Review
Department of Food Science, School of Food Engineering, State University of Campinas (UNICAMP), Rua Monteiro Lobato, 80, ZIP code 13083-862, Campinas, SP, Brazil
Department of Biological Sciences, School of Science and Letters, So Paulo State University (UNESP), Rua Dom Antonio, 2100, ZIP code 19806-380, Assis, SP, Brazil
a r t i c l e
i n f o
Article history:
Received 27 June 2012
Accepted 20 November 2012
Keywords:
Lignocellulosic materials
Chemical and enzymatic hydrolysis
Xylanases
Xylo-oligosaccharides
a b s t r a c t
Currently, there is worldwide interest in the technological use of agro-industrial residues as a renewable
source of food and biofuels. Lignocellulosic materials (LCMs) are a rich source of cellulose and hemicellulose.
Hemicellulose is rich in xylan, a polysaccharide used to develop technology for producing alcohol, xylose, xylitol and xylo-oligosaccharides (XOSs). The XOSs are unusual oligosaccharides whose main constituent is xylose linked by 14 bonds. The XOS applications described in this paper highlight that they are considered
soluble dietary bers that have prebiotic activity, favoring the improvement of bowel functions and immune
function and having antimicrobial and other health benets. These effects open a new perspective on potential applications for animal production and human consumption. The raw materials that are rich in hemicellulose include sugar cane bagasse, corncobs, rice husks, olive pits, barley straw, tobacco stalk, cotton stalk,
sunower stalk and wheat straw. The XOS-yielding treatments that have been studied include acid hydrolysis, alkaline hydrolysis, auto-hydrolysis and enzymatic hydrolysis, but the breaking of bonds present in these
compounds is relatively difcult and costly, thus limiting the production of XOS. To obviate this limitation, a
thorough evaluation of the most convenient methods and the opportunities for innovation in this area is
needed. Another challenge is the screening and taxonomy of microorganisms that produce the xylanolytic
complex and enzymes and reaction mechanisms involved. Among the standing out microorganisms involved
in lignocellulose degradation are Trichoderma harzianum, Cellulosimicrobium cellulans, Penicillium janczewskii,
Penicillium echinulatu, Trichoderma reesei and Aspergillus awamori. The enzyme complex predominantly
comprises endoxylanase and enzymes that remove hemicellulose side groups such as the acetyl group. The
complex has low -xylosidase activities because -xylosidase stimulates the production of xylose instead
of XOS; xylose, in turn, inhibits the enzymes that produce XOS. The enzymatic conversion of xylan in XOS
is the preferred route for the food industries because of problems associated with chemical technologies
(e.g., acid hydrolysis) due to the release of toxic and undesired products, such as furfural. The improvement
of the bioprocess for XOS production and its benets for several applications are discussed in this study.
2012 Elsevier Ltd. All rights reserved.
Contents
1.
2.
3.
4.
5.
Introduction . . . . . . . . . . . . . . . . . . . . . .
Chemical structure of xylo-oligosaccharides . . . . . . . .
Health benets of xylo-oligosaccharides . . . . . . . . .
Soluble bers as substitutes for antibiotics in feed . . . . .
Technologies for obtaining xylo-oligosaccharides . . . . .
5.1.
Pretreatments for xylan extraction . . . . . . . . .
5.1.1.
Autohydrolysis . . . . . . . . . . . . . .
5.1.2.
Alkaline and acid pretreatments . . . . . .
5.2.
Enzymatic treatment of pre-hydrolyzed LCM residues
5.2.1.
Xylanases . . . . . . . . . . . . . . . .
5.3.
Enzymatic technology for XOS production . . . . .
. .
. .
. .
. .
. .
. .
. .
. .
and
. .
. .
. . . . . . . .
. . . . . . . .
. . . . . . . .
. . . . . . . .
. . . . . . . .
. . . . . . . .
. . . . . . . .
. . . . . . . .
XOS production
. . . . . . . .
. . . . . . . .
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
76
76
77
77
78
78
78
79
80
80
82
Corresponding author at: So Paulo State University (UNESP), Rua Dom Antonio, 2100, ZIP code 19806-380, Assis, SP, Brazil. Tel./fax: +55 1833025848x5716.
E-mail addresses: anafbio@yahoo.com.br (A.F.A. Carvalho), poliva@assis.unesp.br (P.O. Neto), douglasfsilva@gmail.com (D.F. da Silva), glaupast@fea.unicamp.br (G.M. Pastore).
0963-9969/$ see front matter 2012 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.foodres.2012.11.021
76
6.
Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 83
Acknowledgments . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 83
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 83
1. Introduction
Currently, one of the greatest challenges is to increase the production of healthy and economically affordable foods that are available
for a large proportion of the world's population. The increase in
food production must be sustainable and avoid the deforestation of
new areas that are currently occupied by forests. In this regard, research into the use of agro-industrial residues for food production
should be intensied because these wastes are underutilized and billions of tons are discarded annually.
Lignocellulosic materials (LCM) represent the most abundant organic residues in the world. Considering only the sugar and ethanol
in Brazil, the estimated production of sugar cane for the 2012/13
season will be 596.6 billion tons (CONAB, 2012). At 135 kg of dried
bagasse per ton of crushed cane (Brienzo, Siqueira, & Milagres,
2009), the total bagasse generated in this season will be approximately 80.5 billion tons.
Cane bagasse is currently used in sugar mills for burning in boilers,
generating steam, heating cane juice and operating the mills, thus
saving energy in the process. Some sugar mills also use cane bagasse
to co-generate electricity. However, some cane bagasse remains that
can be used for other purposes, especially to produce products with
high added value.
Sugar cane bagasse has 2429% hemicellulose, represented by
1-arabino-(4-0-methyl-D-glucurono)-D-xylan, as well as cellulose,
lignin, minerals, waxes and other compounds (Brienzo et al., 2009;
Gottschalk, Oliveira, & Bom, 2010; Pandey, Soccol, Nigam, & Soccol,
2000; Rodrigues et al., 2010; Song & Wei, 2010). Hemicellulose is a
heteropolysaccharide with xylan as the major carbohydrate and
mainly comprises polymers of xylose, arabinose, galactose, mannose
and glucose (Garrote, Domnguez, & Paraj, 1999).
