polymer for nutraceutical carrier applications (Hassani et al., 2020). A
research group synthesized quercetin-loaded mucoadhesive chitosan-
arabic gum nanoparticles via the ionic gelation method at a low pH of
3.5. They reported a promising encapsulation efficiency of 94.8–98%
and systematic time-lapse release of quercetin at 4.42–8.89% up to 8 h.
The adhesive property of nanoparticles provided an efficient delivery
system for targeted absorption of quercetin (Kim et al., 2019). Similarly,
EGCG was created by forming a nanocomplex using zein and arabic
gum. The results of the experiments indicated a great amount of EGCG
release on gastric digestion, indicating sustained release of EGCG from
the nanocomplex and increased the EGCG bioavailability (Jin et al.,
2022).
4.2. Proteins as nanocarriers for nutraceutical delivery
Different types of protein biopolymers have been studied to act as
biomaterials for nanocarrier fabrication. Protein polymers from natural
sources offer the ability to self-assemble into a different structure based
on their encoding sequence, and these sequences can further be modified
through recombinant engineering techniques. Also, protein nanocarriers
can be prepared by combining other proteins or polysaccharides for
different applications. Protein polymer nanocarriers impart stability to
nutraceuticals in both in vitro and ex vivo environments (Liu et al.,
2022), as most nutraceuticals are not stable in their free forms.
4.2.1. Silk fibroin
Silk fibroin is a natural protein-based biopolymer obtained from the
glands of arthropods of the animal phylum, which includes silkworms,
spiders, scorpions, bees, and mites. Its types differ by structure and
properties based on the source species. Natural silk is biodegradable and
biocompatible, and has been widely used in biomedical and food ap­
plications in the form of a variety of three-dimensional structures such as
three-dimensional scaffolds, biofilms, fibers, and hydrogels. Natural silk
is biodegradable and biocompatible. It showed the excellent quality of
cell adhesion and proliferation at the sites. It is liable to a vast range of
chemical modifications due to the presence of many active sites such as
arginine–glycine–aspartic acid sequence and tyrosine residue. These
active sites showed good cross-linking properties with many bio­
conjugates; therefore, silk protein polymer greatly attracts researchers
for the fabrication of nanocarriers for nutraceutical encapsulation
B.S. Paliya et al.
(Subia et al., 2014).
Silk fibroin nanoparticles and their capability of increasing quercetin
loading efficiency through adsorption and the releasing pattern of
quercetin were successfully synthesized (Lozano-Pérez et al., 2017). The
silk-quercetin nanoparticles showed high encapsulating efficiency and
sustain release of nutraceuticals throughout the experiment in both
simulated gastric environment and phosphate buffer. Another study
used adsorption to create resveratrol-loaded silk fibroin nanoparticles
(Lozano-Perez et al., 2014). The synthesized nanocomposite showed
immunomodulatory and anti-inflammatory activities. Similarly,
curcumin-silk fibroin nanoparticles were fabricated using a solution-
enhanced dispersion of supercritical CO2 technique to enhance the
antifungal activity of quercetin against Candida albicans (Xue et al.,
2019). They also reported a remarkable improvement in the solubility of
quercetin compared to native quercetin.
4.2.2. Collagen and gelatin
Collagen and gelatin are natural biopolymers produced abundantly
in the human and animal bodies. This protein-based fibrous polymer is
the main structural component of the extracellular matrix and is pri­
marily located in connective tissues, forming major parts of tendons,
ligaments, and skin. Collagen’s biological characteristics, such as being
biocompatible, biodegradable, and exerting low antigenicity, make it a
suitable material for nanoparticle delivery systems in pharmaceutical
and food applications. Like other proteins, collagens are flexible enough
to be configured for size and shape during nanoparticle synthesis.
Collagen nanoparticles demonstrated large surface area, loading ca­
pacity, absorption capacity, and tissue adhesion. Past research indicated
that collagen resembled the microenvironment of therapy. It was a few
tumors, which facilitates the effective infiltration of collagen nano­
particles into that area for cancer therapy studied that heat-treated
collagen fiber acts as a natural alternative to synthetic emulsifiers in
the preparation of functional foods, particularly acidic foods (Bealer
et al., 2020). Curcumin nanoparticles were combined with collagen and
chitosan in various ratios to create a nano-scaffold (Rezaii et al., 2019).
Collagen-based curcumin nanoparticles were evaluated for wound
healing properties on full-thickness punched wounds in male WinStar
rats. Interestingly, they found collagen-chitosan scaffolds efficiently
accelerated wound healing compared to blank collagen-chitosan scaf­
folds. The study suggested the implication of protein nanocarriers for
nutraceutical ingredients for therapeutic purposes. One research group
tried to enhance resveratrol’s bioavailability by encapsulating resvera­
trol in glutathione-coated collagen nanoparticles. A recent study showed
a therapeutic effect of resveratrol in epilepsy complications, but its
clinical applications are limited due to resveratrol’s low bioavailability.
The fabricated resveratrol nanoparticles showed activity in cognitive
impairment prevention in mice and demonstrated enhanced control of
epilepsy seizures compared to native resveratrol. Also, the nano-
resveratrol at the concentration of 0.4 mg/kg outperformed than that
of native resveratrol at 40 mg/kg (Siddiqui et al., 2022).
Gelatin is a thermally denatured and disintegrated derivative of type
I collagen. Gelatin, like collagen, demonstrated biocompatibility,
biodegradability, and abundance availability. Gelatin is also considered
an excellent nanomaterial due to its low carcinogenicity and antige­
nicity. Gelatin nanoparticles can efficiently cross semipermeable
blood–brain barriers and successfully deliver drugs and nutraceutical
molecules. Moreover, gelatin nanoparticles can easily be blended with
other polymers such as proteins and polysaccharides which further
improve their therapeutic properties.
