Process Biochemistry 41 (2006) 1913–1923

www.elsevier.com/locate/procbio

Review
Advances in the manufacture, purification and applications
of xylo-oligosaccharides as food additives and nutraceuticals
Andrés Moure *, Patricia Gullón, Herminia Domı́nguez, Juan Carlos Parajó
Chemical Engineering Department, University of Vigo (Campus Ourense), As Lagoas, 32004 Ourense, Spain
Received 16 March 2006; received in revised form 4 May 2006; accepted 10 May 2006

Abstract
Recent developments on the manufacture, purification and biological effects of xylo-oligosaccharides are reviewed. Xylo-oligosaccharides
(XO) can be produced by chemical and/or enzymatic methods from a variety of xylan-containing raw materials, and then refined by
physicochemical treatments. Considered as food ingredients, xylo-oligosaccharides have favourable technological properties and cause prebiotic
effects derived from their ability to modulate the intestinal function. Besides the effects related to their effects in the large bowel, a range of
additional biological activities have been reported for XO. Other topic discussed include the utilization of XO in synbiotic preparations, their
technological properties and market perspectives.
# 2006 Elsevier Ltd. All rights reserved.

Keywords: Xylo-oligosaccharides; Xylanases; Hydrothermal treatments; Refining; Biological effects

Contents

1. Biomass, xylan and xylan hydrolysis products. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1913

2. Autohydrolysis of lignocellulosic materials for producing xylo-oligosaccharides . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1915
3. Refining of XO . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1915
4. Intestinal action of XO . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1917
5. Other properties of XO. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1918
6. Other biological effects of XO . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1920
7. Economic and market considerations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1920
Acknowledgement . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1920
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1921

1. Biomass, xylan and xylan hydrolysis products
Biomass from plant material is the most abundant and
widely spread renewable raw material for the sustainable
production of clean and affordable biofuels, biopower, and
high-value bioproducts. Lignocellulosic materials (LCM) from
forest, agriculture, set-aside lands, industry or urban solid
wastes, mainly made up lignin, cellulose and hemicelluloses,
are potential feedstocks for chemical utilization.
* Corresponding author. Tel.: +34 988 3870082; fax: +34 988 387001.
E-mail address: amoure@uvigo.es (A. Moure).
1359-5113/$ – see front matter # 2006 Elsevier Ltd. All rights reserved.
doi:10.1016/j.procbio.2006.05.011
LCM are heterogeneous and present a complex chemical
nature. Their integral benefit can be achieved by chemical
fractionation, following the ‘‘biomass refining’’ philosophy [1],
based on the selective separation of the main components to
yield a variety of high added-value bioproducts.
The most abundant hemicellulosic polymers are xylans,
made up of xylose units. Xylans represent an immense resource
of biopolymers for practical applications, accounting for 25–
35% of the dry biomass of woody tissues of dicots and lignified
tissues of monocots, and occur up to 50% in some tissues of
cereal grains. The structure of xylans depends on the source
considered. The most common xylans are made up of a main
backbone of xylose linked by b-1!4 bonds, where the
1914 A. Moure et al. / Process Biochemistry 41 (2006) 1913–1923

Table 1
Recent literature on hydrothermal processing of xylan-containing feedstocks
Raw material Processing Objectives of the work References
Crop residues (corncobs) Two-step aqueous treatment Manufacture of XO by hydrothermal [27]
(100–140 8C and below 200 8C, processing and further refining
respectively)
Sugarcane bagasse Hydrothermal processing Assessment of the effects of the [28]
at 200 8C solid to liquor ratio
Eucalyptus globulus wood Non-isothermal hydrothermal Kinetics of hemicellulose decomposition [29]
treatments up to 224 8C into oligomers and monomers
E. globulus wood Hydrothermal treatment under optimal Comparative evaluation of substrates for [30]
conditions for XO production XO production and product characterization
E. globulus wood One- or two-stage hydrothermal Manufacture of XO by hydrothermal [31]
treatments for optimal XO yield processing and further refining
Aspen wood Aqueous treatments in microwave Product purification and structural [32]
