Fish Sci (2012) 78:169176

DOI 10.1007/s12562-011-0423-y

ORIGINAL ARTICLE

Food Science and Technology

Salt- and pH-induced functional changes in protein concentrate

of edible green seaweed Enteromorpha species
Ganesan Kandasamy Suresh Kumar Karuppiah
Peddi Venkata Subba Rao

Received: 31 December 2010 / Accepted: 5 October 2011 / Published online: 11 November 2011
The Japanese Society of Fisheries Science 2011

Abstract Protein concentrates (PCs) were extracted from

three edible green seaweed species of Enteromorpha
(E. compressa, E. linza, and E. tubulosa) and were studied
for their functional properties with respect to salt and pH.
The protein content in the PC was found to be 60.35 2.0,
53.83 0.70, and 33.36 1.04% in E. compressa,
E. tubulosa, and E. linza respectively. The minimum
nitrogen solubility was observed at pH 4.0 in all three PCs.
The water-holding and oil-holding capacities in the three
PCs ranged from 1.22 0.06 to 1.53 0.07 ml H2O/g PC
and from 1.05 0.07 to 1.34 0.10 ml oil/g PC respectively. Foaming capacity and stability were found to be
pH-specific. They varied significantly with pH and NaCl
concentration (P \ 0.05). The inexpensive source of protein concentrate from Enteromorpha species could be
incorporated into value-added food products.
Keywords Enteromorpha  Protein concentrate 
Functional properties  Salt  pH

Introduction
The use of plant proteins as functional ingredients in foods
depends mainly on the benefits that they can produce [1],
and so their use in the formulation of new food products or
in conventional foods has been a focus of much research in
recent years. In order to use plant proteins as food ingredients, their physicochemical and functional properties

G. Kandasamy  S. K. Karuppiah  P. V. Subba Rao (&)

Marine Biotechnology and Ecology Discipline, Central Salt
and Marine Chemicals Research Institute, Council of Scientific
and Industrial Research, CSIR, Bhavnagar 364 002, India
e-mail: cultivation.pvsrao@gmail.com

must be evaluated [2]. The demand for relatively inexpensive sources of proteins that are incorporated into
value-added food products is increasing worldwide. Much
of the research is focused on various sources of plant
proteins [2, 3] that may help in increasing the nutritional
value of food products at low cost. Functional properties of
food proteins are important in food processing and food
product formulation. Some of these properties are water/oil
holding, emulsification, foam formation, viscosity, and
gelation. The basic requirements for a protein to be considered as a good foaming agent are the ability to (1)
adsorb rapidly at the air-water interface during bubbling,
(2) undergo rapid conformational change and rearrangement at the interface, and (3) form a cohesive viscoelastic
film via intermolecular interactions. The first two criteria
are essential for good foam ability, whereas the third one is
important for the stability of the foam [4, 5].
Seaweeds belonging to the Chlorophyta (e.g., Ulva) and
Rhodophyta (e.g., Porphyra) contain a substantial amount
of protein (1047% DW) with potential for human and
animal nutrition (e.g., as functional food and fish feed) [6].
Furthermore, edible Chlorophyta species have 1622.1%
protein content as a percentage of dry matter [7]. The green
marine alga Enteromorpha has the greatest potential for
commercial exploitation because of its abundant availability and varied chemical composition, besides its quality, and accumulation of basic nutrients for other living
organisms [8, 9]. It has been included in a great variety of
dishes, including raw salads, soups, cookies, meals, and
condiments [10]. In view of this, a preliminary investigation was aimed at determining the influence of pH and salt
strength on the functional performance of protein concentrates of Enteromorpha compressa, E. linza, and E. tubulosa for considering their use as ingredients in food
industry.

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Fish Sci (2012) 78:169176

Materials and methods

Nitrogen solubility

Sample preparation

Nitrogen solubility of the protein concentrate at 2% w/v was

determined by a modified method of Were et al. [12], and for
the determination of the pH-dependent solubility profile for
pH values in the range 212, NaOH (0.5 M) or HCl (0.5 M)
was used. The slurries were mixed for 1 h using a magnetic
stirrer before centrifugation at 5,4279g for 20 min. The
nitrogen content in supernatants was determined by the
micro-Kjeldahl method. Triplicate determinations were
carried out, and the solubility profile was obtained by plotting averages of protein solubility (%) against pH, and the
percent soluble protein was calculated as follows:

Enteromorpha compressa, E. linza, and E. tubulosa

plants were collected during February 2006 from Okha
(2228.650 N and 6904.010 E), Gujarat. Fresh plants were
thoroughly washed with distilled water, and epiphytes were
removed. All cleaned seaweeds were dried at 60C in a hot
air oven for 10 h. All samples were dried to constant
weight. The dried samples were pulverized by using a
grinder and passed through a screen with an aperture of
0.5 mm. The milled seaweed samples were then stored in
air-tight plastic bags, which were in turn kept in desiccators
at room temperature prior to seaweed protein concentrate
(PC) extraction.