Xylan is also a heteropolysaccharide with a backbone formed by
xylose homopolymer subunits (Nabarlatz, Ebringerov, & Montan,
2007; Rodrigues et al., 2010; Saha, 2003; Santos, Sarrouh, Rivaldi,
Converti, & Silva, 2008).
In other LCM residues such as tobacco stalk, cotton stalk, sunower stalk and wheat stalk, the main chemical composition is 1921%
xylan, 3344% cellulose and 2029% lignins (Akpinar, Erdogan, &
Bostanci, 2009a,b). Reports on the chemical composition of corncobs,
one of the most important LCMs studied for xylo-oligosaccharides
(XOS) production, suggest the presence of glucan 3135%, lignin
1014% and a high xylan content (3035%) (Garrote, Dominguez, &
Paraj, 2002; Tan et al., 2008; Yang, Xu, Wang, & Yang, 2005).
Samanta, Senani, et al. (2012) found similar results for corncobs:
27.7% cellulose, 38.8% xylan, 9.4% lignin, 75.9% neutral detergent
ber and 37.1% acid detergent ber.
In recent years, there has been increasing interest in new prebiotics such as XOS. XOSs are produced by hydrolyzing xylan. Commercialized as a white powder, XOSs are oligomers containing two
to ten xylose molecules linked by 14 bonds (Mkelinen et al.,
2010). XOSs are considered non-digestible oligosaccharides (NDOs),
non-cariogenic in humans and have important biological properties.
They are used as dietary sweeteners in low-calorie diet foods and for consumption by individuals with diabetes (Choque Delgado, Tamashiro,
Junior, Moreno, & Pastore, 2011; Rivero-Urgell & Santamaria-Orleans,
2001; Vzquez et al., 2000).
Some prebiotics such as fructo-oligosaccharides and mannan oligosaccharides are currently widely used in feed (for pigs, chickens, cattle
and pets), replacing the use of antibiotic growth promoters (AGPs).
Due to the overuse of antibiotics, virulent strains of pathogens are
X1 = xylose
OH
HO
77
OH
HO
HO
OH
X2 = xylobiose
HO
OH
O
OH
HO
HO
OH
O
O
HO
OH
HO
OH
OH
n = 1:
n = 2:
n = 3:
n = 4:
n = 5:
X3 = xylotriose
X4 = xylotetraose
X5 = xylopentaose
X6 = xylohexaose
X7 = xyloheptaose
2007). Some studies indicate that 4 g of XOS per day for 3 weeks improved the intestinal microbiota among people who are above
65 years old (Chung et al., 2007).
XOS also have favorable technological features, including stability
at acidic pH, heat resistance, the ability to achieve signicant biological effects at low daily doses, low calorie content, non-toxicity and
other properties that have yet to be studied (Vzquez et al., 2000).
4. Soluble bers as substitutes for antibiotics in feed
The use of antibiotic growth promoters (AGPs) in animal production began half a century ago when residues of chlortetracycline production were added to chicken feed. These residues were added to
serve as a source of vitamin B12, but they caused a growth stimulation that was far too large to be explained as only a vitamin effect
(Brezoen, Van Haren, & Hanekamp, 1999). This observation was
quickly extended to other antibiotics and other animal species, leading to widespread adoption of AGP inclusion in feeds. In recent decades, considerable amounts of antibiotics have been used in animal
production, as both therapeutics and growth promoting agents
(Falco et al., 2007).
Antibiotics have been widely used in animal production, not only
to treat or prevent infection in farm animals but also for low
dose-long term administration, usually given in feed, to help animals
gain weight more quickly and increase the productivity and yield in
animal production. However, overuse of antibiotics encourages resistance and the production of more virulent strains of pathogens. Consequently, beginning in 2006, the European Union banned the use of
growth promoting antibiotics (GPAs) in farming (Falco et al., 2007;
Wallace, 2007).
Mannan oligosaccharides (MOS) act mainly by preventing colonization more than by stimulating benecial microorganisms, and they are
considered important prebiotics. Many pathogens have mbriae that
Table 1
Xylo-oligosaccharide benets for the healthy human.
XOS Benets
References
Modulation of the colon microbiota increasing the number of bidobacteria and Lactobacillus.
78
or more desirable properties than the more established oligosaccharides. Other studies have been showed the XOS ability in the growth
of some benecial bacteria. The growth of Lactobacillus brevis and
their capability to utilize XOS from corncob as carbon and energy
source was proved (Moura et al., 2007).
XOSs were obtained by autohydrolysis of rice husk and puried
by nanoltration and ion exchange processing. They were evaluated
in a medium for promoting the growth of Bidobacterium adolescentis
CECT 5781, Bidobacterium longum CECT 4503, Bidobacterium
infantis CECT 4551 and Bidobacterium breve CECT 4839. All of them
grew on XOS medium, but the specic growth rate of B. adolescentis
(0.58 h 1) was higher than the ones determined for B. longum,
B. infantis and B breve (0.37, 0.30 and 0.40 h 1, respectively). The percentage of total XOS by B. adolescentis was 77% in 24 h of culture and
the highest percentage of consume corresponded to xylotriose
(90%), followed by xylobiose (84%), xylotetraose (83%) and
xylopentaose (71%) (Gulln et al., 2008). On the other hand, XOS
shows an ability for suppressing the growth of Clostridium and bacteriostatic action against Vibrio anguillarum (Izumi & Azumi, 2001).
5. Technologies for obtaining xylo-oligosaccharides
5.1. Pretreatments for xylan extraction
Various pretreatments have been used for the complete extraction
of hemicellulose to make xylan for enzymatic reactions. The pretreatments include thermal and chemical methods (Aachary & Prapulla,
2009).
Before starting the pretreatment, the raw material can be prepared by washing with ethanol or ethyl acetate to remove impurities
and other compounds (e.g., waxes and pectin) and make the later
XOS-producing steps easier (Brienzo et al., 2009; Vzquez et al.,
2005). Furthermore, alcohols and ketones can be used in the recovery
of soluble xylan or to concentrate the released XOS (Akpinar et al.,
2009b; Vzquez et al., 2005).
The current methods used for pretreatments are autohydrolysis,
acid or alkaline pre-hydrolysis and enzymatic hydrolysis.