oven at 180 8C characterization
Corncobs Hydrothermal treatment under optimal Comparative evaluation of substrates for [30]
conditions for XO production XO production and product characterization
Corncobs Isothermal hydrothermal treatments at Study of fractionation effects [33]
temperatures in the range 145–190 8C
Corncobs Hydrothermal treatments at Kinetics of hemicellulose decomposition [34]
temperatures in the range 150–190 8C into oligomers and monomers
Barley hulls Hydrothermal treatment under optimal Comparative evaluation of substrates for [30]
conditions for XO production XO production and product characterization
Barley hulls Aqueous treatments under optimized Manufacture of XO by hydrothermal [35]
conditions for the production of oligomers processing and further refining
Barley hulls Non-isothermal hydrothermal treatments Kinetics of hemicellulose decomposition [36]
up to 220 8C and byproduct characterization
Brewery spent grains Hydrothermal treatments at temperatures Oligomer generation for further [37]
in the range 150–190 8C manufacture of fermentation media
Brewery spent grains Hydrothermal treatments at temperatures Kinetics of hemicellulose decomposition [38]
in the range 150–190 8C into oligomers and monomers
Almond shells Hydrothermal treatments at temperatures Kinetics of hemicellulose decomposition [25]
in the range 150–190 8C into oligomers and monomers
Corn stover Hydrothermal treatments for fractionation Optimization of the enzymatic digestibility [16]
(optimal temperature, 190 8C) of spent solids from autohydrolysis
Corn stover Aqueous fractionation at 200 8C in flow- Comparative evaluation of reaction [22]
and batch-reactors technologies
Corn fiber Hydrothermal processing at 160 8C for Manufacture of oligosaccharide solutions [39]
hemicellulose solubilization for further conversion into monosaccharides
with a macroreticular strong cation exchanger
Rice hulls Hydrothermal treatment under optimal Comparative evaluation of substrates for [30]
conditions for XO production XO production and product characterization
Rice hulls Non-isothermal hydrothermal treatments Kinetics of hemicellulose decomposition [40]
up to 225 8C into oligomers and monomers
Rice hulls Aqueous treatments under optimal Manufacture of XO by hydrothermal [41]
conditions for the production of oligomers processing and further refining
Rice hulls Hydrothermal treatments in a Manufacture of water-soluble compounds [19]
percolation reactor at temperatures from hemicelluloses
in the range 140–220 8C
Rice hulls Two-step aqueous treatment Production of high-DP xylan oligosaccharides [42]
(120 and 198 8C, respectively) for and water-soluble lignins
hydrolysis of hemicellulose
and susceptible lignin
Flax shive Aqueous treatments in microwave oven Structural characterization of [43]
for partial depolymerization and hemicellulose-derived products
solubilization of hemicelluloses
Wheat straw Steam explosion at temperatures in the Characterisation of degraded [44]
range 200–220 8C hemicellulosic polymers
Bamboo Hydrothermal processing in a percolation Optimization of XO production [45]
reactor at temperatures in the
range 175–180 8C
 A. Moure et al. / Process Biochemistry 41 (2006) 1913–1923
structural units are often substituted at positions C2 or C3 with
arabinofuranosyl, 4-O-methylglucuronic acid, acetyl or phe-
nolic substituents [2].
2. Autohydrolysis of lignocellulosic materials for
producing xylo-oligosaccharides
Xylo-oligosaccharides (XO) are produced from xylan-
containing LCM by chemical methods (for example, by
autohydrolysis with water or steam or in media catalyzed with
externally added mineral acids), direct enzymatic hydrolysis of
a susceptible substrate [3–8] or a combination of chemical and
enzymatic treatments [6,9–13].
The state of the art on the manufacture and applications of XO
until 2000 was reviewed previously [14]. This work aims to
provide an overview of the recent developments in the same field,
devoting a particular interest to processes involving at least one
aqueous treatment (with hot, compressed water or steam) to
cause the degradation of xylan to soluble carbohydrates.
When the aqueous processing of xylan-containing LCM
(autohydrolysis or hydrothermal treatment) is carried out under
suitable operational conditions, the hemicellulosic chains are
progressively broken down by the hydrolytic action of
hydronium ions (generated from water autoionization and
from in situ generated organic acids), yielding soluble products
(mainly oligosaccharides), and leaving both cellulose and
lignin in solid phase with little chemical alteration.
