Food Sci. Biotechnol.
19(6): 1551-1557 (2010)
DOI 10.1007/s10068-010-0220-x
RESEARCH ARTICLE
Enzymatic Extracts from Edible Red Algae, Porphyra tenera, and
Their Antioxidant, Anti-acetylcholinesterase, and Anti-inflammatory
Activities
Mahinda Senevirathne, Chang-Bum Ahn, and Jae-Young Je
Received: 2 July 2010 / Revised: 3 August 2010 / Accepted: 13 August 2010 / Published Online: 31 December 2010
© KoSFoST and Springer 2010
Abstract Enzymatic extracts from Porphyra tenera were
prepared using 4 proteases (Protamex, Neutrase, Flavourzyme,
and Alcalase) and 7 carbohydrases (AMG, Celluclast,
Dextrozyme, Maltogenase, Termamyl, Promozyme, and
Viscozyme), and biological activities of the enzymatic
extracts from P. tenera were determined as antioxidant,
anti-acetylcholinestrase (AChE), and anti-inflammation.
The Alcalase and Maltogenase extracts showed higher 1,1-
diphenyl-2-picrylhydrozyl (DPPH) scavenging activities
compared to the other extracts. At the 2.5 mg/mL, 94.38%
(Alacalase extracts), and 80.13% (Maltogenase extracts)
scavenging capacities were observed. The Alcalase and
Maltogenase extracts were also showed strong reducing
power, ferrous ion chelating, and hydrogen peroxide (H2O2)
scavenging capacities. In addition, 2 enzymatic extracts
effectively protected hydroxyl radical-induced DNA damage.
In the case of AChE inhibition, the Flavourzyme (99.32%
inhibition) and Viscozyme extracts (82.68% inhibition)
were observed. All enzymatic extracts showed no cytotoxicity
in RAW264.7 macrophages, and all enzymatic extracts
effectively inhibited lipopolysaccharides (LPS)-induced
nitric oxide (NO) production in RAW264.7 macrophages.
These results suggest that the enzymatic extracts from P.
tenera would be useful as an ingredient for functional
foods.
Keywords: Porphyra tenera, enzymatic extract,
acetylcholinesterase, antioxidant, anti-inflammatory activity
Mahinda Senevirathne, Chang-Bum Ahn, Jae-Young Je ( )
School of Food Technology and Nutrition, Chonnam National University,
Yeosu, Jeonnam 550-749, Korea
Tel: +82-61-659-3416; Fax: +82-61-659-3419
E-mail: jjy1915@chonnam.ac.kr
Introduction
Edible seaweeds such as Porphyra tenera have long been
included in Asian-pacific menus, which include Korea.
They provide an excellent source of bioactive compounds
such as carotenoids, dietary fibre, protein, essential fatty
acids, vitamins, and minerals (1). In addition, polysaccharides
such as sulfated galactans, xylans, mannans, and cellulose,
which constitute the main cell wall components of red
seaweeds, have attracted attention as biologically active
resources and nutraceuticals (2). Marine algae, like other
photosynthesizing plants, have antioxidant potentials to
protect them from hazardous effects, including damage to
DNA, proteins, and lipids by ultraviolet (UV) light or
oxidation by reactive oxygen species (ROS), as they are
exposed to sunlight and high-oxygen concentrations in the
sea (3). Synthetic antioxidants such as butylated hydroxy-
anisole (BHA), butylated hydroxytoluene (BHT), and tert-
butylhydroquinone (TBHQ) are known as commercially
available antioxidants. However, they have been reported
to be carcinogenic according to some studies (4). Hence,
researchers are focusing their attention on the development
of alternative safer antioxidants from natural sources
including seaweeds.
Dietary flavonoid intake is shown to have a huge beneficial
effect against the risk of dementia (5). The possible
mechanism put forwarded for the beneficial effects of fruits
and vegetables is the potentiation of antioxidant defenses,
including enzymatic and non-enzymatic antioxidants by
plant polyphenolic constituents and other antioxidant
nutrients (6). Furthermore, oxidative stress is associated
with membrane dysfunction, neuronal protein misfolding,
glial cell activation, and cell death, which cause normal
ageing and certain neurodegenerative diseases (7). Hence,
the most promising approach for treating Alzheimer’s
1552
disease is to enhance the acetylcholine level in the brain
using types of acetylcholinestarase (AChE) inhibitors (8).
