Ulvan and its biological activity

Presented at 1st
International Conference on Biodiversity, Food Savety and Health, Yogyakarta 22-23th
Nopember 2016
1
1
1. INTRODUCTION
Potential green seaweed as a source of
active compounds is very high, one of which is
ulva that produce polysaccharide sulfate is
ulvan. Ulvan was produced by green algae
belonging to Ulvales such as Ulva and
Enteromorpha sp.), the seaweed that growing
rapidly, especially in the eutrophicated coastal
areas. Various research reports on bioactivity
ulvan shows that this compound is potential to
be used as a source of natural ingredients for
humans. Thus the need for reviews farther what
it ulvan, seaweed producing, as well as the
potential biological activity and ulvan extraction
of the seaweed.
As one of the countries with the longest
coastline in the world, Indonesia is a potential
source for the production of green seaweed
including Ulva sp. Indonesia have for many
years been a major producer of seaweed in the
world, especially for red algae Capaphycus sp
and Gracilaria sp.
Nevertheless the use of green seaweed
such as Ulva is still low. The information about
seaweed producer ulvan in Indonesia and its
utility are still not widely reported, so this paper
will try to dig up this information.
The aim of this paper is to review what is
ulvan, extraction, functional properties,
biological activity and potential sources of
seaweed producing ulvan in Indonesia.
Utilization of seaweed producing ulvan in
Indonesia and how to maintain the sustainability
of its resources also discussed in this paper.
2. Ulvan and Its extraction
Ulvan is a sulfated polisaccharides that
containing in green seaweed Ulvales, such as
Ulva sp and Entheromorpha sp. It is mainly
consisted of disaccharide repeating sequences
composed of sulfated rhamnose and glucuronic
acid, iduronic acid, or xylose (Robic et al,
2009). The two major repeating disaccharides
are aldobiuronic acids for type A ulvan,
ulvanobiuronic acid 3-sulfate (A3s) for type B
ulvan, and ulvanobiuronic acid 3-sulfate (B3s)
(Fig. 1). Partially sulfated xylose residues at O-2
Ulvan from Green Seaweed and its Biological Activity :
a review
Subaryono1)
1). Research Centre for Marine and Fisheries Product Competitiveness and Biotechnology
Jl. KS Tubun Petamburan VI, Tanah Abang, Jakarta Indonesia
Email: yono_ipn@yahoo.co.id
Abstract
A review about ulvan derived from green seaweed and its biological activity has been carried out. This
paper. This paper review of what is ulvan, extraction, functional properties, biological activity and potential
sources ulvan seaweed in Indonesia. Also discussed in this paper the use of seaweed producing ulvan in
Indonesia and how to maintain the sustainability of its resources. Ulvan is a sulfated polysaccharides that
water-soluble can represents about 8−29% of the algae dry weight. The extraction of ulvan was conducted in
water with or without acid. The biological activity of ulvan was reported as antioxidant, anticoagulant,
antiviral, and anticancer. Research on ulvan from Indonesian seaweed is still very limited. Although, the
distribution of seaweed species producer ulvan in Indonesia was spread out. Ulva sp.
were found in some areas, especially in the south waters of the Java, Sumatra and Nusa Tenggara island.
Ulva have been used as chips seaweed on the south coast of Java, especially in Gunung Kidul Yogyakarta.
Management and collection techniques of natural seaweed needs to be improved to maintain the
sustainability seaweed resources. Ulva seaweed farming also needs to be done, so that no longer rely on
natural products.
Key words: biological activity, marine resource, seaweed, ulva, ulvan,