The pomegranate ellagitannins nutraceuticals were created by
creating gelatin nanoparticles in three different mass ratios (1:5, 5:5,
and 7:5) (Li et al., 2011). Ellagitannins are composed of punicalagin A
(16.6% w/w), punicalagin B (32.5% w/w), ellagic acid (1% w/w), and a
small amount of ellagic acid. Only punicalagin anomers associated with
gelatin were reported during nanoparticle synthesis, and particles were
effective in inducing late-stage necrosis and apoptosis. Direct mixing of
11
Food Chemistry 428 (2023) 136709
protein polymer and nutraceutical solutions yielded a gelatin nano­
carriers for loading tea catechin (Chen et al., 2010). The developed
nanoparticles had a mean size of 200 nm and exhibited potential anti­
oxidant activity. An independent research group improved the water
dispersibility of buriti oil by incorporating it with gelatin nanoparticles.
This increased bioavailability of oil containing gelatin nanoparticles
showed enhanced antimicrobial properties as compared to native oil
(Castro et al., 2020).
4.2.3. Soy protein
Soy protein is one of the most abundant types of plant protein ob­
tained from soybeans. It is a globular protein with two main subunits,
glycinin and conglycinin. Soy protein is biocompatible, biodegradable,
and has high strength but low water stability. Subsequently, soy protein
is lowly immunogenic and available at a low cost. Since soy protein is a
polymer of amino acid monomers, its nanoparticles also facilitate the
loading of highly hydrophobic drugs. Due to its aqueous solubility, it can
be used for oral drug delivery of the drugs and food bioactives.
An attempt has been made to synthesize curcumin-loaded soy pro­
tein nanoparticles (Bealer et al., 2020). The systematic steps of prepa­
ration included dispersion, desolation of soy protein, curcumin
incorporation, cross-linking and evaporation of the liquid portion. The
investigation of each step revealed their role in particle size and shape.
The mean size of successfully synthesized curcumin-loaded nano­
particles was ranged from 220 to 286 nm. Later, soy protein with cel­
lulose nanocarrier (SPI-CNC) was developed (Wang et al., 2020). They
reported that the SPI-CNC complex could be a promising nanocarrier
agent for the delivery of curcumin and other hydrophobic nutraceutical
compounds. In another study, a ternary nanoassembly comprising soy
protein, pectin, and tea polyphenol was fabricated (Jin et al., 2018). The
cross-linking between all ingredients was achieved through photo­
catalysis, which also modulated the binding affinities among all in­
gredients. The study suggested that such nanoassemblies could be used
as an alternative agent for enhancing the physiochemical stability of
nutraceuticals and food bioactive compounds. Nanogels through self-
assembly among soy protein, soy polysaccharide, and folic acid were
prepared (Ding & Yao, 2013). They used high-pressure homogenization
to disintegrate soy protein bonds, which facilitates binding between soy
protein and all components at pH 4.0. After that, heat treatment was
used to process protein denaturation, gelation, and the fabrication of
nanogels complexes with folic acid-loaded soy protein/polysaccharide.
The resulting nanogels contained highly stable folic acid molecules after
exposure to oxygen, heat, and light.
4.2.4. Corn zein
Corn protein is made up of zein protein (20 kDa), which is found in
the cytoplasm of corn endosperm. Zein protein is amphiphilic and has
three forms (α, β and γ) depending on solubility and molecular weight.
Zein protein has gained some recent advances for serving as a promising
natural polymer for the development of nanocarrier applications in the
pharmaceutical, medical, and food industries due to its unique proper­
ties such as biocompatibility, biodegradability, physical strength, good
fiber content, and the capability of forming films (Bealer et al., 2020).
The presence of protein hydrophobicity in zein nanoparticles protects
the encapsulated drug from an acidic environment of the gut.
Zein nanoparticles have successfully been utilized for carrying
nutraceuticals such as vitamins and proteins (Hurtado-López & Murdan,
2006). Curcumin-loaded zein nanoparticles were designed to ameliorate
the bioavailability of curcumin in Winstar rats, according to blood
plasma samples tested after oral administration (Brotons-Canto et al.,
2021). Subsequently, a novel zein nanocarrier loaded with resveratrol
was evaluated for improving its bioaccessibility and anti-inflammatory
activity (Liu et al., 2021). To stabilize zein nanoparticles, they used
microbial transglutaminase as a cross-linking agent in conjunction with
sodium caseinate. The results of the synthesized nanoparticle treatment
indicated anti-inflammatory activity with lower toxicity. EGCG, was
B.S. Paliya et al.
successfully loaded onto zein protein nanoparticles (Chen et al., 2022;
Liu et al., 2021; Xie et al., 2021). In another study, the zein-lecithin-
EGCG complex was fabricated using the anti-solvent coprecipitation
method. The fabricated nanocomplex demonstrated excellent stability
with sustained release of EGCG, suggesting alternate uses of zein
nanocarriers in food industries.
4.2.5. Wheat gluten
Gluten protein is a natural biopolymer and a by-product of wheat
starch preparation. Gluten protein can be considered an alternative
natural polymer source due to its biodegradability and abundant sour­
ces. Gluten protein is a combination of monomeric gliadin (single chain
polypeptides of 28 to 55 kDa) and glutenin (polymers of 100 kDa to 10
MDa) proteins. Former proteins impact viscosity and later give elasticity
a physical character. Gluten protein showed some important charac­
teristics such as heat and water stability, fiber formation, and biocom­
patibility. A novel nutraceutical was developed by incorporating
ascorbic acid into gliadin nanoparticles (Voci et al., 2022). This
formulation imparts chemical stability to ascorbic acid and shows
improved antioxidant properties as compared to the free native form.
The study results proved that gliadin protein can be utilized for nutra­
ceutical ingredient delivery and stability in fortified foods. Another
study showed that gliadin and soluble soy protein nanoparticles
encapsulated in curcumin demonstrated excellent stability, antioxidant
activity, and bioaccessibility (Guo et al., 2022).