5.1.1. Autohydrolysis
Autohydrolysis involves the deacetylation of xylan by the thermal
hydrolysis of hemicellulose to produce acetic acid (Garrote et al.,
1999, 2002; Kabel et al., 2002). The products obtained from LCM by
this process contain a variety of undesirable components such as lignin, monosaccharides, furfural, and others, thereby requiring further
purication (Zhu, Wu, Zhang, Li, & Gao, 2006).
XOS produced by autohydrolysis contains a signicant proportion
of compounds with a high degree of polymerization (Nabarlatz et al.,
2007). This treatment can be used as a rst step in the fractionation
processes for LCM.
After prehydrolysis, the LCM can be separated into two fractions.
The soluble and liquid phase is rich in hemicellulose, and it can be
decomposed into valuable products, such as soluble oligosaccharides,
sugars and aldehydes. The precipitated solids are high in cellulose and
lignin, and they can be separated for further processing to enable a
variety of possible applications, including the production of cellulose
pulp (Caparrs, Ariza, Garrote, Lpez, & Daz, 2007) and fermentable
sugars or fuel (Yez et al., 2006).
When LCM containing xylan is submitted to autohydrolysis under
mild operating conditions, xylo-oligosaccharides (XOSs) are the main
products of the hydrolysis of hemicellulose. To obtain XOS with a degree of polymerization of 26 xylose units, the products released by
autohydrolysis require extra treatment, e.g., enzymatic or acid hydrolysis (Moure et al., 2006). Moreover, autohydrolysis requires specialized equipment that needs to be operated at high temperatures (Zhu
et al., 2006).
79
the reaction time was long, and probably allowed the degradation
of xylan to xylose under these conditions.
The effect of different acid concentrations (1.22%4.90% H2SO4) on
XOS production from xylans of tobacco stalk (TSX), cotton stalk
(CSX), sunower stalk (SSX) and wheat stalk (WSX) was evaluated
(Akpinar et al., 2009b). Reducing sugar production rates increased
with time and acid concentration.4.9% sulfuric acid at 100 C produced mainly monosaccharides rather than oligosaccharides. The
best conditions for acid hydrolysis were 2.45% H2SO4 in 30 min of reaction time at 100 C. However even under these condition the XOS
yield was low for tobacco (13%), cotton stalk (7.5%), sunower stalk
(12.6%) and wheat straw (10.2%).
More recently, Otieno and Ahring (2012b) obtained high yield of
XOS from switchgrass (Panicum virgatum 84.1%), morning light
(Miscanthus sinensis 64.9%) and cane bagasse (92.2%) using rapid
thermo-acid process (0.1% H2SO4 at 145 C for 1 h). The content
(19.7%) and quality of XOS (70% X2 to X4) from cane bagasse was
also interesting and xylose content was not high (5.9%) when it was
compared with enzymatic hydrolysis (22.5%) (Brienzo et al., 2010).
The rapid thermo-acid treatment of sugarcane bagasse seems to be
also more interesting for the XOS yield (92%) when compared to
the yield of 3940% by enzymatic method (Brienzo et al., 2010;
Carvalho, Oliva-Neto, & Pastore, 2012a,b). Furthermore, the rst
treatment is simpler in the hydrolysis step. The major impediment
of the acid treatment is the production of toxic components (furfural
and HMF) that require a greater degree of purication.
The type and chemical structure of the substrate and the enzyme
specicity are important factors in the enzymatic hydrolysis of
xylan to produce XOS.
The XOS yield from enzymatic hydrolysis using xylan extracted
from tobacco stalk by alkaline pretreatment (11.4%) was slightly
lower than from acid hydrolysis. However, two of the major disadvantages of acid hydrolysis are the production of considerably high
amounts of monosaccharides and the formation of undesirable toxic
furfural (Akpinar et al., 2010). These last products make purication
steps more difcult and expensive in addition to decrease in the
XOS yield. If the importance of pretreatment and the limitation of
acid hydrolysis is to be proven, the best procedure is to use alkaline
and thermal hydrolysis. An extraction from corncob using 2% and
12% alkali and steam hydrolysis improved the xylan recovery
from12.5% to 63.2% (for potassium hydroxide) and 14.4% to 83.5%
(for sodium hydroxide), respectively. These results showed the importance of higher alkali concentrations in hydrolysis, with increased
xylan recovery at increased alkali levels. Xylan obtained by this
Fig. 2. Some alternative chemical pretreatments and acid or enzymatic treatments for XOS production from lignocellulosic residues.
80
Table 2
Sources of LCM residues, pretreatments for xylan extraction, and different treatments for XOS production.
LCM residue
Pretreatment
xylan
extraction
Corncob
Alkali pretreat.
Corncob
Acid pretreat.
Corncob
Steam cooked
Palm oil
Autohydrolysis
Tobacco stalk
Alkali pretreat.
Sunower stalk
Alkali pretreat.
Cotton stalk
Alkali pretreat.
Wheat straw
Alkali pretreat.
Tobacco stalk
Alkali pretreat.
Corncob
Alkali pretreat.
Cane bagasse
Alkali pretreat.
Corncob
Alkali pretreat.
Corncob
Alkali pretreat.
Corncob
Alkali pretreat.
Triploid Populus tomentosa Alkali pretreat.
Cane bagasse
Alkali pretreat.
Reference
Reaction time XOS production XOS
Treatment XOS production Xylan
yield (%)
production
extraction for XOS
(g/l)
production
yield (%)
Enzymatic hydrolysis
Enzymatic hydrolysis
Enzymatic hydrolysis
Enzymatic hydrolysis
enzymatic hydrolysis
Enzymatic hydrolysis
Enzymatic hydrolysis
Acid hydrolysis
Enzymatic hydrolysis
Enzymatic hydrolysis
Acid hydrolysis
Acid hydrolysis
Acid hydrolysis
Enzymatic hydrolysis
Enzymatic hydrolysis
40.8
39.2
40.0
21.8
18.9
21.3
20.6
20.0
17.9
86.0
83.5
83.5
83.5
85.0
24 h
24 h
24 h
24 h
24 h
24 h
24 h
24 h
30 min
8h
96 h
15 min
30 min
60 min
14 h
8h
81.0
52.0
77.0
15.9
13.8
12.9
60.0
37.1
86.6
82.4
68.5
36.8
5.8
2.7
3.7
3.1
2.8
2.7
1.8
1.5
6.7
5.7
0.65
0.79
0.88
4.0
1.7
Table 3
Fungal producers of xylanases and enzymatic activities on different substrates and fermentation process.
