Advances in Colloid and Interface Science 274 (2019) 102039

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Advances in Colloid and Interface Science

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

Historical perspective

Structure and functions of oleosomes (oil bodies)

Constantinos V. Nikiforidis
Biobased Chemistry and Technology, Wageningen University and Research, Bornse Weillanden 9, P.O. Box 17, 6708WG Wageningen, the Netherlands

a r t i c l e i n f o a b s t r a c t

Article history: Oleosomes are natural oil droplets, abundant in plants and more specifically in seeds, composing 20–50 wt% of
20 September 2019 their mass. The structure of oleosomes is the mechanism that seeds developed to safely store energy in the
Available online 17 October 2019 form of triacylglycerols and use it during germination. For this, the phospholipid/protein membrane that covers
and protects the triacylglycerols has been wisely developed during evolution to grant them extreme stability
Keywords:
against physical and chemical stresses. The remarkable property-performance relationships of oleosomes have
Emulsions
Oil droplets
generated a lot of interest to incorporate them in oil-in-water emulsions and take advantage of their sophisti-
Oil bodies cated membrane. However, the structure-function relationship of the molecular components in the oleosome
Oleosomes membrane is still not well understood and requires more attention in order to take complete advantage of
Natural carriers their potential functions. The aim of this review is to give insights into the architecture of the oleosomes and
to discuss the exploitation of their properties in advanced and broad applications, from carrying and protecting
sensitive molecules to bio-catalysis.
© 2019 The Author. Published by Elsevier B.V. This is an open access article under the CC BY license (http://
creativecommons.org/licenses/by/4.0/).

Contents

1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1
2. Oleosomes in plants. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
3. Oleosome extraction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
4. The architecture of oleosome membrane . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
5. Structure-function relationship of oleosome membrane components. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3
6. Advanced applications . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3
7. Future research. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
8. Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
Acknowledgments . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5

1. Introduction interface is stabilized by low molecular weight surfactants and/or

Plant-derived lipids, which are mostly triacylglycerols and smaller
amounts of polar lipids are extensively exploited in numerous applica-
tions. They are mostly extracted from oilseeds (i.e. soybeans, sunflower
seeds, rapeseeds) through pressing and/or organic solvent extraction.
Their uses include bulk oil for cooking, biofuels, biobased chemicals
[1] and as building blocks of soft matter, like emulsions and gels [2]. In
the cases that the oil is used in oil-in-water emulsions as the dispersed
phase, it is then homogenised to form oil droplets, while the oil/water
E-mail address: costas.nikiforidis@wur.nl.
proteins.
On the other hand, the oil in eukaryotes is already in the form of
emulsified droplets, stabilized by a unique protein/phospholipid mem-
brane. Those natural oil droplets that can be found in plants have a di-
ameter of up to a few microns called oleosomes (or oil bodies) [3,4].
The same motives can also be found in mammalian adipocytes where
the oil droplets are in the nanoscale range called lipid droplets [5,6].
The formation of these natural droplets, with an oil core and a hydro-
philic surface, is the way that nature solved the issue of the immiscibility
of liquids with different polarities.
Soft matter scientists should be inspired by these natural structures
and use their membranes as prototypes when designing synthetic oil
droplets. Or, especially for plant-derived oleosomes they could be