Solubility%
Amount of nitrogen in supernatant sample

 100
Amount of nitrogen in protein concentrate

Extraction of protein concentrate

Water-holding capacity

Protein concentrate (PC) from each of the three species of

Enteromorpha was extracted using the modified method as
described by Wong and Cheung [11]. In brief 150 g of
seaweed powder was suspended in deionized water (1:20
w/v). Then the suspension was gently stirred overnight at
35C. After incubation, the suspension was centrifuged at
5,000 rpm for 20 min. The supernatant was collected, and
the pellet was resuspended in de-ionized water in the
presence of 0.5% (v/v) 2-mercaptoethanol. Then the pH of
the mixture was adjusted to 12 with 1 M NaOH. The
mixture was gently stirred at room temperature for 2 h
before centrifugation under the same conditions narrated
above. The second supernatant was collected and combined
with the previous supernatant. The combined supernatant
was stirred at 0 4C, and its pH was adjusted to 7 before
precipitation with solid ammonium sulphate. The extraction procedure mentioned above was repeated five times on
the residue. Seaweed PCs were precipitated from the
supernatant by slowly adding solid ammonium sulphate
with stirring until an 85% saturation (60 g/100 ml) was
reached. Then the mixture was allowed to stand for 30 min
before centrifugation under conditions mentioned above.
The pellet (PC) obtained was dialyzed against distilled
water until the total dissolved solutes (TDS) (mg/l) of
dialysate, measured by its conductivity, were similar to
those of the distilled water. Then the retentates containing
the seaweed PCs were freeze-dried, ground to powder, and
stored in air-tight bags in desiccators before evaluation of
their protein quality.
The percentage of crude protein content of PCs was
calculated by multiplying the nitrogen content by a factor
of 6.25, which was determined by Perkin-Elmer PE 2400
series II CHNS/O elemental analyzer (Waltham, MA,
USA).

Water-holding capacity (g H2O/g PC) was determined

using the method of Beuchat [13]. Protein concentrate of
0.5 g was weighed into a preweighed 15-ml centrifuge
tube. Then 10 ml of distilled water was added and mixed
using a vortex mixer (Tarson, India) at the highest speed
for 2 min. After the mixture was thoroughly wetted, samples were allowed to stand at room temperature for 30 min,
then centrifuged at 8059g for 20 min. The supernatant was
decanted, and the centrifuge tube containing sediment was
weighed. Water-holding capacity was calculated. Triplicate
samples were analyzed.

123

WHCg H2 O=g PC

W2  W1
W0

where W0 is the weight of the dry sample (g), W1 the

weight of the tube plus the dry sample (g), and W2 weight
of the tube plus the sediment (g).
Oil-holding capacity
Oil-holding capacity (ml/g PC) was determined using the
method of Chakraborty [14]. One gram of protein was
weighed into preweighed 15-ml centrifuge tubes and thoroughly mixed with 10 ml of groundnut oil using a Vortex
mixer. Samples were allowed to stand for 30 min. The protein-oil mixture was centrifuged at 8059g for 20 min.
Immediately after centrifugation, the supernatant was carefully poured into a 10 ml graduated cylinder, and the volume
was recorded. Triplicate samples were analyzed.
Foaming capacity and stability
The foaming capacity and stability were studied according
to the method of Coffman and Garcia [15]. Protein

Fish Sci (2012) 78:169176

171

Foaming capacity%

V2  V1
 100
V1

E.compressa

30

Nitrogen solublity (%)

concentrate (25 mg) was mixed with 25 ml of distilled

water at different pH values and NaCl concentrations, and
the mixture was whipped using a vortex mixture for 10 min
at room temperature and transferred to a measuring cylinder. The volume increase is expressed as percent foaming
capacity. The foam stability was determined by measuring
the decrease in volume of foam as a function of time up to
a period of 30 min.