As a delivery system for food-grade -carotene, gliadin protein
nanoparticles were incorporated into pickering emulgels. This emulgel
formulation was found to be -carotene stable in a stimulated gastrin
environment, with good fluidity and thermal stability (Cheng et al.,
2020). It was investigated how to overcome the degradation problem of
α-tocopherol or vitamin E by encapsulating it in gliadin nanoparticles.
Experimental results indicated significant nutraceutical release behav­
iour for nanoparticles with improved stability (Duclairoir et al., 2002).
The stability of gliadin and glutenin nanoparticles is typically lower,
although these nanoparticles can be stabilized using a variety of anionic
and cationic polysaccharides. A group of researchers have successfully
used arabic gum to enhance the stability of the gliadin nanoparticles
(Wu et al., 2018).
5. Fabrication of polysaccharide-protein nanoconjugates
Protein-polysaccharide nanoassemblies are efficient tools for con­
structing highly potential and economically viable carrier systems with
highly acceptable biosafety and biocompatibility. Biological systems
contain a diverse structural array of protein and polysaccharide poly­
mers allows for different combinations to develop tailored protein-
polysaccharide nanocomposites incorporating bioactive nutraceuticals.
Thus protein-polysaccharide nanoconjugates can be used to build ver­
satile delivery systems (Wei & Huang, 2019). The protein-
polysaccharide nanocarrier system has some advantages over other
delivery systems. For example, proteins and polysaccharides reduce the
astringency of several bioactive polyphenols and nutraceutical com­
pounds through the masking effect. So, a nutraceutical carrier system
can be constructed using protein-polysaccharide polymers containing
nutraceutical molecule without compromising food taste, which is the
major factor for any food product. Protein allows multiple sites for the
attachment of a variety of molecules, and its response to numerous
stimuli can be altered in a controlled setting. Subsequently, poly­
saccharides’ strength, stability, and solubility offer the fabrication of
more sophisticated nano-delivery systems having the collective advan­
tages of both polymers.
The complexes of protein and polysaccharide are formed due to
strong hydrophobic interactions and electrostatic bonding between
oppositely charged molecules. The initially formed complexes further
aggregate to reduce free energy until they attain a certain size and
shape. Then, in solution, a discrete liquid-coacervate droplet formed due
12
Food Chemistry 428 (2023) 136709
to two macroscopic liquid phase separations that coexisted with a dilute
liquid phase. The phase coexists with a solvent sub-phase at equilibrium
(Cooper et al., 2005). Protein-polysaccharide nanoassemblies are far
more stable than a protein-based nanocarrier because proteins precipi­
tate at an isoelectric pH point and polysaccharides prevent this protein
aggregation through steric hindrance. Moreover, the incorporation of
polyphenols with protein nanocarriers causes acceleration of protein
aggregation and precipitation, while the protein-polysaccharide nano­
complex potentially reduces the coprecipitation of both protein and
polyphenols. Overall, the complex structure increases the strength of the
protein material while blending with the polysaccharide. The protein-
polysaccharide complex exhibited important physiochemical proper­
ties such as emulsifiers and stabilizers for processed food items. The
polysaccharide and protein complex allows the construction of an
amphiphilic nanoconjugate that can be strongly anchored at the
water–oil interface due to hydrophobic regions of proteins that allow
viscoelastic layer formation along with polysaccharide regions (non-
adsorbing), leading to enhanced steric stabilization and imparting gel­
ling behaviour to the mixture. Furthermore, amphiphilic properties
allow for the encapsulation and stability of several non-polar phytonu­
traceuticals delivering them to the target cell and improving their
bioavailability.
The protein-polysaccharide nanocomposites have been reported for
a vast range of applications in the fields of food industries, biomedicine,
and green chemicals. The complex allows the formation of a variety of
gels, fibers, and films according to required applications. Protein-
polysaccharide nanocomposites offer numerous opportunities to
improve the biological properties of nutraceutical ingredients, phar­
maceuticals, and bioactive molecules. Emulsions, hydrogels, microcap­
sules, and nanomaterials are examples of protein-polysaccharide
biomaterials systems that are widely used for bioactive molecule de­
livery. So, current studies are trying to explore the applications of
protein-polysaccharide nanoconjugates to transport common plant-
derived nutraceuticals such as food polyphenols, vitamins, caroten­
oids, and flavonoids (Sun et al., 2021; Sun et al., 2022).
5.1. Types of protein-polysaccharide nanoconjugates
In biological systems, many combinations of proteins and poly­
saccharides occur naturally through electrostatic and coacervation
phenomena. Such complexes activate different biological functions in
associated biopolymeric systems (Cooper et al., 2005). Among the
various mechanisms studied, protein-polysaccharide interactions
exhibited primarily two types of interactions: non-covalent and covalent
bonding. A covalent bond-based interaction was achieved through a
Maillard-type reaction that occurred under controlled physiochemical
parameters like pH reaction time, moisture, and temperature. The con­
jugates obtained through covalent bonding show good heat stability.
Non-covalent interactions are characterized by the hydrogen bonding,
hydrophobic, Van-der Waals, and electrostatic interactions (Gentile,
2020). However, specific numbers of polysaccharides show proper
attachment to protein polymers because of the steric hindrance offered
by macromolecules. In this regard, unfolded casein allows more poly­
saccharide interactions than folded proteins (ovalbumin and lysozyme).
5.2. Maillard-type protein-polysaccharide nanoconjugates
French chemist Louis Maillard published the first description of the
Maillard reaction in 1912. A sequence of non-enzymatic browning re­
actions called the Maillard reaction take place spontaneously under
controlled pH, reaction duration, temperature, and moisture conditions.
Covalent bonds between the reducing ends of carbohydrates and amino
groups of proteins, particularly between the lysine residue’s -amino
group and the carbonyl group of reducing carbohydrates, are where it all
begins. There are typically three stages to the Maillard reaction: the
initial, intermediate, and final. The three stages are correlative and can
B.S. Paliya et al.
all take place at the same time.
