Enzyme and Microbial Technology 41 (2007) 733739

Enhanced production of cellulase-free thermostable xylanase by

Bacillus pumilus ASH and its potential application in paper industry
Bindu Battan a , Jitender Sharma a, , Saurabh Sudha Dhiman a , Ramesh Chander Kuhad b
aDepartment of Biotechnology, Kurukshetra University, Kurukshetra 136119, India
b Department of Microbiology, University of Delhi South Campus, New Delhi 110021, India
Received 21 February 2007; received in revised form 15 June 2007; accepted 15 June 2007

Abstract
A very high level of cellulase-free, thermostable xylanase has been produced from newly isolated strain of Bacillus pumilus under submerged
fermentation in a basal medium supplemented with wheat bran (2%, w/v) pH 8.0 and at 37 C. After optimization of various production parameters,
an increase of nearly 13-fold in xylanase production (5407 IU/ml) was achieved. The produced xylanase is stable in neutral to alkaline pH region
at 70 C. The suitability of this xylanase for use in the bioleaching of eucalyptus Kraft pulp was investigated. A xylanase dose of 5 IU/g of oven
dried pulp of 10% consistency exhibited the optimum bleach boosting of the pulp at pH 7.0 and 60 C after 180 min of treatment. An increase of
5% in brightness along with an increase of 21% and 28% in whiteness and fluorescence respectively, whereas 18% decrease in the yellowness of
the biotreated pulp was observed. Enzyme treated pulp when subjected to chemical bleaching, resulted in 20% reduction in chlorine consumption
and up to 10% reduction in consumption of chlorine dioxide. Also a reduction of about 16% in kappa number and 83% in permanganate number,
along with a reduction in COD value and significant improvement in various pulp properties, viz. viscosity, tensile strength, breaking length, burst
factor, burstness, tear factor and tearness were observed in comparison to the conventional chemical bleaching.
2007 Elsevier Inc. All rights reserved.

Keywords: Thermostable; Xylanase; Paper and pulp; Biobleaching

1. Introduction In the process of paper production, pulping is a step in which

Xylanases (endo-1,4--d-xylanohydrolase; EC 3.2.1.8), the
xylan degrading enzymes, have been reported mainly from bac-
teria [13], fungi [2], actinomycetes [4] and yeast [5,6]. The
main reason for investigating its production by bacteria and
fungi is its wide variety of biotechnological applications such
as bleaching of paper pulp, increasing the brightness of pulp,
in food processing, poultry feeds, degumming of plant fibers,
etc. [7]. Xylanases have been widely reported to improve the
effectiveness of conventional bleaching chemicals in removing
lignin from hardwood and softwood kraft pulps [810]. Since
the last two decades the bleaching of pulp has become an issue
of great concern primarily because of the environmental haz-
ards caused by the release of the adsorbable organic halogens
and due to increasing public awareness. This increased envi-
ronmental awareness has become a significant driving force for
imposing strict regulations.
Corresponding author. Tel.: +91 1744 239239.
E-mail address: jksharmakuk@rediffmail.com (J. Sharma).
fibres are broken apart and most of the lignin is removed. The
residual lignin is then removed by a multistep bleaching pro-
cess [11]. Chemical pulping which is frequently used around
the world is mainly achieved by sulphate (kraft) process. The
kraft process consists of cooking the wood chips at high tem-
peratures and at high alkaline pH, in which part of the xylan is
dissolved in the pulping liquor; while short-chain xylan precipi-
tates in a more or less crystalline form on the surface of cellulose
microfibrils. This process removes around 95% of the total lignin
but the 5% residual lignin gives paper a brownish colour and so
the paper needs to be bleached [12]. The chemicals used for
bleaching process have resulted in the release of large amounts
of chlorinated organic compounds that are known to have toxic,
mutagenic and carcinogenic effects. In response to government
and environmental protection groups, paper industries are cur-
rently changing practices to reduce organic pollutant loads on the
environment [1315]. Thus, the use of enzymes for biobleach-
ing has provided an alternative and cost-effective method for
process change.
The use of microbial enzymes for bleaching of paper and
pulp is now gaining momentum. The microbial enzymes have