E.linza

E.tubulosa

25
20
15
10
5
0
2

10

12

pH

where V1 is the volume of protein solution before

whipping, and V2 is the volume of protein solution after
whipping.
Foam stability%
Volume after standing  Volume before whipping

 100
Volume before whipping

Fig. 1 Nitrogen solubility of protein concentrates as function of pH

in Enteromorpha species

The total protein content of three species of Enteromorpha

biomass was determined and calculated on the basis of dry
weight. The protein content was found to be highest in
E. tubulosa (19.09 0.91%) followed by E. compressa
(17.48 0.41%) and E. linza (12.5 1.26%). A higher
percentage of recovered protein concentrate of Enteromorpha was noted in E. compressa (6.48%) followed by
E. tubulosa (6.16%) and E. linza (5.71%). The total nitrogen
and protein contents in protein concentrate (PC) of three
species of Enteromorpha are presented in Table 1. The
protein content in PC was found to differ among the species

of Enteromorpha. The protein content in PC obtained (w/w,

dry weight) was 60.35 2.01, 53.83 0.70, and 33.36
1.04% respectively for E. compressa, E. tubulosa, and
E. linza. Nitrogen content (%) PC of Enteromorpha species
ranged from 5.34 0.17 to 9.66 0.03%. The nitrogen
solubility at different pH conditions (212) differed (Fig. 1).
Thus, the nitrogen solubility (%) was found to range from
14.96 0.35 to 26.38 0.88 in E. compressa, from
10.87 1.12 to 20.31 1.66 in E. linza, and from
13.60 0.85 to 25.41 1.94 in E. tubulosa. The minimum
nitrogen solubility of 10.87 1.12, 13.6 0.85, and
14.96 0.35 was recorded respectively for E. linza, E. tubulosa, and E. compressa at pH 4. There was not much
variation in nitrogen solubility in three PCs of Enteromorpha
at alkaline pH. The water- and oil-holding capacities of PCs
of Enteromorpha species are presented in Table 2. The
water-holding capacity of protein concentrate of E. compressa was found to be higher (1.53 0.07 g water/g PC)
than that of E. tubulosa (1.32 0.11 g water/g PC) and
E. linza (1.22 0.06 g water/g PC). High water absorption
of proteins helps to reduce moisture loss in packaged bakery
goods. Also it is required to maintain freshness and moist
mouth feel of baked foods. The oil-holding capacity of
protein concentrate of E. compressa, E. linza, and E. tubulosa was found to be 1.34 0.10, 1.05 0.07, and
1.08 0.04 ml oil/g, respectively.

Table 1 Percent nitrogen and protein content in Enteromorpha protein concentrate

Table 2 Water- and oil-holding capacities of protein concentrate

(PC) of Enteromorpha species

Source of PC

Nitrogen
content (%)

Protein
content (%)

Source of PC

Water-holding
capacity (g H2O/g PC)

Oil-holding capacity
(ml oil/g PC)

E. compressa

9.66 0.03

60.35 .2.01

E. compressa

1.53 0.07

1.34 0.10

E. linza

5.34 0.17

33.36 1.04

E. linza

1.22 0.06

1.05 0.07

E. tubulosa

8.61 0.11

53.83 0.70

E. tubulosa

1.32 0.11

1.08 0.04

Statistical analysis
The data reported in all tables are mean with standard
deviation of triplicate observations and were subjected to
one-way analysis of variance (ANOVA) using SPSS 7.5.

Results
Protein content, nitrogen solubility,
and water/oil-holding capacity

Values are expressed as mean standard deviation for triplicate

determination

Values are expressed as mean standard deviation for triplicate

determination

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Values are expressed as mean standard deviation for triplicate determinations. Values in the same column with different letters are significantly different (P \ 0.05)