Presented at 1st
Nopember 2016
2
2
can also occur in place of uronic acids. In
addition, glucuronic acid can branch at O-2 of
rhamnose 3-sulfate (Ray and Lahaye 1995). Low
proportions of galactose, glucose, mannose, and
protein are also generally found in ulvan.
Ulvan content in seaweed generally reach
8-29% of the seaweed dry weight. Ulvan
represents the major biopolymeric fraction of the
cell wall of Ulva, and it is believed to control the
osmolar stability of the cell and to maintain a
suitable environment and protection of the cell
(Paradosi et al, 2002). As is often the case for
cell wall polysaccharides, also ulvans are
complexed with protein moieties in their native
state.
To date, it is difficult to accurately
determine the sugar composition of ulvan
because of refarctory to acid hydrolysis of its
aldobiouronic linkage and iduronic acid is
partially destroyed during acid hydrolysis
(Lahaye and Robic, 2007). Ulvan compounds
consisting of approximately 20% rhamnose,
12% clucuronic acid, and 12% sulfat. other
simple sugars that compose the ulvan were
approximately 4.4 % xylose, 3% iduronic acid,
2% glucose and 1.5% galactose (Robic et al,
2009).
The extraction of ulvan from green
seaweed can be conducted using some methods.
Paradossi et al (1999) extracted ulvan from Ulva
rigida using Mili Q water. The seaweed was
dispersed in 1 l of MilliQ water, and was heated
and stirred for 1 h at 90–100°C. After cooling, a
viscous opaque supernatant was decanted from a
solid residue. The pellet was re-extracted as
above and the supernatants were pooled and
precipitated in 1 vol. of a mixture water:ethanol
1:1 (v:v). The supernatant had a gellish looking,
depending on the amount of algal raw material
dispersed. Yhis method get a yield of 7% of dry
seaweed.
Robic et al (2009) extracted ulvan from
Ulva armoricana and Ulva rotundata using
solution of sodium oxalat. Ulvans were extracted
in 0.05 M sodium oxalate in deionized water
(dry solid content 6.8% w/w) by refluxing for 2
h at 85°C. The suspension was filtered and the
residues were extracted by deionized water (dry
initial solid content 6.8% w/w) for 1 h at room
temperature and filtered. The combined extracts
were concentrated by ultrafiltration using
membrane with Mw 10 kDa (Amicon) and
diafiltered against five volumes of deionized
water and then freeze-dried.
h
Figure 1. Structure of repeating disaccharides in ulvan from Ulva sp.
a. A3s b. B3s
c. U3s d. U2’s3s
Source: Robic et al, 2009
a b
c d

Presented at 1st
Nopember 2016
3
3
Kanno et al. (2014) extracted Ulva
pertusa using hot water, 0.05 M sulfuric acid,
0.1 M sodium oxalat and 1 mg cellulose in
phosphate buffer. Ten grams of dried Ulva in
300 ml extracting solvent was ground with an
electric mixer. The mixture was stirred at the
temperature of 20-1000
C. The residue was
filtered through nonwoven fabric and filter paper
under reduced pressure. The filtrate was
dialyzed against a solution of 0.1 mol/L of
CaCl2 to remove precipitated nonsulfated uronic
polysaccharides. The solution was subsequently
dialyzed against pure water. The dialyzed
solution was subjected to diethylaminoethyl-
cellulose column chromato-graphy with a NaCl
gradient. The fraction containing ulvan was
dialyzed against pure water. This was followed
by freeze-drying that yielded ulvan as a beige-
colored solid with yield as in table 1.
Jaulneau et al. (2010) extracted ulvan
from Ulva spp. using hot water with high
pressure. As much as 100 g of water-washed
algae were grinded and were autoclaved for 2
hours at 900
C (1.97 atm) in 1 l distilled water
and the resulting extract was filtered through a
nylon mesh (80 μm porosity) on a intered glass
funnel (G2 porosity). The filtrate was
lyophilized. In this extract, the compounds of
high molecular mass were precipitated with 2.5
volumes of ethanol for 48 hours at −200
C. The
supernatant and pellet were separated by
filtration and lyophilized.
Rahimi et al (2016) extracted ulvan using
ultrasound-assisted extraction, and get about
8.36% yield. The milled sample (20 g) was
treated with 80 % ethanol (200 mL) with
constant mechanical stirring overnight at room
temperature. The supernatants were then
clarified by centrifugation at 100
C and 6000×g
for 10 min. The sediment was rewashed with
EtOH under the same conditions, rinsed with
acetone, centrifuged at 100
C and 6000×g for
10min, and then dried at room temperature in a
fume hood. The dried samples (20 g) were
extracted with distilled water and sonicated with
an ultrasound bath (Soner, 206H, Taiwan; 53
KHz; 180 W) at the temperature of 30–1000
C,
the ratio of water to raw material 10:1 to 80:1
(mL g–1) and pH 4–9 for 10–70 min. The
extracts were centrifuged at 100
C and 10,000
rpm for 10 min, and the supernatants were
collected. The supernatants were concentrated
by evaporation under reduced pressure at 600
C
to approximately 50 mL. EtOH (99 %) was
added into the supernatant to obtain a final
EtOH concentration of 70 % and then kept at
40
C overnight. The precipitate was obtained by
centrifugation at 10 °C and 9400×g for 10 min,
washed with EtOH (99 %), which was followed
by acetone, and then dried at room temperature.
2. Biological activity of Ulvan
Kaeffer et al (1999) studied the citotoxic
activity of ulvan on normal and cancer cell. The
effects of ulvan were examined on the adhesion,
proliferation and differentiation of normal or
tumoral colonic epithelial cells cultured in
conventional or rotating culture conditions. It
has showed that sulfated ulvans (MW < 5,000)
inhibited the Caco-2 cell proliferation/
differentiation program by inducing a low cell
reactivity to Ulex europeaus-1 lectins in defined
or serum-supplemented media but were inactive
on normal colonocytes. In conclusion, this
compound could be a source of oligosaccharides
with a bioactivity, a cytotoxicity or a
cytostaticity targeted to normal or cancerous
epithelial cells.
Morelli et al (2016) also found that ulvan
is potential used as in-situ hydrogel forming
Table 1. Extraction methods and the yield of ulvan obtained
Reagents Temperature
(o
C)
Time (hours) Yield (%)
Hot water 100 2 3.71
0.05M Sulfuric acid 20 24 3.49
0.1M Sodium oxalate 20 24 3.06
1mg Of cellulose in pH 6.4
phosphate buffer
45 144 0.98
Source: Kanno et al, 2014