https://doi.org/10.1016/j.cis.2019.102039
0001-8686/© 2019 The Author. Published by Elsevier B.V. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
2 C.V. Nikiforidis / Advances in Colloid and Interface Science 274 (2019) 102039
extracted in their native form and be used as such for natural oil-in-
water emulsions [7]. The big advantages of using these droplets are
that: they are completely natural and derived in simple steps from
plant sources, the intensive bulk oil extraction omitted; no additional
emulsifiers are needed, no homogenization step is required, and their
sophisticated membrane designed through thousand years of evolution
opens new paths for advanced applications.
2. Oleosomes in plants
The extraction of intact and native oleosomes requires knowledge of
their exact location in the plant matrix and their interactions with sur-
rounding molecules. Oleosomes are present in many plant tissues, like
roots [8] and leaves [9], however, they are most abundant in seeds
and nuts [10]. They are biosynthesized in seed embryo cells by lipid bio-
synthetic enzymes and accumulate in dry seed cells [6,11]. The
oleosome core is comprised of triacylglycerols that are used as an en-
ergy and carbon source during seed germination and seedling growth.
To be able to disperse the triacylglycerols in cells and also protect
them from degradation due to environmental stresses, a protective
membrane is placed around them. The formed natural oil droplets
with a typical diameter of 0.2 to 2.0 μm and the membrane is a sophis-
ticated mixture of a monolayer of phospholipids and anchored unique
proteins (Fig. 1) [4,12,13]. The exact molecular composition of the
oleosome membranes can vary between different plant tissues [10],
however their functional performance is following similar patterns [7].
In the dense cell environment, native oleosomes are fully com-
pressed and packed. Due to the high compression, the oleosome mem-
brane cannot be visualised. However, after seed soaking in aqueous
media and subsequent swelling of the cells (Fig. 2) the oleosome parti-
cles are becoming discrete [13]. This observation indicates that under
compression oleosomes have an irregular disc-like shape, while when
given available space they transform to spherical droplets. Apparently,
the molecules of the oleosome membrane interact synergistically and
form a responsive and flexible membrane with the ability to self-
shape [13].
3. Oleosome extraction
The oleosome surface is comprised of the phospholipid polar heads
and the hydrophilic domains of the interfacial proteins. Due to the hy-
drophilic character and the electrokinetic potential, oleosomes follow
the same extraction rules with proteins and are extracted aqueously
at a pH range between 7 and 9 [14]. However, due to the larger diameter
of the oleosome particles, compared to protein molecules, the diffusion
kinetics of oleosomes are lower [15]. After their extraction, the
oleosomes are obtained as natural oil-in-water emulsions or as dense
oleosome creams [12,16].
To promote the extensive use of oleosomes at the industrial level it is
essential to set up an easy extraction process that does not affect the
oleosome membrane and its stability. Therefore, efforts towards a sus-
tainable oleosome extraction in high yields are being made [17–19].
Up to today, oleosome extraction yields of up to 95 wt% have been re-
ported, showing that it is possible to compete for the bulk oil extraction,
Fig. 1. Oleosome structure. The components of the illustration are not on-scale.
Fig. 2. Hazelnut cells after soaking for 48 h. Adapted with permission from Capuano
et al. [64].
where pressing and organic solvent extraction is applied [20]. Big scale
extraction processes have also been proposed [21], however, the pro-
cess is not optimized yet and there is still room for improvement in
order to become competitive with the current approach of producing
oil-in-water emulsions.
An important decision when extracting oleosomes is to define
whether pure oleosomes or mixtures of oleosomes and storage proteins
are needed. Oleosomes, are in close contact with other intracellular ma-
terial, like proteins (Fig. 2), which is co-extracted together with the
oleosomes [22]. For some applications, especially foods, like salad dress-
ings, where proteins are already used to adjust the system's macro-
scopic properties, having mixtures of oleosomes and plant storage
proteins can be beneficial [23,24]. In other applications though, maybe
pure oleosomes are needed [25]. Extrinsic material, like proteins, may
interact with the oleosome interface and affect, positively or negatively,
their behaviour and properties, like the interactions with surrounded
material, their impact on the rheological properties of the system and
their physical and chemical stability [22,26,27]. To obtain pure
oleosomes, a couple of washing steps are required, which include pH
adjustment and centrifugation in order to remove the extrinsic material
form the oleosome interface [28].
4. The architecture of oleosome membrane
The oleosome membranes are the only membranes that consist only
by a monolayer of phospholipids anchored with proteins (Fig. 3)
[29–31]. The phospholipid layer has a thickness of about 0.9 nm and
composes about 2 wt% of the total mass of the seed oleosomes [32].
There are several groups of phospholipids present, however the most
dominant are phosphatidylcholine (PC) and phosphatidylserine (PS)
[10]. As 1H NMR analysis has shown for sunflower oleosomes, the
2
8 nm
H2O
3 nm
9 nm
TAGs 5 nm
Proteins Phospholipids
Fig. 3. Proposed model of oleosome membrane conformation. The fold of the hydrophobic
region is embedded in the oleosome core, while the hydrophilic domains are exposed in
the aqueous phase. The components of the illustration are not on-scale.
 C.V. Nikiforidis / Advances in Colloid and Interface Science 274 (2019) 102039
fatty acid fraction is dominated by palmitic, linoleic and oleic acids in a
2:1:0.25 ratio [33]. The high amount of saturated fatty acids is an indica-
tion that the oleosome membrane is a thermotropic crystalline
mesophase, however, further research is required towards this
direction.
The proteins present on the oleosome membrane are oleosins,
caleosins, and steroleosins [6,34]. Oleosins are the most abundant,
with molecular masses between 15 and 20 kDa [10,35]. Oleosins have
an extended central hydrophobic domain, which according to molecu-
lar modeling forms two α-helical hairpin structures separated by a
turn region [30]. The N-terminal (N) and C-terminal (C) domains are ex-
pected to facilitate association with the phospholipid polar heads. This
unique architecture reminds the structure of surfactants with a polar
head and a lipophilic chain, providing oleosins with a significant interfa-
cial activity [35,36]. Experimental data derived by a structural proteo-
mics approach support the proposed model and confirm that the
hydrophobic central region of rapeseed oleosins is pinned into the triac-
ylglycerol core (Fig. 3) [37]. Besides the fact that the membrane polar
lipids and proteins are anchored on the oleosome interface due to
