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Bioresource Technology 159 (2014) 280285

Contents lists available at ScienceDirect

Bioresource Technology
journal homepage: www.elsevier.com/locate/biortech

Characterization of agarophytic seaweeds from the biorenery context


Ravi S. Baghel, C.R.K. Reddy , Bhavanath Jha
Discipline of Marine Biotechnology and Ecology, CSIR Central Salt and Marine Chemicals Research Institute, Bhavnagar 364002, Gujarat, India
Academy of Scientic and Innovative Research (AcSIR), New Delhi, India

h i g h l i g h t s

g r a p h i c a l a b s t r a c t

 Characterization of agarophytic

seaweed biomass for commercial


products.
 Investigated seaweeds contained
good amount of industrially
important pigments.
 Appreciable amount of macro and
micro-minerals were estimated.
 Investigated seaweeds contained
good quantity of protein and
cellulose.
 Low lipid contents and higher agar
contents were determined.

a r t i c l e

i n f o

Article history:
Received 19 December 2013
Received in revised form 18 February 2014
Accepted 22 February 2014
Available online 3 March 2014
Keywords:
Agarophytes
Agar
Cellulose
Pigments
Protein

a b s t r a c t
The major seaweed components such as natural colorants (R-phycoerythrin (R-PE), R-phycocyanin
(R-PC)), minerals, proteins, lipids, cellulose and agar which are of considerable commercial value were
estimated in 15 different agarophytic seaweeds. R-PE and R-PC contents ranged from 138.33 17.67
to 1039.43 27.65 lg/g and 50.26 6.63 to 818.2 48 lg/g on fresh weight (fw) basis, respectively.
Appreciable amounts of both macro-minerals (K, Na, Ca, Mg) and micro-minerals (Fe, Zn, Se, Mn) were
estimated. The total lipid and protein contents were 0.65 0.06% to 1.53 0.07% and 4.75 0.5% to
19.31 3.5% on dry weight (dw) basis respectively while cellulose and agar contents varied from
3.7 0.13% to 12.20 0.45% and 9.17 0.62% to 25.23 0.50% dw, respectively. The overall nding of this
study enable the selection and value addition of agarophytic feedstock for biorenery.
2014 Elsevier Ltd. All rights reserved.

1. Introduction
Marine macroalgae are harvested worldwide from both cultivated farms and wild stocks, and used as feedstock for food,
phycocolloids, phytosupplements (soil additives, fertilizers),
pharmaceuticals, nutraceuticals and cosmetics (Hanisak, 1998;
McHugh, 2003). Phycocolloids are considered as one of the major

Corresponding author at: Discipline of Marine Biotechnology and Ecology, CSIR


Central Salt and Marine Chemicals Research Institute, Bhavnagar 364002, Gujarat,
India. Tel.: +91 278 256 5801/256 3805x6140; fax: +91 278 256 6970/256 7562.
E-mail address: crk@csmcri.org (C.R.K. Reddy).
http://dx.doi.org/10.1016/j.biortech.2014.02.083
0960-8524/ 2014 Elsevier Ltd. All rights reserved.

commercial seaweed extracts with a market value over US$ 1 billion. Among the phycocolloid yielding seaweeds harvested and
processed worldwide, agarophytes constitute about 72,000 dry
tonnes from which ca. 9600 tonnes of agar produced annually with
a market value over US$173 million (Bixler and Porse, 2011).
Agarophytes are typically the red algae that include many commercially exploited genera Gracilaria, Gelidium and Gelidiella. The
members belonging to the genera of Gracilaria and Gelidium are
the major agar producers in the world, while Gelidiella acerosa is
the preferred source of agar in India (McHugh, 2003). Farming of
Gracilaria is being carried out in several countries including the
Philippines, Chile, China, Korea, Indonesia, Namibia, Vietnam and
Argentina at a commercial scale (McHugh, 2003). Some of the

