LWT - Food Science and Technology 48 (2012) 30e36

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LWT - Food Science and Technology

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Functional characterization of gelatin extracted from bones of red snapper

and grouper in comparison with mammalian gelatin
R. Jeya Shakila*, E. Jeevithan, A. Varatharajakumar, G. Jeyasekaran, D. Sukumar
Department of Fish Processing Technology, Fisheries College and Research Institute, Tamil Nadu Veterinary and Animal Sciences University, Tuticorin 628 008, India

a r t i c l e i n f o a b s t r a c t

Article history: The average yields of fish gelatin extracted from the bones of red snapper (Lutjanus campechanus) and
Received 29 December 2011 grouper (Epinephelus chlorostigma) were 9.14 and 13.66 g/100 g, respectively. The protein contents of the
Received in revised form bone gelatin ranged from 78.5 to 82.36 g/100 g. The pH of the extracted gelatins were acidic (pH 4.3e4.6)
29 February 2012
with an absorption maxima at 214 nm. The viscosities were higher than mammalian gelatin with values
Accepted 4 March 2012
13e18 cP. Melting temperatures were higher (26
C) and gelling temperature were lower (16
C) than
that of mammalian gelatin with 21
C, 22
C respectively. The bloom strengths were comparable with
Keywords:
mammalian gelatin with values of 7.5e7.7 N. In fish gelatin, foaming abilities/stabilities and fat
Fish gelatin
Gel strength
binding capacity (FBC) were higher than mammalian gelatin. The water holding capacity of grouper bone
Viscosity gelatin being similar to mammalian gelatin. SDS protein-patterns of the bone gelatins of two species did
Foam ability not show variations. The a, b and g chains showed that the triple helical structure were not totally
SDS-PAGE destroyed in the fish gelatin. The physical and functional properties for fish bone gelatin suggested that
Mammalian gelatin their qualities were similar to mammalian gelatin and suitable for the food and packaging applications.
Ó 2012 Elsevier Ltd. All rights reserved.

1. Introduction Extraction of fish gelatin from several fish such as cod

Gelatin is a mixture of peptides and proteins produced by partial
hydrolysis of collagen. The quality of gelatin depends on its physi-
cochemical properties, rheological properties and severity of the
manufacturing method. In the food industry, gelatin is used as an
ingredient to improve the elasticity, consistency and stability of
foods. They are used in jelly production, confectionary, edible films,
encapsulation, fruit juice clarification, dairy processing, soups, etc.
Most commercial gelatins are currently sourced from beef bone
and hide, pig skin and bones. It was reported that 41 g/100 g of the
gelatin produced in the world is sourced from pig skin, 28.5 g/100 g
from bovine hides and 29.5 g/100 g from bovine bones. In some
countries, the use of mammalian gelatin is restricted, for reasons
such as transmission of the bovine spongiform encephalopathy
(BSE) and for religious reasons (Gilsenan & Ross-Murphy, 2001).
Fish, therefore, forms an important source of gelatin, but its
commercial production is very low to about 1 g/100 g of the annual
world gelatin production (Jamilah & Harvinder, 2002). Fish gelatins
are mainly obtained from skins, bones, fins, scales and swim
bladder of fish.
(Gudmundsson & Hafsteinsson, 1997), hake (Gomez-Guillen
et al., 2002), red and black tilapia (Jamilah & Harvinder, 2002)
has been reported. Production of fish gelatin is considered as
a better way of utilization of the processing wastes from the
fishing industry (Gomez-Guillen et al., 2002; Muyonga, Cole, &
Duodu, 2004a,b). Red snapper (Lutjanus campechanus) is the
highly valued food fish of tropical waters with good market.
Grouper (Epinephelus chlorostigma) is yet another food fish of
good export potential. The processing wastes viz. bones and fins
constitute about 50 g/100 g of their total body weight. On an
average 30 tonnes of processing wastes are generated by this
company every month. Gelatin extraction from the bones would
be therefore an effective way of utilization of these wastes. With
this background, the present study was undertaken to extract
the gelatin from the bones of these fishes, characterize and
understand the functional properties in comparison with
mammalian gelatin.
2. Materials and method
2.1. Raw material