0141-0229/$ see front matter 2007 Elsevier Inc. All rights reserved.
doi:10.1016/j.enzmictec.2007.06.006
734 B. Battan et al. / Enzyme and Microbial Technology 41 (2007) 733739
high specificity for their substrates, the reaction conditions are
mild and there is no substrate loss due to chemical modifications.
Several criteria are essential for choosing the suitable xylanase
producing microorganism for biobleaching of pulp. It is essen-
tial that the enzyme should be active at neutral and alkaline pH
and should be thermostable. Interest in cellulase-free xylanase
has been of more recent origin following the concept of utiliz-
ing xylanase in pulp and paper industry to produce rayon grade
paper pulps or superior quality dissolving pulps [16]. Any cel-
lulase activity will have serious economic implications in terms
of cellulose loss, degraded pulp quality, and increased effluent
treatment cost.
Not much literature has been reported on the high-level pro-
duction and process economy of utilizing xylanolytic enzymes.
Wide scale industrial applications of xylanase require their cost-
effective production to make the process economically viable.
Thus, the aim of our studies was to reduce the cost of xylanase
production by optimizing the fermentation medium and to study
its application in the pulp and paper industry. In the present inves-
tigation, we report very high level production of a thermostable
and cellulase-free xylanase using agro-residues in submerged
fermentation from a newly isolated strain of Bacillus pumilus
that has been potentially applied in the selective hydrolysis of
the hemicellulose component in the pulp and paper industry.
2. Materials and methods
2.1. Microorganism
B. pumilus was isolated from a soil sample collected from sanitary landfill
using xylan agar medium (pH 7.0) at a temperature of 37 C. Its ability to pro-
duce xylanase was qualitatively confirmed when it formed transparent digestion
halos on birchwood xylan agar plates on treatment with Congo red followed
by washing with 1 M NaCl. The organism was identified as B. pumilus ASH
from the Institute of Microbial Technology (IMTECH), Chandigarh, India on
the basis of its morphological, physiological and biochemical characteristics. It
has been deposited at MTCC (Microbial Type Culture Collection), Chandigarh,
India and has been given accession number 7411. The culture was maintained
and stored at 4 C on nutrient agar medium
2.2. Xylanase production under submerged fermentation
The enzyme production was studied in Erlenmeyer flask (250 ml) contain-
ing 50 ml of basal medium having (g/l): yeast extract, 5.0; MgSO4 7H2 O, 0.2;
KH2 PO4 , 1.0 supplemented with wheat bran, 5.0 and peptone, 5.0; pH 8.0. The
flasks were inoculated with 2% (O.D. 0.5) of the overnight grown inoculum
and incubated at 37 C under shaking conditions (200 rpm). The enzyme was
harvested by centrifuging at 10,000 g for 15 min. The supernatant was treated
as enzyme and was assayed for xylanase activity. The optimization studies were
also performed by altering the fermentation conditions and composition of this
basal medium under optimum shaking conditions (200 rpm).
2.3. Xylanase assay
Birchwood xylan and carboxymethyl cellulose were used as the assay sub-
strates for xylanase and cellulase, respectively. The reaction mixture for each
enzyme assay contained 490 l of 2% of respective substrates prepared in phos-
phate buffer, pH 7.0 and 10 l of appropriately diluted enzyme and was incubated
at 60 C for 10 min. Both undiluted enzyme and a series of dilutions were assayed
against carboxymethyl cellulose for cellulase activity at different pH values rang-
ing from pH 6 to 9. The enzyme activity was determined by measuring the release
of reducing sugars during the enzyme substrate reaction using Millers method
[17]. One unit of enzyme activity was defined as the amount of enzyme that cat-