The spent solids from autohydrolysis are suitable for further
utilization, for example enzymatic hydrolysis of cellulose [15–
22], utilization for feed or fuel, or as a construction material
[23]. In this field, it can be noted that multiproduct processes
from lignocellulosic materials (according to the ‘‘biorefiney
approach’’) present better economic perspectives, and decrease
greenhouse gas emissions in agreement with the objectives of
the Kyoto Protocol [24].
The main soluble products from autohydrolysis reactions are
usually referred as ‘‘oligosaccharides’’, even if they may
present a broad distribution of molecular weights [25]. The
term ‘‘oligosaccharide’’ is usually reserved for DP in the range
3–10 [26], whereas fractions of higher molecular weight can
been considered as ‘‘dietary fiber’’. However, the definition of
this latter term has seen several revisions since it was proposed
in 1953, and a variety of definitions are available based either
on analytical determination by specific methods or on their
biological effects, leading to some confusion between the
concepts of ‘‘oligosaccharide’’ and ‘‘dietary fiber’’. For
example, the definitions of dietary fiber by the American
Association of Cereal Chemists in 2000 assumes that
oligosaccharides makes part of dietary fiber, whereas the
National Academy of Science classified in 2002 the ‘‘isolated,
non-digestible carbohydrates’’ as ‘‘functional fiber’’, including
oligosaccharides and compounds with higher molecular weight
[26]. Considering that the only chemical difference between
polymeric and oligomeric xylan-degradation products is the
DP, and that xylobiose (DP2) has been considered as an
oligosaccharide in food studies [14], the whole set of soluble
fragments from xylan hydrolysis (with a broad range of DP, the
1915
major fractions corresponding usually to DP
20) has been
denoted as xylo-oligosaccharides (XO) or ‘‘substituted
oligosaccharides’’, particularly in studies where the distribution
of molecular weights was unknown.
Recent studies have been reported on the manufacture of XO
by chemical processing of a variety of feedstocks, including
crop residues [27], sugarcane bagasse [28], hardwoods [29–32],
corncobs [30,33,34], barley hulls [30,35,36], brewery spent
grains [37,38], almond shells [25], corn stover and corn fiber
[16,22,39], rice hulls [19,30,40–42], flax shive [43], wheat
straw [44] and bamboo [45]. Table 1 summarizes information
on this topic.
In autohydrolysis treatments, XO behave as typical reaction
intermediates, and their maximum concentration is achieved
under medium-severity conditions. The molecular weight
distribution depends on both the substrate employed and the
reaction conditions. Treatments of increased severity lead to
decreased degrees of polymerization, but also to increased
decomposition of XO into xylose. Kinetic studies dealing with
XO production from a variety of substrates have been recently
reported [25,29,34,36,37,40].
XO obtained by autohydrolysis present a rich substitution
pattern, conserving the major structural features of the native
xylan [46,47]. For example, only a part of the ester groups are
split in hydrothermal treatments, in comparison with the
saponification caused by processes involving alkaline stages.
Detailed structural information has been reported for XO
obtained from several sources, using techniques such as high-
performance anion-exchange chromatography (HPAEC),
matrix-assisted laser desorption/ionization time-of-flight mass
spectrometry (MALDI-TOF MS), reversed phase chromato-
graphy (RPC), LC–MS and electro- and nanospray-MS, 1H and
13
C NMR homonuclear and heteronuclear two-dimensional
techniques and SEC [32,43,46–54], allowing the identification
and quantification of XO substituents, as well as their locations
in the xylopyranose rings.
3. Refining of XO
When XO are produced by water or steam treatments, a
variety of other compounds (including monosaccharides, acetic
acid, products derived from the extractive and acid-soluble
lignin fractions of the feedstock, furfural from pentose
dehydration, soluble inorganic components of the feedstock,
and protein-derived products) appear in the reaction media
[35,41].
In order to produce food-grade XO, the autohydrolysis
liquors have to be refined by removing both monosaccharides
and non-saccharide compounds, to obtain a concentrate with an
XO content as high as possible. The usual purity of commercial
XO lies in the range 75–95%. Purification of XO obtained by
enzymatic processing of substrates containing susceptible
xylan is facilitated by the previous chemical processing of the
lignocellulosic raw material as well as by the specific action of
xylanases.