* Corresponding author. Tel./fax: þ91 461 2332354. Bones of red snapper (L. campechanus) and brown spotted
E-mail address: jeyashakila@gmail.com (R. Jeya Shakila). grouper (E. chlorostigma) were obtained from a private fish

0023-6438/$ e see front matter Ó 2012 Elsevier Ltd. All rights reserved.
doi:10.1016/j.lwt.2012.03.007
 R. Jeya Shakila et al. / LWT - Food Science and Technology 48 (2012) 30e36
processing plant, M/s Kondia Seafoods, Tuticorin, India and
brought to the laboratory and held frozen at 20
C until use.
2.2. Gelatin extraction
Gelatin extraction was carried out as per the method of
Ninan, Jose, and Abubacker (2011) with slight modification.
Bones were washed with tap water to remove superfluous
material and treated twice with NaOH (0.2 g/100 g) at 1:6 ratio
(g: mL) for 45 min to remove the non-collageneous protein.
They were washed thoroughly with tap water and again treated
twice with H2SO4 (0.2 mL/100 mL) at 1:6 ratio (g: mL) for 45 min
to increase swelling as well as to remove salts. They were then
washed thoroughly with water and treated twice with citric acid
(1 g/100 g) at 1:6 ratio (g: mL) for 45 min; and washed with tap
water thoroughly. The final extraction was carried out with
distilled water at 1:1 ratio (g: mL) at 45
C for 24 h. The extract
was filtered through Whattman No.4 filter paper under vacuum,
lyophilized (Alpha 2, Martin Christ, Germany) and used for
characterization. The yield of the gelatin was calculated using
the following formula (Binsi, Shamasundara, Dileepa, Badiib, &
Howell, 2009).
Weight of dried gelatinðgÞ  100
Yield ¼
Weight of the raw sampleðgÞ
2.3. Biochemical composition
The biochemical composition of the bones and bone gelatins
were determined by the standard AOAC methods (1995). Mois-
ture content was determined by oven drying of samples at
105
C for 20e24 h. Total crude protein content of gelatin was
determined using the Kjeldahl method using Kel plus Nitrogen
analyzer (Pelican Equipments, Chennai). Ash content was
determined by ashing dried material in a muffle furnace set at
550
C for 24 h. Total lipids were extracted from 2 g dried sample
using Soc plus extraction apparatus (Pelican Equipments,
Chennai).
2.4. pH
Gelatin, 1.0 g dissolved in 100 mL distilled water and
measured in a pH meter at 25
C (Cheow, Norizah, Kyaw, &
Howell, 2007).
2.5. Absorbance spectrum
Absorption maxima of gelatin solution, 1 g/100 mL was
measured at 200, 280, 350, 400, 500, 600 and 800 nm by the UV/
Vis. Spectrophotometer (Model V-530, Jasco, Japan).
2.6. Viscosity
The viscosity was measured as per the method (Cho, Jahncke,
Chin, & Eun, 2006). Gelatin solution, 6.67 g/100 mL was
measured for the viscosity (cP) in the Brookfield Digital Viscom-
eter (Model DV E, Brookfield Eng Lab Inc., Middleboro, MA)
Volume of foamðmLÞ þ Volume of liquidðmLÞ
Foam formation abilityðFAÞ ¼
Initial volume of solutionðmLÞ
31
equipped with a No. 1 Spindle at 60 rpm at ambient temperature
(30  2
C).
2.7. Melting point
The melting point was determined as per the procedure of
Wainewright (1977). Gelatin solution, 6.67 g/100 mL was taken
in the test tubes (12  75 mm), covered with parafilm and