R.S. Baghel et al. / Bioresource Technology 159 (2014) 280285

agarophytic species particularly Gracilaria corticata which grows


abundantly along different coastal regions of India is largely remained as untapped resource due to its inferior quality agar in
terms of gel strength. The potential utilization of this and others
for bioenergy could be realized only if their potential for other
components having high commercial value are recovered. Till date,
agarophytic seaweeds are used for only commercial production of
agar. The earlier studies have largely restricted to agar extraction
that too from a limited number of species only. Also, no attempt
made to explore their potential for extraction of other value added
products of commercial importance. While recovering agar, the
agarophytic species could be utilized for extraction of a number
of biochemical components such as natural colorants (pigments),
proteins, lipids, minerals and cellulose leading to an effective utilization of total biomass. Recently certain seaweeds have been
investigated for natural pigments R-phycoerythrin and R-phycocyanin content from Porphyra spp. (Sampath-Wiley and Neefus,
2007), Grateloupia turuturu (Denis et al., 2009), Palmaria palmate
(Dumay et al., 2013) and Gracilaria crassa (Baghel et al., 2014). Earlier reports have demonstrated the agarophytes as a rich source of
macro-elements (Na, K, Ca, and Mg) over the terrestrial vegetables
besides containing appreciable amount of protein (Kumar et al.,
2011). Apart from that, algal lipids are rich sources of PUFAs that
exhibit anti-hypercholesterolemic, antioxidant, anticancer, antidiabetic, antihypertensive, anti-inammatory activities and thus
agarophytic algae could also be a promising raw materials for lipid
products that can be utilized in pharmaceutical and functional food
industry (Kumari et al., 2013). Moreover certain seaweeds are reported to have a good quantity of cellulose content (Siddhanta
et al., 2009). Recently Kumar et al. (2013) characterized higher content of cellulose (holocellulose 60%) in the residual pulp of Gracilaria verrucosa after agar extraction and subsequently used it as a
feedstock for bioethanol production. The cellulose recovered as a
by-product from agar producing algal species could be an alternative feedstock for bioenergy. Therefore, seaweeds are nowadays
gaining the global attention as an alternative renewable feedstock
for biofuels. To date, commercial biofuels production depends on
rst generation substrates such as sugarcane and corn. But biofuel
production from such sources led to an intense debate Food
verses Fuel. Further, the second generation biofuels from lignocellulosic biomass has so far remained unviable due to lack of costeffective and environment friendly downstream process. In view
of the above stated inherent constraints associated with rst and
second generation biofuel feedstock, an attention has been turned
to marine macroalga as a possible feedstock for bioenergy because
of their higher productivity, no competition with agricultural land,
no agricultural inputs, amenability for efcient depolymerisation,
etc. It has been evident from the earlier investigations that processing of feedstock for bioethanol/biobutanol per se is not economical
and thus, led to a biorenery approach value adding the feedstock
through recovery of a series of other major components in an integrated manner. The major gains of biorenery is that the high production cost of biofuel can be compensated by co-production of
other value added products such as proteins, lipids, pigments, minerals, cellulose, polysaccharides from biofuel feedstock.
The previous investigations have recently demonstrated recovery of at least two or three products in an integrated manner from
different seaweed feedstocks such as agar and bioethanol from G.
verrucosa (Kumar et al., 2013), nutrient rich liquid fertilizer and
bioethanol from Kappaphycus alvarezii (Khambhaty et al., 2012),
pigment and agar from Gracilaria lemaneiformis (Niu et al., 2013),
and fuel intermediates, agricultural nutrients and pure water from
K. alvarezii seaweed (Mondal et al., 2013). However, the studies on
recovery of all major seaweed components are scarce. It is this context, a comprehensive effort was made to characterize the red seaweed biomass for all major components aiming at their possible