The search for natural anti-inflammatory compounds
from oriental herbs or seaweeds and their potentials for
treating various inflammatory diseases have received
enormous attention among researchers. The activation of
macrophages plays a key role in inflammatory responses
against pathogens. Macrophages can kill or reduce the
harmful effects of pathogens by releasing various pro-
inflammatory mediators such as ROS, reactive nitrogen
species, cycloxygenase-2, and proinflammatory cytokines
(9). The overproduction of these inflammatory mediators
has been implicated in many inflammatory diseases such
as rheumatoid arthritis, atherosclerosis, chronic hepatitis,
and inflammatory brain diseases (10). Hence, the control of
activated macrophages may have therapeutic potential for
the treatment of inflammatory diseases.
The objectives of the present study were to evaluate
enzymatic extracts from P. tenera for their antioxidant and
anti-AChE activities and to assess the protective effects of
the extracts against lipopolysaccharides (LPS)-induced
nitrite production in RAW264.7 macrophage cells.
Materials and Methods
Materials The carbohydrases (AMG 300L, Celluclast
1.5L FG, Dextrozyme, Maltogenase, Promozyme, Viscozyme
L, and Termamyl) and proteases (Alcalase 2.4L FG,
Flavourzyme 500 MG, Neutrase 0.8L, and Protamex) were
purchased from Novo Co. (Bagsvaerd, Denmark). Mouse
macrophage RAW264.7 cells (ATTC No.: TIB-71) were
obtained from the American Type Culture Collection
(USA). Dulbecco’s modified Eagle’s minimal (DMEM)
essential medium, penicillin/streptomycin, fetal bovine
serum (FBS), and other materials required for culturing the
cells were purchased from Gibco BRL, Life Technologies
(USA). Bacterial LPS was purchased from Sigma-Aldrich
(USA). All other chemicals used in this study were of
analytical grade.
Proximate composition The moisture, ash, protein, and
lipid contents of Porphyra tenera were determined using
AOAC (11), while carbohydrates content was calculated
by subtracting the combined contents of moisture, lipid,
ash, and protein from 100.
Preparation of enzymatic extracts Enzymatic hydrolysis
was performed as described by Heo et al. (12). Briefly,
60 mL of buffer solution was added to 2 g of ground
sample, and then 40 mg (or 40 µL) of each enzyme was
mixed to give enzyme/substrate ratio as 500:1. Enzymatic
hydrolysis was performed for 8 hr, and then the enzymatic
M. Senevirathne et al.
extracts were clarified by centrifugation at 5,000×g for
20 min. Finally, the enzymatic extracts from P. tenera were
obtained after filtering the supernatant and were lyophilized
and stored at −20oC until use.
Determination of total polyphenolic (TPC) and total
flavonoid contents (TFC) The TPCs of the enzymatic
extracts from P. tenera were evaluated using the Folin-
Ciocalteau method as described by Singleton et al. (13).
Briefly, 40 µL of P. tenera (1 mg/mL) was mixed with 200
µL of Folin-Ciocalteau reagent and 1,160 µL of distilled
water for 3 min, followed by 600 µL of 20%(w/v) sodium
carbonate. The mixture was shaken for 2 hr at room
temperature, and a 200 µL aliquot was added to each well
of a 96-well microplate. Absorbance was measured at
the wave length of 720 nm using a microplate reader
(SpectraMax® M2/M2e; Molecular Devices Corp., CA,
USA). The concentration of phenolic compounds was
estimated using a calibration curve traced with gallic acid
as a polyphenol reference.
TFC was determined using the method described by
Meda et al. (14) with minor modifications. In brief, 5 mL
of 2%(w/v) aluminium trichloride was mixed with the
same volume of the enzymatic extracts from P. tenera.
Absorbance readings were taken at 415 nm after 10 min,
against a blank sample consisting of 5 mL of sample
solution and 5 mL of distilled water without aluminium
trichloride. The total flavonoid contents were determined
using a standard curve of quercetin (0-50 µg/mL).
DPPH scavenging assay The 1,1-diphenyl-2-picrylhydrozyl
(DPPH) radical scavenging effects of the enzymatic
extracts from P. tenera were measured according to the
method described by Blois (15) with slight modifications.
Briefly, 100 µL of DPPH solution (1.5×10−4 M) was mixed
with or without the enzymatic extracts (100 µL), after
which the mixture was incubated at room temperature for
30 min. The absorbance of the mixture was then
determined at a wave length of 517 nm.
