Bioresource Technology 304 (2020) 123019

Contents lists available at ScienceDirect

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

Review

Arabinofuranosidases: Characteristics, microbial production, and potential T

in waste valorization and industrial applications
Vikram Poriaa, Jitendra Kumar Sainia, Surender Singha,b, , Lata Nainb, Ramesh Chander Kuhadc,d
⁎

a
Department of Microbiology, Central University of Haryana, Mahendergarh, Haryana PIN-123031, India
b
Division of Microbiology, Indian Agricultural Research Institute, New Delhi PIN-110012, India
c
Central University of Haryana, Mahendergarh, Haryana PIN-123031, India
d
Lignocellulose Biotechnology Laboratory, Department of Microbiology, University of Delhi South Campus, New Delhi PIN-110021, India

ARTICLE INFO ABSTRACT

Keywords: Alpha-L-arabinofuranoside arabinofuranohydrolase (ARA), more commonly known as alpha-L-arabinofur-

Arabinofuranohydrolase anosidase (E.C. number 3.2.1.55), is a hydrolytic enzyme, catalyzing the cleavage of alpha-L-arabinose by acting
Arabinoxylans on the non-reducing ends of alpha-L-arabinofuranosides, alpha-L-arabinans containing (1,3)- and/or (1,5)-linked
Arabinogalactans arabinoxylans and arabinogalactans. ARA functions as debranching enzyme removing arabinose substituents
Accessory enzymes
from arabinoxylan and arabinoxylooligomers, thereby, boosting the hydrolysis of arabinoxylan fraction of
Biomass
Hemicellulose
hemicellulose and improving bioconversion of lignocellulosic biomass. Previously, comprehensive information
on this enzyme has not been reviewed thoroughly. Therefore, the main aim of this review is to highlight the
important properties of this interesting enzyme, microorganisms used for its production, and enhanced pro-
duction using genetic engineering approach. An account on synergism with other biomass hydrolyzing enzymes
and various industrial applications of this enzyme has also been provided along with an outlook on further
research and development.

1. Introduction reasons for renewed interests in the various accessory/auxiliary enzyme

Globally there is a growing trend to use sustainable resources for the
production of energy, fuels, and other chemicals. Lignocellulose re-
presents a wealth of resources for such processes, and each of its
components can be utilized in a biorefinery approach. Enzymatic hy-
drolysis of the polysaccharides derived from cell walls of lignocellulosic
feedstock is a significant step during biomass bioconversion (Gielkens
et al., 1997). Due to the presence of multiple components, complete
valorization of the lignocellulosic biomass often becomes difficult.
Therefore, much of the recent research is focused on enhancing the
efficiency of the biomass conversion process. Significant efforts are now
being put in efficiently digesting the hemicellulosic components of the
cell wall polysaccharides, to improve the overall biomass to sugar yield.
This is mainly achieved by increasing the cellulolytic enzyme accessi-
bility towards the cellulosic substrates. Moreover, the pentose sugars
(e.g., xylose) released by the saccharification of the hemicelluloses
could also be potentially converted to biofuels or biochemicals in a
biorefinery approach. Hemicellulose degradation has beneficial effects
on the bioconversion of cellulose by decreasing cellulase inhibition
resulting in enhanced sugar yields from biomass. These are some of the
that help in the complete hydrolysis of biomass (Zhou et al., 2019).
Hemicellulose is the second most abundant plant polymer after
cellulose (Tsujibo et al., 2014). Hemicellulose is a heteropolymer con-
sisting mainly of pentose sugars (xylose and arabinose) linked through
β-1,4 linkages with side chains of hexose sugars (glucose, galactose, and
mannose) and, 4-O-methyl glucuronic acid, and galacturonic acid re-
sidues. Xylan is the major plant hemicellulose made up of D-xylopyr-
anoside residues joined through β-1,4 bonds forming the main chain.
Xylan is categorized into arabinoxylan, xyloglucans, and glucuronox-
ylan, etc., based on substitutions present in the xylan backbone
(Sjostrom, 1993). Xylan backbone is commonly found to be substituted
by acetic acid, arabinose, glucuronic acid, ferulic acid, p-coumaric acid,
or 4-O-methyl glucuronic acid (Ordaz-Ortiz and Saulnier, 2005). Sugar
residues present in xylan are modified depending on the plant species
they are originating from, for example, xylan from birch wood contains
acetylated sugar (D-xylose) residues. Moreover, branching of the xylose
chain can be induced by linking of L-arabinofuranose to O at position 2
or 3 or through linking of D-glucuronic acid and 4-O-methyl-D-glu-
curonic acid residues to O at position 2 (Gielkens et al., 1997). Arabi-
noxylan is formed by polymerization of β-1,4-xylopyranosyl residues

⁎
Corresponding author at: Department of Microbiology, Central University of Haryana, Mahendergarh 123031, Haryana, India.
E-mail address: ssriari@gmail.com (S. Singh).