281

recovery in an integrated manner. Thus, the present investigation


was accomplished to determine the pigments, minerals, proteins,
lipids, agar and cellulose content from different agarophytic seaweeds from Indian coast.
2. Methods
2.1. Collection of algal samples
Macroalgal samples were collected during low tide periods during 20112013 from different sites along the Gujarat and Tamil
Nadu coast, India (Table 1). The sample were transported to the
laboratory under cool conditions and washed with sterile seawater
to remove undesired foreign particles and epiphytes. Approximately 500 mg of algal tissues for each sample were frozen and
stored at 40 C for the determination of pigment content and rest
of the material was shade dried for the determination of other biochemical products.
2.2. Quantication of pigments
For the quantication of pigments, R-phycoerythrin (R-PE) and
R-phycocyanin (R-PC), 100 mg of frozen algal tissues were homogenized in liquid nitrogen with mortar and pestle. The homogenate
were added to 0.8 ml of 0.1 M phosphate buffer (pH 6.8) and incubated overnight at 4 C. After incubation, homogenized samples
were vortexed and centrifuged at 15,000g for 10 min, 4 C. The
supernatant were collected and residues were added with 0.2 ml
of phosphate buffer followed by vortexing. Homogenate were centrifuged at 15,000g for 10 min, 4 C and supernatants were collected. The absorbances were recorded at 564, 618 and 730 nm
using a dual-beam UVvisible spectrophotometer. The contents
of R-PE and R-PC were calculated according to Sampath-Wiley
and Neefus (2007) as follows:

R  PC 0:154A618  A730
R  PE 0:1247A564  A730  0:4583A618  A730
2.3. Mineral composition analysis
For mineral composition, 100 mg grounded dried samples were
treated with 10 ml of conc. HNO3 overnight (Santoso et al., 2006).
Thereafter, 2.5 ml of conc. HClO4 and 250 ll of H2SO4 were added
to the samples followed by heating until no white smoke was emitted. 100 ml of 2% HCl was added in the digested sample and ltered with a 0.22-lm membrane lter. The samples were
analyzed using inductively coupled plasma atomic emission spectroscopy (Perkin-Elmer, Optima 2000, USA).
2.4. Total protein and lipid content determination
The organic nitrogen content were quantied with dry ne
grounded sample using the instrument, Elementar Analysensysteme GmbH vario MICRO cube, calibrated using sulfanilamide as
a reference standard. The total protein contents were estimated
by multiplying the nitrogen content by a factor of 6.25 (Loureno
et al., 2002). Lipids were extracted from 500 mg dry algal samples
following the method of Bligh and Dyer (1959) and lipid content
were determined by the gravimetric method.
2.5. Cellulose extraction and characterization
Cellulose was extracted from the seaweeds samples following
the method reported by Mihranyan et al. (2004). 10 g of dried algal

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R.S. Baghel et al. / Bioresource Technology 159 (2014) 280285

Table 1
Description of collection sites of different agarophytic seaweeds.
Species

Collection site

Location

Gelidium micropterum

Veraval
Old Mandapam Jetty
Veraval
Valinokkam
Veraval
Adri
Nochiyoorani
Veraval
Old Mandapam Jetty Valinokkam

N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N

Gelidium pusillum
Gelidiella acerosa

Gelidiopsis variabilis

Gracilaria edulis
Gracilaria salicornia
Gracilaria dura

Gracilaria corticata

G. corticata v. cylindrica

Gracilaria folifera
Gracilaria textorii
Gracilaria fergusonii
Gracilaria crassa
Gracilaria debilis
Gracilaria verrucosa

Krusadai Iceland
Jegathapattinam
Veraval
Rameswaram
Veraval
Adri
Okha
Veraval
Adri
Nochiyoorani
Veraval
Adri
Krusadai Iceland
Veraval
Krusadai Iceland
Veraval
Veraval
Adri
Rameswaram
Pamban
Okha
Okha