alyzes the release of 1 mol of reducing sugars from the respective substrates
in one min under the standard assay conditions.
All the experiments were carried out independently in triplicates and the
results presented here are the mean of the three.
2.4. Chemicals and pulp samples
Birchwood xylan, carboxymethyl cellulose, 3,5-dinitrosalicylic acid were
purchased from Sigma Chemical Co. (St. Louis, MO, USA). All other chemicals
used were of highest purity available commercially. Pulp used for biobleach-
ing was hardwood pulp obtained from Ballarpur Industries Limited (BILT),
Yamunanagar, Haryana, India. Agro residues were procured locally.
2.5. Parametric optimization of xylanase production
The optimization for xylanase production by B. pumilus was carried out for
the following parameters:
1. Incubation period: submerged fermentation (SmF) was carried out for
1830 h.
2. Inoculum age and size: a 1224-old culture at a level of 14% was used as
inoculum.
3. Temperature: xylanase production at temperature ranging from 30 to 55 C
was studied.
4. pH: basal medium of pH ranging from 5 to 10 was used for xylanase pro-
duction.
5. Carbon source and the concentration of the selected carbon source: various
carbohydrates and agricultural residues/byproducts were tested as sole car-
bon source for xylanase production and the effect of different concentrations
of the selected carbon source on xylanase production was studied.
6. Effect of nitrogen source and the concentration of the selected nitrogen source
on xylanase production: production medium devoid of any nitrogen source
was supplemented with different inorganic and organic nitrogen sources like
peptone and yeast extract and the effect of various concentrations of the
selected nitrogen source on xylanase production was studied.
7. Effect of various surfactants, detergents and other additives: different addi-
tives like EDTA and SDS at a concentration of 0.2 (w/v) and Tween 80, Tween
20, glycerol, Triton X, olive oil and oleic acid at a concentration of 0.2 (v/v)
were supplemented in the medium and the effect of various concentrations
of the selected additive was studied.
After optimization of these parameters, xylanase production was carried out
under optimized nutritional and fermentation conditions for maximum yield of
the enzyme to be applied in pulp and paper industry.
2.6. Optimization of enzyme dose and other reaction parameters
for biobleaching
The optimization of enzyme dose and retention time for biobleaching
was carried out by treating moistened unbleached pulp with varying doses of
xylanase, ranging between 0 and 12.5 IU/g for variable time intervals starting
from 30 min to 3 h. The optimum pH and temperature for efficient biobleaching
were determined by carrying out the enzymatic treatment at 10% (w/v) pulp
consistency in transparent plastic bags under different pH values ranging from
6 to 10 and at different temperatures varying from 55 to 70 C.
2.7. Biobleaching of kraft pulps (ECDED1 D2 )
Pulp used for bleaching was a mixture obtained from woods of six differ-
ent trees, namely eucalyptus, poplar, eucalyptus rulla, small vaneer, bamboo,
and debarka bamboo hardwood (DBH). Fifty grams extensively washed oven
dried kraft pulp of 10% consistency was treated with xylanase (5 IU/g) in plas-
tic bags under optimum conditions. Then xylanase treated pulp was exposed to
CDED1 D2 bleaching sequence to obtain pulp with different properties. There-
after, various experiments were conducted to measure the reduction in chlorine
 B. Battan et al. / Enzyme and Microbial Technology 41 (2007) 733739
consumption to obtain same amount of brightness in both xylanase-treated and
control (untreated). In these experiments, all the parameters and treatment con-
ditions were kept the same except for the dose of chlorine treatment given at the
chlorine and chlorine dioxide (CD) stage.