heated in a water bath at 60
C for 15 min. The solution was
cooled immediately in ice-chilled water and matured at 10
C for
16e18 h. Five drops of indicator containing 75 mL of chloroform
and 25 mL of methyl red (0.2 g/100 mL) were placed on the
surface of the gel, placed in a water bath at 10
C and heated at
the rate of 0.2
C per min. The temperature at which the drops
began to move freely down the gel was taken as the melting
point.
2.8. Gelling temperature and gelling time
Gelling temperature and gelling time were determined by the
method of Muyonga et al. (2004a,b). Gelatin solution, 10 g/100 mL
was taken in the standard Borosil R test tubes (12 mm  75 mm)
and placed in warm water bath maintained at 40
C. The bath was
then cooled slowly at the rate of 0.2
C per min. A thermometer was
inserted into the solution and lifted out at 30 s intervals. The
temperature at which the solution no longer dripped from the tip of
the thermometer was recorded as the gelling temperature. Setting
time was determined in the same way but the gelatin solution was
transferred to a water bath maintained at 10
C. A rod was inserted
in the solution and raised at intervals of 15 s. The time at which the
rod could not detach from the solution was recorded as the gelling
time.
2.9. Bloom strength
Bloom strength of the gelatin gel was determined by the
method of Cheow et al. (2007). Gelatin solution, 2 g/100 mL was
taken in a beaker with dimension of 4.8 cm diameter and 1.8 cm
height. Solution was heated at 65
C for 25 min, cooled to room
temperature and allowed to mature at 10
C for 16e18 h. Bloom
strength was determined by Universal Testing Machine (TA plus
Texture Analyzer, Lloyd Instruments, U.K) equipped with
a 12.7 mm diameter flat-face cylindrical teflon-coated plunger
at a cross-head speed at 0.5 mm/s. The maximum force (g)
was recorded as the plunger penetrated 4 mm into the
gelatin gel.
2.10. Foam formation ability/foam stability
Foam formation ability (FA) and foam stability (FS) of gelatin
were determined by the procedure of Cho et al. (2004). Gelatin
solution, 1 g/100 mL was taken in a beaker and swollen at 60
C.
Foam was prepared by homogenizing the solution continuously by
using a magnetic stirrer for 30 min. The solution was then poured
into a 100 mL measuring jar. The FA and FS were calculated by the
following formula.
32 R. Jeya Shakila et al. / LWT - Food Science and Technology 48 (2012) 30e36
Volume of foam þ Volume of liquidðinitialÞ
Foam stabilityðFSÞ ¼
Volume of foam þ Volume of liquidðafter 30 minÞ
2.11. Water holding capacity and fat binding capacity
Water holding capacity (WHC) and fat binding capacity (FBC) of
gelatin were determined as per the procedure of Cho et al. (2004).
Gelatin, 10 mg was taken in a centrifuge tube, to which 0.5 mL
distilled water or 0.1 mL of sunflower oil was added to measure
WHC and FBC, respectively. The solution was held at room
temperature for 1 h, mixed in a vortex mixer for 5 s at a time
interval of 15 min and then centrifuged at 450 g for 20 min
(Universal 32 R, Andreas-Hettich, Germany). The upper portion was