Strains
Xylanase activity
Substrate
Fermentation
Reference
Cellulosimicrobium cellulans
Penicillium janczewskii
P. janczewskii
P. janczewskii
Penicillium janthinellum
P. janthinellum
Trichoderma reesei RUT-C30
Aspergillus awamori 2B.361 U2/1
A. awamori
A. awamori
Malbranchea ava MTCC 4889
Thermomyces lanuginosus ATCC 46882
0.7
15.4
5.8
3.1
55.3
23.8
10 U/ml
25 U/ml
4.4
100 U/g
15,000 U/g
5098 U/g
Cane bagasse
Wheat bran
Oat bran
Cane bagasse
Pretreated corncob
Pretreated cane bagasse
Pretreated cane bagasse
Pretreated cane bagasse
Grape marc
Wheat bran
Rice straw
Cellulose bagasse
SmF
SmF
SmF
SmF
SmF
SmF
SmF
SmF
SSF
SSF
SSF
SSF
U/ml
U/ml
U/ml
U/ml
U/ml
U/ml
U/g
81
-4-O-Me-GluUA
Acetylxylan
Endo-1,4--xylosidase
-D-glucuronidase
-Xylose
-L-arabinofuranosidase
-araf.
Xylo-oligosaccharides
pcou./fer.
p-coumaric acid or
-Xylosidase
Fig. 3. Schematic structure of hemicellulose and the sites of xylanase reactions. The main chain consists of xylose residues with -1.4 linkages (Sunna & Antranikian, 1997).
82
enzymes of this group are only active on substrates containing D-xylose and long chains of xylo-oligosaccharides, they have low catalytic
versatility and the products of their action can be hydrolyzed by the
enzymes of family 10 (Biely et al., 1997). The family 10 xylanases produce XOSs that are smaller than those produced by xylanases from
family 11 (Maslen, Goubet, Adam, Dupree, & Stephens, 2007).
According to Song and Wei (2010), the demand for enzymes is
growing faster than ever, and this demand is a force driving research
on xylanases and cellulases. However, the production costs and low
yields are important problems for industrial applications. The incorporation of cheap sources (such as sugarcane bagasse, wheat straw
and corncob) in the culture media for the growth of microorganisms
and enzyme production should help to decrease the production costs
for the enzyme complexes. Consequently, several processes have
been and are being developed to use agro-industrial residues to generate products such as ethanol, proteins and enzymes using microorganisms (Lakshmi et al., 2009). Moreover, the bioconversion of these
substrates may help reduce the environmental impact caused by the
accumulation of waste (Camassola & Dillon, 2009).
Xylanolytic enzymes are highlighted due to their potential applications in the bioconversion of lignocellulose to sugars (Lakshmi et
al., 2009). They also have potential applications in industries in the
clarication of juices and vegetable oil extracts, as aids in the digestive acquisition of nutrients by pigs and birds that use cereal-based
diets, to improve our for bakery food products, as bleaching agents
in pulp and paper and in the saccharication of agricultural, industrial
and food wastes (Beg, Kapoor, Mahajan, & Hoondal, 2001; Moure
et al., 2006; Polizeli et al., 2005; Subramanian & Prema, 2002;
Techapun, Poosaran, Watanabe, & Sasaki, 2003). Furthermore,
xylanases can be used to produce high-value xylo-oligosaccharides
(Vzquez et al., 2000).
5.3. Enzymatic technology for XOS production
After the pretreatment of the LCM, the extracted xylan must be
treated by enzymatic hydrolysis (Ai et al., 2005; Akpinar et al.,
2009a; Brienzo et al., 2010; Carvalho et al., 2012a,b; Yang et al.,
2011) or an acid treatment (Akpinar et al., 2010; Rodrigues et al.,
2010). According to Sun et al. (2004), acid hydrolysis is not
recommended for this step, due to the production of undesirable
products such as furfural and hydroxymethylfurfural and the large
amount of monosaccharides released.
Currently, enzymatic hydrolysis is considered the best option to
produce XOS for the food industry (Aachary & Prapulla, 2009). Production of XOS is carried out by xylanases hydrolyzing -1,4 xylan
bonds. The enzymatic complex must have a low activity of exoxylanases (-xylosidases) to attenuate the production of xylose,
which inhibits XOS production (Vzquez et al., 2002). Moreover, to
increase XOS production, an endo-xylanase, which hydrolyses -1,4
bonds in the main chain of xylan, producing -anomers of XOS, and
enzymes that remove side groups must be used (Aachary &
Prapulla, 2009; Akpinar et al., 2010).
In the enzymatic process of oligosaccharide production, the enzyme can be added directly to the reaction medium (Akpinar et al.,
2010), immobilized (Jiang et al., 2004), produced in situ via microbial
fermentation or immobilized inside the biomass (Dorta, Cruz,
Oliva-Neto, & Moura, 2006; Oliva-Neto & Meno, 2009; Santos et al.,
2008). Fractionation of XOSs with different degrees of polymerization
can easily be accomplished by ultraltration to remove oligosaccharides with undesirable degrees of polymerization (Akpinar et al.,
2010).
Catalysis using immobilized and free endo-xylanases from Bacillus
halodurans for XOS production from corncob xylan was compared.
The conditions of the enzymatic reaction were as follows: 50 C,
pH 8.0, 12.8 U/g xylan and 2% of substrate in 24 h. The free enzyme
was more efcient in XOS production than the immobilized one.
The free enzyme converted xylan to higher-level oligomers with a degree of polymerization greater than 4, as well as 32.5% xylobiose and
xylotriose, suggesting that free xylanase worked on both longer (insoluble) and shorter (soluble) xylan chains. On the contrary, the
immobilized xylanase converted xylan to XOS with shorter lengths
and 25.2% of the mixture composed by xylobiose and xylotriose,
suggesting that the immobilized enzyme was more limited to acting
on shorter xylans (Lin, Tseng, & Lee, 2011).
In addition, other aspects inuence XOS yield and the composition
of oligosaccharides produced by enzymatic reactions, such as
xylanase type, xylan composition, the type of pretreatment, time
and other parameters of the reaction (Table 2).
Corncob has high potential to produce endo-xylanases and XOS.