Presented at 1st
Nopember 2016
4
4
systems for biomedical applications. The
preparation of ulvan-based hydrogels displaying
thermogelling behaviour. ulvan was provided
with thermogelling properties by grafting
poly(N-isopropylacrylamide) chains onto its
backbone as thermosensitive component. The
thermogelling properties of the copolymer were
investigated by thermal and rheological
analyses. Sol–gel transition of the copolymer
was found to occur at 30–310
C.
Fraction of sulfated polisaccharides from
Ulva Ulva pertusa was also has
immunostimulating activity (Tabarsa et al.,
2012). Methanol was used as a sulfate acceptor
for the removal of sulfates from the
polysaccharides. When desulfation was carried
out at 120 °C, the sulfates were removed upto
90.1% from the F2 fraction without considerable
backbone degradation. The GC–MS analysis as
well as NMR spectra revealed that the backbone
of the polysaccharides was mainly composed of
α-(1→4)-l-rhamnopyranosyl, β-(1→4)-d-glucu-
ronosyl, β-(1→2)-l-rhamnopyranosyl, and β-
(1→4)-d-xylopyranosyl residues with branches
at O-2 position of rhamnose. In line with the
study, Rahimi et al. (2016) also found that
ulvans have macrophage-stimulating capacity,
indicating their potential value for health
Ulvan also used in bone tissue
engineering and as polymeric component of
bone cement (Dash et al, 2014; Barros et al,
2013). Successful modification of UV cross-
linked ulvan scaffolds was revealed by 1
H NMR.
The presence of the mineral formation was
evidenced by Raman spectroscopy and XRD
techniques. Investigations of the morphology
confirmed the homogeneous mineralization
using ALP. The MC3T3 cell activity clearly
showed that the mineralization of the
biofunctionalized ulvan scaffolds was effective
in improving the cellular activity. Mechanical
and in vitro bioactivity tests indicate that the
inclusion of CMU in the cement formulation
enhances its mechanical performance, generates
non-cytotoxic cements and induces the diffusion
of Ca and/or P-based moieties from the surface
to the bulk of the cements.
Rahimi et al. (2016) stated that ulvan was
one of the components that has good antioxidant
activity. The extracted polysaccharides exhibited
appreciable 2,2-diphenylpicrylhydrazyl (DPPH)
radical scavenging and reducing power as well
as macrophage-stimulating capacity indicating
their potential value for health and food
industry. Qi et al. (2005) have prepared ulvans
of different molecular weights from Ulva
pertusa using sulfur trioxide/N,N-
dimethyflormamide (SO3-DMF) in formamide,
and their antioxidant activities investigated. The
results showed that low molecular weight ulvans
have a strong antioxidant activity. Ulvan may
also modulate lipid metabolism in rats and mice.
Similar to that report, Costa et al. (2010) also
reported the antioxidant activities of green
seaweed extract.
Govindan et al. (2012) studied
anticoagulant activity of ulvan, and showed that
this compound has anticoagulant activities. The
anticoagulant activity of the purified
polysaccharide was studied by prolongation of
APTT Ex Vivo using rat model. The sulphated
polysaccharide prolonged the APTT in a dose
dependent manner and can be developed as a
promising anticoagulant agent.
The antiviral activity of ulvan was studied
by Aguilar-Briseño et al. (2015). The ulvan
antiviral activity was tested using syncytia
formation, exhibiting an IC50 of 0.1 μg/mL;
ulvan had a better anti cell-cell spread effect
than that previously shown for fucoidan, and
inhibited cell-cell fusion via a direct effect on
the F0 protein. The mixture of ulvan and
fucoidan showed a greater anti-spread effect
than SPs alone, but ulvan antagonizes the effect
of fucoidan on the viral attachment/entry. Both
SPs may be promising antivirals against
paramyxovirus infection but their mixture has no
clear synergistic advantage
3. Seaweed producing ulvan in Indonesia and
its utilization.
Indonesia as a country with a vast beach
has a huge potency as a producer of seaweed,
including species that contain ulvan. some types
such as Ulva sp. and Enteromorpha sp. found in
some areas in Indonesia. Ulva commonly found
in waters with a rocky bottom, and is widely
spread in Indonesian waters. This seaweed
corals settled or associated with other types.
Ulvan many found and dominate marine life in
Ambon (Litaay, 2014). ulva also found in the
waters of Nusa Tenggara, Bali waters, and