removed and the tube was drained for 30 min on a filter paper by
tilting to a 45
angle. The WHC and FBC were calculated by the
following formula:
Wt: of the content of the tube; g ðafter drainingÞ
WHC or FBC ðg=100 gÞ ¼
Wt: of the dried gelatin; g
2.12. Electrophoretic analysis
Protein pattern of the gelatin was analyzed using sodium
dodecyl sulfate poly-acrylamide gel electrophoresis (SDS-PAGE)
according to the method of Laemmli (1970) with slight modifi-
cation. Gelatin, 200 mg was dissolved in 10 mL of 5 g/100 mL
SDS, kept in water bath at 100
C for 30 min and centrifuged for
8500 rpm for 5 min. The supernatant was then mixed with
sample buffer (1:1) contained TriseHCl (pH 6.8), 2-mercaptoe-
thanol, sucrose, bromophenol blue and 5 g/100 mL SDS. Then,
20 ml was loaded along with the standard high MW protein
marker (Fermentas Lifesciences, Germany). Electrophoresis was
carried out at 50 mA initially and then at 100 mA. Gel was
stained with 0.05 g/100 mL Coomassie blue R-250 for visuali-
zation of bands.
2.13. Statistical analysis
The whole experiment was repeated twice. The average mean
values were calculated from three determinations of each experi-
ment and expressed with standard deviations.
Fig. 1. Biochemical composition of red snapper and grouper bones.
3. Results and discussion
3.1. Biochemical composition of fish bones
Moisture content of the red snapper and grouper bones were
around 49 g/100 g (Fig. 1). Bones in general had lower moisture
content than the skins (Binsi et al., 2009). The protein content
ranged between 13 and 15 g/100 g on wet weight basis, with
grouper having higher protein than red snapper. The same range of
protein was observed in bone of young and adult Nile perch
(Muyonga et al., 2004b). The ash contents were very high (19e22 g/
100 g) compared to protein in the bones. Such higher ash content of
39.1 g/100 g was also recorded from the bones of Nile perch by
Muyonga et al. (2004b). The fat percentage was also very high
(13e14 g/100 g) in the bones. This was unique to the fish species
harvested in tropical waters. Variations in the chemical composi-
tion among the fishes and their parts occur due to the differences in
their feed intake, migratory behavior, spawning changes, and
seasonal variations. Taxonomical as well as structural variations
also contribute for the compositional variations in the skin, bone,
fin and scale of the fish species.
3.2. Yield
Yields of gelatin from the bones of red snapper and grouper
were 9.14 and 13.66 g/100 g, respectively. Several authors have
reported different gelatin yields from different species and their
parts. From the bones of Nile perch, 6.1e11.5 g/100 g gelatin yield
was obtained by Muyonga et al. (2004b). Most of the other reports
are pertaining to the yields from the skin of fishes that ranged from
4 to 16 g/100 g (Binsi et al., 2009; Gomez-Guillen et al., 2002;
Jamilah & Harvinder, 2002; Jongjareonrak, Benjakul, Visessanguan,
Prodpran, & Tanaka, 2006; Ninan, Jose, Abubacker, Mathew, &
Geethalakshmi, 2009). The reason for the vast differences in the
 R. Jeya Shakila et al. / LWT - Food Science and Technology 48 (2012) 30e36 33

