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Fig 2. Protein Concentration During Fermentation at Different pH
3.2. Effect of pH and Temperature on xylanase activity
Experimental results of the inluence of various pHs and temperatures were shown in Fig.3. The
optimum activity of xylanase was observed at pH 7 and 70
o
C.
Fig 3. Effect of Temperature and pH of Xylanase Activity at Different pH
3.3. Effect of Temperature and pH on Enzyme Stability
Thermal and pH stability of the enzyme was observed in the absence of substrate at various
temperatures and pHs. The experimental results were shown in Fig 4 and Fig 5. Fig 4presentsthe relative
residual activity, whereas Fig 5 presentage actual activity of the enzyme. The enzyme was the most
stable at pH 7 and 60°C. After incubating the enzyme at pH 7 and 60°C for 105 minutes the remaining
activity was still 209 Uml 60.
Fig. 5. Stability of Xylanase Enzyme at Different pH and Temperature
Fig 6. Stability of Xylanase Enzyme at Various pHs and Temperatures
3.4.Electroforesis and Zymogram
Electrophoretic studies of the xylanase crude extracts using SDSPAGE and zymograms showed
the presence of multiple protein bands. The molecular weight of the enzyme was approximately 27.67 kDa.
3.5. Thin Layer Chromatography TLC
Thin Layer Chromatography was showed that the xylanase hydrolysed xylan into xylobiose, xylotriose,
xylotetraose and xylopentaose Fig. 7.
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Fig. 7. Thin Layer Chromatogram of Xylan Hydrolyzed by The XylanaseE-7 : Xylanase
Product at pH 7; E-9 : Xylanase Product at pH 9, S: Xylooligosaccharide Standards: X1: Xylose; X2:
Xylobiose; X3: Xylotriose; X4: Xylotetraose; X5: Xylopentaose; X6: Xylohexaose.
Discussion
Based on experimental data, B. halodurans CM1 produced extracellular thermophilic xylanolytic
enzyme. The optimal production of the enzyme was at 55
o
C, pH 10 after 21 hours fermentation with 100rpm agitation. The optimum activity of the enzyme was
observed at pH 7, 70
o
C. Some research has been reported that the activity of xylanolytic enzyme
ranging from pH 5-10, with the optimum temperature range 50-75
o
C depending on the species.The optimum activity of Bacillus sp. AQ-1 was at pH 7, 60
o
C [18], Bacillus licheniformis I-5 at pH 7, 50
o
C [26], Bacillus sp. NCIM 59 at pH 6 to 7.5, 25
o
C [16], Bacillusspat pH 7, 55
o
C [28], Bacillussp JB99 at pH 10, 45
o
C [13]. As shown in Fig. 3, the xylanolytic enzyme
produced by B. halodurans CM1 was optimally active at pH 7, 70
o
C. Similar results were observed that optimal temperature of xylanase produced by Bacillus
sp TAR-1 was at 75
o
C at pH 7 and 70
o
C at pH 9.0 [14]. Based on the experimental results xylanase used in this
study was a thermophilic xylanolytic enzyme which might have potential for pulp bio-bleaching
p
rocesses. This data is also supported by the stability of enzymes
which stable at pH 7 and temperature 60
o
C. The study shows that after incubating the enzyme at pH 7 and 60°C
for 105 minutes the remaining activity was still 209 Uml 60. The optimal temperature and stability of
this enzyme are comparable to thermostable xylanase produced extracellularly by Bacillus thermoleovoran
K-3d dan
B. lavothermus LB3A that has been reported stable for 2 hours at pH 7, temperature 70
o
C [20]. The other study reported that the xylanolitic enzyme
from thermophilic strain Clostridium absonum CFR- 702 was stable up to 60
o
C Xyl-I and 50
o
C Xyl- II [19].In biobleaching process using xylanase, the
most important parameters are enzyme stability at higher pH and temperature. Biobleaching process
using xylanase from Streptomyces thermoviolaceus was optimized and found to be most effective at
65
o
C and pH 6.0, with the crude enzyme preparation blended to a pulp concistency of 5 [9]. The study
of xylanase from Bacillus sp for biobleaching of kraft pulp has been evaluated at 60
o
C at pH 9.0.[1]. Thermoactivecellulase-free xylanase produced from
alkaliphilic Bacillus sp that was stable at 60
o
C for more than 60 minutes was having potential for bio-
bleachingprocess.[1]. Xylanase can also used for deinking process, conditions in a previous research,
the deinking reaction was allowed for 30 minutes at 11 consistensy, pH 7.0 and 50
o
C with continuous slow mixing [17].