Presented at 1st
Nopember 2016
5
5
along the south coast of the Java and Sumatra
island.
Ulva not yet cultivated and still harvested
from the wild. In Gunung Kidul, this seaweed
has been utilized as a seaweed chips (Figure 2).
Ulva lactuca is a type that is used in these chips
product, and nutritional value test showed this
product to contain 18.7% water, 14.9% protein,
12:04% fat, 50.6% carbohydrates, and 0.2%
fiber (Anonimous, 2016).
Because ulva not been cultivated, so to
meet the needs of this seaweed at Gunung Kidul,
this seaweed should be drawn from the other
waters such as Pemeungpeuk of West Java, and
Lampung. To keep the seaweed resource
conservation, the management of harvesting
should be regulated so that still leaves of
seaweed in order to continue to grow. On the
other hands, to meet the needs of increasingly
high seaweed, the cultivation need to be
developed. With the cultivation of seaweed,
needs of seaweed are not fully rely on the
natural supply.
Figure 2. Ulva chips traded on the tourism
beach location at Gunung Kidul
CONCLUSSIONS
Ulvan is a sulfated polysaccharides that
water-soluble can represents about 8−29% of the
algae dry weight. The extraction of ulvan was
conducted in water with or without acid, or
using high temperautre with pressure. The
biological activity of ulvan was reported as
antioxidant, anticoagulant, antiviral, and
anticancer. Research on ulvan from Indonesian
seaweed is still very limited. Although, the
distribution of segaweed species producer ulvan
in Indonesia was spread out. Ulva sp. were
found in some areas, especially in Ambon, Bali,
the south waters of the Java, Sumatra and Nusa
Tenggara island. Ulva have been used as chips
seaweed on the south coast of Java, especially in
Gunung Kidul Yogyakarta.
ACKNOWLEDGEMENT
Our thanks to the PUI team of R & D
centers for marine and fisheries products
competitiveness and biotechnology Jakarta that
has funded me to participate in this conference
REFERENCES
Aguilar-Briseño JA et al. 2015. Sulphated
polysaccharides from Ulva clathrata
and Cladosiphon okamuranus
seaweeds both inhibit viral
attachment/entry and cell-cell fusion,
in NDV infection. Mar Drugs
13(2):697-712.
Anonimous. 2016. Ulva Chips and benefits.
http://www.biodiversitywarriors.org/ke
ripik-selada-laut-unik-dan-menyehat-
kan.html. acessed Nopember 6th 2016.
Barrosa, AAA., Alvesa, A., Nunesc, C.,
Coimbrac, MA., Piresa, RA., and
Reis, RL. 2013. Carboxymethylation
of ulvan and chitosan and their use as
polymeric components of bone
cements. Acta Biomaterialia 9
(11):9086–9097.
Costa, LS. et al. 2010. Biological activities of
sulfated polysaccharides from tropical
seaweeds. Biomedicine &
Pharmacotherapy 64: 21–28.
Dash, M., Samal, SK., Bartoli, C., Morelli, A.,
Smet, PF., Dubruel P., and Chiellini, F.
2014. Biofunctionalization of Ulvan
Scaffolds for Bone Tissue Engineer-
ing. ACS Appl. Mater. Interfaces 6 (5):
3211–3218.
Jaulneau, V. 2010. Ulvan, a Sulfated
Polysaccharide from Green Algae,