Fig. 2. Biochemical composition of red snapper and grouper bone gelatin.

yields among the fish species could be due to the loss of collagen freshwater carps (4.01e4.88) (Ninan, Abubacker et al., 2011; Ninan,
during extraction, leaching during washing steps or incomplete Jose et al., 2011) and bigeye snapper (6.44) (Binsi et al., 2009) have
hydrolysis of the collagen (Jamilah & Harvinder, 2002) and differ- been reported. This was mainly because of the different pretreat-
ences in collagen content, their composition in the skin, bone as ments employed during the extraction involving both alkaline and
well as their matrix (Jongjareonrak et al., 2006). The yield was also acid treatments.
influenced with an increase in extraction temperature and decrease
in extraction pH as well as alkali and acid concentrations (Ninan 3.5. Absorption maxima
et al., 2009). In our study, the critical factor responsible for the
difference in the yields among the species was mainly due to their Absorption maxima of the grouper and red snapper fish gelatins
variation in protein content rather than the extraction procedures. were 214 nm, but slightly lower than mammalian gelatin (Table 1).
Commercial gelatin generally showed an absorption maximum of
3.3. Biochemical composition of fish gelatin 230 nm and that of the bovine collagen in the range between 210
and 230 nm (Zhang, Guoying, & Bi Shi, 2006). The absorption
The moisture contents of bone gelatins were in the range of property of the gelatin solution was important as it influences the
4.10e6.24 g/100 g (Fig. 2) and the low moisture was due to barrier properties of the gelatin films particularly to the UV and
lyophilization instead of oven drying. Such lower moisture contents visible light.
were also observed in bigeye snapper skin gelatin, 4.80 g/100 g and
in rohu and common carp skin gelatin, 8.10e8.48 g/100 g (Binsi 3.6. Viscosity
et al., 2009; Ninan, Abubacker et al., 2011; Ninan, Jose et al.,
2011). The protein content ranged between 78 and 82 g/100 g The viscosity of grouper bone gelatin was higher (18.5 cP) than
with grouper having 4 g/100 g higher than red snapper. Such levels red snapper and mammalian bone gelatins (Table 1). Commercial
of protein were also recorded by Muyonga et al. (2004b) in Nile mammalian gelatin viscosity normally ranges between 2.0 and
perch bone. There are many reports on the protein content in skin 7.0 cP and even up to 13.0 cP for specialized ones. The viscosity
gelatins of different fishes ranging from 78.1 to 94.6 g/100 g (Binsi reported for Nile perch bone gelatin was similar to our findings
et al., 2009; Jongjareonrak et al., 2006; Jongjareonrak et al., 2010; (Muyonga et al., 2004b). Variations in the viscosities among fish
Ninan, Abubacker et al., 2011; Ninan, Jose et al., 2011). The bone species and their parts were recorded with giant catfish skin gelatin
gelatin had high ash contents (6.58e10.32 g/100 g) mainly because having 31.3 cP and rohu and common carp gelatin having 6.06 and
of higher mineral content in the raw material. Muyonga et al. 5.97 cP, respectively (Jongjareonrak et al., 2010; Ninan, Abubacker
(2004b) also reported higher ash contents from Nile perch bone et al., 2011; Ninan, Jose et al., 2011). There existed a relationship
gelatins (8.4e11.2 g/100 g), similar to our observation. However, between viscosity and pH of the gelatin solution. Jamilah and
fish skin gelatins contained very low level (0.33e5.9 g/100 g) of ash Harvinder (2002) stated that the effect of pH on viscosity is
(Cho et al., 2006; Haug, Draget, & Smidsrød, 2004; Jongjareonrak minimum at the isoionic point of gelatin and maximum at pH 3 and
et al., 2006). The recommended maximum limit for ash content is 10.5. The same was also confirmed in red snapper and grouper,
only 2.6 g/100 g for edible gelatin but not given specific for skin or which showed higher viscosities at pH 3 and pH 10 (Fig. 3). The
bone gelatin. The fish bone gelatin in general contained lower variation was therefore mainly due to the differences in the pH of
protein than skin gelatin due to the presence of higher ash and fat the extracted gelatin rather than the species or their parts.
contents. It had indicated that inadequate leaching process and
high mineral contents in bones were responsible for high ash
contents (Ninan, Abubacker et al., 2011; Ninan, Jose et al., 2011). An Table 1
Physical properties of gelatin.
additional step is therefore required to remove the mineral as well
as fat contents from the bone gelatin during extraction. Properties Red snapper Grouper Mammalian gelatin
pH 4.65  0.15a 4.31  0.18a 6.18  0.25b
Absorption maxima 214  0.52a 214  0.97a 216  0.66a
3.4. pH
Viscosity (cP) 15.30  0.26a 18.50  0.30b 13.80  0.16c
Melting temp (C) 26.00  0.07a 25.00  0.03a 21.00  0.10b
The pH of the bone gelatins of red snapper and grouper were Gelling temp (C) 16.00  0.12a 16.00  0.18a 22.00  0.23b
4.65 and 4.31, respectively (Table 1). Slight variation in the pH value Gelling time (s) 26.00  0.25a 24.00  0.34a 16.00  0.16b
was observed among the fish species. Wide variations in the pHs of All values are mean  standard deviation.
skin gelatin of cod (2.7e3.9) (Gudmundsson & Hafsteinsson, 1997), Different superscripts in the same row indicate significant differences (P < 0.05).
34 R. Jeya Shakila et al. / LWT - Food Science and Technology 48 (2012) 30e36

Table 2
Functional properties of red snapper and grouper bone gelatin in comparison with
mammalian gelatin.