https://doi.org/10.1016/j.biortech.2020.123019
Received 30 November 2019; Received in revised form 9 February 2020; Accepted 11 February 2020
Available online 12 February 2020
0960-8524/ © 2020 Elsevier Ltd. All rights reserved.
V. Poria, et al.
having side chains of α-L-arabinofuranosyl residues linked through α-
(1,3 or 1,2) bonds.
Degradation of xylan requires the action of multiple enzymes, in-
cluding xylanases and β-xylosidases. Xylanases are extracellular en-
zymes responsible for the breakdown of the β-1,4-xylosidic bonds
during hydrolysis of the xylan to release xylo-oligomeric sugars. These
xylooligomers can be further broken down into simpler sugars by the
action of β-xylosidases at their non-reducing ends (Tsujibo et al., 2014).
Side branching of L-arabinofuranosyl units is linked to the backbone of
D-xylose residues at the C-2 position. The complete breakdown of
arabinoxylans requires the combined action of different microbial en-
zymes, mainly arabinosidases and endo-β-xylanases (Crous et al., 1996).
For removal of substituents from xylan backbone, a variety of deb-
ranching enzymes secreted by several microorganisms, such as glu-
curonidases, α-L-arabinofuranosidases (ARA), acetyl xylan esterase, are
required (Gielkens et al., 1997). Terminal non-reducing α-L-1,2-, α-L-
1,3-, and α-L-1,5-arabinofuranosyl residues are hydrolyzed by ARA;
while the endo-1,5-α-L-arabinofuranosidic bonds from arabinans con-
taining L-arabinose in the backbone chain are hydrolyzed by endo-1,5-
α-L-arabinases. Endo-1,5-α-L-arabinases do not have terminal non-re-
ducing L-arabinofuranosyl residue hydrolyzing activity (Margolles-
Clark et al., 1996).
The conversion of arabinoxylan into its constituents requires the
combined action of different enzymes, which includes; endo-1,4-β-xy-
lanase, which act on xylan and convert it to xylo-oligosaccharides; β-
xylosidases, which releases D-xylose by hydrolyzing xylo-oligo-
saccharides; and α-L-arabinofuranosidase, which acts on arabinose side
chains. Access to the xylosidic linkage of xylan backbone to endo-1,4-β-
xylanase and β-xylosidases is restricted by the arabinose side residues
and, therefore, arabinofuranosidase play an important role during xylan
hydrolysis (Sun et al., 2012). Both arabinose and arabino-oligo-
saccharides find many useful applications in food technology, bioe-
nergy production, and nutrition research (Hong et al., 2009). Plant
hetero-polysaccharides such as arabinogalactans, arabinoxylans, and
arabinans contain L-arabinose residues in furanose form (Margolles-
Clark et al., 1996).
The accessory enzymes, ARAs have remained neglected due to lack
of understanding about their definite role in enhancing biomass con-
version. Some studies have supported (Alvira et al., 2011), while others
denied (Gao et al., 2011; Zhang et al., 2011) their importance in im-
proving the lignocellulose conversion process. Hence, there is a need to
revisit this important group of enzymes. Therefore, the main objective
of this review is to provide comprehensive and up to date information
about characteristics, production and their role in biomass valorization,
and other industrial biotechnological applications of ARAs.
2. Classification
Glycosidic bonds linking alcohol to the anomeric carbon of arabi-
nose residue are hydrolyzed by arabinosyl hydrolases. ARAs come
under arabinosyl hydrolases, which are a class of glycoside hydrolase
group of enzymes (Fritz et al., 2008). ARAs can be classified into dif-
ferent categories depending upon its mechanism, substrate-specificity,
amino acid sequence, and specific activity.
2.1. Classification based on substrate specificity and mode of action
Substrate specificity based classification is the most straightforward
classification of glycoside hydrolases, which uses the Enzyme
Commission (EC) number for a given enzyme as per the International
Union of Biochemistry and Molecular Biology (IUBMB) convention
(Henrissat, 1991). In this classification system, O-glycoside hydrolases
are given the EC number 3.2.1 (representing the enzymes which hy-
drolyze O-glycosyl bonds, the fourth number denoting the substrate
specificity and sometimes molecular mechanism). Glycoside hydrolases
follow a general acid catalysis mechanism to hydrolyze the O-glycosyl
2
Bioresource Technology 304 (2020) 123019
linkages. In this mechanism, two amino acids participate in single/
double displacement reactions, which cause inverted or retained con-
figurations, respectively, at the anomeric C of glycoside undergoing
hydrolysis (Henrissat, 1991). Depending upon their substrate specificity
(arabinose anomeric configuration) and how ARA acts on its polymeric
substrate (exo- or endo- acting), these enzymes are classified under
glycoside hydrolases into three different groups (Fritz et al., 2008): α-L-
arabinofuranoside hydrolases (E.C. 3.2.1.55), β-L-arabinosidases (E.C.
3.2.1.88) and endo-1,5-α-L-arabinanases (E.C. 3.2.1.99).
The ARAs are categorized into three types. Type-A hydrolyzes α-1,5-
L-arabinofuranoside and arabino-oligomers but doesn’t hydrolyze
polymers. These are also able to hydrolyze synthetic p-nitrophenyl
(PNP) α-L-arabinofuranosides. Branched (arabino-oligosaccharides),
linear (L-arabinan) polymers, as well as synthetic PNP-α-L-arabinofur-
anoside, are hydrolyzed by the type-B enzyme which causes the release
of xylose and L-arabinose. Third type enzymes are highly specific for
natural substrates having arabinosidic bonds and are known as (1 → 4)-
β-D-arabinoxylan arabinofuranohydrolases (AXH) (Contesini et al.,
2017; Sakamoto et al., 2011).
AXH type is further characterized and classified into three sub-
groups having different mechanisms. The first type is active on S2/ara
(arabinose residues joined to C-2 of the xylose residues having single
substitution) and S3/ara (arabinose residues joined to C-3 of the xylose
residues having single substitution) in arabinoxylan and is termed as
AXH-m. It has been reported in Aspergillus awamori, Bifidobacterium
adolescentis, and Bacillus subtilis. The second sub-type hydrolyzes D3/
ara (terminal arabinose moieties joined to C-3 of the xylose residues
having double substitutions) branches of the arabinoxylan and is
termed as AXH-d3. It has been reported in B. adolescentis and Humicola
insolens. The third subtype acts on both types of xylose residues, which
have one and two substitutions in arabinoxylan and is known as AXH-
md (Sakamoto et al., 2011).
According to Beldman et al., (1997), enzymes that degrade arabi-
nans are categorized as exo- or endo-acting enzymes. Enzymes de-
grading polysaccharides containing (1 → 5)-α-L-arabinan chain at in-
ternal sites are classified according to the enzyme nomenclature as
(1 → 5)-α-L-arabinan (1 → 5)-α-L-arabinanohydrolases. Enzymes that
act in exo-manner to release L-arabinose from polymeric substrates
(e.g., arabinoxylans, arabinans, and arabinogalactans or from arabino-
oligosaccharides and artificial substrates having p-nitrophenyl group)
are systematically named as α-L-arabinofuranoside arabinofuranohy-
drolases (ARAs).
2.2. Classification based on amino acid sequence
Amino acid sequencing of ARAs has revealed that these belong to
the glycoside hydrolase class of proteins. According to the CAZY data-
base (www.cazy.org), ARAs are grouped under class glycoside hydro-
lase (EC 3.2.1.-). The enzymes in the glycoside hydrolase class hydro-
lyze the glycosidic linkage between carbohydrate-carbohydrate or
carbohydrate-noncarbohydrate moieties. Depending upon the degree of
sequence similarity of amino acids, different glycoside hydrolases have
been classified into different families. These families include GH2, GH3,
GH43, GH51, GH54, and GH62 (Shi et al., 2013; Thakur et al., 2019).
Table 1 summarizes the characteristics of ARA enzymes belonging to
different GH families.
GH2 family ARAs belong to the GH-A clan of glycoside hydrolase
family. These enzymes cleave glycosidic bonds with retaining me-
chanism, and their catalytic domain has (β/α)8 3D structure. Glutamate
acts as a nucleophile and proton donor (Shi et al., 2014). In GH2 family,
ARAs are grouped along with β-galactosidase (EC 3.2.1.23), β-D-ga-
lactofuranosidase (EC 3.2.1.146), β-mannosidase (EC 3.2.1.25); β-xy-
losidase (EC 3.2.1.37), β-glucuronidase (EC 3.2.1.31), mannosylglyco-
protein endo-β-mannosidase (EC 3.2.1.152), α-L-arabinopyranosidase
(EC 3.2.1.-), exo-β-glucosaminidase (EC 3.2.1.165), and β-galactur-
onidase (EC 3.2.1.-).
V. Poria, et al.
Table 1
Characteristics of ARA enzymes classified on the basis of amino acid sequences.
Glycoside Hydrolase (GH) Family
GH2 GH3
Clan A –
Stereochemistry Retention of anomeric Retention of anomeric
configuration configuration
Catalytic Domain (β/α)8 3D structure – 5-fold β-propeller 3D
Nucleophile (Base) Glutamate Aspartate
Proton Donor Glutamate Glutamate (for hydrolases)/
Histidine (for phosphorylases)
GH3 family ARAs have catalytic action similar to those belonging to
the GH2 family; however, Aspartate is the catalytic nucleophile (base)
in their case. Glutamate acts as a catalytic proton donor for hydrolases,
and histidine acts as a catalytic proton donor for phosphorylases. Some
ARAs belonging to family GH43 show both α-xylosidase and α-L-ara-
binofuranosidase activities (Shi et al., 2013). In the GH3 and GH43
families, ARAs are grouped along with other β-acting enzymes involved
in lignocellulose hydrolysis. Some of the ARAs of GH43 have associated
α-xylosidase activities also (Shi et al., 2013) and release arabinose from
O-3 of the di-substituted xylose moieties (De La Mare et al., 2013).
GH43 ARAs break glycosidic linkages with inverting type of mechanism
(Shallom et al., 2002) and cause the release of (1 → 3) α-L-linked
arabinofuranosyl side chains from di-substituted xylose residues and
function only on arabinoxylan (Sørensen et al., 2006). These enzymes
possess catalytic domain (CD) with 5-fold β-propeller three-dimen-
sional (3D) structure, aspartate and glutamate being the catalytic nu-
cleophile (base) and catalytic proton donor, respectively.
The GH51 type ARAs release (1 → 2) and (1 → 3)-α-L-arabinofur-
anosyl residues from mono-substituted xylopyranosyl residues in ara-
binoxylan (Bouraoui et al., 2016) and hydrolyze glycosidic-linkages
while retaining the anomeric configuration (Shallom et al., 2002) via
double displacement mechanism. Their action involves two glutamate
residues, one acting as a proton donor and the other as a base. Their CD
has (β/α)8 barrel structure and proton donating and nucleophilic glu-
tamates are on β-4 and β-7 strands, respectively (Fritz et al., 2008).
GH51 enzymes mostly originate from bacteria (Bouraoui et al., 2016).
Few enzymes in this family also possess endoglucanase activity. In a
study, an ARA gene from Xanthomonas axonopodis was amplified and
expressed in Escherichia coli. This GH51 family ARA was able to effi-
ciently hydrolyze both mono- and di-substitutions at the terminal or
internal xylopyranosyl units of arabinoxylan. This enzyme has its active
site in a (β/α)8-barrel, which is associated with the β-sandwich domain.
The N- and the C-terminal regions of the barrel are put together by β-
sheets of β-sandwich. Therefore, the β-sandwich domain provides sta-
bility to the catalytic domain. Two cysteine residues, Cys80 and
Cys186, are present, which contribute to the high thermostability of
this enzyme by forming a disulfide bridge (Dos Santos et al., 2018).
In the GH54 family, ARAs are placed together with β-xylosidases
and act upon their target linkages with retaining mechanism (Shallom
et al., 2002), possessing type-B substrate specificity (De La Mare et al.,
2013). Their CD is similar to GHs of clan-B possessing β-sandwich fold
with glutamate and aspartate as nucleophilic and proton donating re-
sidues, respectively. The ARAs belonging to the GH-F clan are the single
group of enzymes classified into the GH62 family. These enzymes hy-
drolyze single-substituted xylose residues present in the arabinoxylans
acting at O-2 and O-3 while releasing arabinose (De La Mare et al.,
2013; Shallom et al., 2002). Some ARAs in family GH43 hydrolyze
mono-substitutions and others are specific for O-3 linked α-L-arabi-
noxylan residues present in di-substituted xylan. α-L-arabinoxylan re-
sidues from mono-substituted xylopyranosyl units are hydrolyzed by
ARAs from GH62 family whereas ARAs found in GH51 and GH54 fa-
milies release α-L-arabinoxylan residues from both mono- and di-
Bioresource Technology 304 (2020) 123019
GH43 GH51 GH54 GH62
F A – F
Inversion of anomeric Retention of anomeric Retention of anomeric Not characterized
configuration configuration configuration
(β/α)8 3D structure β-sandwich fold 3D –
structure structure
Aspartate Glutamate Glutamate –
Glutamate Glutamate Aspartate –
substituted xylopyranosyl residues (Dos Santos et al., 2018).
3. Microorganisms producing α-L-arabinofuranosidase
Microorganisms play a significant role in bioconversion of plant-
based polymers, including cellulose, hemicellulose, and pectin, etc., by
secreting different hydrolytic and/or debranching enzymes. Many mi-
croorganisms, including fungi, bacteria, and actinomycetes have been
reported to secrete ARAs, along with other major lignocellulolytic en-
zymes (Maheswari and Chandra, 2000; Yang et al., 2016). However,
large scale production of ARAs requires improved microbial strains
with high production capabilities. Few plants have also been reported
for ARA production (Chávez Montes et al., 2008).
3.1. Bacteria producing α-L-arabinofuranosidase
Bacteria are an important group of microorganisms that secrete
enzymes involved in the degradation of different plant polysaccharides.
These bacteria include genus Alicyclobacillus, Bacillus, Bifidobacterium,
Geobacillus, Paenibacillus, Pseudomonas, Thermotoga, etc. (Table 2).
Laere et al. (1997) isolated ARA of B. adolescentis which showed the
ability to hydrolyze the di-substituted xylose residues in arabinoxylan
to release arabinosyl residues. Tuncer (2003) studied the production
and characteristics of ARA by Thermomonospora fusca. This enzyme was
categorized into AXH-d3 class of arabinofuranohydrolase. Yang et al.
(2015) isolated a bi-functional enzyme from Alicyclobacillus sp. A4
showing both ARA and endo-xylanase activities and characterized this
enzyme under family GH51. Hoffmam et al., (2013) reported an ARA
from mesophilic bacteria B. subtilis belonging to family GH51. Bifido-
bacterium longum produces ARA with bifunctional activity on both α-L-
arabinopyranosides and β-D-galactopyranosides (2011). This ARA was
cloned, overexpressed, and the recombinant enzyme was fully char-
acterized for biochemical and kinetic parameters, revealing differences
in these properties when compared to properties of the already char-
acterized non-recombinant enzymes of B. breve and C. cellulovorans.
Bacterial ARAs are generally produced by the submerged fermentation
process, but Khandeparkaer and Jalal (2015) have also reported the use
of solid-state fermentation process for the production of a 97 kDa en-
zyme from Arthrobacter sp., which was fully characterized after its
purification.
3.2. Actinomycetes producing α-L-arabinofuranosidases
Actinobacteria are known to produce a significant amount of many
industrially important enzymes (Singh et al., 2014). Many researchers
have cloned the arabinofuranosidase from pure cultures of bacteria/
fungi (Gielkens et al., 1997; Yang et al., 2016), but only a few reports
are available on arabinofuranosidase production from actinobacteria.
Compared to fungi, actinobacteria have a faster growth rate and being
prokaryotic; they are more amenable to genetic manipulation. Unlike
most bacteria, the enzymes produced by most of the actinobacteria are
extracellular, thus require minimal downstream processing.
3
V. Poria, et al. Bioresource Technology 304 (2020) 123019

Table 2
Important characteristics of microbial α-L-arabinofuranosidase.
S. No. Microorganism GH Family of enzyme Molecular weight Optimum pH Optimum temperature Reference

Bacteria
1. Alicyclobacillus sp. A4 51 56.7 kDa 6.0 60 °C (Yang et al., 2015)
2. Anoxybacillus kestanbolensis 51 58 kDa 5.5 65 °C (Canakci et al., 2008)
3. Bacillus pumilus 51 60 kDa 7.0 55 °C (Degrassi et al., 2003)
4. Bacillus stearothermophilus – 114 kDa 7.0 70 °C (Bezalel et al., 1993)
5. Bacillus subtilis 51 59.5 kDa 6.5 35–50 °C (Hoffmam et al., 2013)
6. Bacillus subtilis 51 58.1 kDa (AbfA) 8.0 50 °C (Inácio et al., 2008)
57.5 kDa (AbfB) 8.0 60 °C
7. Bifidobacterium longum H-1 51 57.43 kDa 4.7 57 °C (Lee et al., 2011)
8. Caldicellulosiruptor saccharolyticus – 58 kDa 5.5 80 °C (Shin et al., 2013)
9. Caldicellulosiruptor saccharolyticus 51 58 kDa 5.5 80 °C (Lim et al., 2010)
10. Geobacillus caldoxylolyticus TK4 51 58 kDa 6.0 75–80 °C (Canakci et al., 2007)
11. Geobacillus vulcani GS90 51 60 kDa 5.0 70 °C (İlgü et al., 2018)
12. Paenibacillus sp. DG-22 51 56.52 kDa 6.0 60 °C (Lee and Lee, 2014)
13. Thermobacillus xylanilyticus 51 56 kDa 5.6–6.2 75 °C (Debeche et al., 2000)
14. Thermotoga petrophila RKU-1 51 57 kDa 6.0 70 °C (dos Santos et al., 2011)
15. Thermotoga thermarum 2 90 kDa 5.5 80 °C (Shi et al., 2014)
16. Thermotoga maritima MSB8 51 55 kDa – 90 °C (Miyazaki, 2005)

Actinomycetes
1. Streptomyces avermitilis NBRC14893 43 53 kDa 6.0 45 °C (Ichinose et al., 2008)
2. Streptomyces chartreusis 51 80 kDa 5.5 55 °C (Matsuo et al., 2000)
43 37 kDa 7.0 50 °C
3. Streptomyces thermoviolaceus OPC-520 62 37 kDa 5.0 60 °C (Tsujibo et al., 2014)
4. Streptomyces sp. PC22 – 79 kDa 6.0 65 °C (Raweesri et al., 2008)
5. Streptomyces lividans 66 – 69 kDa 6.0 60 °C (Manin et al., 1994)