Experimental studies using 233 U/g endo-xylanase from Aspergillus
oryzae MTCC 5154 and alkali-pretreated corncob (6%) at pH 5.4 and
50 C for 14 h, efciently exposed the xylan to the action of
endo-xylanases. Under these conditions, a high yield of XOS (79.7%),
corresponding 10.1 mg/ml, was obtained, and xylobiose (70.5%) was
the major component of the XOS mixture produced (Aachary &
Prapulla, 2009).
The pretreated xylan from triploid Populas tomentosa was converted
into XOS by a crude xylanase from Pichia stipitis. This enzyme converted
36.8% of the xylan to XOS, equivalent to 3.95 mg/ml, and 95% of the total
oligosaccharides were xylobiose, xylotriose and xylotetraose (Yang et
al., 2011).
XOS was produced by the catalysis of xylan extracted from corncob pretreated by mild alkali conditions (extraction yield of 17.9%)
using a partially puried xylanase from Aspergillus foetidus MTCC
4898. The enzymatic hydrolysis produced 60% (6.7 mg/ml) xylooligosaccharides, mainly xylobiose and xylotriose, after reaction of
8 h, with 20 U of xylanase/g xylan at 45 C (Chapla et al., 2012).
Moreover, the agroindustrial residue of palm oil was pretreated by
autohydrolysis and the solid residue obtained was hydrolyzed using
xylanases from Trichoderma viride. The sugar yield was: 13.9%
xylobiose, 1.97% xylotriose, 1.63% xylotetraose and 25.6% xylose
(Sabiha-Hanim et al., 2011).
The hydrolysis of different xylans was compared for XOS production using 200 U/g of xylanase from A. niger at 50 C over 24 h.
Under these conditions, the best production was with xylan from tobacco stalk (XOS yield of 13.8%), with a xylobiose yield of 7.95%
(1.59 mg/ml), a xylotriose yield of 5.9% (1.18 mg/ml) and yields of
other XOSs in lesser amounts (Akpinar et al., 2009a). The same
work compared the xylanases from different microorganisms. XOS
yield produced by xylanases from A. niger was greater than xylanases
from Trichoderma longibrachiatum on extracted xylan, with T.
longibrachiatum producing large amounts of monosaccharides.
The use of the endoxylanase from Trichoderma viridae (Sigma,
USA) for hydrolysis of xylan from grass extracted by alkaline hydrolysis produced approximately 2.8 mg/ml of XOS, xylobiose (1.7 mg/ml)
and xylotriose (1.1 mg/ml). The maximum yield of xylobiose (11%)
was obtained at pH 5.0 at 45 C, 87 U/g xylan in 10.1 h, and xylotriose
(7%) at pH 5.0 at 40 C and 66 U/g xylan in 16.5 h. (Samanta, Jayapal,
et al., 2012).
The maximum XOS production using xylan from sugar cane bagasse extracted by alkali and reacted with a commercial endoxylanase from T. viridae (Sigma, USA) was observed at 40 C, pH 4.0
with a dose of 13.3 U/g in 8 h. Under these conditions, 1.15 mg/ml
of xylobiose and 0.57 mg/ml of xylotriose were obtained. A response
surface analysis predicted the ideal conditions to be41 C, pH 4.9, enzyme dose 15.6 U/g and reaction time 19.2 h for maximizing
xylobiose yield, while for maximum xylotriose yield, the conditions
were 40 C, pH 4.13, dose 29.4 U/g and an incubation time of 18.1 h
(Jayapal et al., 2013).
However, the enzymatic hydrolysis with the xylan extracted from
sugar cane bagasse pretreated with alkali and peroxide showed a
maximum XOS conversion of 37.1%, after 96 h with 2.6% substrate,
83
Ai, Z., Jiang, Z., Li, L., Deng, W., Kusakabe, I., & Li, H. (2005). Immobilization of Streptomyces olivaceoviridis E-86 xylanase on Eudragit S-100 for xylo-oligosaccharide production. Process Biochemistry, 40, 27072714.
Akpinar, O., Erdogan, K., Bakir, U., & Yilmaz, L. (2010). Comparison of acid and enzymatic hydrolysis of tobacco stalk for preparation of xylo-oligosaccharides. Food
Science and Technology, 43, 119125.
Akpinar, O., Erdogan, K., & Bostanci, S. (2009a). Enzymatic production of xylooligosaccharide from selected agricultural wastes. Food and Bioproducts Processing,
87, 145151.
Akpinar, O., Erdogan, K., & Bostanci, S. (2009b). Production of xylo-oligosaccharides by controlled acid hydrolysis of lignocellulosic materials. Carbohydrate Research, 344, 660666.
Antoine, A. A., Jacqueline, D., & Thonart, P. (2010). Xylanase production by Penicillium
canescens on soya oil cake in solid state fermentation. Applied Biochemistry and
Biotechnology, 160, 5062.
Badhan, A. K., Chadha, B. S., & Saini, H. S. (2007). Purication of the alkalophilic
xylanases from Myceliophthora sp. IMI387099 using cellulose-binding domain as
an afnity tag. World. Journal Microbiology Biotechnology, 24, 973981.
Balakrishnan, H., Dutta-Choudhary, M. D., Srinivasan, M. C., & Rele, M. V. (1992).
Cellulase-free xylanase production from an alkalophilic Bacillus species. World
Journal of Microbiology and Biotechnology, 8, 627631.
Bastawde, K. B. (1992). Xylan structure, microbial xylanases, and their mode of action.
World Journal Microbiolology Biotechnology, 8, 353368.
Beg, Q. K., Kapoor, M., Mahajan, L., & Hoondal, G. S. (2001). Microbial xylanases and their
industrial applications: A review. Applied Microbiology and Biotechnology, 56, 326338.
Belancic, A., Scarpa, J., Peirano, A., Diaz, R., Steiner, J., & Eyzaguirre, J. (1995). Penicillium
purpurogenum produces several xylanases: Purication and properties of two of
the enzymes. Journal of Biotechnology, 41, 7179.
Bhat, M. K. (1998). Oligosaccharides as functional food ingredients and their role in improving the nutritional quality of human food and health. Recent Research Developments in Agricultural & Food Chemistry, 2, 787802.
Biely, P. (1985). Microbial xylanolytic systems. Trends in Biotechnology, 3, 286290.
Biely, P., Vransk, M., Tenkanen, M., & Kluepfel, D. (1997). Endo--1,4-xylanase families: differences in catalytic proprieties. Journal of Biotechnology, 57, 151166.