powder were defatted with repeated extraction with 150 ml MeOH


for a period of eight days at room temperature. The defatted algal
powder were added with 150 ml acetate buffer containing 3.6 g
NaClO2 and kept for bleaching at 60 C for 6 h. The bleached algal
samples were washed with water until the pH reached 7. The
washed algal were treated with 60 ml of 0.5 M NaOH solution at
60 C overnight. The alkali treated algal materials were washed
with water till neutrality, ltered and dried at room temperature.
The dried materials were re-suspended in 40 ml hydrochloric acid
(5% v/v) and were heated up to boiling and resultant slurry were
kept overnight at ambient temperature (30 C), followed by water
washing for removing the excess acid, ltered and dried to get cellulose. Cellulose yields were calculated based on initial seaweed
samples used for extraction process. Celluloses of the greatest
yields (%) (Gelidium pusillum) was selected for characterization.
The Fourier transform infrared (FT-IR) spectra of the cellulose samples were recorded on a Perkin-Elmer Spectrum GX FTIR (USA)
instrument.
2.6. Agar extraction, characterization and analysis of physical
properties
The native agar from Gracilarials were extracted following the
method of Meena et al. (2008) while native agar extraction from
Gelidium micropterum, G. pusillum and G. acerosa involved pretreatment of 0.5% acetic acid for 1 h. Agar of the greatest yields (%) (G.
pusillum) was selected for characterization. The Fourier transform
infrared (FT-IR) spectra of the cellulose samples were recorded
on a Perkin-Elmer Spectrum GX FTIR (USA) instrument. Physical
properties (gel strength, gelling temperatures) were determined
in a 1.5% agar solution. The gel strength g/cm2 was measured using
Gel Tester (Kiya Seisakusho, Ltd., Tokyo, Japan). Gelling temperatures of gel samples were measured following the method of
(Meena et al., 2008).

20 54.870 ; E 70 20.830
9 17.140 ; E 79 09.530
20 54.870 ; E 70 20.830
9 09.930 ; E 78 38.070
20 54.870 ; E 70 20.830
20 57.580 ; E 70 16.760
9 15.940 ; E 79 01.980
20 54.870 ; E 70 20.830
9 17.140 ; E 79 09.530
9 09.930 ; E 78 38.070
9 14.220 ; E 79 12.300
9 57.500 ; E 79 11.610
20 54.870 ; E 70 20.830
9 18.540 ; E 79 20.360
20 54.870 ; E 70 20.830
20 57.580 ; E 70 16.760
22 27.060 ; E 69 4.020
20 54.870 ; E 70 20.830
20 57.580 ; E 70 16.760
9 15.940 ; E 79 01.980
20 54.870 ; E 70 20.830
20 57.580 ; E 70 16.760
9 14.220 ; E 79 12.300
20 54.870 ; E 70 20.830
9 14.220 ; E 79 12.300
20 54.870 ; E 70 20.830
20 54.870 ; E 70 20.830
20 57.580 ; E 70 16.760
9 19.270 ; E 79 17.710
9 16.230 ; E 79 11.300
22 28.440 ; E 69 03.580
22 28.440 ; E 69 03.580

2.7. Statistical analysis


All the analyses were performed in triplicate (except mineral
composition which was analyzed in duplicates) and the mean values were recorded.
3. Results and discussion
3.1. Pigments content
Fifteen different agarophytic seaweeds belonging to three different families Gelidiaceae, Gracilariaceae, and Lomentariaceae
were evaluated for their pigment R-phycoerythrin (R-PE) and Rphycocyanin (R-PC) potentials (Fig. 1). Pigments contents were
presented in Table 2. Among the investigated algal species, R-PE
contents ranged from 138.33 17.67 to 1039.43 27.65 lg/g fw.
Highest R-PE content 1039.43 27.65 lg/g fw was recorded for
G. micropterum followed by Gelidiopsis variabilis (935.79 33) and
G. pusillum (887.30 98 lg/g fw). Lowest R-PE content
138.33 17 mg/g fw was recorded for Gracilaria Salicornia.
R-phycicynain (R-PC) contents ranged from 50.26 6.63 lg/g to
818.2 48 lg/g fw. Phycobiliproteins, R-PE and R-PC possess excellent spectroscopic properties, high absorption coefcients, and
high quantum yields which make them useful for wide range
applications in biomedical research and diagnostics as a uorescent probes (Ayyagari et al., 1995). The various synthetic dyes
widely used in food and cosmetics as colorants that are considered
as harmful to health and personal care. It is important to have a
non toxic pigment as an alternative to synthetic toxic pigments.
Phycobiliproteins are used as natural colorants for food and cosmetics (Pangestuti and Kim, 2011). The non toxic nature of natural
pigments along with various biological activities such as antioxidant, anticancer, anti-inammatory, anti-obesity, anti-angiogenic
and neuroprotective activities make them suitable alternatives to

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Table 2
Estimated pigments content (lg/g fw) in different agarophytic seaweeds (Mean SD,
n = 3).

