Food Chemistry 131 (2012) 639644

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Food Chemistry
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A review of the progress in enzymatic concentration and microencapsulation

of omega-3 rich oil from sh and microbial sources
Jaroslav A. Kralovec a, Shuocheng Zhang a, Wei Zhang a, Colin J. Barrow b,
a
b

Ocean Nutrition Canada Ltd., 101 Research Drive, Dartmouth, NS, Canada B2Y 4T6
School of Life and Environmental Sciences, Deakin University Geelong, VIC 3217, Australia

a r t i c l e

i n f o

Article history:
Received 14 February 2011
Received in revised form 6 June 2011
Accepted 30 August 2011
Available online 19 September 2011
Keywords:
Microencapsulation
Complex coacervation
Omega-3 fats
EPA
DHA
Lipase
Functional food

a b s t r a c t
Technology continues to evolve for the concentration and stabilisation of omega-3 fatty acids for delivery
into food and beverage products. The use of lipases for selective concentration of EPA and DHA, or for
re-esterication reactions, is important in the production of omega-3 concentrates. Enzymatic strategies
require robust enzymes that can be immobilised and multiply re-used. Novel and mild processing methods are particularly important for providing oils with good sensory properties, which are required for successful use as functional food ingredients. Although in some cases good quality oils can be used directly in
some foods, such as margarine, many foods require that microencapsulated and stabilised omega-3 oils
be used. This is particularly important when the oils are preconcentrated. There are a number of industrially used microencapsulation methods, but the most widely used are complex coacervates and spray
dried emulsions. Fish oil is still the most widely used source of long-chain omega-3 fatty acids for addition to food, although algal oil is the primary source of DHA for infant formula use in North America. Algal
oil is still signicantly more expensive than sh oil for most applications, although many groups are
improving both the cost and quality of omega-3 oil from algal sources. In particular, Thraustochytrid
and Schizochytrid strains are a promising source of both DHA and EPA, and with further improvement
could be used to provide varying ratios of these omega-3 fats. In this short review we will describe some
of the current research in omega-3 fat concentration and microencapsulation, with particular emphasis
on the use of lipases for concentration and complex coacervation for microencapsulation.
Crown Copyright 2011 Published by Elsevier Ltd. All rights reserved.

1. Introduction
Omega-3 fats are long chain polyunsaturated fats containing
methylene-separated double bonds starting from the third carbon
atom counted from the methyl-terminus. The presence of bisallylic methylene groups and all double bonds being in the cis-conguration makes these molecules prone to structural changes, particularly oxidation, isomerisation and polymerisation.
Omega-3 fats are ingredients used in dietary supplements,
healthy foods, and pharmaceutical products. These bioactive fatty
acids have well established health benets and are primarily derived
from sh oil. The main bioactive omega-3 fatty acids are cis5,8,11,15,17-eicosapentaenoic acid (EPA) and cis-4,7,10,13,16,19docosahexaenoic acid (DHA) (Fig. 1). EPA and DHA are essential
components of healthy nutrition and have been shown clinically
to decrease the risk of coronary heart disease, partly through an ability to reduce serum triglyceride levels and help prevent secondary
heart attack. EPA and DHA are precursors to anti-inammatory
Corresponding author.
E-mail address: cbarrow@deakin.edu.au (C.J. Barrow).

mediators and have demonstrated benets for the prevention of

inammatory mediated disorders including allergy, diabetes,
Alzheimers disease and related neurodegenerative diseases (Lavie,
Milani, Mehra, & Ventura, 2009).
2. The production of omega-3 concentrates from sh oil
Fish oil is the most abundant and the cheapest source of EPA
and DHA. The best omega-3 terrestrial sources (seeds of ax,
perilla, kiwifruit and chia) are very rich in a-linolenic acid (ALA)
but have very low levels of EPA and DHA. The major health benet
of the consumption of ALA is that it converts to EPA and DHA.
However, in the human body ALA is converted to EPA and DHA
at an efciency of only 510% for EPA and 15% for DHA (Davis &
Kris-Etherton, 2003). Most sh oils do not contain more than a
30% combined level of EPA and DHA. For example, the primary
sources of most commercially used omega-3 sh oil is anchovy
(Engraulis ringens) and sardine (Sardinops sagax sagax) oils that
contain
1522% of EPA and 915% of DHA. This natural oil is normally
known as 1812 triglyceride (TG) sh oil. However, oils of