Blaut, M. (2002). Relationship of prebiotics and food to intestinal microora. European
Journal of Nutrition, 41, 1116.
Botella, C., Ory, I., Webb, C., Cantero, D., & Blandino, A. (2005). Hydrolytic enzyme production by Aspergillus awamori on grape pomace. Biochemical Engineering Journal,
26, 100106.
Brezoen, A., Van Haren, W., & Hanekamp, J. C. (1999). Emergence of a debate. AGPs and
public health. Human health and antibiotic growth promoters (AGPs): Reassessing
the risk. Heidelberg Appeal Nederland Foundation, 131.
Brienzo, M., Carvalho, W., & Milagres, A. M. F. (2010). Xylo-oligosaccharides production
from alkali-pretreated sugarcane bagasse Using xylanases from Thermoascus
aurantiacus. Applied Biochemistry and Biotechnology, 162, 11951205.
Brienzo, M., Siqueira, A. F., & Milagres, A. M. F. (2009). Search for optimum conditions
of sugarcane bagasse hemicellulose extraction. Biochemical Engineering Journal, 46,
199204.
Camassola, M., & Dillon, A. J. P. (2009). Biological pretreatment of sugar cane bagasse
for the production of cellulases and xylanases by Penicillium echinulatum. Industrial
Crops and Products, 29, 642647.
Campbell, J. M., Fahey, G. C., & Wolf, B. W. (1997). Selected indigestible oligosaccharides affect large bowel mass and fecal short-chain fatty acids, pH and microora
in rats. The Journal of Nutrition, 127, 130136.
Caparrs, S., Ariza, J., Garrote, G., Lpez, F., & Daz, M. J. (2007). Optimization of Paulownia fortunei autohydrolysisorganosolv pulping as a source of xylooligomers and
cellulose pulp. Industrial and Engineering Chemistry Research, 46, 623631.
Carvalho, A. F., Oliva-Neto, P., & Pastore, G. M. (2012a). Enzymatic production of
xylooligosaccharide from alkali-pretreated sugarcane bagasse. 16th World Congress
of Food Science and Technology, Foz do Iguau, Brazil.
Carvalho, A. F., Oliva-Neto, P., & Pastore, G. M. (2012b). Production of xylooligosaccharides by hemicellulose extracted from sugar cane bagasse. Congress of
Microbiology. Santos, Brazil.
Chapla, D., Pandit, P., & Shah, A. (2012). Production of xylooligosaccharides from
corncob xylan by fungal xylanase and their utilization by probiotics. Bioresource
Technology, 115, 215221.
Choque Delgado, G. T., Tamashiro, W. M. S. C., Junior, M. R. M., Moreno, Y. M. F., &
Pastore, G. M. (2011). The putative effects of prebiotics as immunomodulatory
agents. Food Research International, 40, 31673173.
Christopher, L., Bissoon, S. S., Singh, S., Szendefy, J., & Szakacs, G. (2005).
Bleach-enhancing abilities of Thermomyces lanuginosus xylanases produced by
solid state fermentation. Process Biochemistry, 40, 32303235.
Christov, L. P., Szakacs, G., & Balakrishnan, H. (1999). Production, partial characterization and use of fungal cellulase-free xylanases in pulp bleaching. Process Biochemistry, 34, 511517.
Chung, Y. C., Hsu, C. K., Ko, C. Y., & Chan, Y. C. (2007). Dietary intake of
xylooligosaccharides improves the intestinal microbiota, fecal moisture, and pH
value in the elderly. Nutrition Research, 27, 756761.
Collins, T., Gerday, C., & Feller, G. (2005). Xylanases, xylanase families and extremophilic
xylanases. FEMS Microbiology Reviews, 29, 323.
CONAB (2012). National company of supply. http://www.conab.gov.br/OlalaCMS/
uploads/arquivos/12_08_10_14_57_19_boletim_cana_portugues_-_agosto_2012_
2o_lev.pdf (Acess in: August 20, 2012)
Dorta, C., Cruz, R., Oliva-Neto, P., & Moura, D. J. C. (2006). Sugarcane molasses and yeast
powder used in the fructooligosaccharides production by Aspergillus japonicus-FCL
119T and Aspergillus niger ATCC 20611. Journal of Industrial Microbiology and
Biotechnology, 33, 10031009.
84
Falco, C. L., Castro-Solla, L., Maertens, L., Marounek, M., Pinheiro, V., Freire, J., et al.
(2007). Alternatives to antibiotic growth promoters in rabbit feeding: A review.
World Rabbit Science, 15, 127140.
Fonseca, A. P., Falco, C. L., Kocher, A., & Spring, P. (2004). Effects of dietary mannan oligosaccharide in comparison to oxytetracycline on performance of growing rabbits.
8th World Rabbit Congress, Puebla, Mxico (pp. 829833).
Garrote, G., Domnguez, H., & Paraj, J. C. (1999). Mild autohydrolysis: An environmentally friendly technology for xylo-oligosaccharide production from wood. Journal of
Chemical Technology and Biotechnology, 74, 11011109.
Garrote, G., Dominguez, H., & Paraj, J. C. (2002). Autohydrolysis of corncob: study of
non-isothermal operation for xylo-oligosaccharide production. Journal of Food
Engineering, 52, 211218.
Gill, H. S., Shu, Q., Lin, H., Rutherfurd, K. J., & Cross, M. L. (2001). Protection against
translocating Salmonella typhimurium infection in mice by feeding the immunoenhancing probiotic Lactobacillus rhamnosus strain HN001. Medical Microbiology
and Immunology, 190, 97104.
Gomes, E., Guez, M. A. U., Martin, N., & Silva, R. (2007). Enzimas termoestveis: fontes,
produo e aplicao industrial. Qumica Nova, 30, 136145.
Gottschalk, L. M. F., Oliveira, R. A., & Bom, E. P. S. (2010). Cellulases, xylanases,
-glucosidase and ferulic acid esterase produced by Trichoderma and Aspergillus
act synergistically in the hydrolysis of sugarcane bagasse. Biochemical Engineering
Journal, 51, 7278.
Grootaert, C., Delcour, J. A., Courtin, C. M., Broekaert, W. F., Verstraete, W., & Wiele, T. V.