Red snapper Grouper Mammalian gelatin

Bloom strength (N) 7.55  0.04a 7.72  0.07a 7.95  0.03b
FA ratio 2.20  0.10a 2.08  0.17a 1.92  0.76b
FS ratio 1.64  0.02a 1.46  0.14a 1.44  0.04a
WHC 197.53  3.42a 247.89  4.10b 242.22  3.81b
FBC 493.90  4.20a 429.57  4.73b 191.39  5.20c

FA-Foam forming ability; FS-Foam stability; WHC-Water holding capacity; FBC-Fat

binding capacity.
All values are mean  standard deviation.
Different superscripts in the same row indicate significant differences (P < 0.05).

to 4.17 N for yellow fin tuna (Ninan, Abubacker et al., 2011; Ninan,

Fig. 3. Viscosity of red snapper and grouper bone gelatin at different pH.
3.7. Melting temperature
The melting points of the red snapper and grouper bone gelatins
were more or less similar (25e26
C), but higher than mammalian
bone gelatin. Muyonga et al. (2004b) recorded 26.5e25.9
C as
melting points for bone gelatins of Nile perch. The melting
temperature of tropical fish gelatins were generally high with
22.5e28.9
C for tilapia skin (Jamilah & Harvinder, 2002) and the
cold water cod fish with 8e10
C (Gudmundsson & Hafsteinsson,
1997). This was related to the lower imino acid content and
decreased proline hydroxylation degree in the cold water fish
gelatin (Gomez-Guillen et al., 2002).
3.8. Gelling temperature and gelling time
Gelling temperatures of fish bone gelatins were 6
C lower than
that of mammalian bone gelatin. Gelling temperatures reported for
Nile perch bone gelatin were 18.5e19.0
C (Muyonga et al., 2004b)
and rohu and common carp skin gelatin were 18.52
C and 17.96
C,
respectively (Ninan, Abubacker et al., 2011; Ninan, Jose et al., 2011).
Gelling temperature of cold water fish gelatin was in general lower
than the warm water fish/mammalian gelatins. This difference was
also related to the imino acid composition of the fishes. The imino
acids were found to stabilize the ordered conformation when
gelatin forms the gel network during gelling (Muyonga et al.,
2004a,b). The lower imino acid content in cold water fishes
reduces the propensity for intermolecular helix formation
(Gilsenan & Ross Murphy, 2000).
Gelling and melting temperatures are also influenced by the
change in ionic strength and pH of gelatin. They decreased with the
increase in ionic strength of >0.5 mol/L, which was probably due to
the reduced electrostatic interaction preventing attractive ionic
inter-chain bridging and gelation of fish gelatin (Haug et al., 2004).
Gelling time required for red snapper bone gelatin was more
(56 s) than grouper and mammalian bone gelatin. Muyonga et al.
(2004b) observed much higher gelling time of 90 s in Nile perch
bone gelatin. Similarly, Ninan, Abubacker et al. (2011) and Ninan,
Jose et al. (2011) also observed higher gelling times in rohu
(106 s) and common carp (103 s) skin gelatins.
Jose et al., 2011).
The hydrogen bond formation between the water molecules and
free hydroxyl groups of amino acid in the gelatin are responsible for
the bloom strength (Babel, 1996, p. 10). The other factors that
contribute for the bloom strength are functional complex interac-
tions with imino groups, the ratio of a-chain, the amount of b-
component and a higher content of free hydroxyl group amino-
acids (Arnesen & Gildberg, 2002). The bloom strength is also
dependent on the isoelectric point of the gelatin solution. To verify
this, bloom strengths of bone gelatin were examined at varying pH
(Fig. 4) and the results showed that it increased with increasing pH
with the maximum at pH 7.0 and later it decreased. Therefore, more
compact and stiffer gels can be formed by adjusting the pH of the
gelatin close to its isoelectric point, where the proteins are more
neutral and the gelatin polymers come closer to each other
(Gudmundsson & Hafsteinsson, 1997).
3.10. FA and FS
The FA and FS ratios of the fish bone gelatin were higher than
mammalian bone gelatin with red snapper bone gelatin having
higher ratios (Table 2). Also, the FA ratios of rohu and common carp
skin gelatin were 2.55 and 2.45 while the FS ratios were 1.83 and
1.91, respectively (Ninan, Abubacker et al., 2011; Ninan, Jose et al.,
2011).
A protein must be capable of migrating rapidly to the airewater
interface, unfolding and rearranging at the interface to express
good foaming ability. To have good adsorption at the airewater
interface, proteins should contain hydrophobic regions (Mutilangi,
Panyam, & Kilara, 1996). A positive correlation exists between
hydrophobicity of unfolded proteins and foaming characteristics.