their amphiphilic character, the interfacial network is probably sup-
ported through electrostatic interactions. The polar lipids on the inter-
face can be negatively charged (i.e. phosphatidylserine, PS) and
interact through electrostatic attractive forces with the basic amino
acid residues of the interfacial proteins [38].
Caleosins have a similar hydrophobic domain to oleosins, however,
the provided models indicate that it is smaller than the one
from oleosins [31,39]. The size of the proteins is between 25 and
35 kDa and their N-terminal region is fairly hydrophilic and contains a
single Ca2+ binding site that is probably exposed to the aqueous
phase [31,40]. As deduced from circular dichroism spectra, caleosin's
secondary structure is dramatically influenced by the polarity of
media, indicating that their properties might change in different
environments [41].
Steroleosins are bigger proteins with molecular sizes above 35 kDa
that have again a hydrophobic anchoring segment at the same length
as caleosins [42]. Moreover, steroleosins have an extended hydrophilic
domain that is homologous to sterol-binding dehydrogenases and
according to the proposed model they are exposed to the aqueous
phase [42].
Small-Angle Neutron Scattering revealed that in the case of
oleosomes derived from rapeseeds, the total membrane thickness that
includes the phospholipids and all three types of proteins is about
9 nm. As it is shown in Fig. 3, the hydrophobic domain of oleosins,
which is the longest amongst the different proteins is about 5 nm, indi-
cating that the outer hydrophilic domain of the oleosome proteins is
about 3 nm high and according to calculations on maize germ oleosins,
they cover an area of about 8.1 nm2 [3,19].
5. Structure-function relationship of oleosome membrane
components
The size of oleosomes in the cells is partly controlled by the relative
amounts of oil to oleosin [3,43]. For example, maize kernels bred for
high oil content and a high ratio of oil to oleosin resulted in larger spher-
ical oleosomes, while low oil to oleosin ratio resulted in smaller
oleosomes [44]. This behaviour is probably linked to the biological
mechanism of oleosome production. Moreover, it has been discovered
that in the case of intracellular cycad seed oleosomes, their stability in
the absence of oleosins is ruled by caleosin molecules [45]. This finding
suggests that caleosins play a major interfacial structural role as well,
and the general oil to protein ratio is the one that determines the size
of oleosomes in the cells.
However, besides proteins, the oleosome membrane phospholipids
certainly play a role in the unique structure and stability of the interface.
Dense crystalline (hard) phases are probably formed due to the satu-
rated aliphatic chains of the phospholipids [46,47]. Furthermore, the
3
crystalline phospholipid platforms are anchored by the hydrophobic
proteins, which probably act as “suspensions” making the membrane
stretchable [48].
A path to understanding the structural role of each component at the
oleosome membrane is to extract each component and investigate the
interfacial activity. Pure oleosins, showed a comparable behaviour on
air-water and oil-water interfaces to the properties of milk proteins, ly-
sozyme and egg yolk apolipoproteins [49,50]. These findings indicated
that despite the strong hydrophobic character of oleosins and even at
the absence of phospholipids a strong elastic network can be formed
at the air/water and oil/water interfaces. When oleosins were mixed
with phospholipids further decreased the interfacial tension of oil/
water interfaces, pointing out that the presence of phospholipids en-
hanced the interfacial activity of oleosins [51]. Moreover, oleosins
were able to be absorbed on oil/water interfaces that were previously
covered with a compressed phospholipid monolayer. Apparently, the
highly hydrophobic domains of oleosins were able to penetrate the
phospholipid monolayer and reach the oil phase [50]. Besides oleosins,
caleosins play also a significant structural role in the oleosome
membrane, however, following their isolation from rapeseed
oleosomes, they exhibited lower interfacial activity comparing to
oleosins. A significant difference was that caleosins couldn't be absorbed
into oil/phospholipid/water interfaces proving little interactions
between caleosins and phospholipids [52].
6. Advanced applications
Oleosomes could be potentially used as such to replace synthetic oil
droplets in emulsion types of applications, from foods to cosmetics and
pharmaceuticals [7,53]. They are superior to the engineered oil droplets
since their natural sophisticatedly designed membrane provides to
oleosomes high stability against environmental stresses. Besides stabil-
ity, it has been reported that oleosome membranes actively interact
with biopolymers in the bulk phase, enhancing the rheological proper-
ties of the emulsions [22,23,27,54]. It is important to state that besides
the hydrophilic nature of the domains of the membrane proteins that
are facing the external side of oleosomes, there are still hydrophobic
batches on the proteins that lead to hydrophobic interactions between
oleosomes and also between oleosomes and extrinsic biopolymers
[22,23,27].
Besides the potential use of oleosomes as natural oil droplets in
emulsions, the high interfacial activity of the molecules at their mem-
brane could be exploited to stabilize the interfaces of foams or emul-
sions. As it has been reported, when soybean oleosomes were inserted
into the aqueous phase of an air/water interface, the oleosomes diffused
immediately to the interface [55]. Fig. 4, presents the steep increase in
surface pressure during this diffusion. It is possible, depending on the
state/stability and concentration of the absorbed oleosomes, that they
might rupture at different time scales. When the interfacial film was fur-
ther compressed soon after rupture of the oleosomes, one to two sharp
changes at the surface pressure was observed. These changes could at-
tributed to the leaking triglycerides and/or the phospholipids and/or
phospholipid-protein mixtures, which are able to lower the surface ten-
sion. The destabilization of oleosomes, when placed on an air/water in-
terface, contradicts the reports that promote the high stability of the
oleosomes [17,18]. To clarify the mechanism that takes place when
oleosomes are placed on an interface, oil-in-water emulsions were pre-
pared with sunflower oleosomes as interfacial stabilizers [56]. With the
aid of confocal microscopy it was shown that initial state of oleosomes
determines their fate on the interface. The smaller oleosomes that
retained their native size during the extraction were stable and were
absorbed intact on the interface following a Pickering stabilization
mechanism. However, the oleosomes that were pre-destabilized due
to the extraction process and coalesced (N5 μm) disintegrate when
were in contact with the oil phase of the emulsion. As a result, fractions
of the oleosome membrane were absorbed on the interface and
4 C.V. Nikiforidis / Advances in Colloid and Interface Science 274 (2019) 102039

