Carbon-Rich Wastes as Feedstocks for Biodegradable Polymer

(Polyhydroxyalkanoate) Production Using Bacteria

Jasmina Nikodinovic-Runic, ... Kevin E. O Connor, in Advances in
Applied Microbiology, 2013
7.2 Bagasse and vinasse
Bagasse is another by-product of sugar industry. It is the
heterogeneous fibrous residue that remains after sugarcane stalks are
crushed for sugar extraction. Typically, from processing 100 tons of
sugarcane in a factory, 30–34 tons of bagasse is obtained (of which
22–24 tons is used in processing and 8–10 tons is saved) (Solomon,
2011; Yadav & Solomon, 2006). Dry bagasse consists of 45%
cellulose, 28% hemicellulose, 20% lignin, 5% sugar, 1% minerals, and
2% ash. Bagasse is similar in component to wood except that it has
high moisture content. Thus, it is currently used as a biofuel and in the
manufacture of pulp and paper products, filler for building materials
and as a substrate for growing mushrooms. Considering the fact that
much more bagasse is available than utilized for the energy
production, further value creation might be possible. As an example,
hemicelluloses and cellulose fractions of bagasse can be hydrolyzed
and utilized by suitable microbial strains for PHA biosynthesis. Very
often hydrolysis is performed using dilute acid and elevated
temperatures (100–130°C), which generate hydrolysates inhibitory
to microorganisms. In the optimization study, highly
tolerant inoculum of Ralstonia eutropha was developed through
directed adaption and evolution to the bagasse hydrolysate (Yu &
Stahl, 2008). On a laboratory scale, using Burkholderia
cepacia and Burkholderia sacchari strains, bagasse hydrolysate
proved to be a better carbon source for the PHB production in
comparison with pure xylose and glucose (Silva et al.,
2004). Burkholderia sacchari IPT 101 on hydrolyzed bagasse reached
62% of PHB cell dry mass at a yield of 0.39 g/g, but lower cell
densities were achieved on hydrolyzed bagasse than on pure sugars
(Silva et al., 2004).
Koller and coworkers recently investigated the integration and the cost
effectiveness of PHB production process into the sugarcane mill (pilot-
scale plant in Sao Paolo, Brazil) and found that embedding of the
industrial-PHB production into a sugar and ethanol factory starting
from the raw material sugarcane makes it possible to achieve a
production price per kilogram PHB that is 4–5 times lower than known
for prior PHB production processes. This cost reduction was enabled
by an extensive utilization of by-products of the sugarcane plant,
especially of bagasse, and strain of Cupriavidus necator DSM 454
(Koller, Hesse, et al., 2010; Koller, Salerno, et al., 2010; Nonato,
Mantelatto, & Rossell, 2001).

Carbon-Rich Wastes as Feedstocks for Biodegradable Polymer

(Polyhydroxyalkanoate) Production Using Bacteria
Jasmina Nikodinovic-Runic, ... Kevin E. O Connor, in Advances in Applied Microbiology, 2013
7.2 Bagasse and vinasse
Bagasse is another by-product of sugar industry. It is the heterogeneous fibrous residue that
remains after sugarcane stalks are crushed for sugar extraction. Typically, from processing 100
tons of sugarcane in a factory, 30–34 tons of bagasse is obtained (of which 22–24 tons is used in
processing and 8–10 tons is saved) (Solomon, 2011; Yadav & Solomon, 2006). Dry bagasse
consists of 45% cellulose, 28% hemicellulose, 20% lignin, 5% sugar, 1% minerals, and 2% ash.
Bagasse is similar in component to wood except that it has high moisture content. Thus, it is
currently used as a biofuel and in the manufacture of pulp and paper products, filler for building
materials and as a substrate for growing mushrooms. Considering the fact that much more
bagasse is available than utilized for the energy production, further value creation might be
possible. As an example, hemicelluloses and cellulose fractions of bagasse can be hydrolyzed
and utilized by suitable microbial strains for PHA biosynthesis. Very often hydrolysis is
performed using dilute acid and elevated temperatures (100–130°C), which generate
hydrolysates inhibitory to microorganisms. In the optimization study, highly
tolerant inoculum of Ralstonia eutropha was developed through directed adaption and evolution
to the bagasse hydrolysate (Yu & Stahl, 2008). On a laboratory scale, using Burkholderia
cepacia and Burkholderia sacchari strains, bagasse hydrolysate proved to be a better carbon
source for the PHB production in comparison with pure xylose and glucose (Silva et al.,
2004). Burkholderia sacchari IPT 101 on hydrolyzed bagasse reached 62% of PHB cell dry mass
at a yield of 0.39 g/g, but lower cell densities were achieved on hydrolyzed bagasse than on pure
sugars (Silva et al., 2004).