0308-8146/$ - see front matter Crown Copyright 2011 Published by Elsevier Ltd. All rights reserved.
doi:10.1016/j.foodchem.2011.08.085

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J.A. Kralovec et al. / Food Chemistry 131 (2012) 639644

cis-5,8,11,14,17-Eicosapentaenoic Acid (EPA)

O
O
O

cis-4,7,10,13,16,19-Docosahexaenoic Acid (DHA)

Fig. 1. Chemical structures of EPA and DHA.

concentration of 5090%, with controlled ratios of EPA and DHA,

are preferred for many supplement and pharmaceutical applications. Therefore, various groups have developed strategies for the
manufacturing of highly concentrated EPA and DHA products.
Fish oil is a mixture of fatty acids (FA) in the TG form with EPA
and DHA as the major long-chain components. Most oils contain
only one EPA or DHA fatty acid per TG molecule. If we assume each
TG molecule contains only one DHA or EPA in an 1812 TG, then the
fatty acids must be removed from the glycerol backbone to enable
concentration beyond 33% of EPA and DHA. Chemical ethanolysis
results in complete removal of all fatty acids from the glycerol
backbone and the formation of ethyl esters (EE), which can then
be concentrated using fractional distillation or urea complexation.
Ethanolysis converts the TG molecules to EEs reducing the average
molecular size to one third and consequently the boiling points are
reduced by about fty percent. For instance, the estimated boiling
point of DHA-TG at atmospheric pressure is 907 65 C, whereas
the boiling point of DHA-EE is only 444 24 C (calculated using
Advanced Chemistry Development (ACD/Labs) Software Solaris V
4.67). The lower boiling point means that EEs are more easily distiled than are TGs. This together with the separation of fatty acids
from the TG glycerol means that EEs are used as starting materials
for concentration of EPA and DHA.
There have been reports that EE concentrates are less bio-available than TG concentrates and there is an argument that TG forms
are natural whereas EE forms are not (Lawson & Hughes, 1988).
Therefore, some regulatory environments demand that nutritional
supplements contain oil in TG form, and so the production of TG
concentrates was achieved early on by converting EE concentrates
back to the TG form by chemical reaction with glycerol. This transesterication was carried out using traditional chemistry involving
the use of strong bases and harsh reaction conditions, resulting in
extensive formation of side-products. The use of lipases as catalysts
offers a milder re-esterication method, resulting in less byproducts and better quality oils. It is well known that lipases catalyse
hydrolysis of lipids at the water lipid interface. However, it is less
known that under anhydrous conditions lipases also effectively
catalyse synthesis of esters (Zaks & Klibanov, 1985). Lipases are
frequently used in the production of pharmaceuticals. Examples
include the use of Candida rugosa lipase in the synthesis of serum
cholesterol reducer lovastatin, Seratia marcescens for the production of coronary vasodilator diltiazem, and Candida antarctica lipase
B (CALB) for the production of anti-inammatory urbiprofen and
pain killer baclofen. In addition, lipases are biodegradable and have
a negligible biological oxygen demand in the waste stream.
Ocean Nutrition Canada Ltd. (ONC) has successfully converted
EE to TG both directly and via an intermediate hydrolysis step
through the free fatty acid (FFA) form (EE-FFA rst and then
FFA-TG) using immobilised CALB. Re-esterication from FFA concentrate was signicantly faster than that from EE and could be
carried out at lower temperatures and gave products in higher
yields. After routinely achieving 90% conversion from FFA to TG