The most widely implicated method for the preparation of Maillard-
type protein-polysaccharide nanoconjugates is either dry heating or wet
heating in an aqueous solution. In this method, the protein-
polysaccharide freeze-dried mixture is heated in a prescribed tempera­
ture range (40–80 ◦ C) and relative humidity (65% or 79%). The mixture
allows for a reaction lasting from hours to weeks. It is a time-consuming
procedure with limited scaling benefits. On the other hand, wet-heating
method is a relatively quick method of heating a protein-polysaccharide
mixture in an aqueous solution to a specific temperature. A limitation of
this method includes a lower degree of grafting, denaturation, and
polymerization of proteins with temperature increases. The stronger
hydrophobic curcumin’s capacity for encapsulation is often positively
connected with a larger degree of grafting with polysaccharides. To
improve the degree of grafting of the resulting soy protein isolate (SPI)-
dextran conjugates, a novel technique known as continuous cyclic re­
action (7 cycles of low-speed agitation at 60 ◦ C for 20 min and water-
bath heating at 83 ◦ C for 10 min) was employed (He et al., 2021).
Plant-derived nutraceuticals appear to have promise health advan­
tages, but their low stability still restricts their use in the food sector. To
improve their ability to be absorbed and metabolized and to provide
greater health advantages, nutraceuticals must become more stable. In
order to preserve the bioactivities of nutraceuticals and improve their
inadequate stability, numerous encapsulating techniques have been
devised. Choosing appropriate edible encapsulating materials based on
biopolymers is crucial in general for getting the desired results. To in­
crease the stability of nutraceuticals for effective application in the food
business, food-grade edible encapsulation materials, particularly for
proteins and polysaccharides, could be used. Food-grade proteins are
appealing for encapsulation of food grade nutraceuticals because of their
improved stability and health advantages. Because of their satisfactory
biocompatibility and biodegradability, polysaccharides and their de­
rivatives are employed as encapsulating materials (Song et al., 2022).
The sodium caseinate-lactose complex, conjugated via the Maillard re­
action, as encapsulating materials used to develop protein-
polysaccharide oil-in-water emulsions and also examined the dura­
bility of fabricated materials (Li et al., 2017). Then, the encapsulation
effectiveness, insolubility, wettability, and microstructure of spray-dried
oil microcapsules were examined. At pH 11, the interactions between
sodium caseinate and lactose were more favourable, with improved
conjugate adsorption at the oil droplet particles and consequently
greater emulsifying characteristics and stability. Controlling the re­
action’s time, pH, water activity, and temperature may stop the pro­
duction of hazardous and antinutritional chemicals. For instance,
biocompatible egg white-galactomannan conjugates were made over the
course of two weeks at 60 ◦ C and 79% relative humidity. In general,
glycated proteins created by the Maillard reaction are less dangerous
than proteins that have undergone chemical modification. Thus,
Maillard-type protein-polysaccharide conjugates may be used in food
items without risk (Sun et al., 2022).
In comparison to crude proteins, Maillard-type protein-poly­
saccharide conjugates often exhibit dramatically increased water solu­
bility. When compared to native SPI, the solubility of SPI-pectin
conjugates was substantially higher (Sun et al., 2022). At both low and
neutral pHs, protein-polysaccharide conjugates have stronger emulsi­
fying abilities than raw proteins, especially at pH levels near the iso­
electric point. Additionally, protein-polysaccharide conjugates’
molecular weight and structure of the polysaccharide, reaction time, and
ratio of protein and polysaccharide are essential parameters for
increasing their emulsifying properties. In comparison to the original
SPI, the SPI-pectin conjugates demonstrated a 3-fold increase in emul­
sifying activity index (Sun et al., 2022).
5.3. Electrostatic type protein-polysaccharide nanoconjugates
Due to the amphiphilic nature of protein molecules, their surface
13
Food Chemistry 428 (2023) 136709
charge depends on the pH of the surrounding solution. Similarly, poly­
saccharides with carboxylate groups show a negative charge at pH levels
above their pKa value (Sun et al., 2022). These electrochemical charges
on the polymer backbone of long chains facilitate the formation of non-
covalent interactions such as hydrogen bonds and hydrophobic in­
teractions. Together, both types of interactions impart stability to protein-
polysaccharide aggregates as well as nanoconjugates. When non-covalent
bonding stabilizes protein-polysaccharide complex in a single-phase sys­
tem, the interacting biopolymer shows associative phase separation and
the formation of the soluble complex “coacervate.” In the case of a non-
interacting biopolymer, the mixture shows segregative phase separa­
tion, and the mixture component distributes uniformly as shown in Fig. 2.
The major factors that play an important role in the stability of electro­
static protein-polysaccharide complexes are pH, ionic strength of the so­
lution, charge distribution and density, biopolymer concentration present
in different mixing ratios, temperature, and shear rate.
Complex coacervatives are a liquid–liquid phase separation of the
biopolymer-rich phase and the solvent-rich phase. Coacervate formation
occurs when two or more oppositely charged biopolymers interact with
each other at a minor scale, leading to the formation of associative
complexes. Subsequently, in association with more interactive molecules
at isoelectric pH, the associative complexes form soluble complexes that
further interact and are held together with each other to form a localized
charged segment known as coacervates. The coacervation is favoured by
the isoelectric pH condition of the solution and can be reversed by dis­
turbing the ionic strength of the solution. Proteins and polysaccharides
are co-soluble substances in solution when pH is higher than the critical
pH. The complexes become less soluble as the pH drops below the critical
pH, and as they begin to assemble into insoluble forms due to charge
neutralisation at the second critical pH, turbidity increases dramatically.
As a result, the stability of the formed coacervation necessitates the
addition of some additional crosslinking agents, such as glutaraldehyde,
which links and binds complex components while also enhancing their
agglomeration and isoelectric bonds, as illustrated in Fig. 3.
Protein-polysaccharide conjugates’ rheological characteristics are
receiving more attention as they are crucial in deciding how protein-
polysaccharide complexes are used in food items. It is necessary to
establish the storage modulus, loss modulus, loss tangent, critical value
of stress at the linear viscoelastic region, fracture stress and strain, and
crossover point in order to fully comprehend the rheological
Fig. 2. Formation of different phases system types between protein and poly­
saccharide polymers.