Koller and coworkers recently investigated the integration and the cost effectiveness of PHB
production process into the sugarcane mill (pilot-scale plant in Sao Paolo, Brazil) and found that
embedding of the industrial-PHB production into a sugar and ethanol factory starting from the
raw material sugarcane makes it possible to achieve a production price per kilogram PHB that is
4–5 times lower than known for prior PHB production processes. This cost reduction was
enabled by an extensive utilization of by-products of the sugarcane plant, especially of bagasse,
and strain of Cupriavidus necator DSM 454 (Koller, Hesse, et al., 2010; Koller, Salerno, et al.,
2010; Nonato, Mantelatto, & Rossell, 2001).
Vinasse is by-product of ethanol production from molasses. It is recalcitrant waste that is
produced at rates 10–15 times faster than ethanol. It has been examined for microbial conversion
into PHA (Bhattacharyya et al., 2012; Pramanik et al., 2012). Sugarcane vinasse contains
significant amounts of sucrose as well as oxalate, lactate, acetate, malate, and pyruvate, which
are ready metabolites to be fed to the tricarboxylic acid cycle. However, vinasse contains a
number of phenolic compounds and their polymers, which are known to be difficult to degrade
biologically and possess antimicrobial and phytotoxic properties (Parnaudeau, Condom, Oliver,
Cazevieille, & Recous, 2008). Recently, vinasse was converted to PHB by an
extremely halophilic archeon Haloarcula marismortui MTCC 1596 in shaken flasks containing
mixture of 10% raw vinasse and 90% pretreated vinasse and 100% pretreated vinasse (activated-
carbon adsorption) achieving 12 and 15 g/l cdw and 23% and 30% of cdw PHB accumulation,
respectively (Pramanik et al., 2012). Poly(3HB-co-3HV) has been obtained using acid
hydrolyzed and activated-carbon pretreated vinasse as carbon source by another halophilic
microorganism, Haloferax mediterranei DSM 1411 that accumulated 19.7 g/l polymer (70%
cdw) under shaken flasks condition (Bhattacharyya et al., 2012).
The production of sugar involves by association other residual streams and process-water quality
management as integral to the factory operations. In the Dutch sponsored project BioTRIP (ref.
to Bioplastics Magazine), a mixed-culture technology for PHA production is being examined for
technical, economic, and stakeholder organizational feasibility. As part of the water quality
management demands for the process effluents a biological water treatment process is being
trialed whereby a PHA-producing activated sludge may be harvested as a by-product.
The biomass is grown on the organic matter found in the process wash waters and the harvested
biomass may then be subsequently used as a resource for PHA production using other available
residual VFA-rich carbon streams that are available on or off-site. The project BioTRIP aims to
address biotechnical, economic as well as stakeholder drivers that would motivate the synergistic
integration of essential residuals and effluent quality management services to the production of
renewable materials such as biopolymers.
View chapterPurchase book
Modification of Cereal Straws as Natural Sorbents for
Removing Metal Ions from Industrial Waste Water
Feng Peng, Run-Cang Sun, in Cereal Straw as a Resource for Sustainable Biomaterials and
Biofuels, 2010
7.2.5.1 Sugarcane Bagasse
Bagasse, an agricultural waste from sugar industry, has been found as low-cost metal adsorbent
[83, 84]. Junior et al. [86] reported the use of succinic anhydride–modified sugarcane bagasse for
the treatment of Cu, Cd, and Pb from aqueous solutions. Sugarcane bagasse consists
of cellulose (43.6%), hemicelluloses (33.5%), and lignin (18.1%) [104]. The presence of these
three biological polymers causes sugarcane bagasse rich in hydroxyl and phenolic groups, and
these groups can be modified chemically to produce adsorbent materials with new properties.
Ngah and Hanafiah [105] reported that the hydroxyl groups in sugarcane bagasse could be
converted to carboxylic groups by using succinic anhydride. The carboxylic groups were later
reacted with three different chemicals mainly NaHCO3, ethylenediamine, and
triethylenetetramine to produce adsorbent materials with new properties which showed different
adsorption capacities for metal ions. It was found that sugarcane bagasse treated with
ethylenediamine and triethylenetetramine showed a remarkable increase in nitrogen content
compared to untreated sample, and triethylenetetramine-modified sugarcane bagasse has a higher
increasing extent. The presence of amide group was also detected in ethylenediamine- and
triethylenetetramine-modified sugarcane bagasses as a result of the reaction between –COOH
and –NH2 groups. Kinetic studies showed that equilibrium time for adsorption of Cu, Cd, and Pb
onto tethylenediamine- and triethylenetetramine-modified sugarcane bagasses were slower than
that for the adsorbent modified with NaHCO3. Triethylenetetramine-modified sugarcane bagasse
was the best adsorbent material for the removal of Cd and Pb as the adsorption capacities for
both the metals were two times higher than the unmodified sugarcane bagasse. This was
probably caused by the higher number of nucleophilic sites introduced in triethylenetetramine-
modified sugarcane bagasse. However, when the sugarcane bagasse was modified with
methanol, the modified adsorbent did not show a good uptake of Cd, and the maximum
adsorption capacity was 6.79 mg·g−1 [87]. In another study, through a fast, effective, and cheap
methodology, it was possible to devise a strategy to introduce chelating functions (carboxylic
acid and amine) to sugarcane bagasse. Modified sugarcane bagasse presented a maximum
adsorption capacity of 139, 313, and 313 mg·g−1 for Cu2+, Cd2+, and Pb2+, respectively [89].