Fig. 6. Confocal Laser Scanning Microscopy analysis of extracted coconut oleosomes

stained red by Nile Red. Adapted with permission from Dave et al. [62].

Fig. 4. Simplified measurement of surface pressure as a function of time and

corresponding scenarios at the air/water interface. Possible scenarios of the oleosome
configuration when exposed to an air/water interface is sketched for illustration.
Adapted with permission from [55]. Copyright (2012) American Chemical Society.
stabilized the oil droplets of the emulsion (Fig. 5). Nonetheless, partial
destabilization of the bigger oleosomes could even have positive impact
on their behaviour as emulsifiers, as the oleosome membrane fractions
contain highly interfacially active molecules [56].
Moreover, the unique conformation of oleosomes could also be
exploited in the niche and advanced applications. For example, the
oleosome membrane prevents the infusion of oxidation promotors
and protects the sensitive fatty acids from oxidation [26,57,58]. This
ability of the oleosomes can be used when extracting and carrying un-
saturated oils from sources like nuts and algae [59–61].
In addition, the oleosome membrane is permeable to different types
of hydrophobic molecules that can be entrapped and protected in the
TAG core. This ability can be used to stain the oleosomes with hydro-
phobic dyes, like Nile Red and visualise them under confocal light (Fig.
6) [62]. But more importantly, another example of encapsulated hydro-
phobic molecules are volatile compounds, like flavours. Oleosomes,
loaded with volatile compounds showed significantly stronger potential
to maintain their headspace volatile concentration in comparison to
synthetic droplets stabilized by phospholipids [25]. Besides volatile
compounds, it has also been proven that astaxanthin, a very sensitive
molecule to oxidation, could be entrapped and protected in the
oleosome core [63]. At the same time, encapsulated astaxanthin
retained its antioxidant activity.
The potential use of oleosomes as carriers of sensitive molecules and
therapeutics can further be supported by the reported data on their sta-
bility during digestion [64–70]. Sunflower oleosomes are digested at a
relatively slow rate in comparison to artificial oil droplets stabilized by
surfactants or whey proteins [70]. During 120 min of in vitro intestinal
digestion of hazelnut oleosomes, only 10–20 wt% of the free fatty
acids of oleosomes were released, making their membrane a good pro-
tective barrier against digestion. Similar data were also obtained in the
case of almond oleosomes [66].
Another significant potential application is the decontamination of
aqueous environments with toxic organic molecules. When oleosomes
were placed in aqueous environments contaminated with organic pes-
ticides, the organic molecules were successfully absorbed in the inner
hydrophobic core of the oleosomes and removed [71]. According to
this research, oleosomes exhibited very good mass transfer properties,
compared to synthetic microcapsules or liquid-liquid extraction tech-
niques. The oleosome proteins exhibit lipase activity, which is activated
during germination to hydrolyse and dispose of triacylglycerols during
germination. This property of the oleosome membrane proteins can
be exploited to use them as substrates for lipolytic enzymes [72]. Fur-
thermore, genetic engineering can also be employed to construct
oleosome based biocatalysts [73]. Organophosphorus hydrolase was