Fungi
1. Aspergillus awamori IFO 4033 51 81 kDa 4.0 60 °C (Kaneko et al., 1998)
62 kDa 4.0 60 °C
2. Aspergillus awamori 62 32 kDa 5.0 50 °C (Kormelink et al., 1991)
3. Aspergillus nidulans 51 88.6 kDa 5.0 70 °C (Damásio et al., 2012)
4. Coprinopsis cinerea 62 41.4 kDa 7.0 45 °C (Hashimoto et al., 2011)
5. Fusarium oxysporum f. sp. Dianthi 54 58 kDa 4.0 50 °C (Chacón-Martínez et al., 2004)
6. Penicillium canescens – 60 kDa 4.7 75 °C (Sinitsyna et al., 2003)
7. Penicillium chrysogenum 62 35 kDa 5.0 40 °C (Sakamoto et al., 2011)
8. Penicillium funiculosum 62 37.1 kDa (Abf62a) 2.2–3.5 and 3.2–7.0 40 °C (De La Mare et al., 2013)
35.4 kDa (Abf62b) 3.2–4.8 40 °C
41.3 kDa (Abf62c) 3.2–4.8 50–52 °C
9. Penicillium purpurogenum 51 70 kDa 5.0 60 °C (Fritz et al., 2008)
10. Penicillium janthinellum 54 64 kDa 5.5 50 °C (Chadha et al., 2017)
11. Phanerochaete chrysosporium 43 83 kDa 5.5 50 °C (Huy et al., 2013)

Streptomyces sp. is reported for the secretion of ARAs. Raweesri et al., (0.049 U/ml) from Streptomyces cuspidosporus using wheat bran and
(2008) produced this enzyme using Streptomyces sp. PC22 and reported xylan as substrate. However, the arabinofuranosidase activity (0.049 U/
its specific activity of 16.34 U/mg using the synthetic substrate. Other ml) was very low in contrast to higher activities of other enzymes like
examples of actinomycetes are S. purpurascens IFO 3389 (Komae et al., CMCase and FPase.
1982), S. coelicolor producing GH62 family enzyme (Maehara et al.,
2014) and S. diastaticus producing 38 and 60 kDa enzymes (Tajana
3.3. Fungi producing α-L-arabinofuranosidase
et al., 1992). The ARAs from different actinomeycetes have been pur-
ified and reported to hydrolyze synthetic substrates in addition to lib-
Many fungi, including ascomycetes as well as basidiomycetes, such
erating arabinose from arabinoxylan and debranched β-1,5-arabinan.
as Penicillium sp., Aspergillus sp., Pleurotus sp., etc. have also been re-
Tsujibo et al., (2014) characterized a gene responsible for ARA
ported to produce ARAs. Penicillium purpurogenum had been reported to
production from S. thermoviolaceus and found it to have a ~ 1.1 kbp
produce a GH51 type of enzyme which is active on synthetic substrate
ORF that coded for 363 amino-acid protein and later overexpressed this
pNP-α-L-arabinofuranoside and showed a specific activity of 1.2 U/mg
gene in E. coli, followed by its purification and characterization. It
(Fritz et al., 2008). Chadha et al., (2017) reported arabinofuranosidase
showed good activity against natural substrates arabinoxylan and oat
production of 212 U/g dry substrate (wheat bran + paddy straw) from
spelt xylan but reduced activity was observed with synthetic substrates.
an efficient hemicellulolytic fungus Penicillium janthinellum. Aspergillus
A modular exo-type ARA having an arabinan binding module had been
nidulans was found to produce this enzyme more efficiently using in-
characterized in S. avermitilis strain NBRC14893 (Ichinose et al., 2008)
organic nitrogen sources and carbon sources containing L-arabinose
having the hydrolytic activities against PNP-α-L-arabinofuranosides but
and crude beet arabinans. Different arabinose containing inducers have
not against other synthetic PNP linked substrates (pNP-α-L-arabino-
been reported in different fungi for producing ARAs. L-arabitol has been
pyranoside/β-d-xylopyranoside/β-d-galactopyranoside). It had been
found to be a good inducer in many fungi. For example, in Penicillium
found to possess an enzymatic activity of 2.92 U/mg. Corn hull arabi-
purpurogenum and Aspergillus terreus, L-arabitol induces the enzyme
noxylan has also been used as a substrate for the production of ARA by
maximally (De Ioannes et al., 2000; Le Clinche et al., 1997). L-arabi-
Streptomyces sp. I10-1, for application in L-arabinose production
nofuranosidase (0.052 U/ml) from Penicillium sp. has been produced on
(Kurakake et al., 2014). The corresponding gene had an ORF of 1.2 kb
optimized media using orange peel as a substrate (Sathishkumar et al.,
coding for 408 amino acids with an estimated 45.2 kDa molecular mass.
2013); whereas wheat bran based SSF process had been used to produce
Maheswari and Chandra (2000) reported arabinofuranosidase activity
the enzyme from A. niger (Patel et al., 2015).