B.S. Paliya et al.
characteristics of protein-polysaccharide conjugates. Protein/poly­
saccharide ratio, pH, and ionic strength all have a substantial impact on
the rheological properties of protein-polysaccharide complex co­
acervates. The effects of various pH levels and protein-polysaccharide
ratios on the rheological characteristics of conjugated rice bran pro­
tein and flaxseed gum. The coacervates had significantly greater struc­
tural strength and loss modulus as measured by the sweep test at 25 ◦ C at
pH 4.0 and a biopolymer ratio of 9:1.
In addition, the architectures of protein-polysaccharide conjugates
are frequently indicated by rheology data. In particular, coacervates
such β-lactoglobulin-pectin coacervates and rice bran protein-flaxseed
gum coacervates with greater storage modulus values than loss
modulus show the development of highly linked gel-like structures.
Additionally, several analytical methods are frequently applied to
examine the microstructure of complicated coacervates. A study sug­
gested that Tremella fuciformis polysaccharide-whey protein isolate
complexes had more organised structures than any of the other two
biopolymers (Hu et al., 2019). When compared to free whey protein
isolate, the complexes of whey protein isolate and T. fuciformis poly­
saccharides include more α-helices.
5.4. Chemical cross-linking and enzymatic cross-linking type
nanoconjugates
Protein-polysaccharide complexes are fabricated through the
chemical cross-linking method using low-toxic and biocompatible cross-
linking agents. This approach can be characterized as short-range and
long-range cross-linkers. Short-range cross-linkers are glutaraldehyde,
genipin, and EDC (1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide)
and considered to be less toxic and more biocompatible than long-range
cross-linkers such as PEG-DBA (poly(ethylene glycol) dibutyraldehyde)
and various other crosslinking agents. To increase the number of con­
jugate yields and improve the biological and physiochemical properties
of biopolymers, the chemical cross-linking agent efficiently forms
numerous cross-links to a variety of proteins and polysaccharides.
Chemical cross-linkers produced comparable stable conjugates, but they
have drawbacks for food applications, for example, glutaraldehyde
consumption causes adverse health effects such as irritation at a certain
level.
In enzymatic cross-linking method, protein-polysaccharide com­
plexes can be achieved by catalyzing the reaction via several oxidase
enzymes working through different mechanisms. Tyrosinase and lac­
case, act on the tyrosine amino acid group presented on polypeptides
and oxidize it into quinone residues. Quinone is a highly reactive species
Fig. 3. Represented complex coacervationtechniques process showing protein-polysacharidenanocomplexformation.
14
Food Chemistry 428 (2023) 136709
that gets stabilized by reacting with nucleophilic substituents present on
the polysaccharide chain, forming a cross-link between protein and
polysaccharide polymers. Enzymatic-based cross-linking for biopolymer
complex synthesis is a reaction specific to initial polymers and requires a
mild reaction to happen. However, the prepared product is less stable
regarding pH sensitivity compared to chemically cross-linked conjugates
(Fig. 4). But the method is highly biocompatible and less toxic (Bealer
et al., 2020).
6. Protein-polysaccharide nanoconjugates for the delivery of
plant-derived nutraceuticals
In the past decade, a vast range of protein-polysaccharide polymer-
based nanocomplexes has been constructed to synthesize core–shell
nanocarriers for the efficient delivery of nutraceuticals. Through a sys­
tematic treatment of alkaline dissolution, acidification, and high-
pressure homogenization, a ternary nanocomplex with the shell of
casein and soy soluble polysaccharide and curcumin in the core in the
water phase was developed. Interestingly, they reported that nano­
particles showed significant curcumin solubilizing power and 97%
loading efficiency. The polysaccharide surface renders the size of
nanoparticles easily dispersible in neutral and acidic solutions and re­
stores their dispensability in water after lyophilization. This ternary
complex carrier nanosystem was proven to be effective for curcumin
loading and its oral delivery and stability (Fan et al., 2021). Carbox­
ymethylated corn fibre gum and nutritious pea protein as encapsulating
polymer shells have been used to create a core–shell ternary nano­
complex for curcumin delivery (Wei et al., 2020). The nanocomplex was
synthesized through hydrophobic and electrostatic interactions. The
resulting nanosystem possesses efficient water dispensability and
loading efficacy. Besides, these particles showed improved thermal and
chemical stability. Researchers have developed a simple method for a
lipid-soluble nutraceutical ingredient curcumin delivery system by
fabricating a protein-polysaccharide nanocomplex (Fan et al., 2021).
They used polysaccharide fucoidan and soybean protein isolate for
core–shell nanoparticle synthesis for curcumin encapsulation through
the electrostatic interaction method in neutral and acidic mediums. The
coacervation method was also used to develop gum arabic (a poly­
saccharide) and whey protein nanofibrils to encapsulate curcumin
through the electrostatic interaction method in neutral and acidic
mediums.
The coacervation method was used to develop gum arabic (a poly­
saccharide) and whey protein nanofibrils to encapsulate curcumin. They
reported that the complex exhibited shear thinning behavior and
B.S. Paliya et al.
Fig. 4. Cross-linking method.
superior surface property, which makes the curcumin-loaded complex
an excellent agent to process functional food preparations such as
emulsions and beverages (Mohammadian et al., 2019). The protein-
polysaccharide-loaded nanocurcumin after UV radiation exposure for
90 min showed 4% and 34% residue remained, indicating improved
photostability as compared to free curcumin. Moreover, nanoassemblies
(insect protein and chitosan) loaded with curcumin exhibited a great
intestinal release profile in a stimulated gastric environment during oral
bioavailability. A hydrogel based on alginate and β-lactoglobulin-pro­
pylene glycol was recently developed for the delivery of curcumin and
probiotics. The hydrogel not only provided encapsulation-based stabil­
ity but also exerted sustained release of nutraceuticals in a stimulated
gastric condition. Further hydrogel-based delivery systems showed
excellent long-term stability during light exposure and storage (Sun
et al., 2022). Chitosan and gum arabic-based nanoassemblies were
prepared as a novel delivery system for curcumin delivery using the
polyelectrolyte complexation method. The fabricated nanoassemblies
showed increased encapsulation efficiency of 90%, curcumin loading of
38%, and a retention rate of 85% higher during storage. The nano­
assemblies improved curcumin stability, delayed its release in activated
GI track, and significantly increased antioxidant activity (Hu et al.,
2017).