Fig. 5. (a) An oil droplet stabilized by sunflower oleosomes. The sample is stained by Nile Blue. Due to signal enhancement the oil phase is shown in black, the continuous aqueous phase in
blue, and the proteins are shown in green and red depending on their concentration. (b) A graphical illustration of the fate of sunflower oleosomes when placed on oil/water interfaces and
their intriguing behaviour upon emulsification. Adapted with permission from Karefyllakis et al. [56].
 C.V. Nikiforidis / Advances in Colloid and Interface Science 274 (2019) 102039
Fig. 7. Schematic illustration of designer oleosomes. Genetically engineered Y. lipolytica
was employed to produce organophosphorus hydrolase decorated oleosomes. Adapted
with permission from Leng et al. [73].
genetically fused with oleosins to create biocatalysts that can degrade
organophosphorus pesticides (Fig. 7). The designer oleosome had a sig-
nificantly improved enzyme activity compared to the free organophos-
phorus hydrolase.
7. Future research
Oleosomes have attracted the interest of both academics and indus-
try in the last decades. Their natural characteristics make them poten-
tially sustainable alternatives to synthetic oil droplets in any system
where oil is dispersed in aqueous environments. Moreover, the unique
conformation of the oleosome membrane that grants oleosomes with
high stability can inspire colloid scientists when designing synthetic
oil droplets. However, the exploitation of all the potentials of the
oleosomes requires a deep understanding of the structural motifs and
the relationship with their performance. The structural dynamics be-
tween the soft and hard phospholipid platforms and also the proteins,
provide the oleosome membrane with stability and stretchability. This
is an area that has not been investigated yet and will provide very inter-
esting data on elasticity and adaptability of oleosomes (Fig. 8). Addition-
ally, the selective permeability of the oleosome membrane, which
makes them great natural carriers of hydrophobic molecules, is yet an-
other scientific field that requires investigation. In general, the
structure-function relationship of the oleosome membranes is still not
well understood and requires more attention in order to take complete
advantage of their potential functions.
Phospholipids Proteins
Fig. 8. Hypothetical conformation of oleosomes under compression.
5
8. Conclusions
Oleosomes are abundant natural oil droplets present in most organ-
isms and can be found in high concentrations is seeds and nuts. They ap-
pear in the cells in sizes between 0.2 and 2.0 μm and are comprised of a
triacylglycerol core surrounded by a dense monolayer of phospholipids
and hydrophobic proteins. Oleosomes can be extracted with the aid of
alkaline aqueous solvents and makes them promising natural replacers
of the synthetic oil droplets that are currently used in emulsion type of
applications. The unique molecular combination in the oleosome mem-
brane equips them with high stability and makes them selectively per-
meable to hydrophobic molecules. It is difficult to produce oil droplets
with such advanced properties. Oleosomes are combining sustainability
aspects with high functionality. The understanding of their properties
and dynamics will provide knowledge on the related biological mecha-
nisms and will lead to their use in advanced applications for colloidal
systems.
Acknowledgments
I would like to thank all the members of the Biobased Functional Ma-
terials Lab for the fruitful discussions and brainstorms. At the time of the
submission of this review, they were: Dimitris Karefyllakis, Juliana
Romero Guzman, Laura Schijven, Eleni Ntone, Simha Sridharan and Dr.
Christos Fryganas.
References
[1] Montero de Espinosa L, Meier MAR. Plant oils: the perfect renewable resource for
polymer science?! Eur Polym J 2011;47:837–52.
[2] Dickinson E. Food emulsions and foams: stabilization by particles. Curr Opin Colloid
Interface Sci 2010;15:40–9.
[3] Tzen JTC, Huang AHC. Surface-structure and properties of plant seed oil bodies. J Cell