4
V. Poria, et al.
3.4. Arabinofuranosidase from extremophiles
Different thermostable ARA have been reported from many ther-
mophilic microorganisms such as Geobacillus caldoxylolyticus TK4
(Canakci et al., 2007), Anoxybacillus kestanbolensis AC26Sari (Canakci
et al., 2008), Caldicellulosiruptor saccharolyticus (Lim et al., 2010),
Thermotoga thermarum (Shi et al., 2014), Thermotoga petrophila (dos
Santos et al., 2011), and Caldicellulosiruptor saccharolyticus (Shin et al.,
2013), etc. The range of optimum pH and temperature of these ARAs is
5.5–6.0 and 65–80 °C, respectively. A hyperthermophilic ARA from
Thermotoga maritima MSB8 has been reported with an optimum tem-
perature of 90 °C (Miyazaki, 2005).
A salt-tolerant ARA was isolated from a recently identified species of
fungi Aspergillus hortai CRM1919. The highest (190 U/L) ARA produc-
tion by this fungus was observed at 35 °C with pH 3.0. On different agro
wastes, the activity of this ARA ranges from 1.14 U/L on sugarcane
bagasse to 34 U/L on orange peel. The optimum activity was observed
at a pH of 4.0 and a temperature of 60 °C. This halotolerant ARA was
also found to be tolerant to ethanol at 10% concentration (Terrone
et al., 2020). A thermostable ARA of GH 51 family was isolated from
Geobacillus vulcani GS90 and was expressed in E. coli BL21. The op-
timum pH and temperature of this ARA were 5 and 70 °C, respectively.
This enzyme was active over a range of pH (4–9) and temperature
(30–90 °C). The highest activity of this enzyme was found to be 200 U/
mg and 526 U/mg with p-nitrophenyl-α-L-arabinofuranoside and sugar
beet arabinan, respectively (İlgü et al., 2018).
A salt-, xylose- and alkali tolerant, bifunctional β-xylosidase/α-L-
arabinofuranosidase belonging to family GH43 was isolated from
Massilia sp. RBM26. This enzyme was also found resistant to various
chemicals. The optimum pH and temperature of this enzyme were 6.5
and 50 °C, respectively (Xu et al., 2019). The resistance of this enzyme
to various chemicals and their salts, xylose, and alkali stability makes it
a potential biocatalyst for the saccharification of lignocellulose, for the
food industry, and industrial processes conducted in seawater. Another
halotolerant bifunctional β-xylosidase/α-L-arabinofuranosidase form
Colletotrichum graminicola was also reported. In presence of 2.5 mol/L
NaCl, this enzyme retained 63% of its activity. The optimum pH and
temperature of this enzyme were 4.5 and 65 °C (Carvalho et al., 2018),
respectively.
4. Characteristics of some α-L-arabinofuranosidase
Some important characteristics of microbially synthesized ARAs,
such as their molecular weight, glycoside hydrolase family, optimum
pH, and temperature, etc. are shown in Table 2. In general, microbial
ARAs have a molecular weight (MW) between 35–114 kDa. Optimal pH
is generally near neutrality in the case of bacterial ARAs, except Ther-
momonospora fusca (Tuncer and Ball, 2003), which have a pH optimum
of 9.0. In the case of fungi, generally, the optimum pH is in the acidic
range. The optimal temperature for microbial ARAs was found to be in
the range 40–90 °C, and most of the thermophilic bacteria have been
found to produce ARAs with higher temperature optima, e.g., T. mar-
itima has been reported to have maximum activity at 90 °C (Miyazaki,
2005). The stability of the enzyme is also a valuable property that can
be increased by immobilizing. In one such study, ARA was immobilized
on various supports (agarose activated by polyethyleneimine polymers,
cyanogen bromide activated Sepharose, DEAE-Sepharose, and Se-
pharose-Q) via ionic adsorption. ARA adsorbed on Sepharose-Q showed
much higher stability than free ARA (Damásio et al., 2012).
5. Metagenomic studies on α-L-arabinofuranosidases from
environmental samples
Many environmental samples have been reported to contain ARA
activities that may be contributed by the microbial communities pre-
sent in those samples. Fortune et al., (2019) characterized three GH51
5
Bioresource Technology 304 (2020) 123019
type ARAs (AFase-D3, AFase-E3, and AFase-H4) from high-temperature
compost metagenome, after their expression in E. coli followed by
purification. The optimum temperature for AFase-D3 was 25 °C, while
the latter two were optimally active at 60 °C, while the optimum pH for
these three purified enzymes were 4.5, 4.0, and 5.0, respectively.
Specific activities and the molecular weight of these enzymes were in
the range of 143–228 U/mg and 55–58 kDa, respectively, and AFase-E3
was the most thermostable of the three. A GH43 type bi-functional
enzyme from a starter mixture of the compost showing ARA and xylo-
sidase activities, with a molecular weight of 59.1 kDa, has also been
reported (Wagschal et al., 2009). Ruminating organisms feed on lig-
nocellulosic biomass and are very efficient degraders of such biomass.
RuXyn1 belongs to family GH43 and is a bifunctional enzyme showing
β-xylosidases and ARA activities, and the second, RuXyn2 belongs to
family GH30 with bifunctional activities similar to RyXyn1. The mo-
lecular weight of these enzymes is 42 and 50 kDa, respectively (Zhou
et al., 2012).
6. Recombinant α-L-arabinofuranosidase
Being the accessory enzymes during hemicellulose hydrolysis, ARAs
are secreted by the indigenous microorganisms along with the other
hemicellulolytic enzymes and helps in complete bioconversion of lig-
nocellulosic biomass (Chimphango et al., 2012; Margolles-Clark et al.,
1996). Therefore, often ARAs are associated with contaminating ac-
tivities of other hemicellulolytic enzymes such as endo-β-xylanase. In
many applications, it is desirable to overproduce ARAs with no side-
activities, and this is possible with the implementation of recombinant
technology. Recombinant microbes can be generated to produce im-
proved enzyme systems using specific medium composition. In one of
the studies, a recombinant mutant Aspergillus niger was created for ex-
tracellular production of xylanase free ARA (Chimphango et al., 2012).
β-xylosidases/α-arabinofuranosidase gene of Paenibacillus woo-
songensis has been heterologously overexpressed in E. coli (Kim and
Yoon, 2010). This recombinant enzyme showed specific activities (U/
mg) of 0.659 and 0.347 on pNP substituted β-xylopyranoside and α-
arabinofuranoside substrates, respectively. Similarly, a bifunctional
gene of Phanerochaete chrysosporium has been overexpressed in Pichia
pastoris (Huy et al., 2013) with up to 1.51-, 1.26- and 1.23-fold in-
creased activities in presence of glucose, xylose, and arabinose, re-
spectively. Cloning and expression of the genes encoding thermostable
ARAs from thermotolerant or thermophilic microbes, such as Geoba-
cillus vulcani (İlgü et al., 2018), Thermotoga maritima (Miyazaki, 2005),
have also been attempted in model microorganism E. coli. Apart from
the bacterial ARAs, fungal ARAs have also been cloned in heterologous
hosts. As an example, researchers overexpressed ARA gene isolated
from Fusarium oxysporum into E. coli (Chacón-Martínez et al., 2004).
The genes encoding for GH62 ARA, pectin esterase, and GH10 endo-1,4-
beta-xylanase from Aspergillus aculeatus were cloned and overexpressed
in Pichia pastoris to study their synergistic action with commercial
Trichoderma reesei cellulase (Laothanachareon et al., 2015). Gene se-
quences of three ARAs (Abf-A, Abf-B, and AXH-d3) from Bifidobacterium
adolescentis have been expressed in E. coli, and fully characterized after
their purification (Lagaert et al., 2010). The two enzymes Abf-A and
Abf-B were classified as GH43 and the third enzyme AXH-d3 was
classified as GH51 type with a molecular weight of 63, 87, and 60 kDa,
respectively. These enzymes had lesser individual activities towards
wheat, rye, and barley arabinoxylans; however, the enzyme cocktail of
these three enzymes had a much better yield of total arabinose from
these natural substrates. This suggests that the recombinant ARAs can
be used to prepare superior enzymatic cocktails for the complete con-
version of lignocellulosic biomass. Another hemicellulolytic cocktail
had been prepared by mixing GH43 and GH51 type ARA from Humicola
insolens and Meripilus giganteus, respectively, with endo-1,4-β-xylanase
and β-xylosidases from H. insolens and T. reesei, respectively (Sørensen
et al., 2007). In addition to improving the expression or yield,
V. Poria, et al.
researchers have also attempted to improve other properties of ARAs
for enhancing their application potential using recombinant DNA
technology. In one such study, chimeric enzyme was prepared by fusing
the ARA gene of T. maritima to family 6 xylan binding domain from C.
thermocellum. This recombinant enzyme not only had increased cata-
lytic activity, but also bound effectively with its substrate which also
helped it to be active on insoluble arabinoxylan. The recombinant en-
zyme successfully improved the rheology as well as the texture of the
wheat flour (Zhou et al., 2019). Fermenter level production of a ge-
netically engineered strain of P. pastoris resulted in reasonably high
extracellular ARA (479.50 ± 12.83U/mg) and high level of synergism
was observed in ARA and xylanase during the catalytic conversion of
corn stover to fermentable sugar (Zheng et al., 2018).
7. Synergistic action of α-L-arabinofuranosidase with other
enzymes
Enzyme synergy is very important during the degradation of com-
plex substrates such as lignocellulosic residues. The synergistic action of
the enzyme occurs when the combined activity of the mixture of dif-
ferent enzymes is significantly higher than the sum of their individual
enzyme activities. The synergistic improvements in the enzyme activ-
ities in the blends can be calculated as % synergism = [(Actual activity
in blend - Theoretically calculated activity)/(Theoretically calculated
activity)] × 100 (Saini et al., 2016). Synergistic action has been well
reported among various cellulases and hemicellulases produced by
many microorganisms such as bacteria and fungi and can be explained
by different mechanisms (Laothanachareon et al., 2015):
a) Downstream enzymes support the hydrolytic activity of upstream
enzymes by hydrolyzing comparatively shorter fragments of the
substrates generated.
b) By increasing accessibility to specific substrates through supportive
action.
c) Through synergistic action of exo-type enzymes attacking terminal
non-reducing as well as the reducing moieties.
d) By altering the substrates physically.
e) The endo/exo effect, in which the endo-type (endocellulases) en-
zymes generate new shorter oligomeric ends for subsequently acting
enzymes (e.g. cellobiohydrolases).
The complete hydrolysis of pectins and hemicelluloses is promoted
by the synergistic actions of α-L-arabinofuranosidases with hemi-
cellulases and pectic enzymes (Damásio et al., 2012). The investigation
of synergistic and sequential action of xylanolytic enzymes produced by
Penicillium sp. on hydrolysis of oat-spelt xylan has revealed the se-
quential order of enzymes as ARA → xylanase → β-xylosidases
(Rahman et al., 2003). The ARAs can act synergistically with cellulases
and are reported to improve the saccharification yield from different
substrates substantially. The ARA and endo-1,4-β-xylanase from A. niger
and A. nidulans, respectively, have been used in combination with
cellulase enzymes, and the resulting enzyme mixture had increased the
enzymatic hydrolysis yield, due to better synergism (Alvira et al.,
2011). Similarly, the mixture of enzymes containing cellulase of T.
harzianum, pectinase of P. echinulatum and ARA of T. reesei, was found