27.5 1.6b

12.5 2.8a
25.8 1.3a

36.3 1.3b
32.5 1.1d

10.0 1.6a
14.3 1.3a

33.3 2.6b
20.0 1.6b

7.5 2.7a
13.3 1.3a

23.3 1.3b
7.5 2.0a

7.5 2.7a
14.2 2.5a

23.3 2.6b

6.4 1.0a
12

5.0 2.1a

10.8 2.9b
10

7.5 1.5b

47.5 1.3e

37.5 2.2d
73.3 0.5e

50.6 1.7d
35.0 1.2e

37.5 0.6f
63.2 1.5f

56.4 1.3e
32.5 0.6c

20.0 2.1b
22.5 1.4b

38.3 1.3c
22.5 1.1b

7.5 1.2a
22.5 1.4b

63.3 1.3d
35.0 2.5e

20.6 2.0c

40.9 2.9d

23.5 5.2c

38.3 1.3c

42.5 1.4c
30.0 1.7c

25.0 1.6b
38.3 1.3c

43.6 1.1d
32.5 1.3c

37.5 1.7d
37.8 1.4c

38.3 1.3c
30.0 0.9d

25.0 1.4c
28.3 1.3b

37.5 2.5c
37.5 2.0f

32.5 1.6d

55.0 2.6e

39.2 2.9d

A
A
B
A

1.5 M NaCl
1.0 M NaCl
0.5 M NaCl
0.1 M NaCl
0 M NaCl
pH

Table 3 Effect of pH and NaCl concentration on foaming capacity (A) and foam stability (B) of protein concentrate of Enteromorpha compressa

The foaming capacity is the volume increase after whipping of PC solution (mg/ml) expressed as a percentage, and
the foam stability is the volume of foam remaining after a
specified time expressed as a percentage of the initial foam
volume. Foaming capacity (FC) of protein concentrate was
investigated under various pH and salt (NaCl) conditions.
The effects of pH and salt concentration on foaming
capacity (FC) and foam stability (FS) of PCs of three
species of Enteromorpha are shown in Tables 3, 4, and 5.
The FC (%) property was found to be pH-dependent. The
FC (%) was found to range from 6.4 1.0 to 55.0 2.6,
15.6 0.9 to 51.8 4.7, and from 19.7 5.5 to
51.2 1.6 for E. compressa, E. linza, and E. tubulosa
respectively at zero ionic strength from pH 2 to 12. As
the pH increased from acidic (pH 2) to alkaline (pH
12), foaming capacity of protein concentrate invariably
decreased in E. compressa. At pH 4.0 and 6.0, the PC of
E. compressa and E. tubulosa had good foaming capacity.
Upon addition of different concentrations of NaCl from 0.1
to 1.0 M, an increase in foaming capacity was observed in
PC of E. compressa at pH 2 (from 37.5 2.5 to
43.6 1.1), pH 4 (from 28.3 1.3 to 38.3 1.3), pH 8
(from 22.5 1.4 to 63.2 1.5), and pH 10 (from
23.3 2.6 to 33.3 2.6). The same trend was also
noticed in E. tubulosa at pH 2 (from 20.3 1.0 to
31.9 2.2), pH 6 (from 47.4 2.1 to 55.9 1.7), and
pH 8 (from 37.2 3.7 to 48.8 1.2). However, foaming
capacity of PC of E. linza was found to increase upon
addition of NaCl concentration from 0.1 to 0.5 M at all pH
values studied except for pH 2. The high concentration of
NaCl solution, i.e., 1.5 M, was found to depress foaming in
all three PCs of Enteromorpha studied. Foaming capacity
was found to be significantly different (P \ 0.05) at 0, 0.1,
and 1.5 M concentrations in PC of E. compressa, while it
was significant at 0.5 M concentration in E. linza and
E. tubulosa. Three species of Enteromorpha, generally
foam stability was found to be better in acidic pH conditions than in alkaline pH conditions in E. compressa and
E. linza, but in E. tubulosa, FS was better in alkaline pH
conditions. It appeared that the foams formed were more
stable in PC of E. compressa and E. linza as the concentration of added salt increased up to 0.5 M. The foam
stability (%) was found to range from 5.0 2.1 to
37.5 2.0, from 4.4 2.0 to 39.0 2.1, and from
11.7 1.7 to 29.2 3.7 in the PCs of E. compressa,
E. linza, and E. tubulsoa, respectively, from pH 2 to 12 at
zero salt (NaCl) concentration. Addition of NaCl from 0.1
to 0.5 M increased foam stability of PC of E. compressa at
pH 2 (from 30.0 0.9 to 32.5 1.3), pH 4 (from
25.0 1.4 to 37.5 1.7), and pH 6 (from 22.5 1.1 to
32.5 0.6). The same trend was also found in PC of