Reducing power The reducing power of the enzymatic
extracts from P. tenera was determined using the method
described by Oyaizu (16) via the reduction of FeCl3. The
enzymatic extracts were mixed with 0.5 mL of 0.2 M
phosphate buffer (pH 6.6) and 0.5 mL of 1%(w/v) potassium
ferricyanide. The mixture was then incubated at 50oC for
20 min. Next, 0.5 mL of 10%(w/v) trichloroacetic acid was
added followed by centrifugation at 1,036×g for 10 min.
Finally, a 0.5 mL aliquot from the upper layer was mixed
with 0.5 mL of distilled water and 0.1 mL of 0.1%(w/v)
FeCl3. The absorbance was measured at a wave length of
700 nm and a higher absorbance value was regarded as
greater reducing power.
Biological Activities of Porphyra tenera
Ferrous ion chelating assay Ferrous ion chelating activity
was measured according to Singh and Rajini (17). The
enzymatic extracts from P. tenera were mixed with
0.1 mM FeCl2 and then allowed to sit at room temperature
for 30 sec. The reaction was initiated by the addition of
0.25 mM ferrozine over 10 min, and then the absorbance
was measured at 562 nm.
Hydrogen peroxide (H2O2) scavenging assay The
H2O2 sscavenging activities of the enzymatic extracts from
P. tenera were determined according to the method
described by Müller (18). Briefly, 100 µL of 0.1 M
phosphate buffer (pH 5.0) along with the enzymatic
extracts were mixed in a 96-well microplate. Next, 20 µL
of 10 mM H2O2 was added and the mixture was incubated
at 37oC for 5 min. Then, 30 µL of 1.25 mM ABTS and
30 µL of peroxidase (1 unit/mL) were added to the
mixture, which was subsequently incubated at 37oC for
10 min. The absorbance was then measured at 405 nm.
Protective effects of enzymatic extracts against hydroxyl
radical-induced DNA damage Hydroxyl radical-induced
DNA damage was evaluated using the method described
by Yeung et al. (19). A reaction was induced by placing the
following reagents (total volume, 12 µL) in an Eppendorf
tube: 0.5 µg of pBR322 DNA, 2 mM FeSO4, and various
concentrations of the enzymatic extracts. Then, the mixture
was incubated at 37oC for 30 min, followed by the addition
of 4 µL of 10 mM H2O2. Next, the mixture was subjected
to 0.8%(w/v) agarose gel electrophoresis, after which the
DNA bands (supercoiled, linear, and open circular) were
stained with ethidium bromide.
AChE inhibition assay The AChE inhibition assay was
conducted via the spectrophotometric method developed
by Ellman et al. (20). Acetylcholine chloride (AChCl) was
employed as the substrate to assay the inhibition of AChE.
The reaction mixtures contained 140 µL of 100 mM
sodium phosphate buffer (pH 8.0), 20 µL of the enzymatic
extracts, and 20 µL of AChE (0.36 U/mL), and were
incubated for 15 min at room temperature. The reactions
were then initiated via the addition of 10 µL of 0.5 mM
dithionitrobenzoicacid (DTNB) and 10 µL of 0.6 mM
AChCl. AChCl hydrolysis was monitored by the formation
of yellow 5-thio-2-nitrobenzoate anions, which resulted
from the reaction of the DTNB with thiocholine released
by the enzymatic hydrolysis of AChCl, at a wavelength of
412 nm for 15 min.
Cytotoxicity assay Cell cytotoxicity was estimated by
the thiazolyl blue tetrazolium bromide (MTT) assay, which
is a test for the normal metabolic status of cells based on
the assessment of mitochondrial activity (21). RAW264.7
1553
cells were seeded in 96-well plates at a concentration of
1.0×105 cells/mL. After 18 hr, the cells were treated with
different concentrations of the enzymatic extracts (50, 100,
250, and 500 mg/mL), and were incubated at 37oC for
another 24 hr. Thereafter, 50 mL of MTT stock solution (2
mg/mL) was added and incubated for 4 hr. Then, the
supernatants were aspirated and the formazan crystals in
each well were dissolved in 150 mL of dimethyl sulfoxide
(DMSO). Absorbance was measured by a microplate
reader at a wavelength of 540 nm.