to enhance the hydrolytic efficiency by 1.16 fold due to better syner-
gism (Delabona et al., 2013). The synergy between genetically en-
gineered hemicellulolytic and pectinolytic enzymes including ARA with
cellulolytic enzymes from T. reesei has also been established by the
enhanced saccharification of rice straw, resulting in more sugar pro-
duction when compared to saccharification without application of ac-
cessory enzymes (Laothanachareon et al., 2015). In this study, the au-
thors have shown that the removal of arabinoside side chains by ARA
enhanced cellulase activity. Evaluation of the recombinant gene cluster
of Paenibacillus sp. encoding ARA and xylanase during sequential or
simultaneous enzyme additions significantly increased xylose release
6
Bioresource Technology 304 (2020) 123019
from different xylan substrates (Shi et al., 2013). The arabinofur-
anosidase enzyme from Streptomyces sp. has been found to possess sy-
nergy with acetyl-xylan esterase while acting on deoiled rice bran to
produce ferulic acid (Uraji et al., 2013). Recombinant and improved
fungal ARA expressed in the yeast efficiently hydrolyzes different lig-
nocellulosic biomasses in synergy with the commercial cellulases re-
covering more than 56% xylose (Marcolongo et al., 2014). Xylan in
sugarcane bagass, which is one of the model lignocellulosic feedstock
for bioethanol production, has arabinosylation on C-2 and/or C-3 and
esterification with ferulic acid at C-5 position. ARA along with feruloyl
esterase, when present in the enzyme mixture as accessory enzymes,
remove such linkages and hydrolyze xylan, which further enhances the
accessibility of cellulase towards lignocellulose. Thus, the substrate
type and its composition have a direct influence on the synergistic ef-
fects of accessory enzymes such as ARA (de Souza Moreira et al., 2017).
The composition of lignocellulosic biomass is directly affected by the
pretreatment method employed. It has been shown that the efficiency of
xylan degrading enzymes gets increased after heat-treatment of xylan
from bagasse and the removal of arabinofuranosyl side chains of xylan
by ARA before enzymatic hydrolysis results in decreased hydrolysis of
xylan by endo-β-1,4-xylanases (Li et al., 2017).
8. Industrial applications of α-L-arabinofuranosidase
ARAs are important enzymes due to their crucial role in the com-
plete bioconversion of hemicelluloses for the synthesis of biofuels and
biochemicals. Their potential applications range from beverage pro-
duction to animal feed processing, including food-processing, biofuel
production, and agro-product manufacturing (Binod et al., 2019;
Canakci et al., 2008; Dos Santos et al., 2018). Many arabinofur-
anosidases that are thermostable have been isolated, which are im-
portant in many industrial processes that need to be carried out at
elevated temperatures. Their main applications include clarification of
the fruit juices, imparting aromatic flavor in wines by hydrolyzing
monoterpenes (Gunata et al., 1989; Michlmayr et al., 2012), increasing
digestibility of feed (Canakci et al., 2008; Fritz et al., 2008), enhancing
digestibility of poultry feed (Ravn et al., 2018), improving quality of
bread texture, increasing biomass sugar hydrolysis in biofuel industries
(Denli and Ercan, 2001; Gunata et al., 1989). Their potential applica-
tions in cancer chemotherapy have also been explored (Chinnathambi
and Lachke, 1995). The summary of the industrial applications of mi-
crobial ARAs is shown in Fig. 1, and the detailed account has been
presented in the following section. For the high production of ARA from
Rhodothermus marinus, the culture medium was optimized on a shake-
flask scale. The maximum activity of ARA (6.599 U/ml), xylanase
(7.499 U/ml), and β-xylosidase (3.899 U/ml) was obtained when
Rhodothermus marinus was cultured in medium containing 3.6 g/l
birchwood xylan and 8.2 g/l yeast extract. Birchwood xylan was the
most effective carbon source for the production of ARA and xylanase,
followed by oat spelt and beechwood xylans, and xylan-rich lig-
nocellulose (Gomes et al., 2000).
Megazyme and Creative Enzyme commercially produce ARAs from
both fungi and bacteria. A GH51 ARA from A. niger (McCleary et al.,
2015), GH62 ARA from A. nidulans (Wilkens et al., 2016) and GH43
ARA from Bifidobacterium adolescentis (Sørensen et al., 2006) are com-
mercial products of Megazyme. Mainly GH43 family ARAs are pro-
duced commercially, but GH62 and GH51 ARAs are also commercially
available.
For expression and production of recombinant ARAs, E. coli is pre-
ferentially used (Kim and Yoon, 2010; Sørensen et al., 2006). However,
P. pastoris has also been used for heterologous expression of ARA from
A. niger through ara codon optimization and N-terminal propeptide
engineering that improved ARA secretion considerably (Zheng et al.,
2018).
V. Poria, et al.
Fig. 1. Important industrial applications of microbial α-arabinofuranosidases.
8.1. Lignocellulosic biomass valorization for production of biofuel and
biochemical
Cellulose, hemicellulose, and lignin are main content in agricultural
wastes. A series of different enzymes can degrade the plant poly-
saccharides. These enzymes include cellulases and hemicellulases.
Cellulose is converted to glucose by cellulases (endoglucanase, cello-
biohydrolase, and glucosidase) (Tathod and Dhepe, 2015). The use of
lignocellulosic biomass for ethanol production has become an im-
portant alternative for biofuel synthesis. For saccharification of dif-
ferent agricultural and forest residues to simple sugars, xylan-degrading
enzymes are essential. Synergistic action of ARAs occurs with the xylan-
degrading enzymes for complete hydrolysis of arabinoxylans by
cleaving α-L-arabinofuranosidic linkages (Greve et al., 1984; Poutanen,
1988). Supplementation of ARAs to other xylanolytic enzymes has
yielded increased sugar content during hydrolysis of various hemi-
cellulose containing biomass, such as oatspelt xylan, maize-cob and
-husk (Raweesri et al., 2008), steam-exploded wheat straw (Alvira
et al., 2011), birchwood xylan (Shi et al., 2013), rye arabinoxylan,
wheat arabinoxylan and debranched arabinan (Chadha et al., 2017)
and sugarcane bagasse (Delabona et al., 2013). In a recent study, the
effect of ARA supplementation on the cellulase hydrolytic action was
affirmed (Xin et al., 2019). The study concludes the inhibitive effect of
xylooligosaccharides on cellulose digestibility and that ARAs: other
hemicellulolytic enzyme ratios need optimum adjustment to bypass
inhibitory effects of arabinoxylooligomers having a lesser degree of
substitution during biomass saccharification.
8.2. Food industry
For bread-making, flour of the grains (such as wheat and rye having
arabinoxylan content of 1.5–3% and 7–8%, respectively) are used
(Shewry et al., 2010), which generally have low water solubility
(Thakur et al., 2019). Water uptake of dough is increased by arabinose
7
Bioresource Technology 304 (2020) 123019
residue present in arabinoxylan, which absorbs around 30% of the
water in the dough. This helps in reducing the dough volume and
changes its texture. Arabinoxylan solubilization in sourdough fermen-
tative processing favorably improves the qualitative features of the
bread (Yang et al., 2017). Bread texture is improved by pentosans,
which are one of the important additives for bread (Yegin et al., 2018).
Pentosans, which are added to wheat flour, are slightly hydrolyzed by
wheat flour enzymes. The action of exo-xylanases, including ARA
(Fessas and Schiraldi, 1998) releases free pentosans from the dough.
The sourdough lactic acid bacterium uses these free pentosans. ARAs
release arabinose during the sourdough fermentation. The activity of
other enzymes is also enhanced by ARAs. Acidity and acetic acid pro-
duction are increased by enhanced availability of soluble carbohy-
drates, thereby, improving the quality of bread (Gobbetti et al., 2000).
Trapped water in arabinoxylan of the bread is released by the action of
ARAs, which enhances its keeping quality by delaying the staling pro-
cess, improving its handling as well as crumb structure, and increasing
volume of the dough (Bosetto et al., 2016; Jiménez and Martinez-
Anaya, 1999). A set of enzymes, xylanases, xylosidase, and arabino-
furanosidase is produced by Thermoascus aurantiacus. After prolonged
incubation, these enzymes released arabinose and xylose during solid-
state fermentation, significantly improving (1.22 fold) the bread-vo-
lume and reducing amylopectin and crumb firmness retrogradation by
17% and 25%, respectively (Oliveira et al., 2014). In a recent study,
chimeric recombinant ARA gene of T. maritima fused to family 6 xylan
binding domain from C. thermocellum, has been used for application in
enhancing rheology and texture of the wheat dough (Zhou et al., 2019).
8.3. Beverage industry
Carbohydrate polysaccharides such as pectin and arabinan are
present in abundant amounts in fruits, and during storage, protein,
polyphenols, and polysaccharides get precipitated, which declines the
quality of fruit juices. ARAs, in combination with pectinolytic enzymes,
V. Poria, et al.
are used to clarify juices in the fruit industry. Arabinose side chains
present in hemicellulose are removed by the addition of ARA, which
helps in arabinan hydrolysis, whereas the pectic substances are digested
by the addition of pectinases (Churms et al., 1983; De Vries et al.,
1982). The solubility of juices is enhanced by the treatment of juice
with enzymes which reduces the haze formation. ARA and xylanases
have been used together for improving the clarity as well as reducing
sugar content and yield of juices of apple, orange, grape, peach, etc.
(İlgü et al., 2018).
The aroma in wine is a result of volatile compounds of the
grapes. The aromatic properties of non-volatile compounds are lost be-
cause of their attachment with carbohydrates as glycoconjugates.
Glycoconjugates are found in the di-glycoside form to which monomeric
pentose and hexose sugar residues are attached (Thakur et al., 2019).
Terpene is commonly found attached to α-L-arabinofuranose and is an
important aromatic molecule. Volatile terpenes and L-arabinose are
produced after the hydrolysis of monoterpenes by ARAs (Belda et al.,
2017; Mateo and Di Stefano, 1997). Recent studies indicate that re-
combinant ARAs can prove fruitful in enhancing the yield of common
fruit juices (İlgü et al., 2018). The application of recombinant enzyme, in
combination with xylanase, can further improve the juice yield.
8.4. Pharmaceutical industry
Hemicellulolytic enzymes can produce arabinoxylan oligomers of
different sizes varying in the arabinose side-chain substitutions. Short-
chain arabinoxylan oligosaccharides, which are in the partially hydro-
lyzed form, are produced from the breakdown of xylan and can promote
the growth of probiotic bacteria (Falck et al., 2018). Arabinoxylan-de-
rived xylooligosaccharides have been proved valuable in supporting
human health, due to their ability to control cholesterol levels and re-
ducing obesity and type II diabetes (Amrein et al., 2003; Lu et al.,
2000). Arabinose, which has an antiglycemic effect, can be produced
with the help of ARAs (Canakci et al., 2008). Excess sucrose utilization
can be prevented by L-arabinose in animals as it inhibits sucrase se-
lectively in the intestine and decreasing glycemic response in a non-
competitive manner when sucrose is ingested. Moreover, it is not ab-
sorbed in the body but has a sweet taste (Ichinose et al., 2008). Xylitol