Orlando et al. [90] used microwave radiation to produce neutral bagasse chelating agents (BCA)
by reaction of urea with the reactive sites present in bagasse such as hydroxyl and carboxylic
groups. The prepared material had a Cu(II) and Hg(II) adsorption capacity of 76.2 and 280.8
mg·g−1, respectively.
Bagasse fly ash, a sugar industry waste, has been converted into an inexpensive and efficient
adsorbent [90]. The product obtained was characterized and utilized for the removal of Zn from
aqueous solutions over a wide range of initial metal-ion concentration (3.06 × 10−4 to 3.06 ×
10−3 M), contact time (24 h), adsorbent dose (5–20 g·L−1), and pH (1.0–6.0). The removal of
Zn(II) was 100% at low concentrations, whereas it was 60–65% at higher concentrations at an
optimum pH of 4.0, using 10 g·L−1 of adsorbent in 6–8 h of equilibration time. The adsorption
capacity of bagasse fly ash for Zn was found to be 13.21 mg·g−1 at pH 4.0 and 30 °C. Hydrogen
peroxide is a good oxidizing agent and used to remove the adhering organic matter on the
adsorbent. The performance of hydrogen peroxide–treated bagasse fly ash for the removal of Pb
and Cr was explored by Gupta and Jain [85]. It was found that hydrogen peroxide–treated
bagasse fly ash was able to remove Cr in a shorter period of time (60 min) when compared with
Pb (80 min). The isotherm study also revealed that the maximum adsorption capacity for Cr was
higher than Pb. However, the recorded values of maximum adsorption capacities for both the
metals were low (2.50 and 4.35 mg·g−1 for Pb and Cr, respectively). The detailed mechanism of
adsorption by the treated bagasse fly ash was not discussed, but it was thought that adsorption
was controlled by film diffusion at lower metal concentration and particle diffusion at higher
concentration of metal ions.
Mohan and Singh [107] studied potentiality of AC derived from bagasse for the removal of
Cd(II) and Zn(II) from aqueous solutions in single as well as multimetal systems. Cd(II)
adsorption was slightly more than Zn(II) and the increased sorption capacity was reported with
increase in temperature. Adsorption on bagasse-based AC occurs through a film-diffusion
mechanism at all concentrations. Using bagasse-based carbon, Ayyappan [88] studied adsorbent
of Pb(II) under batch adsorption. Desorption of Pb(II) from sorbed carbon was achieved by
eluting with 0.1 M HNO3. Carbon was retrieved by washing with 0.1 M CaCl2 solution and
reused. Similar studies were also carried out with Cr [88].
View chapterPurchase book
Plant Genetic Engineering Towards the Third Millennium
G. Selman-Housein, ... F. Miranda, in Developments in Plant Genetics and Breeding, 2000
Sugarcane Bagasse
The bagasse is the main residue of the sugarcane industry representing, by weight, almost 30 %
(hundreds of millions of tons per year world-wide) of the sugarcane agricultural product,
therefore bagasse utilization is important for both economical and environmental
considerations. Sugarcane bagasse is a good low cost raw material for paper production or
animal feed but, as for other plants, the cell wall structural polymer, lignin, rich in p-coumaryl
subunits, has a negative effect on digestibility (Leng, 1991) and paper pulping properties (Chiang
et al., 1988).