Fig. 1. Absorbance spectra of crude aqueous extract of different agarophytic


seaweeds. R-PE, R-phycoerythrin; R-PC, R-phycocyanin.

synthetic toxic pigments in food, cosmetics and pharmaceutical


application (Naidu et al., 1999; Pangestuti and Kim, 2011).
Screened algal species showed good content of R-PE and R-PC.
Therefore, these agarophytic seaweeds could be an industrial
source of these pigments.
3.2. Mineral composition
ICP analysis of the extracts from different samples for minerals
revealed that agarophytic seaweeds contained high amounts of
the macrominerals (K, Na, Ca and Mg) as well as appreciable quantity of trace elements (Fe, Mn, Mo, Se, Zn and Ni) needed in human
nutrition (Table 3). The members of Gracilariaceae contained high
amount of K ranged from 7910 29.69 to 15,033 69.29 mg/100 g
dw, followed by Na ranged from 2208 8.48 to 5217 52.32 mg/
100 g dw, which were in accordance with the values reported by
Kumar et al. (2011). The member of Gelidiaceae and Lomentariaceae
showed comparatively low content of K which ranged from
831.9 10.60 to 5665.5 3.53 mg/100 g dw. Moreover all the
agarophytes invested in this study contained good amount of nutritionally important macrominerals Ca (243.2 1.831643.5
11.45 mg/100 g dw) and Mg (237.1 12.02855.3 14.00 mg/
100 g dw). Further, Ca level was comparable to those of lettuce
(177.4 mg/100 g dw) and cabbage (368 mg/100 g dw) while Fe content for the most of the seaweeds (8.8 2.5470.8 0.28 mg/100 g
dw) were even higher than spinach (23.3 mg/100 g dw) (USDA,
2001). However, Mn, Mo, Se, Zn and Ni were detected in small
amounts. Recent studies showed an effective application of different
seaweeds minerals rich extract as a liquid fertilizer on various crops
such as Triticum aestivum var. Pusa Gold and wheat (Kumar and
Sahoo, 2011; Shah et al., 2013). The process for fractionation of minerals rich liquid extract without destroying the main product was
successfully demonstrated by Eswaran et al. (2005). As the agarophytic seaweeds contained good quantity of plant nutritionally
important macro and microminerals which could make them a suitable source of commercial liquid fertilizer for various agricultural
and vegetable crops.

Species

R-phycoerythrin (R-PE)

R-phycocyanin (R-PC)

Gelidium micropterum
Gelidium pusillum
Gelidiella acerosa
Gelidiopsis variabilis
Gracilaria edulis
Gracilaria salicornia
Gracilaria dura
Gracilaria corticata
G. corticata v. cylindrica
Gracilaria folifera
Gracilaria. textorii
Gracilaria fergusonii
Gracilaria crassa
Gracilaria debilis
Gracilaria verrucosa

1039.43 27.65
887.30 98.95
684.15 38.36
935.79 33.03
373.25 13.48
138.33 17.67
383.71 19.89
657 7.93
380 26.85
752.66 3.78
193.33 25.71
264.33 14.01
237.45 11.15
249.33 28
684 10

269 12.16
445.06 42.92
595.98 43.12
818.2 48.40
276.68 23.37
50.26 6.63
180.18 24.05
323.66 10.69
268.33 30.02
611.66 3.21
54 3.4
216.66 52.54
237.16 9.24
205.66 39.92
315 6.4