is derived from L-arabinose released by the action of ARA (Sakakibara
et al., 2009). These sugar alcohols have a low glycemic index, are non-
cariogenic, enhance calcium and B-vitamin uptake, and improve dental
health and, therefore, can be used as low-calorie sweeteners with ad-
ditional benefits (Grembecka, 2015).
Additionally, ARAs can be helpful in mycobacterial disease control.
Mycobacterium tuberculosis and Mycobacterium leprae, which are the
causative agent of tuberculosis and leprosy, respectively, contain four
macromolecules lipoarabinomannan (Shewry et al., 2010), mycolic
acids (MA), arabinogalactan (AG), and peptidoglycan (PG) which
constitute their cell envelope. Mycobacterial cell wall arabinans and
galactans consist of D-arabinofuranose and D-galactofuranose, and the
development of a new anti-mycobacterium drug can be focused on the
degradation of these sugars (Asmarani et al., 2016; Shewry et al.,
2010). Similarly, α-L-arabinofuranosidase is very important in cancer
chemotherapy (Chinnathambi and Lachke, 1995).
8.5. Paper and pulp industry
Arabinose side group removal by the action of ARAs increases pulp
delignification as these side chains restrict the activities of enzymes
used for bleaching (Bezalel et al., 1993; Numan and Bhosle, 2006).
Purified ARA from Bacillus stearothermophilus increases the delignifica-
tion of softwood kraft pulp when used in combination with xylanase
(Bezalel et al., 1993), resulting in up to 19.2% lignin removal.
8
Bioresource Technology 304 (2020) 123019
8.6. Animal feed industry
The digestibility of animal feed can be increased by using endo-xy-
lanases and ARA synergistically (Graham and Inborr, 1991). Arabinosyl
side residues present in hemicellulose affect the digestibility of feed-
stuffs as these side residues restrict the actions of different enzymes
(Greve et al., 1984; Numan and Bhosle, 2006). ARA cleaves arabinosyl
side residues and promote the action of other hydrolyzing enzymes
(Cozannet et al., 2017). A trifunctional enzyme showing endo-xylanase,
ARA, and acetyl xylan esterase activity was isolated from Acidothermus
cellulolyticus 11B. This enzyme released the highest amount of arabinose
from the wheat straw as compared to arabinose released from beech-
wood, birchwood, and oat spelt xylan. This enzyme can be beneficial
for industrial applications as it possesses three different activities,
which can reduce the use of multiple enzymes (Shahid et al., 2018).
9. Future prospects
In order to address the impending energy challenge, there has been
keen interest in developing low-cost fuels like bioethanol and other
feedstock chemicals. Carbohydrate-active enzymes or CAZymes, in-
cluding cellulases, hemicellulases, and other auxiliary enzymes, make
up one-third of the total cost to produce cellulosic biofuels.
Lignocellulolytic cocktails are effective in achieving high theoretical
glucose yield, but despite their efficiency, the yield of xylose in the total
saccharified biomass is generally low, which indicates the lack of these
enzymes in completely degrading hemicelluloses. Therefore, accessory
enzymes like ARAs can prove crucial in biomass hydrolyzing enzyme
cocktails to increase the efficiency of conversion of xylan to xylose. The
ARA helps in degrading lignocellulosic biomass completely into
monomeric sugar or its basic reducing units by acting in tune with
many other carbohydrate-active enzymes. Other than lignocellulose
deconstruction, ARA has wide applications in the food industry, animal
feed, therapeutic, and pharmaceutical industries. Further research
should focus on exploring the application potential of these enzymes in
the health sector, where xylooligosaccharide can be synthesized as
prebiotics. Moreover, bioprospecting of more effective and efficient
ARA for advanced cellulosic biofuel production need to be explored for
effective conversion of biomass to simple sugars, by preparing biomass-
degrading enzyme cocktails enriched with ARA. Thermostable ARAs are
needed for many applications as high temperatures result in increased
reaction velocities, reduced risk of contamination, and high substrate
solubility. Therefore, thermostable ARAs from extremophiles have
considerable industrial interests. Whole-genome sequencing, tran-
scriptomic and proteomic studies have the potential to help identify
novel hydrolytic/accessory genes or isozymes and mechanisms of in-
ducing expression of genes of the potential accessory enzymes.
Research efforts in this direction will further improve the enzyme
cocktail efficiencies, thereby improving the economics of the produc-
tion of cellulosic ethanol.
10. Conclusion
ARAs act on glycosidic bonds that link alcohol and anomeric carbon
in arabinose residues during hemicellulose degradation. The synergistic
action of ARAs and cellulases is required for the complete deconstruc-
tion of lignocellulose. Supplementation of ARAs with other hydrolases
is known to increase saccharification efficiency, which can be instru-
mental in reducing the bioethanol production cost. Despite many in-
dustrial applications, ARAs have not gained the attention of industry
due to lack of availability of hyper producing strains. There is a need to
explore the diverse niches and metagenomes for isolation of ARA pro-
ducing microorganisms or genes for cloning with characteristics sui-
table for different industries.
V. Poria, et al.
Credit authorship contribution statement
Vikram Poria: Writing - original draft. Jitendra Kumar Saini:
Writing - original draft. Surender Singh: Conceptualization, Writing -
review & editing. Lata Nain: Writing - review & editing. Ramesh
Chander Kuhad: Conceptualization, Supervision.
Declaration of Competing Interest
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influ-
ence the work reported in this paper.
Acknowledgments
Authors VP, JKS, and SS thank Central University of Haryana for
administrative and infrastructural support. Author JKS acknowledges
SERB-DST, GOI for financial support (ECR/2016/000929/LS). Authors
SS and JKS acknowledge DBT, GOI for financial support under
Twinning Programme for NER (BT/PR25530/NER/95/1239/2017).
References
Alvira, P., Negro, M.J., Ballesteros, M., 2011. Effect of endoxylanase and alpha-L-arabi-
nofuranosidase supplementation on the enzymatic hydrolysis of steam exploded
wheat straw. Bioresour. Technol. 102 (6), 4552–4558.
Amrein, T.M., Gränicher, P., Arrigoni, E., Amadò, R., 2003. In vitro digestibility and
colonic fermentability of aleurone isolated from wheat bran. LWT – Food Sci.
Technol. 36 (4), 451–460.
Asmarani, O., Fanani, M.Z., Puspaningsih, N.N.T., 2016. Biochemical potential of α-L-
arabinofuranosidase as anti-tuberculosis candidate. Procedia Chem. 18, 82–89.
Belda, I., Ruiz, J., Esteban-Fernández, A., Navascués, E., Marquina, D., Santos, A.,
Moreno-Arribas, M.V., 2017. Microbial contribution to wine aroma and its intended
use for wine quality improvement. Molecules (Basel, Switzerland) 22 (2), 189.
Beldman, G., Schols, H.A., Pitson, S.M., Searle-van Leeuwen, M.J.F., Voragen, A.G.J.
1997. Arabinans and arabinan degrading enzymes. In: Advances in Macromolecular
Carbohydrate Research, Vol 1, (Ed.) R.J. Sturgeon, pp. 1–64.
Bezalel, L., Shoham, Y., Rosenberg, E., 1993. Characterization and delignification activity
of a thermostable α-l-arabinofuranosidase from Bacillus stearothermophilus. Appl.
Microbiol. Biotechnol. 40 (1), 57–62.
Binod, P., Papamichael, E., Varjani, S., Sindhu, R., 2019. Introduction to green biopro-
cesses: industrial enzymes for food applications. In: Parameswaran, B., Varjani, S.,
Raveendran, S. (Eds.), Green Bio-processes: Enzymes in Industrial Food Processing.
Springer Singapore, Singapore, pp. 1–8.
Bosetto, A., Justo, P.I., Zanardi, B., Venzon, S.S., Graciano, L., dos Santos, E.L., de Cássia
Garcia Simão, R., 2016. Research progress concerning fungal and bacterial β-
Xylosidases. Appl. Biochem. Biotechnol. 178 (4), 766–795.
Bouraoui, H., Desrousseaux, M.-L., Ioannou, E., Alvira, P., Manaï, M., Rémond, C.,
Dumon, C., Fernandez-Fuentes, N., O’Donohue, M.J., 2016. The GH51 α-l-arabino-
furanosidase from Paenibacillus sp. THS1 is multifunctional, hydrolyzing main-chain
and side-chain glycosidic bonds in heteroxylans. Biotechnol. Biofuels 9 (1), 140.
Canakci, S., Belduz, A.O., Saha, B.C., Yasar, A., Ayaz, F.A., Yayli, N., 2007. Purification
and characterization of a highly thermostable α-l-Arabinofuranosidase from
Geobacillus caldoxylolyticus TK4. Appl. Microbiol. Biotechnol. 75 (4), 813–820.
Canakci, S., Kacagan, M., Inan, K., Belduz, A.O., Saha, B.C., 2008. Cloning, purification,
and characterization of a thermostable alpha-L-arabinofuranosidase from
Anoxybacillus kestanbolensis AC26Sari. Appl. Microbiol. Biotechnol. 81 (1), 61–68.
Carvalho, D.R.D., Carli, S., Meleiro, L.P., Rosa, J.C., Oliveira, A.H.C.D., Jorge, J.A.,
Furriel, R.P.M., 2018. A halotolerant bifunctional β-xylosidase/α-l-arabinofur-
anosidase from Colletotrichum graminicola: Purification and biochemical character-
ization. Int. J. Biol. Macromol. 114, 741–750.
Chacón-Martínez, C.A., Anzola, J.M., Rojas, A., Hernández, F., Junca, H., Ocampo, W.,
Del Portillo, P., 2004. Identification and characterization of the α-l-arabinofur-
anosidase B of Fusarium oxysporum f. sp. dianthi. Physiol. Mol. Plant Pathol. 64 (4),
201–208.
Chadha, B.S., Monga, A., Oberoi, H.S., 2017. α-L-Arabinofuranosidase from an efficient
hemicellulolytic fungus Penicillium janthinellum capable of hyddrolyzing wheat and
rye arabinoxylan to arabinose. J. Microbiol. Biotechnol. Food Sci. 6 (5), 1132–1139.
Chávez Montes, R.A., Ranocha, P., Martinez, Y., Minic, Z., Jouanin, L., Marquis, M.,
Saulnier, L., Fulton, L.M., Cobbett, C.S., Bitton, F., Renou, J.-P., Jauneau, A., Goffner,
D., 2008. Cell wall modifications in Arabidopsis plants with altered α-l-arabinofur-
anosidase activity. Plant Physiol. 147 (1), 63–77.
Chimphango, A.F.A., Rose, S.H., van Zyl, W.H., Görgens, J.F., 2012. Production and
characterisation of recombinant α-l-arabinofuranosidase for production of xylan
hydrogels. Appl. Microbiol. Biotechnol. 95 (1), 101–112.
Chinnathambi, S., Lachke, A.H., 1995. An extracellular α-l-arabinofuranosidase from
Sclerotium rolfsii. World J. Microbiol. Biotechnol. 11 (2), 232–233.
Churms, S.C., Merrifield, E.H., Stephen, A.M., Walwyn, D.R., Polson, A., van der Merwe,
K.J., Spies, H.S., Costa, N., 1983. An L-arabinan from apple-juice concentrates.
9
Bioresource Technology 304 (2020) 123019
Carbohydr. Res. 113 (2), 339–344.
Contesini, F.J., Liberato, M.V., Rubio, M.V., Calzado, F., Zubieta, M.P., Riano-Pachon,
D.M., Squina, F.M., Bracht, F., Skaf, M.S., Damasio, A.R., 2017. Structural and
functional characterization of a highly secreted alpha-l-arabinofuranosidase (GH62)
from Aspergillus nidulans grown on sugarcane bagasse. Biochim. Biophys. Acta
Proteins Proteom. 1865 (12), 1758–1769.
Cozannet, P., Kidd, M.T., Montanhini Neto, R., Geraert, P.-A., 2017. Next-generation non-
starch polysaccharide-degrading, multi-carbohydrase complex rich in xylanase and
arabinofuranosidase to enhance broiler feed digestibility. Poult. Sci. 96 (8),
2743–2750.
Crous, J.M., Pretorius, I.S., van Zyl, W.H., 1996. Cloning and expression of the α–L-ara-
binofuranosidase gene (ABF2) of Aspergillus niger in Saccharomyces cerevisiae. Appl.
Microbiol. Biotechnol. 46 (3), 256–260.
Damásio, A.R.D.L., Pessela, B.C., Segato, F., Prade, R.A., Guisan, J.M., Polizeli,
M.D.L.T.M., 2012. Improvement of fungal arabinofuranosidase thermal stability by
reversible immobilization. Process Biochem. 47 (12), 2411–2417.