in the laboratory with multiple re-use of the biocatalyst, the reaction was gradually scaled-up to manufacturing using proprietary
packed enzyme bed reactors. A plant assembly of four reactors allows us to manufacture up to 7500 kg of re-esteried TG per day
(unpublished results). Other researchers have used lipases to
incorporate EPA and DHA into food grade oils. For example, Hamam and Shahidi used a variety of lipases, including CALB, to successfully incorporate levels of between about 30% and 60%, EPA,
DPA or DHA into high-laurate canola oil (Hamam & Shahidi,
2006). These types of structured lipids containing both mediumchain and omega-3 fatty acids could become tailored functional
food ingredients with specic bioactivity, bioavailability and stability proles. A recent study shows that correctly immobilised lipases can be used with organic solvents to improve specicity. By
cross-linking immobilised Rhizomucor miehei lipase (RML) with
polyfunctional polymers these researchers were able to used 2propanol to increase EPA to DHA selectivity of this enzyme, resulting in a product with 22:1 ratio of EPA to DHA (Fernandez-Lorente
et al., 2011).
Despite success with enzymatic EE to TG conversion, there have
been questions about how much structural resemblance the
re-esteried TG share with the original natural 1812 TG oils. Unfortunately, CALB is not regioselective and not surprisingly a regiospecic analysis demonstrated differences between natural 1812
TG and its re-esteried (1812 TG) counterpart (Fig. 2). That is, natural 1812 TG has more DHA at position 2 with a close to equal
distribution of EPA across all three positions of glycerol, while
re-esteried concentrate has a statistical distribution of all fatty
acids across the three positions on glycerol. Of course, all our TG
concentrates made by re-esterication of EE concentrates contain
60% or more EPA and DHA and thus there will always be differences between natural 1812 TG oils and TG concentrates such as
4020 TG or 0555 TG, simply because these oils contain twice as
much of DHA and EPA, so that these long-chain fatty acids replace
almost a third of FFA residues in the starting oil. Shahidi and
co-workers have recently shown that chemical randomisation of
EPA, DPA and DHA in both seal blubber oil and menhanden oil
results in more even distribution of these omega-3 fats among
the terminal sn-1,3 and middle sn-2 position versus the natural
oils. This in term appears to lead to decreased oil stability, with increased omega-3 at the more exposed sn-1,3-positions, although
results were partly confounded due to modication in levels of
a-tocopherol antioxidant particularly in the seal oil (Wang,
Reyes-Suarez, Kralovec, & Shahidi, 2010).
To conserve the original positional distribution of FA residues in
the natural triglycerides we developed an alternative concentration process. In the rst step the starting 1812 TG oil was hydrolysed in the presence of Thermomycetes lanuginosus lipase. The goal
was to remove shorter chain FAs while retaining EPA and DHA
on the glycerol backbone. Using this strategy the degree of hydrolysis was in 4555% range. After hydrolysis and stripping FFA off
using distillation, the EPA and DHA content of the glyceride portion

J.A. Kralovec et al. / Food Chemistry 131 (2012) 639644

641

Fig. 2. 13C NMR spectra of starting 1812 TG oil and the oil after hydrolysis by lipase form Thermomycetes lanuginosus (TL 100) followed by reassembling back to triglyceride,
in the presence of immobilised CALB (Novozym 435) under anhydrous conditions.

was increased to 23% and 15%, respectively. In the second step the
generated free hydroxyl groups of the produced diglycerides were
re-esteried with a number of suitable sources of EPA and DHA in
the presence of CALB. For instance, 4020 FFA concentrate when
used as EPA and DHA donor boosted the levels of EPA and DHA
to 28% and 17%, respectively, thus increasing EPA and DHA content
of the starting 1812 TG by about 50%. Both, the hydrolytic and
esterication steps were monitored and analysed using several
standard and modied methods. In the case of reactions consuming or generating FFA, a simple determination of acid values by
alkalimetric titration was used. The positional distribution of the
original oil remained conserved after hydrolysis followed by reassembly, without concentration, as assessed by NMR.
Although the importance of positional distribution of fatty acid
residues in omega-3 oils and their concentrates is a matter of
debate, we developed a strategy for retention of positional distribution based on saturate removal, saturate removal followed by
re-esterication. The oils obtained by selective enzyme hydrolysis
were shown to have superior stability, superior sensory proles,
minimum polymer levels, trans-isomers and migration of double
bonds. However, it should be noted that the positional retention
strategy has to date produced concentrates with signicantly lower EPA and DHA levels than those made by EE to TG conversion
(45% vs. 60% EPA and DHA level). The lower levels of EPA and
DHA may be an important reason for better stability and sensory
proles. The use of lipases has the potential to enable the production of more stable oils with good sensory properties, and having
FA distributions more representative of the natural TG oils.