(Kumar et al., 2011). However, seaweeds could not be a conventional source of energy since their lipid content is low, but their
PUFA contents can be as high as those of terrestrial vegetables
(Darcy-Vrillon, 1993; Kumari et al., 2010). Since seaweeds lipid
contained good amounts of PUFA, could be useful for nutraceuticals. Protein contents for investigated seaweeds were ranged between 4.75 0.5% and 19.31 3.5% dw (Table 4) which were
accordance to previously reported protein contents for different
seaweeds (McDermid and Stuercke, 2003; Gressler et al., 2010;
Kumar et al., 2011; Narasimman and Murugaiyan, 2012). Most of
the Gracilarials showed protein contents ranged between 4% and
10% dw except G. corticata (15.08 1.42% dw) and G. foliifera
(15.16 3.51% dw). G. pusillum, G. micropterum, G. variabilis and
G. acerosa (19.31 3.5%, 18.45 3.70%, 17.62 1.86% and
14.93 1.89%) were showed comparatively higher protein contents. Such higher protein content recorded in these seaweeds
indicates their possible utilization for commercial protein production to full human and animal nutritional requirements.
3.4. Cellulose content
The yield of cellulose were ranged from 3.7 0.13% to
12.20 0.45% dw (Table 4). The highest yield of cellulose obtained
from G. pusillum, (12.20 0.45%) followed by G. variabilis
(11.38 0.62%), G. micropterum (10.63 0.41%) and G. acerosa
(9.77 0.23). Lowest cellulose contents were recorded from Gracilaria dura (3.7 0.13%) and G. foliifera (3.77 0.10%). The cellulose
yields (3.712%) obtained from agarophytic seaweeds were
comparable to cellulose yields (213%) previously reported by
Siddhanta et al. (2009, 2011) for different seaweeds. The algal
cellulose could be used for bioenergy production.
The FT-IR spectra of cellulose obtained from G. pusillum is presented in Supplementary Fig. 1. The characteristic bands of cellulose were found to be the similar as earlier reported FT-IR
spectra for cellulose in the literature (Siddhanta et al., 2011). The
characteristics bands of cellulose were (KBr, mmax, cm1) at 3428
(OH stretching), 2923 (CH str), 1628 (bound H2O), 1434 (CH
bending) and 1022 (COC bending) found (Supplementary Fig. 1).
3.5. Agar contents and agar physical properties

3.3. Total lipid and protein content


The total lipid contents varied with species and ranged between
0.65 0.06% and 1.53 0.07% dw basis (Table 4). The present study
shown that the investigated seaweeds contained low amounts of
lipid content which are in accordance to the previous reports (ranged 0.57 0.063.03 0.21%) published for different seaweeds

The agar yields for the investigated seaweeds were ranged from
9.17 0.62% to 25.23 0.50% dw (Table 5) which were in accordance with yields reported earlier for different seaweeds in literature (Mehta et al., 2010; Meena et al., 2008; Prasad et al., 2007).
Highest yield of agar recorded from G. pusillum (25.23 0.50%) followed by G. dura (25.15 0.78%) and G. acerosa (24.5 0.7%).

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R.S. Baghel et al. / Bioresource Technology 159 (2014) 280285

Table 3
Major minerals and trace elements (mg/100 g dw) determined by atomic absorption spectrophotometry in different agarophytic seaweeds (Mean SD, n = 2).

K
Na
Ca
Mg
Fe
Mn
Mo
Ni
Se
Zn

K
Na
Ca
Mg
Fe
Mn
Mo
Ni
Se
Zn

G. micropterum

G. pusillum

G. acerosa

G. variabilis

G. edulis

G. salicornia

G. dura

G. corticata

831.9 10.60
1473.6 6.22
484.4 3.67
399.8 2.26
25.8 0.01
3 0.01
1.2 0.84
0.7 0.14
16.4 0.56
7.4 0.28

1515.1 28.14
361.3 6.92
727.1 23.61
237.1 12.02
47 0.84
4.8 0.01
1.2 0.28
1 0.01
15.1 0.14
5.1 1.27

5665.5 3.53
2394.5 113.84
377.85 86.05
607.25 122.68
64.55 2.89
5.1 0.01
0.75 0.07
1 0.14
0.7 0.01
24.55 1.76