Modification of Cereal Straws as Natural Sorbents for

Removing Metal Ions from Industrial Waste Water
Feng Peng, Run-Cang Sun, in Cereal Straw as a Resource for
Sustainable Biomaterials and Biofuels, 2010
7.2.5.1 Sugarcane Bagasse
Bagasse, an agricultural waste from sugar industry, has been found
as low-cost metal adsorbent [83, 84]. Junior et al. [86] reported the
use of succinic anhydride–modified sugarcane bagasse for the
treatment of Cu, Cd, and Pb from aqueous solutions. Sugarcane
bagasse consists of cellulose (43.6%), hemicelluloses (33.5%),
and lignin (18.1%) [104]. The presence of these three biological
polymers causes sugarcane bagasse rich in hydroxyl and phenolic
groups, and these groups can be modified chemically to produce
adsorbent materials with new properties. Ngah and Hanafiah [105]
reported that the hydroxyl groups in sugarcane bagasse could be
converted to carboxylic groups by using succinic anhydride. The
carboxylic groups were later reacted with three different chemicals
mainly NaHCO3, ethylenediamine, and triethylenetetramine to produce
adsorbent materials with new properties which showed different
adsorption capacities for metal ions. It was found that sugarcane
bagasse treated with ethylenediamine and triethylenetetramine
showed a remarkable increase in nitrogen content compared to
untreated sample, and triethylenetetramine-modified sugarcane
bagasse has a higher increasing extent. The presence of amide group
was also detected in ethylenediamine- and triethylenetetramine-
modified sugarcane bagasses as a result of the reaction between –
COOH and –NH2 groups. Kinetic studies showed that equilibrium time
for adsorption of Cu, Cd, and Pb onto tethylenediamine- and
triethylenetetramine-modified sugarcane bagasses were slower than
that for the adsorbent modified with NaHCO3. Triethylenetetramine-
modified sugarcane bagasse was the best adsorbent material for the
removal of Cd and Pb as the adsorption capacities for both the metals
were two times higher than the unmodified sugarcane bagasse. This
was probably caused by the higher number of nucleophilic sites
introduced in triethylenetetramine-modified sugarcane bagasse.
However, when the sugarcane bagasse was modified with methanol,
the modified adsorbent did not show a good uptake of Cd, and the
maximum adsorption capacity was 6.79 mg·g−1 [87]. In another study,
through a fast, effective, and cheap methodology, it was possible to
devise a strategy to introduce chelating functions (carboxylic acid and
amine) to sugarcane bagasse. Modified sugarcane bagasse
presented a maximum adsorption capacity of 139, 313, and 313
mg·g−1 for Cu2+, Cd2+, and Pb2+, respectively [89]. Orlando et al. [90]
used microwave radiation to produce neutral bagasse chelating
agents (BCA) by reaction of urea with the reactive sites present in
bagasse such as hydroxyl and carboxylic groups. The prepared
material had a Cu(II) and Hg(II) adsorption capacity of 76.2 and 280.8
mg·g−1, respectively.
Bagasse fly ash, a sugar industry waste, has been converted into an
inexpensive and efficient adsorbent [90]. The product obtained was
characterized and utilized for the removal of Zn from aqueous
solutions over a wide range of initial metal-ion concentration (3.06 ×
10−4 to 3.06 × 10−3 M), contact time (24 h), adsorbent dose (5–20
g·L−1), and pH (1.0–6.0). The removal of Zn(II) was 100% at low
concentrations, whereas it was 60–65% at higher concentrations at an
optimum pH of 4.0, using 10 g·L−1 of adsorbent in 6–8 h of
equilibration time. The adsorption capacity of bagasse fly ash for Zn
was found to be 13.21 mg·g−1 at pH 4.0 and 30 °C. Hydrogen peroxide
is a good oxidizing agent and used to remove the adhering organic
matter on the adsorbent. The performance of hydrogen peroxide–
treated bagasse fly ash for the removal of Pb and Cr was explored by
Gupta and Jain [85]. It was found that hydrogen peroxide–treated
bagasse fly ash was able to remove Cr in a shorter period of time (60
min) when compared with Pb (80 min). The isotherm study also
revealed that the maximum adsorption capacity for Cr was higher than
Pb. However, the recorded values of maximum adsorption capacities
for both the metals were low (2.50 and 4.35 mg·g−1 for Pb and Cr,
respectively). The detailed mechanism of adsorption by the treated
bagasse fly ash was not discussed, but it was thought that adsorption
was controlled by film diffusion at lower metal concentration and
particle diffusion at higher concentration of metal ions.
Mohan and Singh [107] studied potentiality of AC derived from
bagasse for the removal of Cd(II) and Zn(II) from aqueous solutions in
single as well as multimetal systems. Cd(II) adsorption was slightly
more than Zn(II) and the increased sorption capacity was reported
with increase in temperature. Adsorption on bagasse-based AC
occurs through a film-diffusion mechanism at all concentrations. Using
bagasse-based carbon, Ayyappan [88] studied adsorbent of Pb(II)
under batch adsorption. Desorption of Pb(II) from sorbed carbon was
achieved by eluting with 0.1 M HNO3. Carbon was retrieved by
washing with 0.1 M CaCl2 solution and reused. Similar studies were
also carried out with Cr [88].