3. Microbial sources of omega-3 concentrates

Two important issues have been raised with regard to the use of
sh oil as a source of EPA and DHA. Firstly, some sh oils contain
man-made pollutants such as methyl mercury, polychlorinated
biphenols (PCBs) and dioxins. Secondly, some sh stocks from
which sh oil is derived, such as tuna, might not be sustainable
due to declining sh stocks. In contrast, microbial oils are sustainable sources of EPA and DHA and their stability can also be more
controllable than that of sh oils. The microbial oils are often less
complex than sh oils, exhibiting a simpler fatty acid proles.
These oils also can have varying ratio of EPA to DHA and often have
natural antioxidants present that can help protection the oils from
oxidative damage during processing.
Various microbial strains could serve as producers of specialty
oils that would meet desired specications. In 1990s, some microbial

sources of omega-3 oils were developed, particularly for the production of DHA for infant formula. DHA and arachidonic acid (20:4 n-6,
AA) are the dominant long-chain fatty acids in breast milk and play
vital roles in neonatal development. As the ratio of AA:EPA:DHA in
mothers milk (2.0:0.2:1.0, w/w) is completely different from that
in sh oils, with certain exceptions such as tuna oil, these sh oils
are unsuitable for the use in infant formulae where high DHA and
low EPA is preferred. Some microbial oils are rich in DHA and contain
little EPA and so can be combined with microbial produced AA to t
requirements for infant formulae. For example, Martek developed
fermentation processes for Crypthecodinium cohnii to commercially
produce microbial oil rich in DHA (4050%), and in collaboration
with DSM produced microbial oils rich in AA using Mortierella alpina.
Recently DSM has purchased Martek and so Martek is now a fully
owned subsidiary of DSM. Currently, Marteks microbial oils are
added to most infant formula in the USA, and are being sold in more
than 60 countries. The use of microalgae oils as sources of fatty acids
for infant formula has been extensively reviewed elsewhere (Behrens & Kyle, 1996; Wright, Coverston, Tiedeman, & Abegglen,
2006). The company also uses a Schizochytrium species to commercially produce microalgal oil high in DHA. Lonza Group, Switzerland
is also a very important player in this eld, as it commercially produces microalgal DHA oil (4550%) using a Ulkenia species (Kiy, Rusing, & Fabritius, 2005). Commercial production processes are
currently under development in Australia, China, India, Japan, Spain,
Norway and Canada (Raghukumar, 2008).
Traditional strain improvement techniques using mutagenesis
have successfully been applied in enhancing microbial PUFA oil
productivity and developing specialty oil strains. Using these
methods, Martek improved the lipid productivity of their DHA
strain by 242% in a 6-yr period. Similarly, Shimizus group succeeded in breading mutant strains of M. alpina for the commercial
production of specialty oils rich in DGLA (C20:3n-6) and other fatty
acids, which also helped to elucidate the PUFA biosynthesis pathway in this microorganism (Sakuradani & Shimizu, 2009). Importantly, strains developed via classic methods are not considered
as GMO. We have recently developed effective methods to efciently trigger mutations and to change the FA proles of Thraustochytrid strains. Using classical strain improvement method, we are
developing modied of Thraustochytrid strains with unique features such as elevated PUFA productivity, the capability to produce
EPA, and the capable of utilising cheap carbon sources such as
glycerol.
We believe that the traditional methods will continue to play a
vital role in various aspects of strain improvement regarding
specialty oils and productivity improvement. However, many

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traditional methods are time-consuming and labour-intensive. As