Purification of autohydrolysis liquors is a complex problem,
and may require multistage processing for reaction and/or
1916 A. Moure et al. / Process Biochemistry 41 (2006) 1913–1923

fractionation. For example, in single-stage autohydrolysis

reactions, a significant part of the dissolved feedstock
corresponds to easily extractable compounds (for example,
waxes, low molecular weight phenolics and soluble inorganic
components), which could be removed by a previous, mild
hydrothermal treatment leaving the hemicellulosic polymers
almost untouched and ready for further hydrolytic conversion
under harsher conditions. Based on this idea, two consecutive
aqueous treatments of increasing severity (the first one
intending the removal of easily extractable compounds, and
the second one intending the selective solubilization of
hemicelluloses) have been performed to obtain either XO with
improved purity or solutions with enhanced susceptibility to
further refining treatments [27,31,42].
Some processing schemes start with a vacuum evaporation
stage in order to both increase the XO concentration and
remove undesired, volatile components.
Solvent extraction is useful for removing non-saccharide
components of autohydrolysis liquors [35,41,31], yielding both
a selectively refined aqueous phase and a solvent-soluble
fraction mainly made up of phenolics and extractive-derived
compounds. Antioxidant properties have been reported for the
ethyl acetate-soluble components of liquors from hydrolytic
processing of biomass [55–57], suggesting the possibility of
achieving an integrated benefit of the several fractions from
autohydrolysis of lignocellulosics (oligosaccharide-containing
aqueous phase from solvent extraction of liquors, antioxidant-
containing organic phase from solvent extraction of liquors and Fig. 1. Scheme of a possible process for utilization of xylan-containing
cellulose-enriched solid phase from autohydrolysis treatments) lignocellulosic materials.
as depicted in Fig. 1.
Solvent precipitation of liquors has been employed for of xylose and arabinose units [59], whereas size-exclusion
refining XO using ethanol, acetone and 2-propanol chromatography has been employed in combination with other
[31,35,41,58]. The degree of purification and the recovery techniques for purification of feruloylated oligosaccharides [62].
yield depends on the solvent employed and on the lignocellu- Jacobs et al. [43] purified hemicellulose-derived products from
lose raw material, which control the XO substitution pattern hydrothermal microwave treatments of flax shive employing ion-
and the possible presence of stabilising, non-saccharide exchange chromatography and/or size-exclusion chromatogra-
components [31]. phy in combination with enzymic processing.
As the presence of even minimal amounts of water limits the Ion exchange has been employed for purification of XO
precipitation of hemicellulose-derived products, solvent [6,12,27,35,41,61,63,64] alone or in multi-step processing,
extraction of freeze-dried autohydrolysis liquors has been looking mainly for desalination and removal of other undesired
carried out using the same solvents employed for precipitation compounds.
[31,35,41]. The best purification effects were achieved with Membranes are gaining importance in XO technology for a
ethanol, but the process showed limited recovery yields [31]. variety of purposes, including generation by enzymatic
Adsorption has been used in combination with other treatments reactions [65,66], refining and concentration. Ultrafiltration
for the refining of XO, intending either the separation of oligo- has been employed as an alternative to ethanol precipitation for
from mono-saccharides [14,59,60] or the removal of undesired isolation of arabinoxylo-oligosaccharides from enzymically
compounds [6,11,12,61]. hydrolyzed arabinoxylan [58], leading to fractions with similar
Chromatographic separation has been carried out for XO degree of polymerization and degree of substitution than the
purification at an analytical level, yielding high purity fractions. precipitated ones. Izumi et al. [8] employed membranes for
For example, samples from hydrothermally treated lignocellu- concentrating enzymatic hydrolyzates of lignocellulose pulp,
losics have been fractionated by anion-exchange chromatogra- and Yuan et al. [11] employed nanofiltration membranes for
phy and size-exclusion chromatography [48,49], whereas concentrating XO obtained by enzymatic breakdown of xylan
chromatographic techniques have been employed for refining from steamed corncobs, whereas concentration and fractiona-
samples before structural characterization of XO, for example by tion of XO by sequential membrane-based steps has been
13
C NMR [7] or MALDI-TOF or nanospray mass spectrometry employed in multistage purification processes [6,12]. Related
[48]. Simulated moving bed, chromatographic separation has membrane studies have been reported for the separation of
also been proposed for purification of oligosaccharides made up glucose from lactose [67], separation of monosaccharide from
 A. Moure et al. / Process Biochemistry 41 (2006) 1913–1923 1917

di- and oligo-saccharides from hydrolyzed starch media [68],
purification of galacto-oligosaccharides [69] and fructo-
oligosaccharides [70,71] by nanofiltration, removal of mono-
saccharides from oligosaccharide fractions of honey [60],
combined ultrafiltration and nanofiltration processing of fructo-
oligosaccharides and inulopolysaccharides [72,73], synthesis
of galactosyl-oligosaccharides from lactose [74], recovery of
milk oligosaccharides [75], separation of pectate oligosacchar-
ides from enzymically hydrolyzed pectate [76], purification of
oligosaccharides from defatted soybean meal [77], purification
of oligosaccharides from milk [78] and production of low-
lactose milk containing oligosaccharides [79].