Similarly, the protein-polysaccharide nanocomplex is implicated in
resveratrol loading and systematic delivery to improve its bioavail­
ability. Zein-chitosan complex was developed via a coacervation tech­
nique for the encapsulation of resveratrol. The optimized ratio for the
highest encapsulation was identified as 1.2: 2 (zein/chitosan). They
reported that the 28/40 kHz dual-frequency ultrasound model exerted
the highest encapsulation efficacy. Furthermore, the size of fabricated
zein-chitosan coacervation complexes was significantly reduced without
affecting their zeta potential (Ren et al., 2019). Electrostatic interaction
between oppositely charged molecules was used to create stabilized and
encapsulated resveratrol with alginate/chitosan-zein nanoparticles and
chitosan-α-lactalbumin nanoparticles. It has observed that hydrogen and
hydrophobic bonding play a major role in particle formation. Protein-
polysaccharide matrix core–shell efficiently enhanced heat, light, and
storage stabilities. UV exposure to the nanocomplex for 200 min showed
retention rates of 44% for free resveratrol and 47% for chitosan-α-lact­
albumin coated resveratrol. While, heat treatment for 300 min at 85 ◦ C
revealed retention rates of 85% for free resveratrol and 86% for chito­
san-α-lactalbumin coated resveratrol (Liu et al., 2020). Further, the
protein-polysaccharide matrix facilitates the sustained release of
resveratrol in gastric digestion. In another study, researchers observed
15
Food Chemistry 428 (2023) 136709
that loading less resveratrol (0.02–100 g) into an oil-in-water emulsion
stabilized by sodium caseinate-corn starch hydrolysate conjugated via
the Maillard reaction and exhibited potential antioxidant activity
(Consoli et al., 2018).
Several studies have reported significant improvements in the bio­
stability and bioavailability of biopolymer-coated bioactive protein
nutraceutical ingredients. For example, entrapped lactoferrin within
whey protein-pectin nanoparticles at a ratio of 2:1 (Raei et al., 2017).
Casein hydrolysate with soybean protein isolate or pectin nanocomplex
was developed to mask astringency and hygroscopicity of soybean
protein. Furthermore, nanoencapsulation also reduced hygroscopicity
values when compared to free hydrolysate (Mendanha et al., 2009). A
food-grade double emulsion was developed by incorporating a bioactive
peptide and polysaccharide complex via the electrostatic interaction
method. The prepared double emulsion showed much improved thermal
stability and more than 90% encapsulation efficiency (Jo & van der
Schaaf, 2021).
Lutein was investigated for encapsulation with a core–shell-based
nanocarrier for its oral delivery. A lutein-loaded nanodelivery system
was developed using rice protease hydrolysate and carboxymethyl cel­
lulose (Li et al., 2020). The heating method was used for nanoparticle
preparation. The prepared nanocomplex efficiently controlled GI
digestion and release of lutein and enhanced lutein uptake by breast
cancer cells, causing cell proliferation inhibition. Also, the bioavail­
ability of biopolymer-coated lutein was reported two times higher than
that of native lutein. A casein-dextrin conjugate-based stabilized lutein-
loaded emulsion was developed and showed a vast range of pH from 3 to
7 (Gumus et al., 2016). This protein-polysaccharide complex prevents
aggregation in a stimulated gastric environment and remains stable due
to the steric repulsion of dextran.
Polysaccharide conjugation with proteins prevents the protein from
precipitating on high-concentration loading of polyphenols, especially
those conjugates prepared via the Maillard reaction. The polyphenol
stability of tea was increased by loading it onto a protein core fabricated
with a gelatin-dextran nanocomplex, prepared through a Maillard re­
action. Epigallocatechin-loaded nanocomplex showed high loading ef­
ficacy of 360% (w/w) of protein, while encapsulation efficiency was
independent of pH change. The nanocomplex demonstrated a compa­
rable cytotoxic effect concerning native free epigallocatechin. Another
study found that encapsulating epigallocatechin with dextran glycosy­
lated casein nanoconjugates improved encapsulation and loading ca­
pacity. The protein-polysaccharide polymer protects epigallocatechin
from degradation in alkaline pH conditions and allows for sustained
B.S. Paliya et al.
release in the intestine (Xue et al., 2014). Naringin, a potential func­
tional and therapeutic flavanone found naturally in citrus fruits, has
narrow bioavailability due to its weak aqueous solubility. An
ovalbumin-carboxymethyl konjac glucomannan nano-delivery system
with a size of about 464 nm was recently fabricated to increase nar­
ingin’s bioavailability (Tang et al., 2022). The authors recorded a 2-fold
higher bioavailability of naringin than that of bulk naringin.
The electrospraying method was used to create protein-cellulose
nanoparticles, and the encapsulation of EGCG in these nanoparticles
was investigated. The fabricated nanoparticles were polydisperse with a
PDI of 1 to 1.5 and varied in size from 180 to 240 nm. Using the HPLC
approach, the encapsulation efficiency of EGCG produced relatively
high values up to 98%, and UV–Vis was employed as a complementary
and practical method. The deterioration of the particles kept with EGCG
under different circumstances was also investigated. When the nano­
particles were held at low relative humidity, the EGCG in them was more
stable. EGCG was shielded from heating, moisture, and dissolution when
it was incorporated into protein-cellulose nanoparticles. The findings
indicate that employing protein-cellulose nanoparticles as EGCG car­
riers has a lot of promise and offer a fresh understanding of the possible
uses of electrospraying as a viable technique for the creation of edible
particles, a result that may be advantageous for encapsulating dietary
nutraceuticals (Paximada et al., 2021). The β-lactoglobulin/gum arabic
(β-Lg-GA)-EGCG nanoconjugates having an average particle size of 133
nm were developed which exhibited an excellent 84.5% encapsulation
efficiency and enhanced antioxidant activity. Following 3 h of digestion,
almost 86% of the EGCG from the β-Lg-GA-EGCG complex was released
in simulated intestinal fluid considerably more quickly than it was in
simulated gastric fluid, demonstrating the effectiveness of the β-Lg-GA-
EGCG nanoconjugates in improving EGCG stability (Gao et al., 2021).
