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DOI 10.1007/s12562-011-0423-y
ORIGINAL ARTICLE
Received: 31 December 2010 / Accepted: 5 October 2011 / Published online: 11 November 2011
The Japanese Society of Fisheries Science 2011
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
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.
123
170
Nitrogen solubility
Sample preparation
Solubility%
Amount of nitrogen in supernatant sample
100
Amount of nitrogen in protein concentrate
Water-holding capacity
123
WHCg H2 O=g PC
W2 W1
W0
171
Foaming capacity%
V2 V1
100
V1
E.compressa
30
E.linza
E.tubulosa
25
20
15
10
5
0
2
10
12
pH
100
Volume before whipping
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
123
123
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
37.5 1.0cd
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
123
174
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
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