De Ioannes, P., Peirano, A., Steiner, J., Eyzaguirre, J., 2000. An α-l-arabinofuranosidase
from Penicillium purpurogenum: production, purification and properties. J. Biotechnol.
76 (2), 253–258.
De La Mare, M., Guais, O., Bonnin, E., Weber, J., Francois, J.M., 2013. Molecular and
biochemical characterization of three GH62 α-l-arabinofuranosidases from the soil
deuteromycete Penicillium funiculosum. Enzyme Microb. Technol. 53 (5), 351–358.
de Souza Moreira, L.R., Corrêa, C.L., Gomes, H.A.R., Midorikawa, G.E.O., Miller, R.N.G.,
Filho, E.X.F., 2017. The role of fungal transcriptome analysis and side-chain hydro-
lyzing enzymes in sugarcane bagasse breakdown. Springer International Publishing,
Cham, pp. 81–106.
De Vries, J.A., Rombouts, F.M., Voragen, A.G.J., Pilnik, W., 1982. Enzymic degradation of
apple pectins. Carbohydr. Polym. 2 (1), 25–33.
Debeche, T., Cummings, N., Connerton, I., Debeire, P., Donohue, M.J., 2000. Genetic and
Biochemical Characterization of a Highly Thermostable α-L-Arabinofuranosidase
from Thermobacillus xylanilyticus. Appl. Environ. Microbiol. 66 (4), 1734.
Degrassi, G., Vindigni, A., Venturi, V., 2003. A thermostable α-arabinofuranosidase from
xylanolytic Bacillus pumilus: purification and characterisation. J. Biotechnol. 101 (1),
69–79.
Delabona, P.D.S., Cota, J., Hoffmam, Z.B., Paixão, D.A.A., Farinas, C.S., Cairo, J.P.L.F.,
Lima, D.J., Squina, F.M., Ruller, R., Pradella, J.G.D.C., 2013. Understanding the
cellulolytic system of Trichoderma harzianum P49P11 and enhancing saccharification
of pretreated sugarcane bagasse by supplementation with pectinase and α-L-arabi-
nofuranosidase. Bioresour. Technol. 131, 500–507.
Denli, E., Ercan, R., 2001. Effect of added pentosans isolated from wheat and rye grain on
some properties of bread. Eur. Food Res. Technol. 212 (3), 374–376.
Dos Santos, C.R., de Giuseppe, P.O., de Souza, F.H.M., Zanphorlin, L.M., Domingues,
M.N., Pirolla, R.A.S., Honorato, R.V., Tonoli, C.C.C., de Morais, M.A.B., de Matos
Martins, V.P., Fonseca, L.M., Büchli, F., de Oliveira, P.S.L., Gozzo, F.C., Murakami,
M.T., 2018. The mechanism by which a distinguishing arabinofuranosidase can cope
with internal di-substitutions in arabinoxylans. Biotechnol. Biofuels 11 223 223.
dos Santos, C.R., Squina, F.M., Navarro, A.M., Oldiges, D.P., Leme, A.F.P., Ruller, R.,
Mort, A.J., Prade, R., Murakami, M.T., 2011. Functional and biophysical character-
ization of a hyperthermostable GH51 α-l-arabinofuranosidase from Thermotoga pet-
rophila. Biotechnol. Lett. 33 (1), 131–137.
Falck, P., Linares-Pasten, J.A., Karlsson, E.N., Adlercreutz, P., 2018. Arabinoxylanase
from glycoside hydrolase family 5 is a selective enzyme for production of specific
arabinoxylooligosaccharides. Food Chem. 242, 579–584.
Fessas, D., Schiraldi, A., 1998. Texture and staling of wheat bread crumb: effects of water
extractable proteins and ‘pentosans'. Thermochim Acta 323 (1), 17–26.
Fortune, B., Mhlongo, S., van Zyl, L.J., Huddy, R., Smart, M., Trindade, M., 2019.
Characterisation of three novel α-L-arabinofuranosidases from a compost meta-
genome. BMC Biotech. 19 (1), 22.
Fritz, M., Ravanal, M.C., Braet, C., Eyzaguirre, J., 2008. A family 51 α-L-arabinofur-
anosidase from Penicillium purpurogenum: purification, properties and amino acid
sequence. Mycol. Res. 112 (8), 933–942.
Gao, D., Uppugundla, N., Chundawat, S.P., Yu, X., Hermanson, S., Gowda, K., Brumm, P.,
Mead, D., Balan, V., Dale, B.E., 2011. Hemicellulases and auxiliary enzymes for im-
proved conversion of lignocellulosic biomass to monosaccharides. Biotechnol.
Biofuels 4, 5.
Gielkens, M.M.C., Visser, J., de Graaff, L.H., 1997. Arabinoxylan degradation by fungi:
characterization of the arabinoxylan-arabinofuranohydrolase encoding genes from
Aspergillus niger and Aspergillus tubingensis. Curr. Genet. 31 (1), 22–29.
Gobbetti, M., Lavermicocca, P., Minervini, F., De Angelis, M., Corsetti, A., 2000.
Arabinose fermentation by Lactobacillus plantarum in sourdough with added pento-
sans and α-L-arabinofuranosidase: a tool to increase the production of acetic acid. J.
Appl. Microbiol. 88 (2), 317–324.
Gomes, J., Gomes, I.I., Terler, K., Gubala, N., Ditzelmuller, G., Steiner, W., 2000.
Optimisation of culture medium and conditions for alpha-l-Arabinofuranosidase
production by the extreme thermophilic eubacterium Rhodothermus marinus. Enzyme
Microb. Technol. 27 (6), 414–422.
Graham, H., Inborr, J., 1991. Application of xylanase-based enzymes in commercial pig
and poultry production. Prog. Biotechnol. 7, 535–538.
Grembecka, M., 2015. Sugar alcohols—their role in the modern world of sweeteners: a
review. Eur. Food Res. Technol. 241 (1), 1–14.
Greve, L.C., Labavitch, J.M., Hungate, R.E., 1984. Alpha-L-arabinofuranosidase from
Ruminococcus albus 8: purification and possible role in hydrolysis of alfalfa cell wall.
Appl. Environ. Microbiol. 47 (5), 1135–1140.
Gunata, Y., Biron, C., Sapi, J., Bayonove, C., 1989. Glycosidase activities in sound and
rotten grapes in relation to hydrolysis of grape monoterpenyl glycosides. Vitis 28,
191–197.
V. Poria, et al.
Hashimoto, K., Yoshida, M., Hasumi, K., 2011. Isolation and characterization of
CcAbf62A, a GH62 α-L-arabinofuranosidase, from the basidiomycete Coprinopsis ci-
nerea. Biosci. Biotechnol. Biochem. 75 (2), 342–345.
Henrissat, B., 1991. A classification of glycosyl hydrolases based on amino acid sequence
similarities. Biochem. J 280 (2), 309.
Hoffmam, Z.B., Oliveira, L.C., Cota, J., Alvarez, T.M., Diogo, J.A., de Oliveira Neto, M.,
Citadini, A.P.S., Leite, V.B.P., Squina, F.M., Murakami, M.T., Ruller, R., 2013.
Characterization of a Hexameric Exo-Acting GH51 α-l-Arabinofuranosidase from the
Mesophilic Bacillus subtilis. Mol. Biotechnol. 55 (3), 260–267.
Hong, M.R., Park, C.S., Oh, D.K., 2009. Characterization of a thermostable endo-1,5-
alpha-L-arabinanase from Caldicellulorsiruptor saccharolyticus. Biotechnol. Lett. 31 (9),
1439–1443.
Huy, N.D., Thayumanavan, P., Kwon, T.-H., Park, S.-M., 2013. Characterization of a re-
combinant bifunctional xylosidase/arabinofuranosidase from Phanerochaete chrysos-
porium. J. Biosci. Bioeng. 116 (2), 152–159.
Ichinose, H., Yoshida, M., Fujimoto, Z., Kaneko, S., 2008. Characterization of a modular
enzyme of exo-1,5-alpha-L-arabinofuranosidase and arabinan binding module from
Streptomyces avermitilis NBRC14893. Appl. Microbiol. Biotechnol. 80 (3), 399–408.
İlgü, H., Sürmeli, Y., Şanlı-Mohamed, G., 2018. A thermophilic α-L-Arabinofuranosidase
from Geobacillus vulcani GS90: heterologous expression, biochemical characteriza-
tion, and its synergistic action in fruit juice enrichment. Eur. Food Res. Technol. 244
(9), 1627–1636.
Inácio, J.M., Correia, I.L., de Sá-Nogueira, I., 2008. Two distinct arabinofuranosidases
contribute to arabino-oligosaccharide degradation in Bacillus subtilis. Microbiology
154 (9), 2719–2729.
Jiménez, T., Martinez-Anaya, M., 1999. Enzymes, a key to improve bread and dough
quality: degradation by products and relationship with quality. In: 17th ICC
Conference of the Cereal Across the Continents, pp. 6–9.
Kaneko, S., Arimoto, M., Ohba, M., Kobayashi, H., Ishii, T., Kusakabe, I., 1998.
Purification and substrate specificities of two α-L-arabinofuranosidases from
Aspergillus awamori IFO 4033. Appl. Environ. Microbiol. 64 (10), 4021–4027.
Khandeparker, R., Jalal, T., 2015. Xylanolytic enzyme systems in Arthrobacter sp. MTCC
5214 and Lactobacillus sp. Biotechnol. Appl. Biochem. 62 (2), 245–254.
Kim, Y.A., Yoon, K.-H., 2010. Characterization of a Paenibacillus woosongensis beta-
Xylosidase/alpha-Arabinofuranosidase produced by recombinant Escherichia coli. J.
Microbiol. Biotechnol. 20 (12), 1711–1716.
Komae, K., Kaji, A., Sato, M., 1982. An α-L-arabinofuranosidase from Streptomyces pur-
purascens IFO 3389. Agric. Biol. Chem. 46 (7), 1899–1905.
Kormelink, F.J.M., Searl-Van Leeuwen, M.J.F., Wood, T.M., Voragen, A.G.J., 1991.
Purification and characterization of a (1,4)-β-d-arabinoxylan arabinofuranohydrolase
from Aspergillus awamori. Appl. Microbiol. Biotechnol. 35 (6), 753–758.
Kurakake, M., Kanbara, Y., Murakami, Y., 2014. Characteristics of α-L-
Arabinofuranosidase from Streptomyces sp I10–1 for Production of L-Arabinose from
Corn Hull Arabinoxylan. Appl. Biochem. Biotechnol. 172 (5), 2650–2660.
Laere, K.M.J.V., Beldman, G., Voragen, A.G.J., 1997. A new arabinofuranohydrolase from
Bifidobacterium adolescentis able to remove arabinosyl residues from double-sub-
stituted xylose units in arabinoxylan. Appl. Microbiol. Biotechnol. 47 (3), 231–235.
Lagaert, S., Pollet, A., Delcour, J.A., Lavigne, R., Courtin, C.M., Volckaert, G., 2010.
Substrate specificity of three recombinant α-L-arabinofuranosidases from
Bifidobacterium adolescentis and their divergent action on arabinoxylan and arabi-
noxylan oligosaccharides. Biochem. Biophys. Res. Commun. 402 (4), 644–650.
Laothanachareon, T., Bunterngsook, B., Suwannarangsee, S., Eurwilaichitr, L.,
Champreda, V., 2015. Synergistic action of recombinant accessory hemicellulolytic
and pectinolytic enzymes to Trichoderma reesei cellulase on rice straw degradation.
Bioresour. Technol. 198, 682–690.
Le Clinche, F., Piñaga, F., Ramón, D., Vallés, S., 1997. α-L-Arabinofuranosidases from
Aspergillus terreus with Potential Application in Enology: Induction, Purification, and
Characterization. J. Agric. Food. Chem. 45 (7), 2379–2383.
Lee, J.H., Hyun, Y.J., Kim, D.H., 2011. Cloning and characterization of α-L-arabinofur-
anosidase and bifunctional α-L-arabinopyranosidase/β-D-galactopyranosidase from
Bifidobacterium longum H-1. J. Appl. Microbiol. 111 (5), 1097–1107.
Lee, S.H., Lee, Y.E., 2014. Cloning, expression, and characterization of a thermostable
GH51 alpha-L-arabinofuranosidase from Paenibacillus sp. DG-22. J. Microbiol.
Biotechnol. 24 (2), 236–244.
Li, H., Wu, H., Xiong, L., Chen, X., Wang, C., Qi, G., Huang, C., Guo, H., Luo, M., Liu, J.,
Long, M., Chen, X., 2017. The hydrolytic efficiency and synergistic action of re-
combinant xylan-degrading enzymes on xylan isolated from sugarcane bagasse.
Carbohydr. Polym. 175, 199–206.
Lim, Y.R., Yoon, R.Y., Seo, E.S., Kim, Y.S., Park, C.S., Oh, D.K., 2010. Hydrolytic prop-
erties of a thermostable α-l-arabinofuranosidase from Caldicellulosiruptor sacchar-
olyticus. J. Appl. Microbiol. 109 (4), 1188–1197.
Lu, Z.X., Walker, K.Z., Muir, J.G., Mascara, T., O'Dea, K., 2000. Arabinoxylan fiber, a
byproduct of wheat flour processing, reduces the postprandial glucose response in
normoglycemic subjects. Am. J. Clin. Nutr. 71 (5), 1123–1128.
Maehara, T., Fujimoto, Z., Ichinose, H., Michikawa, M., Harazono, K., Kaneko, S., 2014.
Crystal Structure and Characterization of the Glycoside Hydrolase Family 62 α-l-
Arabinofuranosidase from Streptomyces coelicolor. J. Biol. Chem. 289 (11),
7962–7972.
Maheswari, M.U., Chandra, T.S., 2000. Production and potential applications of a xyla-
nase from a new strain of Streptomyces cuspidosporus. World J. Microbiol. Biotechnol.
16 (3), 257–263.
Manin, C., Shareek, F., Morosoli, R., Kluepfel, D., 1994. Purification and characterization
of an α-L-arabinofuranosidase from Streptomyces lividans 66 and DNA sequence of the
gene (abfA). Biochem. J 302 (2), 443–449.
Marcolongo, L., Ionata, E., La Cara, F., Amore, A., Giacobbe, S., Pepe, O., Faraco, V.,
2014. The effect of Pleurotus ostreatus arabinofuranosidase and its evolved variant in
10
Bioresource Technology 304 (2020) 123019
lignocellulosic biomasses conversion. Fungal Genet. Biol. 72, 162–167.
Margolles-Clark, E., Tenkanen, M., Nakari-Setälä, T., Penttilä, M., 1996. Cloning of genes