Song and colleagues summarized the previous studies on the stabi­
lization of anthocyanin using protein-polysaccharide polymers (Song
et al., 2022). The high encapsulation effectiveness (over 96%) attained
by the combination of xanthan gum and carboxymethyl starch improved
the stability of blueberry anthocyanins. Blackberry juice and Lactoba­
cillus acidophilus may be co-encapsulated with gum arabic and malto­
dextrin to protect anthocyanins and L. acidophilus. It has also been
demonstrated that alginate-pectin hydrogel particles may successfully
encapsulate 116% of the anthocyanins in blueberries (Song et al., 2022).
In addition, vitamins are crucial micronutrients and nutraceutical
supplements that are required for the regulation of a number of bio­
logical processes. Recent studies established that the nanoencapsulation
of vitamins minimized their loss during processed food preparation and
food storage. Such functional foods showed better target delivery of
vitamins and sustained release in the biological environment. The soy-
protein and polysaccharide conjugate nanogels were prepared to
enhance the stability and systematic delivery of nutraceutical folic acid.
Polysaccharide-encapsulating biomaterials provide superior water dis­
persibility, even in acidic conditions. Folic acid in nanogels indicates fast
release at pH 7.0. More importantly, folic acid-loaded nanogels
demonstrated excellent physical stability to heat, light, and oxygen in
acidic environments (Ding & Yao, 2013). In a gastric acidic environ­
ment, lipid-soluble vitamin D3 is highly susceptible to degradation. To
overcome this problem, ovalbumin and a high-methoxyl pectin nano­
complex were prepared as delivery carriers for vitamin D3. The fabri­
cation of the nanocomplex was facilitated by hydrogen bonding and
electrostatic and hydrophobic interactions. In vitro sustained release
testing of complex loaded vitamins in gastric acidic conditions revealed
11% release in 60 min. Whereas vitamin release from the nanocomplex
was significantly enhanced 98% within 120 min in the intestinal envi­
ronment. Additionally, it demonstrated pH responsiveness to the sus­
tained release of loaded vitamins in a different section of the digestive
tract, which unquestionably increases the bioavailability of sensitive
vitamins (Xiang et al., 2020).
A subsequent study revealed that the addition of sodium alginate
further enhanced the vitamin D3 stability in ovalbumin–pectin
16
Food Chemistry 428 (2023) 136709
nanocomplex system (Gao et al., 2021). Similarly, a β-carotene precur­
sor of vitamin A and a powerful antioxidant are prone to early degra­
dation and poor dispensability. Several methods have been employed to
develop sophisticated delivery systems for β-carotene. Among all
methods, protein-polysaccharide complex-stabilized oil–water nano­
emulsion proved to be the best of all. For example, the ovalbumin-
dextran complex demonstrated excellent emulsifying stability over a
wide pH levels ranged from 3 to 10, a high salt concentration (150 mM
NaCl), and temperatures up to 90 ◦ C (Sun et al., 2021). Recent studies on
protein-polysaccharide nanocomposite-based emulsions for β-carotene
stabilization which include whey protein hydrolysate-pectin and pea
protein-high methoxyl pectin colloidal delivery systems, demonstrated
better β-carotene stability over a wide pH range.
Minerals are essential micronutrients that perform specific biological
functions required to maintain human health. But the prevention tech­
nique used for the fortification of such minerals often masks their effect
and alters the physical properties of food. To improve mineral iron
stability in food, cold-set hydrogel incorporated with an iron-loaded
nanocomposite of whey protein isolate and gellan gum was formu­
lated. The formulation showed a high encapsulation efficiency of 94% of
the mineral iron. However, when the formulation was evaluated for
preparation at different concentrations of biopolymer ratio and iron, the
encapsulation efficiency drastically varied. But in all cases, encapsulated
iron showed up to 89% release in gastric conditions. The acidic release
of iron was due to electrostatic repulsion between iron and complex
bonds, facilitated by the net cationic charge on proteins in an acidic pH
(Kazemi-Taskooh & Varidi, 2021).
7. Challenges and future perspectives
Research has shifted to a protein-polysaccharide nanoconjugate-
based nutraceutical delivery system because of its superior properties,
which have shown to be successful in delivering bioactive components.
However, a number of problems with the development and use of de­
livery systems remain and will need to be resolved in the future. First,
nutraceutical compounds may become less bioavailable (Dong et al.,
2021). Effectiveness of nutraceuticals is based on their real bioavail­
ability. However, throughout the production and distribution of the
complete system, there may be loss or alteration that reduces the
bioavailability of the nutraceutical compounds. Because of this, it is
essential to increase the bioavailability of nutraceutical compounds
using workable methodologies based on their unique features. The
protection of encapsulated nutraceuticals can be improved by, for
example, choosing the right packing materials, using crosslinking agents
to create a more compact particle structure, or including antioxidants
(Bealer et al., 2020). Furthermore, the efficiency of protein-
polysaccharide conjugate-based delivery systems, particularly those
based on carrageenan, might be impacted by multi-component in­
teractions. For instance, it is necessary to examine the performance of
delivery systems in actual food systems and confirm their suitability as
delivery carriers in order to determine whether the food added to the
carrageenan-based nutraceutical delivery system has an unpleasant
taste or whether multi-component interactions can reduce the nutri­
tional value of the bioactive ingredients (Aga et al., 2021).