(2007). Microbial metabolism and prebiotic potency of arabinoxylan oligosaccharides in the human intestine. Trends in Food Science & Technology, 18, 6471.
Gulln, P., Moura, P., Esteves, M., Girio, F. M., Domnguez, H., & Paraj, J. C. (2008).
Assessment on the fermentability of xylo-oligosaccharides from rice husks by probiotic bacteria. Journal of Agricultural and Food Chemistry, 56, 74827487.
Hinz, S. W. A., Pouvreau, L., Joosten, R., Bartels, J., Jonathan, M. C., & Wery, J. (2009).
Hemicellulase production in Chrysosporium lucknowense C1. Journal Cereal Science,
50, 318323.
Hughes, S. A., Shewry, P. R., Li, L., Gibson, G. R., Sanz, M. L., & Rastall, R. A. (2007). In
vitro fermentation by human fecal microora of wheat arabinoxylans. Journal of
Agricultural and Food Chemistry, 55, 45894595.
Izumi, K., & Azumi, N. (2001) Xylooligosaccharide compositions useful as food and feed
additives. Japan Patent JP, 2, 001, 226,409.
Jaskari, J., Kontula, P., Siitonen, A., Jousimies-Somer, H., Mattila-Sandholm, T., &
Poutanen, K. (1998). Oat -glucan and xylan hydrolysates as selective substrates
for Bidobacterium and Lactobacillus strains. Applied Microbiology and Biotechnology, 49, 175181.
Jayapal, N., Samanta, A. K., Kolte, A. P., Senani, S., Sridhar, M., Suresh, K. P., et al. (2013).
Value addition to sugarcane bagasse: Xylan extraction and its process optimization
for xylooligosaccharides production. Industrial Crops and Products, 42, 1424.
Jiang, Z., Zhu, Y., Li, L., Yu, X., Kusakabe, I., Kitaoka, M., et al. (2004). Transglycosylation
reaction of xylanase B from the hyperthermophilic Thermotoga maritima with the
ability of synthesis of tertiary alkyl -D-xylobiosides and xylosides. Journal of Biotechnology, 114, 125134.
Jorgensen, H., Morkeberg, A., Kristian, B. R. K., & Olsson, L. (2005). Production of cellulases and hemicellulases by three Penicillium species: Effect of substrate and evaluation of cellulose adsorption by capillary electrophoresis. Enzyme Microbiology
Technology, 36, 4248.
Kabel, M. A., Carvalheiro, F., Garrote, G., Avgerinos, E., Koukios, E., Paraj, J. C., et al.
(2002). Hydrothermally treated xylan rich by-products yield different classes of
xylo-oligosaccharides. Carbohydrate Polymers, 47, 4756.
Lakshmi, G. S., Rao, C. S., Rao, R. S., Hobbs, P. J., & Prakasham, R. S. (2009). Enhanced
production of xylanase by a newly isolated Aspergillus terreus under solid state
fermentation using palm industrial waste: A statistical optimization. Biochemical
Engineering Journal, 48, 5157.
LeBlanc, A. M., Castillo, N. A., & Perdigon, G. (2010). Antiinfective mechanisms induced
by a probiotic Lactobacillus strain against Salmonella enterica serovar Typhimurium
infection. International Journal of Food Microbiology, 138, 223231.
Leite, R. S., Daniela, A. B., Eduardo, D. S., Denis, S., Eleni, G., & Roberto, S. (2007). Production of cellulolytic and hemicellulolytic enzymes from Aureobasidium pulluans on
solid state fermentation. Applied Biochemistry and Biotechnology, 137140, 281288.
Licht, T. R., Ebersbach, T., & Frkir, H. (2012). Prebiotics for prevention of gut infections. Trends in Food Science & Technology, 23, 7082.
Lin, Y. S., Tseng, M. J., & Lee, W. C. (2011). Production of xylooligosaccharaides using
immobilized endo-xylanase of Bacillus halodurans. Process Biochemistry, 46,
21172121.
Mkelinen, H., Forssten, S., Saarinen, M., Stowell, J., Rautonen, N., & Ouwehand, A. C.
(2010). Xylo-oligosaccharides enhance the growth of bidobacteria and Bidobacterium lactis in a simulated colon model. Benecial Microbes, 1, 8191.
Maslen, S. L., Goubet, F., Adam, A., Dupree, P., & Stephens, E. (2007). Structure elucidation of arabinoxylan isomers by normal phase HPLCMALDI-TOF/TOF-MS/MS.
Carbohydrate Research, 342, 724735.
Moura, P., Barata, R., Carvalheiro, F. P., Grio, F., Loureiro-Dias, M. C., & Esteves, M. P.
(2007). In vitro fermentation of xylo-oligosaccharides from corn cobs autohydrolysis by Bidobacterium and Lactobacillus strains. LWT Food Science and
Technology, 40, 963972.
Moura, P., Cabanas, S., Loureno, P., Grio, F., Loureiro-Dias, M. C., & Esteves, M. P.
(2008). In vitro fermentation of selected xylo-oligosaccharides by piglet intestinal
microbiota. LWT Food Science and Technology, 41(10), 19521961.
Mouro, J. L., Pinheiro, V., Alves, A., Guedes, C. M., Pinto, L., Saavedra, M. J., et al. (2006).
Effect of mannan oligosaccharides on the performance, intestinal morphology and
cecal fermentation of fattening rabbits. Animal Feed Science and Technology, 126,
107120.
Moure, A., Gulln, P., Domnguez, H., & Paraj, J. C. (2006). Advances in the manufacture, purication and applications of xylo-oligosaccharides as food additives and
nutraceuticals. Process Biochemistry, 41, 19131923.
Mussatto, S. I., & Mancilha, I. M. (2007). Non-digestible oligosaccharides: A review.
Carbohydrate Polymers, 68, 587597.
Nabarlatz, D., Ebringerov, A., & Montan, D. (2007). Autohydrolysis of agricultural
by-products for the production of xylo-oligosaccharides. Carbohydrate Polymers,
69, 2028.
Narang, S., Sahai, V., & Bisaria, V. S. (2001). Optimization of xylanase production by
Melanocarpus albomyces IIS68 in solid state fermentation using response surface
methodology. Journal of Bioscience and Bioengineering, 91, 425427.