Foaming capacity and foam stability

37.5 1.0cd

Fish Sci (2012) 78:169176

32.5 0.6c

172

39.0 2.1d

11.7 1.5b

20.0 4.0ab

17.3 2.3a

21.0 2.0ab

20.7 3.1ab

32.7 3.1c

29.3 1.2bc

26.0 2.5b

15.0 2.0a

21.6 1.2b

37.9 1.1c

51.7 1.4d

16.1 1.0a

41.5 3.4c

56.0 4.2d

27.3 2.3ab

43.0 4.6c

33.3 4.4bc

18.5 3.4a

0.5 M NaCl

29.1 2.7c

17.2 1.6a

31.6 2.1cd

44.1 0.6e

34.8 1.7d

22.9 1.3b

16.7 1.2b

21.3 1.2b

29.3 1.2c

33.3 3.1c

10.0 2.0a

6.7 1.1a

1.0 M NaCl

28.1 1.6cd

25.3 1.2bc

30.9 0.6d

23.7 1.2b

19.3 1.6a

18.6 1.7a

35.3 3.0c

34.7 1.2c

22.7 2.3b

19.3 2.3b

18.5 1.2b

8.7 1.2a

1.5 M NaCl

20.3 1.5a

51.2 1.6c

10

12

19.4 4.2b

29.2 3.7c

20.8 3.7ab

11.7 1.7a

27.3 3.1c

16.7 1.5ab

68.8 3.7e

22.3 0.6a

37.2 3.7b

47.4 2.1c

55.8 0.7d

20.3 1.0a

0.1 M NaCl

7.3 1.2ab

5.3 2.3a

9.3 1.1b

16.7 1.1c

4.7 1.2a

5.3 1.2a

36.5 2.2c

19.3 1.6a

46.7 1.6d

47.8 1.9d

32.2 2.2c

26.4 1.0b

0.5 M NaCl

25.2 3.4bc

36.6 2.4d

16.7 2.3ab

32.6 1.3cd

20.7 1.3ab

14.8 3.4a

38.6 2.2c

20.7 0.6a

48.8 1.2d

55.9 1.7e

44.8 1.7d

31.9 2.2b

1.0 M NaCl

3.3 1.2a
6.0 2.01ab

11. 3 3.1bc

9.3 3.1abc

15. 3 1.2c

22.7 3.1d

37.9 1.1ab

27.7 13.4a

41.8 0.6b

50.0 1.1b

37.8 1.1ab

30.7 1.3a

1.5 M NaCl

Values are expressed as mean standard deviation for triplicate determinations. Values in the same column with different letters are significantly different (P \ 0.05)

45.0 2.0c

19. 7 5.5a

25.6 3.4ab

31.9 2.7b

pH

0 M NaCl

Table 5 Effect of pH and NaCl concentration on foaming capacity (A) and foam stability (B) of protein concentrate of Enteromorpha tubulosa

Values are expressed as mean standard deviation for triplicate determinations. Values in the same column with different letters are significantly different (P \ 0.05)

39.4 4.2cd

12

7.0 2.0ab

25.4 4.8ab

51.8 4.7d

10

20.3 2.5c

15.6 0.9a

4.4 2.0a

22.8 1.6c

24.2 4.0ab

33.3 5.7bc

0.1 NaCl

0 M NaCl

pH

Table 4 Effect of pH and NaCl concentration on foaming capacity (A) and foam stability (B) of protein concentrate of Enteromorpha linza

22.7 2.1b

23.3 2.5b

13.3 1.6a

8.7 0.9a

12.6 1.6a

4.7 1.1a

25.3 2.8c

15.1 1.6b

35.4 2.2d

39.3 1.3d

27.0 0.6c

9.0 1.0a

Fish Sci (2012) 78:169176

173

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E. tubulosa at pH 2 (from 5.3 1.2 to 14.8 3.4), pH 4

(from 4.7 1.2 to 20.7 1.3), and pH 6 (from 16.7 1.1
to 32.6 1.3). The increase in foam stability, however,
was seen in PC of E. linza only at pH 2 (from 16.1 1.0 to
22.9 1.3) and pH 6 (from 37.9 1.1 to 44.1 0.6). FS
was found to be significantly different (P \ 0.05) at 0, 1.0,
and 1.5 M concentrations in PC of E. compressa, while it
was significant at 1.5 M concentration in E. linza.