Biol 1992;117:327–35.
[4] Tzen JTC. Integral proteins in plant oil bodies. ISRN Botany 2012;2012:16.
[5] Olzmann JA, Carvalho P. Dynamics and functions of lipid droplets. Nat Rev Mol Cell
Biol 2019;20:137–55.
[6] Murphy DJ. The biogenesis and functions of lipid bodies in animals, plants and mi-
croorganisms. Prog Lipid Res 2001;40:325–438.
[7] Nikiforidis CV, Matsakidou A, Kiosseoglou V. Composition, properties and potential
food applications of natural emulsions and cream materials based on oil bodies.
RSC Adv 2014;4:25067–78.
[8] Jayaram S, Bal AK. Oleosomes (lipid bodies) in nitrogen-fixing peanut nodules. Plant
Cell Environ 1991;14:195–203.
[9] Shimada TL, Hara-Nishimura I. Leaf oil bodies are subcellular factories producing an-
tifungal oxylipins. Curr Opin Plant Biol 2015;25:145–50.
[10] Tzen JTC, Cao YZ, Laurent P, Ratnayake C, Huang AHC. Lipids, proteins, and structure
of seed oil bodies from diverse species. Plant Physiol 1993;101:267–76.
[11] Shimada TL, Hayashi M, Hara-Nishimura I. Membrane dynamics and multiple func-
tions of oil bodies in seeds and leaves. Plant Physiol 2018;176:199–207.
[12] Nikiforidis CV, Karkani OA, Kiosseoglou V. Exploitation of maize germ for the prep-
aration of a stable oil-body nanoemulsion using a combined aqueous extraction-
ultrafiltration method. Food Hydrocoll 2011;25:1122–7.
[13] Nikiforidis CV, Kiosseoglou V, Scholten E. Oil bodies: an insight on their microstruc-
ture—maize germ vs sunflower seed. Food Res Int 2013;52:136–41.
[14] Tzen JTC, Peng CC, Cheng DJ, Chen ECF, Chiu JMH. A new method for seed oil body
purification and examination of oil body integrity following germination. J Biochem
1997;121:762–8.
[15] Matsakidou A, Mantzouridou FT, Kiosseoglou V. Optimization of water extraction of
naturally emulsified oil from maize germ. LWT Food Sci Technol 2015;63:206–13.
[16] Nikiforidis CV, Scholten E. High internal phase emulsion gels (HIPE-gels) created
through assembly of natural oil bodies. Food Hydrocoll 2015;43:283–9.
[17] Chen BC, McClements DJ, Gray DA, Decker EA. Physical and oxidative stability of pre-
emulsified oil bodies extracted from soybeans. Food Chem 2012;132:1514–20.
[18] Iwanaga D, Gray DA, Fisk ID, Decker EA, Weiss J, McClements DJ. Extraction and
characterization of oil bodies from soy beans: a natural source of pre-emulsified soy-
bean oil. J Agric Food Chem 2007;55:8711–6.
[19] Nikiforidis CV, Kiosseoglou V. Aqueous extraction of oil bodies from maize germ
(Zea mays) and characterization of the resulting natural oil-in-water emulsion. J
Agric Food Chem 2009;57:5591–6.
[20] Gunstone DF. Production and trade of vegetable oils. In: Gunstone DF, editor. Vege-
table oils in foo technology: composition, properties and uses. Oxford, UK: Blackwell
Publishing Ltd; 2011. p. 1–24.
[21] Kapchie VN, Hauck CC, Wang H, Murphy PA. Process improvement for semipurified
oleosomes on a pilot-plant scale. J Food Sci 2011;76:C60–C853.
[22] Nikiforidis CV, Kiosseoglou V. Physicochemical stability of maize germ oil body
emulsions as influenced by oil body surface-xanthan gum interactions. J Agric
Food Chem 2010;58:527–32.
6 C.V. Nikiforidis / Advances in Colloid and Interface Science 274 (2019) 102039
[23] Nikiforidis CV, Biliaderis CG, Kiosseoglou V. Rheological characteristics and physico-
chemical stability of dressing-type emulsions made of oil bodies-egg yolk blends.
Food Chem 2012;134:64–73.
[24] Karefyllakis D, Octaviana H, van der Goot AJ, Nikiforidis CV. The emulsifying perfor-
mance of mildly derived mixtures from sunflower seeds. Food Hydrocoll 2019;88:
75–85.
[25] Fisk ID, Linforth RST, Taylor AJ, Gray DA. Aroma encapsulation and aroma delivery by
oil body suspensions derived from sunflower seeds (Helianthus annus). Eur Food
Res Technol 2011;232:905.
[26] Karkani OA, Nenadis N, Nikiforidis CV, Kiosseoglou V. Effect of recovery methods on
the oxidative and physical stability of oil body emulsions. Food Chem 2013;139:
640–8.
[27] Nikiforidis CV, Donsouzi S, Kiosseoglou V. The interplay between diverse oil