Presented at 1st
Nopember 2016
6
6
Activates Plant Immunity through the
Jasmonic Acid Signaling Pathway.
Journal of Biomedicine and
Biotechnology Vol. 2010, Article ID
525291, 11 pages.
Kaeffer, B., Bénard, D. Lahaye, M., Blottière,
HM., and Cherbut, C. 1999.Biological
properties of Ulvan, a New Source of
Green Seaweed Sulfated Polysac-
charides, on Cultured Normal and
Cancerous Colonic Epithelial Cells.
Planta Med 65(6): 527-531.
Kanno, K., Fujita, Y., Honda, S. Takahashi, S
and Kato, S. 2014. Urethane foam of
sulfated polysaccharide ulvan derived
from green-tide-forming chlorophyta:
synthesis and application in the
removal of heavy metal ions from
aqueous solutions. Polymer Journal
1:1–6.
Litaay, C. 2014. Distribution and diversity of
macroalgae communities in the
Ambon Bay. Jurnal Ilmu dan
Tekonologi Kelautan Tropis 6 (1):
131-142.
Morelli, A. Betti, M., Puppi, D., and Chiellini.
2016. Design, preparation and
characterization of ulvan based
thermosensitive hydrogels. Carbohyd-
rate Polymers. 2016. 136:1108–1117.
Paradossi, G., Cavalieri, F., and Chiessi, E.
2002. A Conformational Study on the
Algal Polysaccharide Ulvan.
Macromolecules 35: 6404-6411.
Paradossi, G., Cavalieri, F., Pizzoferrato, L. And
Liquori, AM. 1999. A physico-
chemical study on the polysaccharide
ulvan from hot water extraction of the
macroalga Ul6a. International Journal
of Biological Macromolecules 25:
309–315
Rahimi, F., Tabarsa, M. And Rezaei, M. 2016.
Ulvan from green algae Ulva
intestinalis: optimization of
ultrasound-assisted extraction and
antioxidant activity. Journal of Applied
Phycology 28, (5): 2979–2990.
Ray, B., and Lahaye, M. 1995. Cell-wall
polysaccharides from the marine green
alga Ulva rigida (Ulvales,
Chlorophyta). Extraction and chemical
composition. Carbohydrate Research
274:251–261
Robic, A., Bertrand, D., Sassi JF., Lerat Y, and
Lahaye, M. 2009. Determination of the
chemical composition of ulvan, a cell
wall polysaccharide from Ulva spp.
(Ulvales, Chlorophyta) by FT-IR and
chemometrics. Journal Applied
Phycocolloid 21:451–456.
Shonima, G M, Thomas, J., Pratheesh P and
Kurup, GM. 2012. Ex vivo
anticoagulant activity of the
polysaccharides isolated from Ulva
fasciata. Int. J. LifeSc. Bt & Pharm.
Res. 1 (3): 194-197.
Tabarsa, M. Lee, SJ., You, SG. 2012. Structural
analysis of immunostimulating
sulfated polysaccharides from Ulva
pertusa. Carbohydrate Research
Volume 361:141–147.
Qi et al. 2005. Antioxidant activity of different
sulfate content derivatives of
polysaccharide extracted from Ulva
pertusa (Chlorophyta) in vitro.
International Journal of Biological
Macromolecules 37 (4): 195–199.

Ulvan and its biological activity

Recommandé

Recommandé

Contenu connexe

Tendances

Tendances (20)

Similaire à Ulvan and its biological activity

Similaire à Ulvan and its biological activity (20)

Dernier

Dernier (20)

Ulvan and its biological activity