microbial genomes can be sequenced in a straightforward way
using modern sequencing technologies, genetic pathway engineering has become a robust technique for target-specic microbial
strain development. Recently, two transformation systems have
been developed for genetic manipulation of M. alpina. One is based
on using a gene gun, and the other one is based on using Agrobaterium tumefaciens as gene delivery system. The latter resulted in
remarkably enhanced transformation efciency and transgene stability. Using these methods, the elongase gene (EL2), a speed limit
enzyme in AA biosynthesis, was overexpressed in M. alpina, which
enhanced AA productivity. These genetic tools enable the development of M. alpina strains for specialty oils rich in n-9, n-7, n-6, n-4,
n-3 or n-1 series PUFAs (Sakuradani & Shimizu, 2009). Through
PUFA pathway engineering, DuPont has recently developed a yeast
strain for commercial production of oils rich in EPA (55% EPA) and
the company is hoping to develop various strains for the production of DHA, ARA or GLA (Sharpe et al., 2009). Additionally, using
a similar strategy, Microbia, Inc. recently has genetically engineered this oleaginous yeast into a carotenoid host for commercial
antioxidant production (Grenfell-Lee, 2009).
At ONC, an isolation programme targeting Thraustochytrids
from various Atlantic locations in Canada has been carried out.
We have isolated and characterised a new strain ONC-T18 of
Thraustochytrium sp., evolutionally most closely related to T. striatum T91-6. Our strain can not only produce up to 80% oil by dry
weight, of which more than 31% are DHA, but also signicant
amounts of carotenoid such as -carotene, astaxanthin, zeaxanthin
and canthaxanthin. When compared to commercial DHA-producting microbial strains, our strain has one of the highest DHA productivities, and has considerable potential as a commercial source of
high DHA oil (Burja, Radianingtyas, Windust, & Barrow, 2006).
We and others have explored Thraustochytrids as producers of
PUFA microalgal oils. Since Thraustochytrids are effective high
yielding lipid producers, efforts are being made to develop transformation systems aimed at engineer strains to improve their
DHA productivity and to enable them to produce high levels of
EPA and carotenoids. Recently, two genetic transformation systems
have been developed for a related Schizochytrium sp. One is based
on using electroporation and the other on using gene gun DNA
delivery method (Ono, Aki, & Kawamoto, 2006; Roessler, Matthews, Ramseier, & Metz, 2007). The PUFA biosynthetic pathways
in this Schizochytrium sp. were elucidated by successfully knocking
out PKS and FAS genes. Moreover, compared with the difculty of
developing oleaginous yeast into EPA or carotenoid production
hosts through multiple steps of genetic engineering, Thraustochytrid strains have natural PUFA and carotenoid biosynthetic pathways with high activity, making them ideal hosts for channeling
metabolic uxes through genetic engineering. Overexpression of
the carotene synthase gene indeed increased carotenoid productivity in a Schizochytrium sp. by about 16 times (Weaver, Metz, Kuner,
& Overton, 2006). Enhancing the levels of antioxidants such as
carotenoids in PUFA producing Thraustochytrid strains may also
simplify the microalgal oil extraction procedures and extend the
shelf life of microbial oils rich in PUFA. However, transformation
systems for other Thraustochytrid strains have not been demonstrated. We have been developing transformation systems for our
novel Thraustochytrid strain and have had some success using a
gene gun.

4. Stabilisation and delivery of omega-3 fatty acid rich oils into

foods
EPA and DHA are unstable and will oxidise quickly leading to
the formation of unpleasant smelling and tasting aldehydes and