2.8. Physical and chemical characterization of kraft pulps
The enzyme treated pulp was thoroughly washed and small sheets were pre-
pared according to TAPPI standard methods [18]. Other pulp properties, viz.
kappa number; estimation of lignin content measured by reaction of pretreated
pulp with acidified potassium permanganate (TAPPI method T 236 cm-85),
brightness(T 452 om-87) of the handsheets was measured as %ISO (Interna-
tional Organization of Standardization) with ISO Colourtech, USA, measured
at 457 nm, P. no. (permanganate no.); estimated by reaction of pretreated pulp
with acidified permanganate solution by using starch and potassium iodide as
an indicator, yellowness, whiteness and fluorescence of pretreated pulp was
measured by ISO Colourtech, USA at 457 nm (T 1216), viscosity; measure of
cellulose chain length estimated by dissolving delignified pulp in cupriethylene-
diamine (CED) through Ostwald viscometer by taking 0.5% of the solution
(TAPPI method T 230 om-94), burst factor (T 403 cm-50), burstness (T 403), ten-
sile strength (T 231 cm-96), breaking length, tear factor, tearness, total chlorine
dioxide consumed and the chemical oxygen demand (COD) of the biobleached
effluent were also determined according to the standard protocols of TAPPI,
Atlanta. Chemical oxygen demand is used as a measure of the oxygen require-
ment of a sample that is susceptible to oxidation by strong chemical oxidants.
Total organic chlorine (TOCl) consumed was determined by the method of Hong
et al. [19].
3. Results and discussion
3.1. Microbial strain and its growth conditions
The isolated bacterial strain is a Gram positive, moderate
thermophile with minimum, optimum and maximum tempera-
ture for growth at 10, 37 and 55 C, respectively. The strain is an
alkalophile being capable of growing at pH values up to 11.0.
3.2. Xylanase production under submerged fermentation
The enzyme profile revealed that the bacterium produced
xylanase and no cellulase activity could be detected. The extra-
cellular levels of xylanase were monitored from 18 to 30 h in
agitated cultures of the bacterium grown in the basal medium.
The enzyme production started after 18 h of incubation and
the production peaked at 26 h (413 IU/ml) and declined there-
after. The time of incubation depends on the growth rate of the
microorganism and its enzyme production pattern. The xylanase
titer was found to increase when the age of inoculum was
increased from 12 to 18 h (436 IU/ml) and increased further
when the inoculum size increased from 1% to 2.5% (526 IU/ml)
and decreased with further increase in inoculum size. Higher
concentration of inoculum is not preferable in industrial fer-
mentations [20].
Optimum incubation temperature for enzyme production was
found to be 37 C (951 IU/ml), which is the optimum growth
temperature of the bacterium while no xylanase production
was observed above 55 C. The bacterium did not produce any
xylanase when grown in medium of pH 4.0 and xylanase pro-
duction started at pH 5.0 peaking in a medium with pH 8.0
(966 IU/ml). Xylanase production by various bacteria and fungi
has been shown to be markedly dependent on pH [2123].
735
Fig. 1. Effect of various carbon sources on xylanase production.
Acidic pH (4.06.0) generally favours fungal xylanases [24]
while higher pH favours bacterial xylanases [25].
Oat spelt xylan and different agricultural residues/byproducts
such as wheat bran, wheat straw, paddy straw, soya bran, bagasse
as well as various carbohydrates were used as sole carbon source
for xylanase production by B. pumilus ASH and the results are
shown in Fig. 1. Among these sources, maximum xylanase titer
was attained in the medium supplemented with oat spelt xylan
closely followed by wheat bran. As the use of purified xylan as
substrate is uneconomical for industrial use while agricultural
residues offer cost-effective alternatives for xylanase produc-