Discussion
The crude protein content of the three Enteromorpha
species was within the range for red and green edible
seaweeds (1047%) as reported by Fleurence [6]. However, the crude protein content of these three Enteromorpha species was found to be higher than that of Ulva
lactuca from northeast of Hong Kong (7.06%) [16].
Extraction of crude protein concentrate from seaweed is
complicated due to the presence of anionic or neutral
polysaccharides as well as phenolic compounds [1719].
In the present study, the percentage of recovery of protein
concentrate was found to be lower as compared to other
seaweed species reported [17]. Ganesan et al. [20] reported
that different solvent extracts of Enteromorpha species
(E. compressa, E. linza, and E. tubulosa) showed a good
amount of phenol content. The higher phenol content in the
Enteromorpha species could be attributed to the lower
recovery of protein concentrate. Minimum nitrogen solubility around pH 4 and 5 was also observed in the PCs of
some other legumes, i.e., black gram [21]. High nitrogen
solubility is required for protein concentrate to be used as
functional ingredients in many foods including beverages,
dressings, coffee whiteners, whipped toppings, confections, etc. [22]. The present study revealed that all PCs of
three species of Enteromorpha contained a considerable
quantity of protein with high nitrogen solubility, and hence
they may be considered for use as ingredients in food
formulations.
Interactions of water and oil with proteins are very
important in food systems because of their effects on the
flavor and texture of foods. Intrinsic factors affecting the
water-binding capacity of food proteins include amino acid
composition, protein conformation, and surface polarity/
hydrophobicity [23]. However, food processing methods
have important impacts on the protein conformation and
hydrophobicity. The mechanism of fat/oil holding/absorption capacity was explained by Kinsella [24] as a physical
entrapment of oil. Fat/oil holding/absorption capacity is a
critical determinant of flavor retention, while fat emulsion
capacity and stability are important attributes of additives
for the stabilization of fat emulsions. Water-holding
capacity values were calculated excluding some soluble

123

Fish Sci (2012) 78:169176

components in the supernatant after centrifugation because

they were meager amount in the supernatant. This might be
due to interaction between protein molecules and trace
amounts of some nonsoluble components that may have
settled down at the bottom of the tube as pellet. Waterholding capacity values ranging from 1.49 to 4.72 (g/g) are
considered critical in viscous foods such as soups and
gravies [25]. High oil absorption/holding is essential in the
formulation of food systems such as sausages, cake batters,
mayonnaise, and salad dressings [22]. As the water-holding
and oil-holding capacities of PCs of all three species of
Enteromorpha are quite adequate, they may be considered
for use to improve the viscous nature of food formulations.
Foaming capacity and foam stability of protein concentrates of Enteromorpha species depend on the concentrations of protein concentrate used because this increases
the viscosity and facilitates the formation of a multilayer,
cohesive protein film at the interface. Some other factors
such as transportation, penetration, and reorganization are
responsible for formation of foam in the medium. Therefore, protein molecules must be capable of migrating at the
air-water interface to show a good foaming property [26].
Besides the protein content, some other molecules with
meager amount such as polysaccharide, and minerals in the
protein concentrate of Enteromorpha could have been
involved in the foaming capacity of PCs of Enteromorpha.
Foaming capacity of winged bean protein concentrate has
been increased upon addition of sucrose, amylose, amylopectin, potato starch, gum arabic, and pectin at 0.25 g/g of
protein to its protein concentrate [27]. Addition of NaCl
improved foaming capacity, which might be due to the
ability of NaCl to aid diffusion and spreading at the
interface of the protein concentrate. The beneficial effect of
low concentrations of NaCl on foams has been reported by
Akintayo et al. [28]. Since foaming capacity appears to be
due to solubilized protein, the differing effects of salt
concentrations may be explained on this basis. As salt
concentrations increased, foams formed were denser as
indicated by decreased specific volume. Formation of foam
requires proteins to solubilize in the aqueous phase and
rapidly unfold to form a cohesive layer of protein around
gas/air droplets [29]. The basic requirements for a protein
to be a good foaming agent are the ability to adsorb rapidly
at the air-water interface during bubbling and the ability to
undergo rapid conformational changes and re-arrangement
at the interface [30]. In the present study, the foaming
capacity of E. compressa was found to be significantly
different (P \ 0.05) at 0, 0.1, and 1.5 M concentrations,
while it was significant at 0.5 M concentration in E. linza
and E. tubulosa. The addition of NaCl concentration at pH
6 gradually improved the foaming capacity of the protein
concentrate of Enteromorpha species with a higher percentage increase. This may be attributed to the fact that