ketones. Various types of antioxidants have been used for the

chemical stabilisation of EPA and DHA, but these are not adequate
to enable sensory stabilisation of these oils for addition to many
foods and beverages. The use of antioxidants to improve the stability of omega-3 containing oils has been reviewed recently (Shahidi
& Zhong, 2010a, 2010b). A large body of work has been done to develop improved antioxidant systems for stabilisation of omega-3
oils in food products. Mechanisms of oxidation are complex and
vary depending upon the food matrix. This is particularly true for
lipid emulsions, where antioxidants are normally exposed directly
to the food matrix components. The mechanisms of lipid oxidation
in food systems has recently been reviewed (Waraho, McClements,
& Decker, 2011) as have methods for evaluating the efcacy of
antioxidants in these systems (Mickael et al., 2010). Lipid emulsions have seen some success in beverage products in particular,
such as milk, where emulsication is a key step in the product process. Even in short shelf-life products such as milk, salad dressing
and yoghurt it is necessary not only to optimise antioxidant combinations for each specic application (Let, Jacobsen, & Meyer,
2007; Nielsen, Klein, & Jacobsen, 2009), but additives, such as chelators are also often required (Hu, McClements, & Decker, 2004). To
ensure adequate protection of a wide range of food and beverage
products against oxidation microencapsulation is required.
Microencapsulation is a unique process that has been used not
only to convert liquids to solids, but also to add functionalities or
improved oxidative stability to ingredients. Its advantages include:
masking the unpleasant avours and odours of the microencapsulated ingredients; protecting ingredients from oxidation and other
unwanted reactions and therefore extending shelf life; controlled
release of ingredients to improve the functionality of food additives and expanding the application range of food ingredients
(Desai & Park, 2005; Gharsallaoui, Roudaut, Chambin, Voilley, &
Saurel, 2007; Gibbs, Kermasha, Alli, & Mulligan, 1999; Gouin,
2004; Madene, Jacquot, Scher, & Desobry, 2006; Thies, 2001).
Microencapsulation techniques can be divided into three classes:
physical processes such as spray drying, spray chilling/coating,
extrusion or uidised bed coating; chemical processes such as
molecular inclusion or interfacial polymerisation; and physicochemical techniques such as single- or multi-core coacervation
and liposome encapsulation. In recent years, microencapsulation
has increasingly become an important technology for the delivery
of numerous nutraceuticals and avour ingredients into food matrix. Table 1 summarises omega-3 concentration and delivery technologies applied to food and beverage applications.
Spray drying is a widely used microencapsulation technique in
the food industry (Drusch, Benedetti, Scampicchio, & Mannino,
2008; Jafari, Assadpoor, He, & Bhandari, 2008). It involves conversion
of liquid oils and avours, in the form of emulsions, into dry powders
using proteins and/or carbohydrate as the matrix materials. However, this technology has limitations that the oil loading level is
low, the level of surface (extractable) oil is high, and air inclusion
in the emulsication process is difcult to avoid leading to particle
ination during the spray drying stage (Keogh et al., 2001). A leading
commercialised spray-dried emulsion product was developed by a
group at CSIRO in Australia. Additional stabilisation and oil loading
was achieved through the use of a casein-based Maillard reaction
product (Kosaraju, Weerakkody, & Augustin, 2009).
Complex coacervation technology was rst developed using
gelatin and gum arabic. This technology was used to make carbonless paper for the printing industry several decades ago. At ONC we
have developed a novel variation of this technology that involves
the addition of a controlled agglomeration step and the formation
of an outer shell that surrounds the agglomerations. This technology has been scaled to manufacturing and is widely used commercially to stabilise and delivery omega-3 oils into foods and
beverages (Barrow, Nolan, & Jin, 2007). The key features of the

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Fig. 3. SEM pictures of the cross-section of a whey gum microcapsule with a

magnication of X2000.

powder products from this technology are its high oil loading level,
normally at least 60%, and its low level of surface oil, normally less
than 0.1%. These levels compare favourably with spray dried powder products which have an oil loading level of typically less than
30% and a surface oil level between 0.2% and 1%. A high loading level means that less shell material is needed to deliver EPA and DHA
so that cost and sensory impact on food are reduced to a minimal
level. The surface oil is essentially unprotected oil and is vulnerable
to rapid oxidation and sensory deterioration. A low level of surface
oil is critical to maintaining the sensory properties of the product
during both powder storage and addition and storage of the fortied food products.
A major disadvantage of complex coacervation technology is
the limitation in the selection of the shell materials. Gelatin has
been the material of the choice for use in complex coacervation
due to its unique gelation properties. However, gelatin has some
limitations, including it not being vegetarian and kosher versions
being relatively expensive. Also, gelatin does not have an ideal

sensory prole for all applications. Alternative shell materials are

needed in some food and beverage applications. In recent years,
a number of studies have been reported on complex coacervation
systems using non-gelatin material such as globular proteins with
anionic polysaccharides, for example, b-lactoglobulin, bovine
serum albumin, egg albumin, soy proteins, pea proteins and whey
proteins with gum Arabic, carrageenan and pectin (De Kruif,
Weinbreck, & De Vries, 2004).
We have developed novel whey protein-gum arabic sh oil systems as a viable option for the commercial production of non-gelatin vegetarian microcapsules (Yan, Zhang, Jin, & Barrow, 2008;
Zhang, Yan, May, & Barrow, 2009). These microcapsules were prepared using a combination of complex coacervation and thermal
crosslinking processes, including spray drying to form a free-owing powder. Chemical or enzymatic crosslinking of whey protein
microcapsules was not required, as it is for gelatin. Instead, a thermal crosslinking process was developed to solidify the microcapsules and prevent their dissolution, essentially by coagulation of
the whey protein. Three types of the microcapsules were produced
with different processing sequences resulting in powder products
with distinguishable features. An example of the cross-section of
one type of these microcapsules is shown in Fig. 3. We found that
processing parameters determined the nal characteristics of the
three types of microcapsules, such as morphology, surface properties and particle integrity, as well as their performances in sensory
evaluations. Sensory stability was similar for each of the three
types of microcapsules, and was determined using an in-house
sensory panel evaluation over a 10 month period.
It is also worth noting that these whey protein and gum arabic
microcapsules performed extremely well in a shelf life study in 2%
ultra high UHT milk, as compared to gelatin microcapsules. UHT
milk treatment involves an ultra-high temperature process.
Though it only lasts a few seconds, this type of heat can be very
destructive to certain types of microcapsules. The fact that the
whey protein and gum arabic microcapsules survived this severe
treatment indicates an ability to resist the negative impact of heat
on the structure and the quality of the microcapsules themselves.
It suggests that since these microcapsules had undergone a