tion therefore, wheat bran was used as the sole carbon source
for further studies. The higher xylanase synthesis in the pres-
ence of wheat bran could be the result of synergistic action
of other hemicellulolytic enzymes secreted by the bacterium
in the presence of these substrates [24,26]. Also, wheat bran has
very low lignin content and contains 28% protein, thus serving
both as a carbon and a nitrogen source [27]. Xylanase activity
increased with the increase in concentration of wheat bran from
0.25% to 2% (w/v) and it declined considerably with further
increase in wheat bran concentration (Fig. 2). This may be due
to the formation of a thick suspension of the substrate which
in turn did not mix freely in shake flasks. Similar situations
have been encountered by others while using high concen-
trations of lignocellulosic as substrate for enzyme production
[28].
Among the various inorganic and organic nitrogen sources
tested (Fig. 3), a combination of peptone, yeast extract and potas-
sium nitrate stimulated the highest xylanase production. Organic
nitrogen sources have been found stimulating the xylanase pro-
duction in Bacillus species [29,30]. Use of peptone as the best
organic nitrogen source and potassium nitrate as the best inor-
ganic nitrogen source in the medium for xylanase production
has been reported earlier [31,32]. The xylanase production was
maximum at a concentration of 0.75% (w/v) of this combina-
tion, i.e. 0.25% (w/v) of each component (Fig. 4) and beyond
this concentration; a slight decrease in the enzyme production
736 B. Battan et al. / Enzyme and Microbial Technology 41 (2007) 733739
Fig. 2. Effect of concentration of optimized carbon source.
was observed. This may be due to the fact that a high concen-
tration of peptone acts as a good inducer for proteases leading
to the increased degradation of other enzymes [33].
The effect of various additives such as metalloenzyme
inhibitor, surfactants and fatty acids on xylanase production
was investigated. Among the additives used, Tween 20, SDS
and EDTA inhibited xylanase production strongly whereas the
maximum increase in xylanase production was observed in the
presence of olive oil at a concentration of 0.2% (v/v) (Fig. 5).
Similar observations have been made by Singh et al. [34].
Such compounds probably increase the permeability of the cell
membrane and cause rapid secretion of the enzymes. After opti-
mization of various production parameters, an increase of nearly
13 fold in xylanase production (5407 IU/ml) was achieved. Thus,
Fig. 3. Effect of various nitrogen sources on xylanase production. P, peptone;
Y.E., yeast extract.
Fig. 4. Effect of concentration of selected nitrogen sources on xylanase produc-
tion.
xylanase was produced in optimized media under optimized
conditions for effective application in the pulp industry.
3.3. Optimization of various parameters for enzyme
controlled treatment
The xylanase produced by B. pumilus is completely cellulase-
free as no cellulase activity could be detected at various pH
values and at different enzyme dilutions. This prompted us
to test the suitability of this xylanase in the pulp and paper
industry. To obtain best results from enzymatic use, enzyme
dosage, retention time and other conditions such as pH and
temperature must be optimized in each case to obtain effective
dispersion of enzyme [11]. Different parameters were optimized
for obtaining the same %ISO brightness as it was achieved in
chemical bleaching. Enzyme extracted from B. pumilus showed
stability over a broad range of pH from 6 to 10 and tempera-
ture from 55 to 70 C. A xylanase comprising two components
active over a pH range of 610 and temperature range of
5060 C has been characterized from an alkalothermophilic
Fig. 5. Effect of various additives on xylanase production.
 B. Battan et al. / Enzyme and Microbial Technology 41 (2007) 733739 737