Multiple purification steps can be necessary for achieving
high-purity XO. Table 2 summarizes information on repre-
sentative processes studied for this purpose.
4. Intestinal action of XO
There is increasing awareness that the human gut microflora
plays a critical role in maintaining host health, both within the
gastrointestinal tract and systemically through the absorption of
metabolites. Although little is known about the individual
species of bacteria responsible for these beneficial activities, it
is generally accepted that the bifidobacteria and lactobacilli

Table 2
Purification processes considered in literature for xylo-oligosaccharide refining
Substrate Method for XO generation Other process stages References
Pulp slurry Enzymatic/acid hydrolysis Concentration (reverse osmosis), [6,12,61]
filtration, ultrafiltration, adsorption,
ion exchange, spray-drying
Birchwood xylan Enzymatic hydrolysis Anion-exchange and size-exclusion [7]
chromatography
Corncobs Steaming and enzymic Flocculation, ion exchange, [11]
hydrolysis nanofiltration, adsorption, evaporation
Corncobs Two-stage hydrothermal Filtration, concentration, adsorption, [27]
treatment ion exchange
E. globulus wood One- or two-step Concentration, solvent (ethyl acetate) [31]
hydrothermal treatments extraction, solvent (ethanol, 2-propanol,
acetone) precipitation
E. globulus wood One- or two-step Solvent (ethyl acetate) extraction, [31]
hydrothermal treatments evaporation, freeze-drying, solvent
(ethanol, 2-propanol, acetone) extraction
Barley husks Hydrothermal treatments Evaporation, ethyl acetate extraction, [35]
solvent (ethanol, 2 propanol, acetone)
precipitation
Barley husks Hydrothermal treatments Evaporation, ethyl acetate extraction, [35]
freeze drying, solvent (ethanol, 2-propanol,
acetone) extraction
Barley husks Hydrothermal treatments Evaporation, ethyl acetate extraction, [35]
ion exchange
Rice husks Hydrothermal treatments Evaporation, ethyl acetate extraction, [41]
solvent (ethanol, 2-propanol, acetone)
precipitation
Rice husks Hydrothermal treatments Evaporation, ethyl acetate extraction, [41]
freeze drying, solvent (ethanol, 2-propanol,
acetone) extraction
Rice husks Hydrothermal treatments Evaporation, ethyl acetate extraction, [41]
ion exchange
Flax shive Hydrothermal microwave Ion-exchange chromatography, [43]
treatments enzymatic processing and/or size-exclusion
chromatography
Wheat Enzymatic hydrolysis Graded ethanol precipitation [58]
Wheat Enzymatic hydrolysis Ultrafiltration with membranes of [58]
different cut-off
Plant tissues Hydrolysis liquors containing Chromatography (simulated [59]
oligo- and mono-saccharides moving-bed system)
Wheat arabinoxylan Enzymatic hydrolysis Anion-exchange and size-exclusion [62]
chromatography
Biomass Steam explosion and further Ion exchange [63]
hydrolysis
Wheat bran Enzymatic hydrolysis Ion exchange [64]
Xylan Enzymatic hydrolysis Membrane reactor for one-step generation [65]
and fractionation of XO
Xylan Enzymatic hydrolysis Membrane reactor for one-step generation [66]
and fractionation of XO
1918 A. Moure et al. / Process Biochemistry 41 (2006) 1913–1923
constitute important components of the beneficial gut micro-
flora [80].