Measurement of nitric oxide (NO) production as a
marker of inflammation The RAW264.7 cells were
cultured in 96-well plates using DMEM and were treated
with different concentrations of the enzymatic extracts for
2 hr. Cellular NO production was induced by adding 1 µg/
mL of LPS as a final concentration followed by incubation
for 24 hr. Next, 50 µL of the supernatant containing NO2−
was mixed with the same volume of Griess reagent, and
further incubated for 15 min (22). Then, the absorbance of
the mixture was measured at a wavelength of 550 nm. The
concentration of nitrite in the media was calculated using a
standard curve obtained for the sodium nitrite dissolved in
DMEM.
Statistical analysis The results are expressed as mean±
standard deviation (SD), and all statistical comparisons
were made using one-way analysis of variance (ANOVA)
followed by Duncan’s test. Differences showing a p-level
of 0.05 were considered statistically significant.
Results and Discussion
Enzymatic extraction, TPC, and TFC of the enzymatic
extracts from P. tenera The proximate composition of
raw P. tenera was analyzed using AOAC methods. On a
dry weight basis, its crude protein and crude carbohydrate
contents were 37.8 and 55.8 g/100 g, respectively (data not
shown). Therefore, we could expect that the enzymatic
hydrolysis of P. tenera using appropriate proteases and
carbohydrases would result in the production of soluble
bioactive materials. To examine this, we employed 7
carbohydrases (AMG 300L, Celluclast 1.5L FG, Dextrozyme,
Maltogenase, Promozyme, Viscozyme L, and Termamyl)
and 4 proteases (Alcalase 2.4L FG, Flavourzyme 500 MG,
Neutrase 0.8L, and Protamex) for the production of
bioactive materials from P. tenera.
A number of studies have shown that the phenolic
contents of plants are related to their antioxidant activities,
and these activities might relate to their redox properties,
which facilitate actions as reducing agents, hydrogen
donors, or singlet oxygen quenchers (22). The TPCs of the
1554 M. Senevirathne et al.

Table 1. Total polyphenolic (TPC) and total flavonoid contents

(TFC) of the enzymatic extracts from P. tenera
TPC TFC
Enzymatic extract 1)
(mg GAE /g of (mg QUE2)/g of
extract) extract)
1)
Alcalase 17.27±0.093) 1.82±0.02
Flavourzyme 15.25±0.27 0.94±0.08
Neutrase 16.11±0.04 0.95±0.04
Protamex 16.84±0.03 1.06±0.00
AMG 09.36±0.32 1.19±0.05
Celluclast 09.15±0.15 1.54±0.05
Dextrozyme 09.50±0.01 1.12±0.03
Maltogenase 09.42±0.11 1.02±0.10
Promozyme 19.52±0.26 1.25±0.06
Termamyl 09.70±0.40 1.00±0.06
Viscozyme 09.40±0.36 1.00±0.06
1)
GAE, gallic acid equivalents
2)
QUE, quercetin equivalents
3)
Values are the mean±SD of 3 determinations.

enzymatic extracts from P. tenera ranged from 9.15±0.15

to 19.52±0.26 mg GAE/g , while their TFCs ranged from
0.94±0.08 to 1.82±0.02 mg QUE/g. A higher TPC was
shown in the Promozyme extract (19.52±0.26 mg GAE/g)
while the Alcalase extract showed the highest TFC
(1.82±0.02 mg QUE/g). Compounds such as water-soluble
polysaccharides, proteins, and organic acids are simultaneously
extracted along with polyphenolic compounds during the
hydrolysis process (23). Also, the enzymatic hydrolysis of
seaweeds may increase the extractable polysaccharides
while releasing polysaccharides-attached polyphenolic
compounds to the medium. The biological activities of
polysaccharides and polyphenolic compounds from the
brown seaweed, Ecklonia cava, were previously studied
(24). The enzymatic hydrolysis process is more reliable
with respect to safety concerns regarding the use of some
organic solvents as extractants in food systems.
Antioxidant activities of the enzymatic extracts from P.
tenera The DPPH assay has been used extensively to
evaluate antioxidant activity because it can accommodate
many samples in a short period and is sensitive enough to
detect active ingredients at low concentrations (25). As
shown in Fig. 1, higher DPPH scavenging activities were
observed in the Alcalase and Maltogenase extracts, in a
dose-dependent manner. The hydrogen donating ability of
antioxidant constituents contributes to their free radical
scavenging nature (26). Siriwardhana et al. (27) reported
that components such as low molecular weight poly-
saccharides, pigments, and proteins and peptides also
influence antioxidant activity. Hence, one could expect that
the reported activities in this study might have been the
synergistic effects of all the constituents in the enzymatic
Fig. 1. Effects of the enzymatic extracts from P. tenera on
DPPH radical scavenging. A, protease extracts; B, carbohydrase
extracts. Values are the mean±SD of 3 determinations; a-cValues
with different letters indicate significant difference within the same
concentration (p<0.05).
extracts in addition to the phenolic contents reported.