3.9. Bloom strength

Bloom strength of grouper bone gelatin was higher (7.55 N) than

red snapper (Table 2). The bloom strength of young and adult Nile
perch bone gelatins was found to be very low, 2.2 N (Muyonga et al.,
2004b). There are reports on the bloom strengths of fish skin
gelatins of other tropical fishes ranging from 1.22 N for sincroaker Fig. 4. Gel strength of red snapper and grouper bone gelatin at different pH.
 R. Jeya Shakila et al. / LWT - Food Science and Technology 48 (2012) 30e36
Additional hydrophobic residues also form a large hydrophobic
sphere on the surface of the protein and improve the foaming
capacity. The differences in FA and FS ratios of the fish bone gelatins
were mainly because of the differences in the hydrophobic amino
acid contents among the species.
The FS ratios indicate the extent of proteineprotein interaction
within the matrix (Mutilangi et al., 1996) and it was found to have
a positive correlation with the molecular weight of peptides.
Variations in FS ratios were also noticed between catfish skin
gelatin and calf skin gelatin by Jongjareonrak et al. (2010) similar to
our observations.
3.11. WHC and FBC
WHC of the red snapper bone gelatin was lower compared to
grouper and mammalian bone gelatins (Table 2). Lower WHC is
mainly related to the lower amounts of hydrophilic amino acids and
lower hydroxyproline content (Ninan, Abubacker et al., 2011;
Ninan, Jose et al., 2011). The FBC of fish bone gelatin was very high
compared to mammalian gelatin, with red snapper having 494 g/
100 g. The degree of exposure of the hydrophobic residues and the
high amount of tyrosine were found responsible for the high FBC
(Ninan, Abubacker et al., 2011; Ninan, Jose et al., 2011).
3.12. Electrophoretic characterization of gelatin
Protein patterns of the bone gelatins of red snapper and grouper
showed three distinct bands a, b, and g chains (Fig. 5). The
b components were more with MW > 200 kDa, while the
a components had MW around 130 kDa. Most of the triple helical
structures of a chain were not been destroyed. There were no MW
fractions of gelatin. However, some authors have observed lower
MW fractions of gelatin, particularly due to high temperature
extraction (Muyonga et al., 2004b). According to Gomez-Guillen
et al. (2002), damage or partial losses of a chains occurring due
Fig. 5. Protein patterns of gelatin from red snapper and grouper bone. M-Molecular
marker; RsBG-red snapper bone gelatin; GBG-grouper bone gelatin.
35
to the heat extraction and gel preparation were responsible for the
lower MW fractions. The same was also confirmed by Binsi et al.
(2009) in bigeye snapper skin gelatin.
Degradation of gelatin to lower MW fractions was responsible
for low viscosity, low melting point, low setting point, high setting
time, as well as decreased bloom strength of gelatin (Normand,
Muller, Ravey, & Parker, 2000; Muyonga et al., 2004b). The fish
bone gelatin extracted did not contain lower MW fractions as the
temperature employed for extraction was low (45
C) and this had
given high bloom strength and foaming ability.
4. Conclusion
The results indicated that bones of red snapper and grouper can
be better utilized for commercial gelatin extraction with good
yields. High viscosity, bloom strength and WHC of bone gelatin find
them a better alternative for mammalian gelatin. Extraction
procedure adopted was found satisfactory as there were no protein
degradation; however slight modification is essential to eliminate
mineral and fat contents in the gelatin extraction. The good func-
tional properties of fish bone gelatin will find them a better place in
food industries as thickeners, gelling and foaming agents. Their
applications in edible film formation, coatings, nanocomposite
films and micro or nano-encapsulation also need further
exploration.
Acknowledgments
The authors thank the Dean, Fisheries College and Research
Institute, Tuticorin for his encouragement and for providing the
necessary facilities to undertake this work. The authors gratefully
acknowledge the financial support from Department of Biotech-
nology, GOI, New Delhi.
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