XO cause prebiotic effects when ingested as part of the diet
(for example, as active ingredients of functional foods) through
the modulation of colonic microflora. From a nutritional point
of view, XO behave as non-digestible oligosaccharides
(NDO’s) (also named short-chain carbohydrates SCC’s) [81],
and show biological effects related to the ones caused by other
oligosaccharides, such as fructo-oligosaccharides, galacto-
oligosaccharides, soybean-oligosaccharides, and isomalto-
oligosaccharides. Recent experimental and review articles
are available on the prebiotic action of various types of
oligosaccharides, including XO [80,82–88].
As NDO’s, XO are not degraded by the low-pH gastric fluid
and digestive enzymes, being metabolised in large bowel. The
colonic fermentation of carbohydrates is a complex problem,
because the end products of the metabolism of given bacterial
species can be used as a substrate by others, and some
microorganisms may grow upon substrates that they are not
able to ferment. The prebiotic character of XO is mainly related
to their ability to specifically stimulate the growth of beneficial
bacteria (bifidobacteria and lactobacilli). An in vitro evaluation
of commercial prebiotic oligosaccharides concluded that XO
increased the number of Bifidobacteria, with comparative
advantage respect to other oligosaccharides [84]. The selective
utilization of XO as a carbon source by beneficial bacteria has
been reported by Crittenden et al. [89], who found that many
Bifidobacterium species and Lactobacillus brevis were able to
grow to high yields using XO, which were also efficiently
fermented by some bacteroides isolates but not by E. coli,
Enterococci, Clostridium difficile or Clostridium perfringens.
The ability of XO with degrees of polymerization in the range
2–10 to increase the number of Bifidobacterium and to suppress
the growth of Clostridium has been claimed [9]. A recent study
[90] reported beneficial effects of XO derived not only from
their ability to increase the populations of bifidobacteria and
lactobacilli, but also from the reduction in concentrations of
secondary bile acids, which exert a negative action on colon and
present a dose-dependent toxic potential related to their co-
mutagenic and tumor-promoting properties. Decreased produc-
tion of secondary bile acids during the digestive process has
been claimed for dietary supplements comprising XO and
physiologically active fatty acids [91].
Besides microbial growth, the colonic XO fermentation
leads to the production of CO2, H2, short chain fatty acids
(acetate, propionate and butyrate, denoted SCFA) and lactate.
These latter may be further metabolised systemically or locally
to provide energy generation for the host. A number of health
effects have been reported for SCFA, including improvement in
bowel function, calcium absorption, lipid metabolism and
reduction of the risk of colon cancer [92], serving as fuel in
different tissues and playing a potential role in the regulation of
cellular processes [93].
Focusing on the effects of XO in gut, studies in rats proved
the ability of feruloyl oligosaccharides for stimulating the
growth of Bifidobacterium bifidum [94], and showed a higher
bioavailability of bound ferulic acid in comparison with the free
compound [95], causing favourable effects on the intestinal
microbiota that may affect the development of precancerous
colonic lesions [96].
Earlier in vitro studies with low-DP, linear XO were
followed by further assays with complex XO to highlight the
effects of molecular weight and structure (including the
presence and type of substituents) on their colonic degradation
patterns. The effects of XO structure on degradability were
assessed by Van Laere et al. [97], who tested the breakdown of
linear and arabinose-containing XO by several strains of
Bifidobacterium, Clostridium, Bacteroides, and Lactobacillus.
The slower fermentation of branched XO respect to linear XO
led to higher butyric acid production, which may result in even
more advantageous effects [98], whereas the presence of
feruloyl substituents may promote the growth of beneficial
bacteria [94]. Kabel et al. [49] studied the fermentation of four
different types of XO (acetylated, linear, substituted with
uronic acids and arabino-XO) by faecal inocula, finding that
linear- and arabino-XO were fermented quickly than the other
substrates during the first 20 h, and that after this adaptation
time, the degradation rate was the same for all the substrates
tested. Fermentation resulted in increased cell material and in
accumulation of relatively high substituted XO with DP in the
range 5–7. SCFA and lactate were formed during the
fermentation and their concentrations increased with the
consumption of total XOS. The fermentation presented two
stages, the first one leading mainly to the formation of acetate
and lactate, and the second one leading to the production of
butyrate and propionate.