body extracts and exogenous biopolymers or surfactants. Food Res Int 2016;
83:14–24.
[28] De Chirico S, di Bari V, Foster T, Gray D. Enhancing the recovery of oilseed rape seed
oil bodies (oleosomes) using bicarbonate-based soaking and grinding media. Food
Chem 2018;241:419–26.
[29] Napier JA, Beaudoin F, Tatham AS, Alexander LG, Shewry PR. The seed oleosins:
structure, properties and biological role. Advances in botanical research. Academic
Press; 2001. p. 111–38.
[30] Alexander LG, Sessions RB, Clarke AR, Tatham AS, Shewry PR, Napier JA. Characteri-
zation and modelling of the hydrophobic domain of a sunflower oleosin. Planta
2002;214:546–51.
[31] Frandsen GI, Mundy J, Tzen JTC. Oil bodies and their associated proteins, oleosin and
caleosin. Physiol Plant 2001;112:301–7.
[32] Purkrtova Z, Jolivet P, Miquel M, Chardot T. Structure and function of seed lipid
body-associated proteins. C R Biol 2008;331:746–54.
[33] Furse S, Liddell S, Ortori C, Williams H, Neylon DC, Scott D, et al. The lipidome and
proteome of oil bodies from Helianthus annuus (common sunflower). J Chem Biol
2013;6:63–76.
[34] Shimada TL, Hara-Nishimura I. Oil-body-membrane proteins and their physiological
functions in plants. Biol Pharm Bull 2010;33:360–3.
[35] Jolivet P, Roux E, d'Andrea S, Davanture M, Negroni L, Zivy M, et al. Protein compo-
sition of oil bodies in Arabidopsis thaliana ecotype WS. Plant Physiol Biochem 2004;
42:501–9.
[36] Lin LJ, Liao PC, Yang HH, Tzen JTC. Determination and analyses of the N-termini of
oil-body proteins, steroleosin, caleosin and oleosin. Plant Physiol Biochem 2005;
43:770–6.
[37] Jolivet P, Aymé L, Giuliani A, Wien F, Chardot T, Gohon Y. Structural proteomics: to-
pology and relative accessibility of plant lipid droplet associated proteins. J Proteo-
mics 2017;169:87–98.
[38] Tzen JTC, Lie GC, Huang AHC. Characterization of the charged components and their
topology on the surface of plant seed oil bodies. J Biol Chem 1992;267:15626–34.
[39] Chen JCF, Tsai CCY, Tzen JTC. Cloning and secondary structure analysis of caleosin, a
unique calcium-binding protein in oil bodies of plant seeds. Plant Cell Physiol 1999;
40:1079–86.
[40] Frandsen G, Müller-Uri F, Nielsen M, Mundy J, Skriver K. Novel plant ca-binding pro-
tein expressed in response to abscisic acid and osmotic stress. J Biol Chem 1996;271:
343–8.
[41] Purkrtova Z, d'Andrea S, Jolivet P, Lipovova P, Kralova B, Kodicek M, et al. Structural
properties of caleosin: a MS and CD study. Arch Biochem Biophys 2007;464:335–43.
[42] Lin L-J, Tai SSK, Peng C-C, Tzen JTC. Steroleosin, a sterol-binding dehydrogenase in
seed oil bodies. Plant Physiol 2002;128:1200–11.
[43] Sarmiento C, Ross JHE, Murphy DJ. Expression and subcellular targeting of a soybean
oleosin in transgenic rapeseed. Implications for the mechanism of oil-body forma-
tion in seeds. Plant J 1997;11:783–96.
[44] Ting JTL, Lee KY, Ratnayake C, Platt KA, Balsamo RA, Huang AHC. Oleosin genes in
maize kernels having diverse oil contents are constitutively expressed independent
of oil contents-size and shape of intracellular oil bodies are determined by the
oleosins oils ratio. Planta 1996;199:158–65.
[45] Jiang P-L, Tzen JTC. Caleosin serves as the major structural protein as efficient as
oleosin on the surface of seed oil bodies. Plant Signal Behav 2010;5:447–9.
[46] Payne G, Lad M, Foster T, Khosla A, Gray D. Composition and properties of the surface
of oil bodies recovered from Echium plantagineum. Colloids Surf B Biointerfaces
2014;116:88–92.
[47] Sato K. Polymorphism of lipid crystals. In: Sato K, editor. Crystallization of lipids.
Wiley Online Library; 2018 [Chapter 2].
[48] Denkov N, Tcholakova S, Lesov I, Cholakova D, Smoukov SK. Self-shaping of oil drop-
lets via the formation of intermediate rotator phases upon cooling. Nature 2015;
528:392−+.
[49] Nikiforidis CV, Ampatzidis C, Lalou S, Scholten E, Karapantsios TD, Kiosseoglou V. Pu-
rified oleosins at air-water interfaces. Soft Matter 2013;9:1354–63.
[50] Roux E, Baumberger S, Axelos MAV, Chardot T. Oleosins of Arabidopsis thaliana: ex-