The SDS-PAGE and zymogram analysis showed the molecular weight of xylanase was 26,7kDa. In
earlier reports showed that xylanase produced by Bacillus spp JB 99 was 16 kDa [13]. Xylanase which
were produced by Bacillus spp reported having molecular masses ranging between 3600-23,000
Daltons [12]. However, in some cases Bacillus sp with Fig 6. Zymogram and SDSPAGE XylanasefromB. Halodurans CMI.Zym: Zymogram; Marker: Low Molecular
Weight Marker of Protein; E-7: Band of Protein Sample at pH 7; E-9: Band of Protein Sample at pH 9
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high molecular mass of 24000 to 145,000 Dalton was also reported [13].
The products of hydrolysis beechwoodxylan was observed as xylobiose, xylotriose, xylotetraose and
xylopentaose which indicated endoxylanase activity. In this study, the enzyme did not produce xylose which
might be due to less incubationtime. In some study, xylose would be formed after 12 hours incubation,
since the initially xylanase cleaved the substrate to liberate xylooligosaccharides and then the resulting
oligosaccharides were probably cleaved to form xylose [11].
Based on this experimental stability data, the xylanase enzyme might be used in pulp and paper
manufacture as an agent for bio-bleaching. The process of lignin removal from chemical pulps to
produced bright or completely white inished pulp is called ‘bleaching’. The most important application of
xylanase enzymes is the prebleaching of kraft pulp. Currently, the most effective application of xylanase
is in prebleaching of kraft pulp to minimize the use
of harsh chemicals in the subsequent treatment stages of kraft pulp. While many applications of enzymes
in paper industries are still in the research and developmental stage, several applications have found
their way into the mills in unprecedented short period of time in the last decade [3]. Several criteria are
essential for choosing a microorganism to produced xylanases. The optimum temperature for xylanase
action ranges between 35 and 60
o
C. In the mills, xylanase pretreatment takes place in the brown stock
high-density storage tanks, in which pulp is present at high temperature approximately 60
o
C and at alkaline pH. Therefore, xylanases that are active and stable at
high temperature and alkaline pH are desirable [6]. Most of the beneicial effects of xylanase prebleaching
can be obtained after only 1-2 hours of treatment [5, 10].
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Optimization of TMP Newsprint Retention System for Retention and Drainage Improvement
Chul-Hun Jung
a
, Jong-Moon Park
b
a
Dept. Forest Product Engineering, Chungbuk National University, Naesudong-ro 52, Cheongju, Chungbuk 361-763, South Korea, 8terra8gmail.com
b
Dept. Forest Product Engineering, Chungbuk National University, Naesudong-ro 52, Cheongju, Chungbuk 361-763, South Korea, jmparkcbu.ac.kr
ABSTRACT
Newsprint stock uses TMP which has wood resin, dissolved and colloidal substances DCS, so they degrade machine runnability, sheet properties, and chemical eficiency. We compared and analyzed two retention
systems, such as PAMBentonite micro particle system and PEOcofactor system. As the TMP ratio rose, PEO cofactor system was more eficient in retention and drainage than the other system. High molecular weight and
non-ionic polymer retention system had less effect on locculation interruption than the traditional electrostatic retention system.
Keywords: newsprint stock, TMP, maicroparticle, PEO, cofactor, retention, drainage
1. Introduction