1159.5 57.27
5217 52.32
1643.5 11.45
855.3 14.00
64.1 0.98
5 0.01
1.8 0.28
2.5 0.14
9.8 0.28
5 0.01

13715 162.63
2208 8.48
672.6 27.71
349.4 14.70
58.3 0.42
11.3 0.14
1.7 0.14
0.8 0.01
9.6 1.97
6.7 0.14

15033 69.29
3732 104.65
881.5 20.22
510.5 12.86
70.8 0.28
5.1 0.14
0.9 0.70
0.8 0.01
10.1 2.40
3.8 0.28

13175 968.73
4483 270.11
253.75 6.71
441.55 8.131
33.55 2.33
9.65 0.07
0.9 0.14
0.35 0.07
1.55 0.91
5.9 0.14

7910 29.69
2680 87.64
310.65 34.011
361.5 36.76
21.65 2.05
12.55 0.21
0.75 0.07
0.9 0.14
1.45 0.07
16 0.56

G. c. v. cylindrica

G. foliifera

G. textorii

G. fergusonii

G. crassa

G. debilis

G. verrucosa

9575 1718.26
2947.5 531.03
336.05 89.30
521.7 85.70
30.4 10.04
9.7 1.41
0.65 0.21
1 0.14
2.45 0.35
10.9 1.41

9491 572.75
3114.5 183.14
243.2 1.83
334.85 16.75
15.7 2.12
4.9 0.14
0.6 0.14
0.95 0.21
0.95 0.21
7.3 1.13

12960 664.68
3150.5 405.17
404.9 24.18
501.5 45.25
31.7 0.84
14.8 8.28
0.7 0.42
0.75 0.21
2.6 0.28
13.25 1.06

11139 1132.78
3015 472.34
255.8 5.09
397.15 49.99
32.25 7.0
5.35 0.49
1.45 1.06
1.15 0.35
1.65 0.21
5.95 1.76

11170 686.94
4105 682.0
255.63 5.76
438.5 7.78
29.7 2.30
8.33 2.28
0.66 0.15
0.366 0.05
ND
6.33 0.75

11480 325.26
4410.5 84.14
506.25 181.93
429.4 140.29
26 2.54
11.2 0.70
0.75 0.07
0.75 0.07
1.7 0.28
8.1 1.13

7924 317.78
4524 213.54
305.9 5.51
498.3 41.01
8.8 2.54
7.4 0.424
1.1 0.14
0.4 0.01
2.25 0.318
4.05 0.07

ND, not detected.

Lowest agar yield obtained from G. fergusonii (9.17 0.62) and


G. textorii (11.18 0.62). G. micropterum was very rst time investigated for their agar potentials contained 23.54 0.52% agar content. The agar contents for members of Gracilaria genus ranged
from 9.17 0.62% to 25.15 0.78%. The FT-IR spectra of agar obtained from G. pusillum is presented in Supplementary Fig. 2. The
characteristic bands of agar at 741, 775 and 889 and 931 cm1
were found to be the similar as earlier reported FT-IR spectra for
agar in the literature (Meena et al., 2011). Due to unavailability
of sufcient algal materials, seaweeds G. debilis, G. verrucosa and
G. variabilis were not investigated for agar contents. However agar
yields for G. debilis 14.8 0.5 dw (Mehta et al., 2010) and 2733%
for G. verrucosa (Kumar et al., 2013) were reported. The highest
gel strength recorded for G. pusillum agar (750 30 g/cm2)
followed by G. micropterum (720 20 g/cm2), G. acerosa (423 15
g/cm2) and G. crassa (290 14 g/cm2) (Table 5). Out of 12 different
species investigated, six species produced low gel strength agar
(P100 g/cm2). The gelling temperatures for studied agar were
ranged from 33 0.5 to 45 0.5 (Table 5).
The overall ndings of this study highlight the scope for
increasing the attractiveness of seaweed biorenery through
development of sustainable downstream processes for recovery
Table 4
Total lipid, protein and cellulose content (% dw) of different agarophytic seaweeds
(Mean SD, n = 3).
Species