Since, the Alcalase and Maltogenase extracts showed the
highest DPPH radical scavenging activities, they were
selected for further antioxidant assays. We also extracted
from P. tenera in the same manner without enzymes.
However, its scavenging activity against DPPH was lower
than those of enzymatic extracts (data not shown).
The reducing powers of the extracts are shown in Fig.
2A, while the Alcalase and Maltogenase extracts showed
absorbances of 0.7 and 0.6, respectively at the 2 mg/mL
concentration. Reducing power can be due to hydrogen-
donating ability, and is generally associated with the
presence of reductones (28). Since, the antioxidant activity
of a constituent is directly related to its reducing power,
this is a reliable method to evaluate the antioxidant
capacities of various compounds. Antioxidant constituents
affect interactions between metals and lipids via the
formation of insoluble metal complexes with ferrous ions
or the generation of steric hindrance. Ferrous chelating
activity reveals the ability of antioxidant constituents to
compete with ferrozine in the reaction medium. This
activity is measured as a decrease in the absorbance of the
red Fe2+/ferrozine complex. As shown in Fig. 2B, the
Alcalase and Maltogenase extracts showed activities of
about 55 and 70%, respectively, at the highest concentration
Biological Activities of Porphyra tenera 1555

Fig. 2. Effects of the enzymatic extracts from P. tenera on reducing power (A), ferrous ion chelating ability (B), and H2O2
scavenging (C). Values are the mean±SD of 3 determinations; a-bValues with different letters indicate significant difference within the
same concentration (p<0.05).

Fig. 3. Protective effects of the enzymatic extracts from P.

tenera on hydroxyl radical-induced DNA damage. Lane 1,
control; lane 2, DNA damage; lane 3-5, Alcalase 1.25, 2, and 4
mg/mL, respectively; lane 6-8, Maltogenase 1.25, 2, and 4 mg/mL,
respectively. OC (open circular) and SC (supercoiled) DNA

used (5.0 mg/mL). The ferrous chelating activity of

antioxidant constituents prevents oxygen-centered radical
generation and consequential oxidative damage. In addition,
ferrous chelating activity plays a significant role in
antioxidant mechanisms since it reduces the concentration
of catalysing transition metals in lipid peroxidation systems
(29). As depicted in Fig. 2C, both the Alcalase and
Maltogenase extracts showed significantly higher activities
for H2O2 scavenging in a dose-dependent fashion. The
Alcalase extract showed 85.05% activity at the highest
concentration used (4.0 mg/mL), while the Maltogenase
extract showed 75.53% scavenging activity at the same
concentration. The H2O2 is relatively unreactive as it is; Fig. 4. Acetylecholinesterase inhibitory activities of the
however, it converts into more reactive radicals such as enzymatic extracts from P. tenera. A, protease extracts; B,
carbohydrase extracts. Values are the mean±SD of 3 determinations;
hydroxyl and singlet oxygen together with superoxide a-d
Values with different letters indicate significant difference within
radicals, which initiate harmful reactions in cells. Hence, the same concentration (p<0.05).
the measurement of H2O2 scavenging activity may be a
useful method to evaluate antioxidant activity.
Conversion of the supercoiled form of plasmid DNA to the this process. Hydroxyl radicals are among the most reactive
open-circular and further linear forms is used as an index of species that can cause damage to DNA and other critical
DNA damage (30). The present study demonstrated that H2O2 cellular components. In line with this notion, catalase and
caused strand breakage in pBR322 supercoiled DNA in the hydroxyl radical scavengers may be effective in blocking the
presence of Fe2+ via the formation of hydroxyl radicals. H2O2 strand breakage of DNA. As shown in Fig. 3, the Alcalase and
and Fe2+-mediated DNA strand breakage are ameliorated by Maltogenase extracts showed strong DNA damage inhibitory
transition metal chelators, antioxidants, radical scavengers, activities in dose-dependent patterns via one or more of the
and catalase enzymes, suggesting the involvement of ROS in antioxidant mechanisms mentioned above.