Table 1
Some current technologies for omega-3 concentration and delivery into food and beverage applications.
Enzymatic
processing
technology

Purpose

Lipase used

Product information

Oil type

Producer

EE or FFA to TG conversion

Candida Antarctica lipase B

(CALB)
CALB
Various

Food grade concentrate

Sardine/Anchovy

Food grade concentrate

Structured Lipids. 3060% EPA,
DPA or DHA
Up to 50% increase in EPA and
DHA levels.
EPA/DHA ratio of 22:1

Sardine/Anchovy
High-laurate canola
oil
Sardine/Anchovy
and Tuna oils.
Sardine/Anchovy oil.

Particle size

Shelf life

Ocean Nutrition
Canada
Pronova
Hamam and Shahidi
(2006)
Ocean Nutrition
Canada
Fernandez-Lorente
et al. (2011)
Producer

4585190 lm

612 months

Clover/Nu-Mega

100% through 20 mesh, >85% 40

mesh, <15% 100 mesh

Surface oil <0.4%,

1 year at <15 C
2 years at 510 C

DSM/Martek

EE or FFA to TG conversion
EPA, DPA and DHA enriched
structured lipids.
Partial Concentration of EPA
and DHA.
Separation of EPA and DHA
Delivery technology

Shell materials

Spray-dried
emulsion
Spray-dried
emulsion
Spray-dried
emulsion
Spray-dried
emulsion
Gravity ow dry
blending

Caseinate, dextrose, glucose

syrup
Cornstarch, gelatin, sucrose

Complex
coacervation

Thermomycetes
lanuginosus lipase
Rhizomucor miehei lipase
(RML)
Loading of EPA/DHA (mg)
per gram of powder
69143, Tuna oil
90, Algal oil

Carbohydrate, protein,
antioxidant
Modied starch, soy protein

170195, Sardine oil

100, Menhaden oil

100 lm

Maltodextrin

65155, Sardine oil

>80% through 20 mesh

612 months at 5
10 C
2 years

Gelatin, polyphosphate

140180, Sardine oil

60 lm, 97% through 100 mesh

One year at 4 C

Lipid Nutrition
(Loders Croklaan)
National Starch/
Omega Protein
Nutri
Pharmaceuticals
Research Inc.
Ocean Nutrition
Canada

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thermal crosslink process when they were manufactured, some

additional heat treatment can enhance rather than decrease their
structural stability, instead of thermally damaging the shell material. Also, we anticipate that the good performance of these new
microcapsules is partly because whey protein is more sensorially
compatible with milk than is gelatin.

5. Future directions
Lipases are successfully used industrially for omega-3 partial
concentration and re-esterication. Improving the EPA and DHA
selectivity of lipases would broaden the range of concentrates that
could be made using enzymatic processing. A major future target is
the development of EPA and DHA selective lipases. Low cost novel
microbial sources of DHA and EPA is another major research target
within the omega-3 eld. We and others are focused on the metabolic engineering of Thraustochytrid strains used, to manipulate
the concentration and ratios of EPA and DHA for food, supplement
and pharmaceutical applications.
Liquid and spray-dried emulsions are being used successfully in
some food and beverage applications for the delivery of omega-3
oils. Complex coacervation has also been shown to be a commercially useful method. Most future work is focused on improving
oil stability to enable incorporation into a wider range of food
products. Also, decreasing the cost of microencapsulation technologies is important to enable commercial success in low cost per
serving food products. We believe that novel and mild processing
technologies, new microencapsulation technologies, and alternative sources of EPA and DHA to sh oil are all important areas of
ongoing and future research and development in the area of omega-3 oils.

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