pression in Escherichia coli, purification, and functional properties. J Agric Food Chem
2004;52:5245–9.
[51] Deleu M, Vaca-Medina G, Fabre JF, Roiz J, Valentin R, Mouloungui Z. Interfacial prop-
erties of oleosins and phospholipids from rapeseed for the stability of oil bodies in
aqueous medium. Colloids Surf B-Biointerfaces 2010;80:125–32.
[52] Purkrtova Z, Le Bon C, Kralova B, Ropers MH, Anton M, Chardot T. Caleosin of
Arabidopsis thaliana: effect of calcium on functional and structural properties. J
Agric Food Chem 2008;56:11217–24.
[53] Gray D. Oil body extraction and uses. Google Patents; 2012.
[54] Kirimlidou M, Matsakidou A, Scholten E, Nikiforidis CV, Kiosseoglou V. Composite
gels structured by a gelatin protein matrix filled with oil bodies. Food Struct 2017;
14:46–51.
[55] Waschatko G, Junghans A, Vilgis TA. Soy milk oleosome behaviour at the air-water
interface. Faraday Discuss 2012;158:157–69.
[56] Karefyllakis D, Jan van der Goot A, Nikiforidis CV. The behaviour of sunflower
oleosomes at the interfaces. Soft Matter 2019;15:4639–46.
[57] Fisk ID, White DA, Lad M, Gray DA. Oxidative stability of sunflower oil bodies. Eur J
Lipid Sci Technol 2008;110:962–8.
[58] Gray DA, Payne G, McClements DJ, Decker EA, Lad M. Oxidative stability of Echium
plantagineum seed oil bodies. Eur J Lipid Sci Technol 2010;112:741–9.
[59] Zhang P, Bari VD, Briars R, Taher ZM, Yuan J, Liu G, et al. Influence of pecan nut pre-
treatment on the physical quality of oil bodies. J Food Qual 2017;2017:9.
[60] Zaaboul F, Raza H, Chen C, Liu Y. Characterization of peanut oil bodies integral pro-
teins, lipids, and their associated phytochemicals. J Food Sci 2018;83:93–100.
[61] Wang ZT, Ullrich N, Joo S, Waffenschmidt S, Goodenough U. Algal lipid bodies: stress
induction, purification, and biochemical characterization in wild-type and Starchless
bemNChlamydomonas reinhardtiib/emN. Eukaryot Cell 2009;8:1856–68.
[62] Dave AC, Ye A, Singh H. Structural and interfacial characteristics of oil bodies in co-
conuts (Cocos nucifera L.). Food Chem 2019;276:129–39.
[63] Acevedo F, Rubilar M, Jofre I, Villarroel M, Navarrete P, Esparza M, et al. Oil bodies as
a potential microencapsulation carrier for astaxanthin stabilisation and safe delivery.
J Microencapsul 2014;31:488–500.
[64] Capuano E, Peltegrini N, Ntone E, Nikiforidis CV. In vitro lipid digestion in raw and
roasted hazelnut particles and oil bodies. Food Funct 2018;9:2508–25016.
[65] Gallier S, Acton D, Garg M, Singh H. Natural and processed milk and oil body emul-
sions: bioavailability, bioaccessibility and functionality. Food Struct Neth 2017;13:
13–23.
[66] Gallier S, Singh H. Behavior of almond oil bodies during in vitro gastric and intestinal
digestion. Food Funct 2012;3:547–55.
[67] Gallier S, Tate H, Singh H. In vitro gastric and intestinal digestion of a walnut oil body
dispersion. J Agric Food Chem 2013;61:410–7.
[68] Makkhun S, Khosla A, Foster T, McClements DJ, Grundy MML, Gray DA. Impact of ex-
traneous proteins on the gastrointestinal fate of sunflower seed (Helianthus
annuus) oil bodies: a simulated gastrointestinal tract study. Food Funct 2015;6:
125–34.
[69] Zheng B, Zhang X, Peng S, Julian McClements D. Impact of curcumin delivery system
format on bioaccessibility: nanocrystals, nanoemulsion droplets, and natural oil bod-
ies. Food Funct 2019;10:4339–49.
[70] White DA, Fisk ID, Makkhun S, Gray DA. In vitro assessment of the bioaccessibility of
tocopherol and fatty acids from sunflower seed oil bodies. J Agric Food Chem 2009;
57:5720–6.
[71] Boucher J, Cengelli F, Trumbic D, Marison IW. Sorption of hydrophobic organic com-
pounds (HOC) in rapeseed oil bodies. Chemosphere 2008;70:1452–8.
[72] Beisson F, Ferté N, Bruley S, Voultoury R, Verger R, Arondel V. Oil-bodies as sub-
strates for lipolytic enzymes. Biochim Biophys Acta (BBA)-Mol Cell Biol Lipids
2001;1531:47–58.
[73] Leng SH, Yang CE, Tsai SL. Designer oleosomes as efficient biocatalysts for enhanced
degradation of organophosphate nerve agents. Chem Eng J 2016;287:568–74.