Lipid

Protein

Cellulose

G.
G.
G.
G.
G.
G.
G.
G.
G.
G.
G.
G.
G.
G.
G.

1.08 0.09
1.32 0.06
1.41 0.04
1.27 0.05
0.98 0.03
1.2 0.04
1.02 0.06
1.53 0.07
1.49 0.10
1.47 0.04
0.76 0.04
0.65 0.06
1.30 0.05
1.26 0.09
1.34 0.06

18.45 3.70
19.31 3.5
14.93 1.89
17.62 1.86
9.83 2.78
4.75 0.5
5.33 0.69
15.08 1.42
9.35 0.62
15.16 3.51
9.56 1.05
8.91 2.42
5.18 0.64
7.06 0.40
7.87 3.25

10.63 0.41
12.20 0.45
9.77 0.23
11.38 0.62
4.88 0.12
4.90 0.15
3.7 0.13
5.27 0.25
6.08 0.19
3.77 0.10
4.62 0.24
4.20 0.21
7.21 0.20
NS
NS

micropterum
pusillum
acerosa
variabilis
edulis
salicornia
dura
corticata
c. v. cylindrica
foliifera
textorii
fergusonii
crassa
debilis
verrucosa

NS, not studied.

Table 5
Yield, gel strength and gelling temperature of agar extracted from different
agarophytic seaweeds (Mean SD, n = 3).
Species

Agar yield
(%)

Gel strength (g/


cm2)

Gelling
temperature(C)

G.
G.
G.
G.
G.
G.
G.
G.
G.

23.54 0.52
25.23 0.50
24.5 0.7
NS
17.83 0.40
15.06 0.61
25.15 0.78
23.62 0.74
18.68 0.86

720 20
750 30
423 15

>100
256 15
250 10
103 5
>100

36 0.5
44 0.5
38.5 0.5

43.5 0.5
45 0.5
33 0.5
40.5 0.5
36 0.5

18.29 0.54
11.18 0.62
9.17 0.62
21.14 0.82
NS
NS

>100
>100
>100
290 14

35.5 0.5
35 0.5
33 0.5
38.75 1

G.
G.
G.
G.
G.
G.

micropterum
pusillum
acerosa
variabilis
edulis
salicornia
dura
corticata
c. v.
cylindrica
foliifera
textorii
fergusonii
crassa
debilis
verrucosa

NS, not studied.

of bulk products from feedstock. The advantage of such downstream process is that it will enable valorization of seaweed biomass for a spectrum of bio-products of high value. As
decolorization and defatting improves the extraction and quality
of various polysaccharides, therefore integrated biorenery approach enable fractionation of these two valuable products i.e. pigment and lipid prior to polysaccharide extraction. Despite several
advantages of seaweed biorenery concept, the eld is yet to gain
the momentum due to several factors including lack of cost effective offshore farming techniques, integrated downstream process,
value addition of fractionated products, etc. The offshore farming
together with setting up small scale bioreneries in coastal areas
can some extent offset the high production cost by reducing the
transportation cost involved in bringing the biomass from its production to processing place.
4. Conclusion
Agarophytic seaweeds investigated in this study were found
rich in natural pigments, minerals, proteins and cellulose content

R.S. Baghel et al. / Bioresource Technology 159 (2014) 280285

apart from agar. Most of these products have historically established global markets with immense economic value. Biomass
characterization carried out in this study will aid in development
of downstream process for sequential recovery of these products
in an integrated manner from feedstock. The viability and sustenance of algal biorenery is largely depends on the biomass valorization technologies employing eco-friendly downstream process.
Acknowledgements
The nancial support received from Project CSC 0116 BioEn is
gratefully acknowledged. Authors would like to acknowledge
Head, Analytical sciences, CSIR-CSMCRI for permitting to use
instrumentation facilities.
Appendix A. Supplementary data
Supplementary data associated with this article can be found, in
the online version, at http://dx.doi.org/10.1016/j.biortech.2014.
02.083.
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