encoding alpha-L-arabinofuranosidase and beta-xylosidase from Trichoderma reesei by
expression in Saccharomyces cerevisiae. Appl. Environ. Microbiol. 62 (10), 3840.
Mateo, J.J., Di Stefano, R., 1997. Description of the β-glucosidase activity of wine yeasts.
Food Microbiol. 14 (6), 583–591.
Matsuo, N., Kaneko, S., Kuno, A., Kobayashi, H., Kusakabe, I., 2000. Purification, char-
acterization and gene cloning of two α-l-arabinofuranosidases from Streptomyces
chartreusis GS901. Biochem. J 346 (1), 9–15.
McCleary, B.V., McKie, V.A., Draga, A., Rooney, E., Mangan, D., Larkin, J., 2015.
Hydrolysis of wheat flour arabinoxylan, acid-debranched wheat flour arabinoxylan
and arabino-xylo-oligosaccharides by β-xylanase, α-l-arabinofuranosidase and β-xy-
losidase. Carbohydr. Res. 407, 79–96.
Michlmayr, H., Nauer, S., Brandes, W., Schümann, C., Kulbe, K.D., del Hierro, A.M., Eder,
R., 2012. Release of wine monoterpenes from natural precursors by glycosidases from
Oenococcus oeni. Food Chem. 135 (1), 80–87.
Miyazaki, K., 2005. Hyperthermophilic α-L-arabinofuranosidase from Thermotoga mar-
itima MSB8: molecular cloning, gene expression, and characterization of the re-
combinant protein. Extremophiles 9 (5), 399–406.
Numan, M.T., Bhosle, N.B., 2006. α-L-Arabinofuranosidases: the potential applications in
biotechnology. J. Ind. Microbiol. Biotechnol. 33 (4), 247–260.
Oliveira, D.S., Telis-Romero, J., Da-Silva, R., Franco, C.M.L., 2014. Effect of a Thermoascus
aurantiacus thermostable enzyme cocktail on wheat bread qualitiy. Food Chem. 143,
139–146.
Ordaz-Ortiz, J.J., Saulnier, L., 2005. Structural variability of arabinoxylans from wheat
flour. Comparison of water-extractable and xylanase-extractable arabinoxylans. J.
Cereal Sci. 42 (1), 119–125.
Patel, H., Chapla, D., Divecha, J., Shah, A., 2015. Improved yield of α-L-arabinofur-
anosidase by newly isolated Aspergillus niger ADH-11 and synergistic effect of crude
enzyme on saccharification of maize stover. Bioresour. Bioprocess. 2 (1), 11.
Poutanen, K., 1988. An α-L-arabinofuranosidase of Trichoderma reesei. J. Biotechnol. 7
(4), 271–281.
Rahman, A.K.M.S., Sugitani, N., Hatsu, M., Takamizawa, K., 2003. A role of xylanase, α-L-
arabinofuranosidase, and xylosidase in xylan degradation. Can. J. Microbiol. 49 (1),
58–64.
Ravn, J.L., Glitsø, V., Pettersson, D., Ducatelle, R., Van Immerseel, F., Pedersen, N.R.,
2018. Combined endo -β-1,4-xylanase and α-L-arabinofuranosidase increases buty-
rate concentration during broiler cecal fermentation of maize glucurono-arabinox-
ylan. Anim. Feed Sci. Technol. 236, 159–169.
Raweesri, P., Riangrungrojana, P., Pinphanichakarn, P., 2008. α-L-Arabinofuranosidase
from Streptomyces sp. PC22: Purification, characterization and its synergistic action
with xylanolytic enzymes in the degradation of xylan and agricultural residues.
Bioresour. Technol. 99 (18), 8981–8986.
Saini, J.K., Singhania, R.R., Satlewal, A., Saini, R., Gupta, R., Tuli, D., Mathur, A., Adsul,
M., 2016. Improvement of wheat straw hydrolysis by cellulolytic blends of two
Penicillium spp. Renew. Energy 98, 43–50.
Sakakibara, Y., Saha, B.C., Taylor, P., 2009. Microbial production of xylitol from L-ara-
binose by metabolically engineered Escherichia coli. J. Biosci. Bioeng. 107 (5),
506–511.
Sakamoto, T., Ogura, A., Inui, M., Tokuda, S., Hosokawa, S., Ihara, H., Kasai, N., 2011.
Identification of a GH62 α-L-arabinofuranosidase specific for arabinoxylan produced
by Penicillium chrysogenum. Appl. Microbiol. Biotechnol. 90 (1), 137–146.
Sathishkumar, T., Nair, D.M., Nithya, R., Priyadharshini, Y., Suprabha, S., Baskar, R.,
2013. Media optimization by central composite design based response surface
methodology of crude [alpha]-L-Arabinofuranosidase from Penicillium Species. Int. J.
Biosci. Technol. 6 (1), 1.
Shahid, S., Tajwar, R., Akhtar, M.W., 2018. A novel trifunctional, family GH10 enzyme
from Acidothermus cellulolyticus 11B, exhibiting endo-xylanase, arabinofuranosidase
and acetyl xylan esterase activities. Extremophiles 22 (1), 109–119.
Shallom, D., Belakhov, V., Solomon, D., Gilead-Gropper, S., Baasov, T., Shoham, G.,
Shoham, Y., 2002. The identification of the acid–base catalyst of α-arabinofur-
anosidase from Geobacillus stearothermophilus T-6, a family 51 glycoside hydrolase.
FEBS Lett. 514 (2–3), 163–167.
Shewry, P.R., Piironen, V., Lampi, A.-M., Edelmann, M., Kariluoto, S., Nurmi, T.,
Fernandez-Orozco, R., Andersson, A.A.M., Åman, P., Fraś, A., Boros, D., Gebruers, K.,
Dornez, E., Courtin, C.M., Delcour, J.A., Ravel, C., Charmet, G., Rakszegi, M., Bedo,
Z., Ward, J.L., 2010. Effects of genotype and environment on the content and com-
position of phytochemicals and dietary fiber components in rye in the
HEALTHGRAIN diversity screen. J. Agric. Food. Chem. 58 (17), 9372–9383.
Shi, H., Zhang, Y., Xu, B., Tu, M., Wang, F., 2014. Characterization of a novel GH2 family
α-l-arabinofuranosidase from hyperthermophilic bacterium Thermotoga thermarum.
Biotechnol. Lett. 36 (6), 1321–1328.
Shi, P., Chen, X., Meng, K., Huang, H., Bai, Y., Luo, H., Yang, P., Yao, B., 2013. Distinct
actions by Paenibacillus sp. strain E18 α-l-arabinofuranosidases and Xylanase in Xylan
degradation. Appl. Environ. Microbiol. 79 (6), 1990–1995.
Shin, K.C., Lee, G.W., Oh, D.K., 2013. Production of ginsenoside Rd from ginsenoside Rc
by alpha-L-arabinofuranosidase from Caldicellulosiruptor saccharolyticus. J. Microbiol.
Biotechnol. 23 (4), 483–488.
Singh, S., Pranaw, K., Singh, B., Tiwari, R., Nain, L., 2014. Production, optimization and
evaluation of multicomponent holocellulase produced by Streptomyces sp. ssr-198. J.
Taiwan Inst. Chem. Eng. 45 (5), 2379–2386.
Sinitsyna, O.A., Bukhtoyarov, F.E., Gusakov, A.V., Okunev, O.N., Bekkarevitch, A.O.,
Vinetsky, Y.P., Sinitsyn, A.P., 2003. Isolation and Properties of Major Components of
Penicillium canescens Extracellular Enzyme Complex. Biochemistry (Moscow) 68 (11),
1200–1209.
Sjostrom, E., 1993. Wood Chemistry: Fundamentals and Applications. Gulf Professional
V. Poria, et al.
publishing.
Sørensen, H.R., Jørgensen, C.T., Hansen, C.H., Jørgensen, C.I., Pedersen, S., Meyer, A.S.,
2006. A novel GH43 α-l-arabinofuranosidase from Humicola insolens: mode of action
and synergy with GH51 α-l-arabinofuranosidases on wheat arabinoxylan. Appl.
Microbiol. Biotechnol. 73 (4), 850–861.
Sørensen, H.R., Pedersen, S., Jørgensen, C.T., Meyer, A.S., 2007. Enzymatic hydrolysis of
wheat arabinoxylan by a recombinant “minimal” enzyme cocktail containing β-xy-
losidase and novel endo-1,4-β-xylanase and α-l-arabinofuranosidase activities.
Biotechnol. Prog. 23 (1), 100–107.
Sun, J., Wen, F., Si, T., Xu, J.H., Zhao, H., 2012. Direct conversion of xylan to ethanol by
recombinant Saccharomyces cerevisiae strains displaying an engineered mini hemi-
cellulosome. Appl. Environ. Microbiol. 78 (11), 3837–3845.
Tajana, E., Fiechter, A., Zimmermann, W., 1992. Purification and characterization of two
alpha-L-arabinofuranosidases from Streptomyces diastaticus. Appl. Environ. Microbiol.
58 (5), 1447–1450.
Tathod, A.P., Dhepe, P.L., 2015. Efficient method for the conversion of agricultural waste
into sugar alcohols over supported bimetallic catalysts. Bioresour. Technol. 178,
36–44.
Terrone, C.C., de Freitas, Montesino, Nascimento, J., Fanchini Terrasan, C.R., Brienzo, M.,
Carmona, E.C., 2020. Salt-tolerant α-arabinofuranosidase from a new specie
Aspergillus hortai CRM1919: Production in acid conditions, purification, character-
ization and application on xylan hydrolysis. Biocatal. Agric. Biotechnol. 23, 101460.
Thakur, A., Sharma, K., Goyal, A., 2019. α-L-arabinofuranosidase: a potential enzyme for
the food industry. In: Parameswaran, B., Varjani, S., Raveendran, S. (Eds.), Green Bio-
processes: Enzymes in Industrial Food Processing. Springer, Singapore. Singapore, pp.
229–244.
Tsujibo, H., Takada, C., Wakamatsu, Y., Kosaka, M., Tsuji, A., Miyamoto, K., Inamori, Y.,
2014. Cloning and Expression of an α-L-Arabinofuranosidase Gene (stxIV) from
Streptomyces thermoviolaceus OPC-520, and Characterization of the Enzyme. Biosci.
Biotechnol. Biochem. 66 (2), 434–438.
Tuncer, M., Ball, A.S., 2003. Purification and partial characterization of α-L-arabinofur-
anosidase produced by Thermomonospora fusca. Folia Microbiol. 48 (2), 168–172.
Uraji, M., Kimura, M., Inoue, Y., Kawakami, K., Kumagai, Y., Harazono, K., Hatanaka, T.,
2013. Enzymatic production of ferulic acid from defatted rice bran by using a com-
bination of bacterial enzymes. Appl. Biochem. Biotechnol. 171 (5), 1085–1093.
Wagschal, K., Heng, C., Lee, C.C., Wong, D.W.S., 2009. Biochemical characterization of a
novel dual-function arabinofuranosidase/xylosidase isolated from a compost starter
mixture. Appl. Microbiol. Biotechnol. 81 (5), 855–863.
Wilkens, C., Andersen, S., Petersen, B.O., Li, A., Busse-Wicher, M., Birch, J., Cockburn, D.,
11
Bioresource Technology 304 (2020) 123019
Nakai, H., Christensen, H.E.M., Kragelund, B.B., Dupree, P., McCleary, B., Hindsgaul,
O., Hachem, M.A., Svensson, B., 2016. An efficient arabinoxylan-debranching α-l-
arabinofuranosidase of family GH62 from Aspergillus nidulans contains a secondary
carbohydrate binding site. Appl. Microbiol. Biotechnol. 100 (14), 6265–6277.
Xin, D., Chen, X., Wen, P., Zhang, J., 2019. Insight into the role of α-arabinofuranosidase
in biomass hydrolysis: cellulose digestibility and inhibition by xylooligomers.
Biotechnol. Biofuels 12 (1), 64.
Xu, B., Dai, L., Zhang, W., Yang, Y., Wu, Q., Li, J., Tang, X., Zhou, J., Ding, J., Han, N.,
Huang, Z., 2019. Characterization of a novel salt-, xylose- and alkali-tolerant GH43
bifunctional β-xylosidase/α-l-arabinofuranosidase from the gut bacterial genome. J.
Biosci. Bioeng. 128 (4), 429–437.
Yang, W., Bai, Y., Yang, P., Luo, H., Huang, H., Meng, K., Shi, P., Wang, Y., Yao, B., 2015.
A novel bifunctional GH51 exo-α-l-arabinofuranosidase/endo-xylanase from
Alicyclobacillus sp. A4 with significant biomass-degrading capacity. Biotechnol.
Biofuels 8 (1), 197.
Yang, W., Jiang, Z., Liu, L., Lin, Y., Wang, L., Zhou, S., 2017. The effect of pentosanase on
the solubilisation and degradation of arabinoxylan extracted from whole and refined
wheat flour. J. Sci. Food Agric. 97 (3), 1034–1041.
Yang, Y., Sun, J., Wu, J., Zhang, L., Du, L., Matsukawa, S., Xie, J., Wei, D., 2016.
Characterization of a Novel alpha-L-Arabinofuranosidase from Ruminococcus albus 7
and Rational Design for Its Thermostability. J. Agric. Food Chem. 64 (40),
7546–7554.
Yegin, S., Altinel, B., Tuluk, K., 2018. A novel extremophilic xylanase produced on wheat
bran from Aureobasidium pullulans NRRL Y-2311-1: Effects on dough rheology and
bread quality. Food Hydrocolloids 81, 389–397.
Zhang, J., Siika-Aho, M., Tenkanen, M., Viikari, L., 2011. The role of acetyl xylan esterase
in the solubilization of xylan and enzymatic hydrolysis of wheat straw and giant reed.
Biotechnol. Biofuels 4 (1), 60.
Zheng, F., Liu, J., Basit, A., Miao, T., Jiang, W., 2018. Insight to improve α-L-arabino-
furanosidase productivity in Pichia pastoris and its application on corn stover de-
gradation. Front. Microbiol. 9, 3016.
Zhou, J., Bao, L., Chang, L., Zhou, Y., Lu, H., 2012. Biochemical and kinetic character-
ization of GH43 β-d-xylosidase/α-l-arabinofuranosidase and GH30 α-l-arabinofur-
anosidase/β-d-xylosidase from rumen metagenome. J. Ind. Microbiol. Biotechnol. 39
(1), 143–152.
Zhou, T., Xue, Y., Zhang, Z., Dong, Y., Gao, R., Li, Y., 2019. Improvement of the
Characteristics of Steamed Bread by Supplementation of Recombinant alpha-L-ara-
binofuranosidase Containing xylan-binding domain. Food Biotechnol. 33 (1), 34–53.