Table 1
Effect of different enzyme dose on pulp biobleaching
Enzyme dose (IU/ml) Brightness ISO (%) P. no. K. no. Yellowness Whiteness Fluorescence

05 31.70 11.8 18.0 13.19 36.24 0.12

7.5 31.78 11.8 18.0 13.20 36.49 0.13
10.0 30.94 11.9 18.0 13.30 39.07 0.15
12.5 30.47 12.1 18.3 13.44 39.80 0.13
Nil 30.2 13.1 19.9

These experiments were performed at pH 7.0, temperature 60 C, at retention time of 180 min and at pulp consistency of 10%.

Table 2
Effect of different retention time on pulp biobleaching
Retention time (min) Brightness ISO (%) P. no. K. no. Yellowness Whiteness Fluorescence

60 29.73 13.5 20.5 13.62 40.41 0.19

120 30.1 12.2 18.5 13.44 38.22 0.10
150 30.65 12.2 18.5 13.36 37.96 0.27
180 31.68 11.9 18.0 13.50 36.23 0.13

All these experiments were performed at constant enzyme dose of 5 IU/ml and at 10% pulp consistency, at pH 7.0 and at temperature 60 C.

Table 3
Effect of different temperatures on pulp biobleaching
Temperature ( C) Brightness ISO (%) P. no. K. no. Yellowness Whiteness Fluorescence

70 29.81 13.3 20.2 13.34 38.93 0.04

60 31.68 11.9 18.0 13.50 36.23 0.13
55 29.97 12.9 19.6 13.45 39.69 0.15

All these experiments were performed at constant enzyme dose of 5 IU/ml and at 10% pulp consistency, at pH 7.0 and at retention time of 180 min.

(AT) Bacillus strain by Dey et al. [35]. After optimization of
various parameters, the optimum enzyme dosage required for
biobleaching by our enzyme was found to be 5 IU/ml at a reten-
tion time of 180 min (Tables 1 and 2). Higher enzyme dose
or longer periods of incubation did not enhance the extent of
biobleaching benefits significantly. The maximum efficiency of
the enzyme in biobleaching was obtained at 60 C and at pH
7.0 (Tables 3 and 4). The optimized conditions for enzymatic
hydrolysis of unbleached kraft pulps are apparently a prerequi-
site for the maximum removal of pentosans and improvement
of other pulp characteristics.
3.4. Biobleaching of kraft pulp
The biobleaching efficiency of cellulase-free xylanase from
B. pumilus was maximum after 180 min at 60 C at pH 7.0 using
a xylanase dose of 5 IU/ml at 10% pulp consistency with 11%
and 7.6% reduction in kappa number and permanganate number
respectively. An increase of 5% in brightness was observed in
case of unbleached pulp after treatment with xylanase enzyme.
Jiang et al. reported the reduction of pulp kappa number by
1.1 point and enhancement in pulp brightness by 5.5 (%ISO)
after treatment of wheat straw pulp with recombinant XynB
at 10 U/g [36]. The biobleaching effects of xylanase treatment
were observed on unbleached kraft pulp at each stage of chem-
ical bleaching. In the ECDED1 D2 process, nearly 47% increase
in brightness by xylanase was achieved after chlorination, 36.2%
after alkali treatment, 33% after D1 stage, 3% after D2 and only
1% after the final step which clearly indicates that the biobleach-
ing effect of xylanase was maximum at the first stage where
the maximum reduction in kappa number and P. no. (80%) was
achieved that was significantly more as compared to the other
stages of either biobleaching or chemical bleaching (Table 5).
Therefore, prebleaching by xylanase can be established as the
most suitable step to facilitate bleach boosting of pulp. The
application of xylanases for improvement in pulp bleaching has
been reported by several workers by using xylanase from Strep-
tomyces sp. QG-11-3 [37], Streptomyces cuspidosporus [38],

Table 4
Effect of different pH on pulp biobleaching
pH Brightness ISO (%) P. no. K. no. Yellowness Whiteness Fluorescence

7 31.68 11.9 18.0 13.50 36.23 0.13

8 30.7 12.1 18.3 13.20 39.96 0.14
9 29.4 13.6 20.6 13.90 39.01 0.17
10 29.93 12.9 19.6 13.67 40.41 0.17

All these experiments were performed at constant enzyme dose of 5 IU/ml and at 10% pulp consistency, at temperature 60 C and at retention time of 180 min.
738 B. Battan et al. / Enzyme and Microbial Technology 41 (2007) 733739