Fish Sci (2012) 78:169176

addition of NaCl at a particular concentration enhances the

protein solubility by weakening the hydrophobic interaction of the protein, while high salt concentration has an
adverse effect on foaming capacity due to the salting effect
of NaCl. The high concentration of NaCl solution, i.e.,
1.5 M, was found to depress foaming in all three PCs of
Enteromorpha studied. These results concur with mucuna
bean protein concentrate (MPC) in which an increase in the
foaming capacity of MPC was reported when the ionic
strength increased progressively from 0.1 to 0.4 M, after
which a decline in the foaming capacity occurred in solutions prepared at higher ionic strengths [31]. The addition
of salt improved the foaming capacity to a maximum value
of 30.5% at 0.4 M NaCl for C. maritima flour due to the
increased protein solubility [32].
The foam stability of protein concentrate of Enteromorpha was found to differ among the species and was pHdependent. An increasing foam stability was noticed at
acidic pH in all three PCs, which could be due to increased
charge density that prevented rapid coalescence of the air
bubbles. The increase in charge density might have stabilized the foams by increasing electrostatic repulsions,
which reduced the rate of coalescence of foam particles.
Foam stability was found to be very firm in E. compressa
and E. tubulosa in which PCs without NaCl addition
exhibited higher foam stability. As the time of standing
progressed, the foam volume decreased, and a similar trend
was also observed in protein isolates of beach pea Lathyrus
maritimus [2]. Kinsella et al. [33] have suggested that, in
foams, the ability to hold water in the protein film surrounding the air particle and the presence of electrostatic
repulsion are important for their stability. Since the PCs of
three species of Enteromorpha exhibited considerable
foaming capacity and foam stability, these may be considered for use in food formulations to improve the value of
food.
Acknowledgments Authors express their gratitude to Director,
Central Salt and Marine Chemicals Research Institute (CSIR),
Bhavnagar for his encouragement and the facilities provided. Senior
author K.G. is thankful to the Council of Scientific and Industrial
Research CSIR, New Delhi for financial assistance in the form of
Senior Research Fellowship.

References
1. McWatters KH, Cherry JP (1977) Emulsification, foaming and
protein solubility properties of defatted soybean, peanut, field pea
and pecan flours. J Food Sci 42:14441447
2. Chavan UD, McKenzie DB, Shahidi F (2001) Functional properties of protein isolates from beach pea (Lathyrus maritimus).
Food Chem 74:177187
3. Tomotake H, Shimaoka I, Kayashita J, Nakajoh M, Kato N
(2002) Physicochemical and functional properties of buckwheat
protein product. J Agric Food Chem 50:21252129