Okazaki, M., Fujikawa, S., & Matsumoto, N. (1990). Effects of xylooligosaccharide on
growth of bidobacteria. Journal of Japan Society of Nutrition and Food Sciences,
43, 395401.
Olano-Martin, E., Gibson, G. R., & Rastall, R. A. (2002). Comparison of the in vitro
bidogenic properties of pectins and pectic-oligosaccharides. Journal of Applied
Microbiology, 93, 505511.
Oliva-Neto, P., & Meno, P. T. P. (2009). Isomaltulose production from sucrose by
Protaminobacter rubrum immobilized in calcium alginate. Bioresource Technology,
100, 42524256.
Oliveira, L. A., Porto, A. L. F., & Tambourgi, E. B. (2006). Production of xylanase and protease by Penicillium janthinellum CRC 87M-115 from different agricultural wastes.
Bioresource Technology, 97, 862867.
Otieno, D. O., & Ahring, B. K. (2012a). The potential for oligosaccharide production
from the hemicellulose fraction of biomasses through pretreatment processes:
Xylooligosaccharides (XOS), arabinooligosaccharides (AOS), and mannooligosaccharides (MOS). Carbohydrate Research, 360, 8492.
Otieno, D. O., & Ahring, B. K. (2012b). A thermochemical pretreatment process to produce xylooligosaccharides (XOS), arabinooligosaccharides (AOS) and mannooligosaccharides (MOS) from lignocellulosic biomasses. Bioresource Technology,
112, 285292.
Pandey, A., Soccol, C. R., Nigam, P., & Soccol, V. T. (2000). Biotechnological potential of
agro-industrial residues I: Sugarcane bagasse. Bioresource Technology, 74, 6980.
Pinheiro, V., Alves, A., Mouro, J. L., Guedes, C. M., Pinto, L., Spring, P., et al. (2004). Effect of mannan oligosaccharides on the ileal morphometry and cecal fermentation
of growing rabbits. Proc.: 8th World Rabbit Congress, Puebla, Mxico (pp. 936941).
Polizeli, M. L. T. M., Rizzatti, A. C. S., Monti, R., Terenzi, H. F., Jorge, J. A., & Amorim, D. S.
(2005). Xylanases from fungi: Properties and industrial application. Applied
Microbiolology Biotechnolology, 67, 577591.
Raimbault, M. (1998). General and microbiological aspects of solid substrate fermentation. Electronic Journal of Biotechnology, 1, 115.
Rastall, R. A., & Maitin, V. (2002). Prebiotics and synbiotics: Towards the next generation. Current Opinion in Biotechnology, 13, 490496.
Rivero-Urgell, M., & Santamaria-Orleans, A. (2001). Oligosaccharides: Application in
infant food. Early Human Development, 65, S43S52.
Rodrigues, T. H. S., Dantas, M. A. A., Pinto, G. A. S., & Goncalves, L. R. B. (2007). Tannase
production by solid state fermentation of cashew apple bagasse. Applied Biochemistry and Biotechnology, 136140, 675688.
Rodrigues, R. C. L. B., Rocha, G. J. M., Rodrigues, D., Izrio Filho, H. J., Felipe, M. G. A., &
Pessoa, A. (2010). Scale-up of diluted sulfuric acid hydrolysis for producing sugarcane
bagasse hemicellulosic hydrolysate (SBHH). Bioresource Technology, 101, 12471253.
Rycroft, C. E., Jones, M. R., Gibson, G. R., & Rastall, R. A. (2001). A comparative in vitro
evaluation of the fermentation properties of prebiotic oligosaccharides. Journal of
Applied Microbiology, 91, 878887.
Sabiha-Hanim, S., Noor, M. A. M., & Rosma, A. (2011). Effect of autohydrolysis and enzymatic treatment on oil palm (Elaeis guineensis Jacq.) frond bres for xylose and
xylooligosaccharides production. Bioresource Technology, 102, 12341239.
Saha, B. C. (2003). Hemicellulose bioconversion. Journal of Industrial Microbiology and
Biotechnology, 30, 279291.
Samanta, A. K., Jayapal, N., Kolte, A. P., Senani, S., Sridhar, M., Suresh, K. P., et al. (2012).
Enzymatic production of xylooligosaccharides from alkali solubilized xylan of natural grass (Sehima nervosum). Bioresource Technology, 112, 199205.
Samanta, A. K., Senani, S., Kolte, A. P., Sridhar, M., Sampath, K. T., Jayapal, N., et al.
(2012). Production and in vitro evaluation of xylooligosaccharides generated
from corncobs. Food and Bioproducts Processing, 90, 466474.
Santos, D. T., Sarrouh, B. F., Rivaldi, J. D., Converti, A., & Silva, S. S. (2008). Use of sugarcane bagasse as biomaterial for cell immobilization. Journal of Food Engineering, 86,
542548.
Senani, S., Sridhar, M., Samanta, A. K., Kolte, A. P., Venugopal, N., & Satish, L. (2009). Selection of lactobacillus as probiotics for use as feed supplement. Proceedings of 13th
Biennial ANSI Conference on Diversication of Animal Nutrition Research in the
Changing Scenario held at Bangalore from 1719 December 2009 (pp. 118).
Sharma, M., & Chadha, B. S. (2011). Chapter 9 Production of hemicellulolytic enzymes for hydrolysis of lignocellulosic biomass. Biofuels, 203228.
Sharma, M., Chadha, B. S., & Saini, H. S. (2010). Purication and characterization of two
thermostable xylanases from Malbranchea ava active under alkaline conditions.
Bioresource Technology, 101, 88348842.
Sharp, R., Fishbain, S., & Macfarlane, G. T. (2001). Effect of short-chain carbohydrates on
human intestinal bidobacteria and Escherichia coli in vitro. Journal of Medical Microbiology, 50, 152160.
Shu, Q., Lin, H., Rutherfurd, K. J., Fenwick, S. G., Prasad, J., & Gopal, P. K. (2000). Dietary
Bidobacterium lactis (HN019) enhances resistance to oral Salmonella typhimurium
infection in mice. Microbiology and Immunology, 44, 213222.
Silva, A. M., Barbosa, F. H., Duarte, R., Vieira, L. Q., Arantes, R. M., & Nicoli, J. R. (2004).
Effect of Bidobacterium longum ingestion on experimental salmonellosis in mice.
Journal of Applied Microbiology, 97(1), 2937.
85