1556
Fig. 5. Effects of the enzymatic extracts from P. tenera on the
viability of RAW264.7 cells. A, protease extracts; B, carbohydrase
extracts; Values are the mean±SD of 3 determinations and are
shown as percent cell viability compared to that of untreated
control cells.
AChE inhibition activities of the enzymatic extracts
from P. tenera In order to evaluate if the enzymatic
extracts from P. tenera could directly inhibit AChE, the
effects of the enzymatic extracts on AChE activity were
tested in vitro and the results are shown in Fig. 4. All the
enzymatic extracts showed percent inhibitions of greater
than 60% against AChE in a dose-dependent manner at the
concentration of 1.0 mg/mL. Moreover, the Flavourzyme
extract showed the highest activity among the protease
extracts while the AMG extract showed the highest activity
among the carbohydrase extracts. It was previously reported
that alkaloids are good AChE inhibitors (31), while Jung
and Park (32) reported flavonoids as AChE inhibitors. One
treatment strategy to enhance cholinergic functions is the
use of AChE inhibitors to increase the amount of
acetylcholine (ACh) available in the synapses between
cholinergic neurons. Hence, the use of proper AChE
inhibitors has attracted particular attention for the treatment
of Alzheimer-type dementia (33). The prevailing view has
been that the efficacy of AChE inhibitors is attained
through their augmentation of ACh-medicated neuron to
neuron transmission. However, AChE inhibitors also
protect cells from free radical toxicity and β-amyloid-
induced injury, and increased production of antioxidants. In
addition, it has been reported that AChE inhibitors directly
M. Senevirathne et al.
Fig. 6. Effects of the enzymatic extracts from P. tenera on the
inhibition of LPS-induced NO production in RAW264.7 cells.
A, protease extracts; B, carbohydrase extracts. Con, negative
control; LPS, lipoplysaccharide treated cells. Values are the mean±
SD of 3 determinations. a-bValues with different letters indicate
significant difference within the same concentration (p<0.05).
inhibit the release of cytokines from microglia and
monocytes (34).
Anti-inflammatory effects of the enzymatic extracts
from P. tenera Prior to examining the anti-inflammatory
effects of the enzymatic extracts from P. tenera, cell
toxicity was first evaluated by the MTT assay, revealing
that all the extracts showed percent cell viabilities greater
than 90% at all concentrations tested (Fig. 5A and 5B). To
determine whether the enzymatic extracts would inhibit the
production of LPS-induced NO, used as a marker of
inflammatory response, RAW264.7 cells were pretreated
with various concentrations of the enzymatic extracts and
then stimulated with LPS (1 µg/mL as a final concentration)
to evoke NO. Untreated RAW264.7 cells secreted basal
levels of NO, while the LPS stimulated cells showed
increased NO production (Fig. 6). The Protamex and
Viscozyme extracts inhibited NO production among the
protease and carbohydrasae extracts, respectively. However,
its activities were decreased with increasing concentration
of the enzymatic extracts. Macrophages actively participate
in inflammatory responses by releasing pro-inflammatory
cytokines such as tumor necrosis factor (TNF)-α,
interleukin (IL)-1β, and interferon-γ (IFN-γ), as well as
Biological Activities of Porphyra tenera
inflammatory factors such as NO and prostagladin E2
(PGE2), that recruit additional immune cells to sites of
infection or tissue injury (10). Hence, inhibitors of
inflammatory molecules have been considered sources for
anti-inflammatory drugs.
Our data suggest that the enzymatic extracts from P.
tenera possess high phenolic content and strong antioxidant
activities. Furthermore, the enzymatic extracts showed
strong inhibition activities against AChE and LPS-induced
NO production in RAW264.7 macrophage cells. In addition,
the enzymatic hydrolysis of P. tenera can provides water
soluble compounds that are not harmful. Hence, this study
suggests that P. tenera could be used as an easily accessible
source of natural antioxidants and anti-inflammatory
agents, and as a possible supplement in the pharmaceutical
industry. Also, the diverse activities shown in this study
reveal the potential for P. tenera to be used in multidis-
ciplinary areas. Future studies are necessary to investigate
the active components in the extracts.
Acknowledgments This research was a part of the
project titled ‘Development of Brain Food Materials from
Seaweeds’ funded by the Ministry for Food, Agriculture,
Forestry, and Fisheries.
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