5. Other properties of XO
Considered as food ingredients, XO show favourable
technological features, including stability in acidic media,
resistance to heat, and ability for offering lower available
energy and for achieving significant biological effects at low
daily intakes. XO are non-cariogenic, save insulin secretion
from the pancreas and stimulate intestinal mineral absorption
[99]. XO can affect bowel habit and are mildly laxative
through stimulation of bacterial growth and fermentation
[81]. A sufficiently high, regular ingestion may cause
diarrhoea due to osmogenic retention of fluid in both the
small and large intestines, which disappears within a few days
because of the increase in population of intestinal microbes
able to readily utilize XO. Although the maximum
permissible dose depends on individual factors, it has been
estimated in 0.12 g/kg body weight for male Japanese adults
[99]. A controlled administration of XO may help to restrain
the growth of pathogenic bacteria, to retard disorders caused
by imbalanced fermentation in colon [100] and to avoid
intestinal disorders such as constipation, inflammatory bowel
disease, diarrhoea and gastritis [101,102]. XO intake has been
found highly effective for the reduction of severe constipation
in pregnant women without adverse effects [103]. Nutritional
infant formulae containing XO have been claimed to have
synergistic effects all along the intestinal tract, improving gut
barrier maturation [104]. XO can be mixed with other
 A. Moure et al. / Process Biochemistry 41 (2006) 1913–1923 1919

Table 3
Recently reported biological activities of XO additional to those related to their modulatory gut effects
Manufacture, substrate Biological effect or application References
Hydrothermal treatment Antioxidant (DPPH-radical scavenging) [42,108]
of rice hulls activity
Enzymatic hydrolysis Antioxidant (DPPH-radical scavenging) [62]
of wheat flour arabinoxylan activity
Enzymatic hydrolysis Antioxidant activity (erythrocyte [64]
of wheat bran hemolysis assay)
Hydrothermal processing Antioxidant activity [109]
of bagassse and enzymatic
processing
Feruloyl xylo-oligosaccharides Protective effect against lipid (LDL) [4]
from enzymatic reactions peroxidation
Active principles of Prevention and treatment of [110]
pharmaceutical preparations oxidative stress
Mixtures of XO and Prevention and control of anemia [102]
tea catechins and arteriosclerosis
Active component of synbiotic Treatment of vaginal and urogenital [111]
preparations with Lactobacillus infections
strains
Active component of Low-glycemic index carbohydrate [112]
pharmaceutical preparations substitute
with flavonoids
Enzymatic hydrolysates Cosmetics [113]
of rice bran
Enzymatic hydrolysates Prevention of atherosclerosis [62]
of wheat flour arabinoxylan
XO with DP 5–20 as active Antihyperlipidemic activity [114]
components of enteral
nutrition preparation
Long chain XO from enzymatic Hypolipemic activity (against cholesterol, [115]
processing of lignocellulosic phospholipid and triglycerides)
materials
Acidic xylo-oligosaccharides Antihyperlipidemic activity [116]
Acidic xylo-oligosaccharides Hair growth stimulation [117]
containing uronic acid residues
Acidic xylo-oligosaccharides from Inhibition of melanin and inhibition of [10]
chemical and enzymatic processing melanoma cell proliferation
of hardwood pulp
Purified acidic xylo-oligosaccharides Treatment of atopic dermatitis [61,118]
from the enzymatic and/or
physicochemical processing of
hardwood pulp
Acidic xylo-oligosaccharides Collagen production enhancer [119]
from the enzymatic processing
of broadleaf pulp
Acidic xylo-oligosaccharides from Active components of moisturizing [120]
enzymatic and/or physicochemical preparations
processing of lignocellulosic materials
Active principles of pharmaceutical Treatment of epithelial covering tissue [121]
preparations
Acidic xylo-oligosaccharides Anti-inflammatory activity [122]
containing uronic acid residues
Chemical–enzymatic processing Therapeutic agents for osteoporosis [123]
of pulp slurry treatment
Acidic xylo-oligosaccharides containing Hyaluronic acid-formation promoters [124]
uronic acid residues from enzymatic
processing of pulps
Acidic xylo-oligosaccharides containing Histamine-release inhibitors [125]
uronic acid residues from enzymatic
processing of pulps
Active components of pharmaceutical Increased absorption and/or bioavailability [126]
preparations of magnesium
Active components of pharmaceutical Prevention of type II diabetes [127]
preparations
Enzymatic processing of algae Cancer cell apoptosis inducers [128]
1920 A. Moure et al. / Process Biochemistry 41 (2006) 1913–1923
Table 3 (Continued )
Manufacture, substrate
Hydrothermal processing of bamboo
Acidic oligosaccharides from enzymatic
processing of birchwood xylan
Enzymatic and acid processing of
hardwood pulps
Acidic xylo-oligosaccharides
containing uronic acid residues
Active principles of pharmaceutical
preparations
Pharmaceutical preparations containing
acidic and neutral XO
Enzymatic processing of rice bran
Active principles of synbiotic
preparations
prebiotics to achieve synergistic effects or make part of
synbiotic preparations together with probiotic microorgan-
isms. Recent patents have been issued on this latter topic
[105–107].