Nanoparticle sizes can be rather enormous. The size of several
developed protein-polysaccharide delivery systems can limit their use in
many commercial products in terms of look, taste, and nutrition, which
can impair consumers’ acceptance. Choosing the right combination of
proteins and polysaccharides as well as crosslinking agents to create a
more compact particle structure, which is beneficial for producing
smaller sizes, is a possible technique (Bealer et al., 2020). Research into
protein-polysaccharide’s metabolic process and modified protein-
polysaccharide nutraceutical delivery systems is also lacking. Studies
have revealed that low gastric pH or the microbial community in the GI
tract do not effectively breakdown polysaccharides, viz., carrageenan,
but other reports suggest that some colonic bacteria can desulfate
B.S. Paliya et al.
carrageenan and only leave a degradable galactan polymer backbone
(Dong et al., 2021). On the exact digestive destiny and metabolic
mechanism of protein-polysaccharide conjugates, however, nothing is
known. To fully understand the metabolic fate of protein-polysaccharide
conjugates in the human body and to build a metabolic process, in-depth
research is required. Additionally, considering the superior properties of
the modified protein-polysaccharide conjugates mentioned above,
whether the modified protein-polysaccharide delivery systems have
more application value than the traditional delivery systems remain to
be further discussed. Furthermore, proteins and polysaccharides have a
rather high viscosity in practical applications; it is still unknown if
lowering their viscosity will make protein-polysaccharide better build­
ing blocks for delivery systems (Dong et al., 2021). In order to extract
proteins and polysaccharides of higher quality for use as encapsulating
materials, the extraction process that is now more effective needs to be
further enhanced.
New information about the creation of biopolymer interfaces that
can take the place of emulsifier layers is provided by protein-
polysaccharide nanoconjugates. For the study of food colloidal struc­
ture, the promise of stabilized interfaces for particles has garnered
considerable interest. Growing interest in complex coacervation is due
to its superior loading capacity, comfortable operating conditions, and
controlled release (Song et al., 2022). The good encapsulation of nu­
traceutical substances is supported by these controlled settings for
polysaccharide-protein complexes, which can improve functional char­
acteristics without enzymatic and chemical changes. A source of pros­
perity and potential cash, the rising food business will no longer be seen
as a low-profit commodity. Utilizing functional bionanomaterials
derived from natural sources to build food structures offers fresh per­
spectives on the creation of future foods with superior sensory qualities
and health advantages while avoiding artificial additions and unhealthy
ingredients (Song et al., 2022). A fascinating and interesting research
area is developing novel edible protein-polysaccharide nanoconjugates
or developing novel nanostructures using underutilized edible bio­
materials for efferent encapsulation and delivery of plant-based
nutraceuticals.
Protein-polysaccharide nanoconjugates have played an important
role in improving the bioactivities and food-related characteristics of
nutraceuticals, making them an attractive candidate for use in industrial
food applications. However, there are still a few issues to take into ac­
count. First, the production of protein-polysaccharide nanoconjugates
may reduce the nutritional value of food because some essential amino
acids, such as lysine, tryptophan, and methionine, are conjugated to
polysaccharides. Second, when the production process (i.e. Maillard
reaction) reaches its final stage, several nano-based products that are
harmful to humans are produced, including advanced glycation end-
products, heterocyclic amines, and acrylamide. Third, the precise
composition and structure of the final products and how they are
metabolized and react in the human body are still unclear at present
because of their complex constituents. Moreover, blocking the bond
between proteins and polysaccharides by crosslinking agents, unclear
mechanism of precise bioactive components, and biosafety issues of
protein-polysaccharide nanoconjugates are still major challenges.
Additional research is required to evaluate the safety and viability of
protein-polysaccharide nanoconjugate applications in the food industry
in light of these difficulties. To produce products with high functional
and biological features, reaction parameters must be properly and
suitably set and carefully regulated. For instance, to increase the reac­
tion rate and decrease the formation of undesirable by-products or
chemicals, other approaches could be used more frequently. The
mechanisms underlying the bioactivities, metabolic processes, and
toxicological characteristics of protein-polysaccharide nanoconjugates
should also be further investigated in order to pinpoint their effects on
human health, which is crucial for determining the application safety of
protein-polysaccharide nanoconjugates. Eventually, in order to purify
and enrich protein-polysaccharide nanoconjugates with particular
17
Food Chemistry 428 (2023) 136709
bioactivities based on their structure and physicochemical properties,
the isolate needs enhanced techniques for detection and separation. This
will enable protein-polysaccharide nanoconjugates to be employed as an
additive or raw material in the industrial manufacturing of food.
8. Conclusions
The multitude of health benefits of nutraceuticals and their role as
therapeutic agents for several chronic and other diseases have evoked
researchers’ interest in exploring more refined methods for nutraceuti­
cal supplement delivery through functional foods. So, current research is
primarily focused on the ingredient encapsulation of nutraceuticals with
biomaterial to improve their stability during oral administration. Bio­
polymers such as polysaccharides, proteins, and protein-polysaccharide
complex-based nanoconjugates have shown great promise in improving
the bioavailability, solubility, gastric stability, adhesion, retention time,
and permeability of many nutraceuticals. Biopolymers have some su­
preme advantages due to their biological origin, biodegradability, and
low toxicity, which make them suitable materials for nutraceutical
carriers. Protein-polysaccharide nanoconjugates are a promising de­
livery system for nutraceutical application. The complex exhibited a
combined biological effect of both polymers and their application,
which could potentially unwrap many prospects for advancement in
health and nutrition.
Declaration of Competing Interest
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influence
the work reported in this paper.
Data availability
No data was used for the research described in the article.
Acknowledgment
The authors would like to acknowledge the Council of Scientific and
Industrial Research, India (OPL-0106). The institutional manuscript
number is CSIR-NBRI_MS/2023/05/02. The authors would like to
acknowledge the institutional research funding and support.
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