Table 5 Table 6
Effect of enzymatic treatment on various pulp properties Comparison of various physical properties of biobleached pulp with chemically
treated pulp
Parameters Control Enzyme
Pulp properties Control Enzyme
100% 100% 90% 80% 70%
Tensile strength (N) 28.4 37.4
Kappa no. 18.2 17.0 17.0 17.0 17.0 Breaking length (m) 31.97 38.70
Cl2 added (%) 4.38 4.38 3.9 3.5 3.1 Burst Factor (lb) 20.1 22.8
Cl2 consumed (%) 4.184 4.183 3.767 3.379 3.054 Burstness (lb) 16 20
Brightness ISO (%) 45.3 48.5 48.0 46.6 45.0 Tear factor (mN) 62 66.4
Alkali added (%) 2.5 2.5 2.5 2.5 2.5 Tearness (mN) 180 207
H2 O2 (%) 0.5 0.5 0.5 0.5 0.5
P. no. 2.4 1.8 1.9 2.2 2.6 These experiments were performed at pH 7.0, 60 C, enzyme dose of 5 IU/ml
Brightness ISO (%) 62.4 64.9 64.2 63.5 62.2 and at pulp consistency of 10%.
D-1 (ClO2 added) (%) 0.9 0.9 0.8 0.7 0.6
ClO2 consumed (%) 0.790 0.489 0.588 0.670 0.787
Brightness ISO (%) 84.1 86.3 84.7 84.4 83.6 the COD% value of the effluent upto 10.62% as compared to
D-2 (ClO2 added) (%) 0.5 0.5 0.4 0.4 0.3 the control. The reduction in COD value is highly appreciable
ClO2 consumed (%) 0.396 0.199 0.289 0.290 0.376
in terms of reduction in environment pollution which is caused
Brightness ISO (%) 86.8 87.7 87.3 87.0 86.1
SO2 added (%) 0.50 0.50 0.50 0.50 0.50 due to the release of the waste effluents from paper industry into
Brightness ISO (%) 87.7 88.6 88.1 87.8 87.2 the surroundings.
Realistic cost estimates and improvement in process eco-
These experiments on bleaching of pretreated pulp were carried out by
ECDED1 D2 process at pH 7.0, temperature 60 C, enzyme dose of 5 IU/ml nomics are the key factors in the commercial success of any
and at 10 CY%. technology and therefore we have attempted to reduce down the
cost of enzyme production by B. pumilus ASH which produces
very high level of xylanase under submerged fermentation in
Bacillus licheniformis 77-2 [12], Chaetomium cellulolyticum a very cost-effective media. The characteristics of enzyme are
[39], Streptomyces sp. strain S38 [40]. well suited for biobleaching of pulp and the enzymatic cocktail
Enzyme treated pulp when subjected to CDED1 D2 steps, is practically cellulase-free to be used as bleach boosting agent
20% reduction in chlorine consumption and upto 10% reduction thereby decreasing the bleaching chemical requirements, thus
in consumption of chlorine dioxide was observed for obtaining can be potentially applied in the pulp and paper industries.
the same %ISO brightness (Table 5). This makes the process
not only economically feasible but also eco-friendly. A chlorine
Acknowledgements
savings in the range of 3035% and 1240% using cellulase-
free xylanase from S. thermoviolaceus [41] and commercial
The authors gratefully acknowledge Ballarpur Industries
xylanase (Ecopulp, Cartazyme NS-10 and Pulpzyme HC) [42],
Limited (BILT), Yamunanagar, Haryana, India for provid-
respectively, have been reported earlier. Beg et al. [37] reported
ing kraft pulp and laboratory facilities. Bindu Battan greatly
a reduction of 8% chlorine by using xylanase from Streptomyces
acknowledges the financial assistance from Council of Scientific
sp. QG-11-3. In this study about 16% reduction in kappa number
and Industrial Research, India in the form of Senior Research
and 83% reduction in permanganate no. with enzymatic treat-
Fellowship during the course of investigation and Saurabh Sudha
ment were observed in comparison to the conventional chemical
Dhiman wishes to thank Kurukshetra University, Kurukshetra
bleaching. Also an increase of 21% and 28% in whiteness and
for University Research Scholarship.
fluorescence and a decrease of 18% in the yellowness of the
biotreated pulp were also observed.
Pretreatment of pulp with xylanase and its subsequent treat- References
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