6. Other biological effects of XO
XO (alone or as active components of pharmaceutical
preparations) exhibit a range of biological activities different
from the prebiotic effects related to gut modulation. Table 3
summarizes representative applications claimed in the last
few years, including antioxidant activity (conferred by
phenolic substituents), blood- and skin-related effects,
antiallergy, antimicrobial, anti-infection and anti-inflamma-
tory properties, selective cytotoxic activity, immunomodu-
latory action, cosmetic and a variety of other properties. It
can be noted that a significant part of the recent
developments has been proposed for acidic oligosaccharides
containing uronic substituents, which can be produced from
hardwoods by a combination of enzymatic and/or chemical
treatments.
Besides biological effects concerning human health, XO
have been employed for phytopharmaceutical [3,4,135] and
feed [9,136] applications.
7. Economic and market considerations
The most important applications of XO in terms of current
and potential market demand correspond to ingredients for
functional foods (for example, in combination with soya
milk, soft drinks, tea or cocoa drinks, nutritive preparations,
dairy products with milk, milk powder and yoghurts, candies,
cakes, biscuits, pastries, puddings, jellies, jam and honey
products, and special preparations for health food for elder
people and children) or as active components of synbiotic
preparations. The origin of the term ‘‘functional food’’ can
be traced to Japan, where the concept of foods designed to be
medically beneficial to the consumer evolved during the
Biological effect or application References
Selective cytotoxicity against acute [129]
lymphoblastic leukemia cells
Antimicrobial activity against [4,7]
Gram-positive
bacteria and against
Helicobacter pylori
Bacteriostatic action against [9]
Vibrio anguillarum
Antiallergy agents [130]
Oral cavity component with [131]
antiplaque effect
Immunomodulating activity [132]
Immunomodulatory action preventing [133]
common cold syndrome
Prevention and treatment of [134]
immune disorders
1980s. The term refers to the practice of fortifying foods with
added ingredients that can confer health effects on the
consumer [137]. The functional food market is growing
rapidly [138], based on the consumers’ awareness of the link
between health, nutrition, and diet as well as on the interest
of food manufacturers due to the increased value that the
added ingredients give to food. This is particularly applicable
to XO, for which a selling price of 2500 yen/kg has been
reported (the highest one among 13 different types of
oligosaccharides) [139]. The same reference reports a total
production of 650 ton of XO per year in Japan, which
accounts for about one-half of the world market. However,
the fastest growth rate is expected for the United States
market. The manufacture of probiotics is growing, but their
long-term exploitation as health promoters is dependent on
several factors, including sound, scientifically proven clinical
evidence of health-promoting activity, accurate consumer
information, effective marketing strategies, and, above all, a
quality product that fulfils consumer expectations [137]. The
modulation of the immune system by oligosaccharides and
their role in the reduction of lifestyle-related diseases as well
as the maintenance and improvement of human health has
been also cited as an area of growing importance [140]. With
the increasing health consciousness among consumers and
the rapid progress of physiologically active functional foods,
the future profile of products containing oligosaccharides
with biological activities seems to be greatly promising
[140].
Acknowledgments
Authors are grateful to the Spanish Ministry of Science of
Technology for the financial support of this work (in the
framework of the Research Project ‘‘Development of
technologies for the integral benefit of industrial bypro-
ducts’’, reference CTQ2005-00745/PPQ, which had partial
financial support from the FEDER funds of the European
Union).
 A. Moure et al. / Process Biochemistry 41 (2006) 1913–1923
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