175
4. Damodaran S (1994) Structurefunction relationship of food
proteins. In: Hettiarachchy NS, Ziegler GR (eds) Protein functionality in food systems. CRC Press, Boca Raton, pp 138
5. Dickinson E, McClements DJ (1995) Molecular basis of protein
functionality. In: Dickinson E, McClements DJ (eds) Advances in
food colloids. Blackie Academic and Professional, London,
pp 2776
6. Fleurence J (1999) Seaweed proteins: biochemical, nutritional
aspects and potential uses. Trends Food Sci Technol 10:2528
7. Mamatha BS, Namitha KK, Amudha S, Smitha J, Ravishankar
RA (2007) Studies on use of Enteromorpha in snack food. Food
Chem 101:17071713
8. Hoppe HA (1979) Marine algae and their products and constituents in pharmacy. In: Hoppe HA et al (eds) Marine algae in
pharmaceutical science. Walter de Gruyter, Berlin, pp 25120
9. Chapman VJ, Chapman DJ (1980) Sea vegetables (algae as food
for man). In: Seaweeds and their uses. Chapman and Hall, London, pp 6297
10. Ohno M (1993) Cultivation of the green algae Monostroma and
Enteromorpha aonori. In: Ohno M, Critchley AT (eds) Seaweed cultivation and marine ranching. Japan International
Cooperation Agency, Japan, pp 715
11. Wong KH, Cheung PCK (2001) Nutritional evaluation of some
subtropical red and green seaweeds. Part II. In vitro protein
digestibility and amino acid profiles of protein concentrates. Food
Chem 72:1117
12. Were L, Hettiarachchy L, Kalapathy U (1997) Modified soy
proteins with improved foaming and water hydration proteins.
J Food Sci 62:821824
13. Beuchat LR (1977) Functional and electrophoretic characteristics
of succinylated peanut flour proteins. J Agric Food Chem
25:25826
14. Chakraborty P (1986) Coconut protein isolate by ultrafiltration.
In: LeMeguer M, Jelen P (eds) Food engineering and process
applications. Elsevier Applied Science, New York, pp 308315
15. Coffman CW, Garciaj VV (1977) Functional properties and
amino acid content of a protein isolate from Mung bean flour. Int
J Food Sci Technol 12:473484
16. Wong KH, Cheung PCK (2000) Nutritional evaluation of some
subtropical red and green seaweeds. Part I. Proximate composition, amino acid profiles and some physico-chemical properties.
Food Chem 71:475482
17. Fleurence J, Le Coeur C, Mabeau S, Maurice M, Landrein A
(1995) Comparison of different extractive procedures for proteins
from the edible seaweeds Ulva rigida and Ulva rotundata. J Appl
Phycol 7:577582
18. Jordan P, Vilter H (1991) Extraction of proteins from material
rich in anionic mucilages: partition and fractionation of vanadatedependent bromoperoxidases from the brown algae Laminaria
digitata and L. saccharina in aqueous polymer two phase system.
Biochim Biophys Acta 1073:98106
19. Ragan MA, Glombitza KW (1986) Phlorotannins, brown algal
polyphenols. In: Round FE, Chapman DJ (eds) Progress in phycological research. Biopress, Bristol, pp 130230
20. Ganesan K, Kumar KS, Subba Rao PV (2011) Comparative
assessment of antioxidant activity in three edible species of green
seaweed, Enteromorpha from Okha, northwest coast of India.
Innov Food Sci Emerg Technol 12:7378
21. Sathe SK, Deshpande SS, Salunkhe DK (1983) Functional
properties of black gram (Phaseolus mungo L.) proteins. Lebensm Wiss Technol 16:6972
22. Chandi GK, Sogi DS (2007) Functional properties of rice bran
protein concentrates. J Food Eng 79:592597
23. Barbut S (1999) Determining water and fat holding. In: Hall GM
(ed) Methods of testing protein functionality. Blackie Academic
and Professional, New York, pp 186225

123

176
24. Kinsella JE (1979) Functional properties of soy protein. J Am Oil
Chem Soc 56:242249
25. Aletor O, Oshodi AA, Ipinmoroti K (2002) Chemical composition of common leafy vegetables and functional properties of
their leaf protein concentrates. Food Chem 78:6368
26. Halling PJ (1981) Protein stabilized foams and emulsions. Crit
Rev Food Sci Nutr 15:155203
27. Sathe SK, Despande SS, Salunkhe DK (1982) Functional properties of winged bean [Psophocarpus tetragonolobus (L.) DC]
proteins. J Food Sci 47:503509
28. Akintayo ET, Oshodi AA, Esuoso AO (1999) Effects of NaCl,
ionic strength and pH on the foaming and gelation of pigeon pea
(Cajanus cajan) protein concentrates. Food Chem 66:5156
29. Tang S, Hettiarachchy NS, Horax R, Eswaranandam S (2003)
Physicochemical properties and functionality of rice bran protein

123

Fish Sci (2012) 78:169176

30.

31.

32.

33.

hydrolyzate prepared from heat-stabilized defatted rice bran with

the aid of enzymes. J Food Sci 68:152157
Fidantsi A, Doxastakis G (2001) Emulsifying and foaming
properties of amaranth seed protein isolate. Colloids Surf B Biointerfaces 21:119124
Adebowale KO, Lawal OS (2003) Foaming, gelation and electrophoretic characteristics of mucuna bean (Mucuna pruriens)
protein concentrates. Food Chem 83:237246
Seena S, Sridhar KR (2005) Physicochemical, functional and
cooking properties of under explored legumes, Canavalia of the
southwest coast of India. Food Res Int 38:803814
Kinsella JE, Damodaran S, German B (1985) Physicochemical
and functional properties of oilseed proteins with emphasis on
soy proteins. In: Alschul AM, Wilke HL (eds) New protein foods.
Academic Press, New York, pp 107179