• Journal of the Korean Wood Science and Technology
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• 제20권3호
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• pp.67-78
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• 1992

The application of white-rot fungi to pulping and bleaching processes has been studied at the Wood Chemistry Laboratory in Kyushu University, cooperatively with the Biotechnology Laboratory of Kobe Steel, Ltd. Some successful results of the studies for a biomechanical pulping process, biobleaching of hardwood and softwood kraft pulp, as well as chlorine free biobleaching of oxygen-prebleached hardwood kraft pulp are dealt with. Biological treatment of the pulp bleaching effluent is also described.

• Journal of the Korean Wood Science and Technology
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• 제32권3호
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• pp.52-58
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• 2004

The use of N-hydroxyphthalimide (NHPI) as a mediator for laccase has proven to be comparable to N-hydroxybenzotriazole (HBT) for the delignification of kraft pulp, and the transformation of a number of industrial dyes. The advantages of NHPI derivatives are the biodegradation of these compounds compared to HBT, which has been shown to be recalcitrant in the environment, and the more reasonable cost of synthetic process.

• Journal of the Korean Wood Science and Technology
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• 제35권6호
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• pp.166-174
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• 2007

효소 전처리에 의한 침엽수 Kraft 펄프의 표백 효율을 살펴보고자 Trichoderma reesei에서 2종류의 xylanase를 cloning하였다. Cloning된 xylanasel과 xylanaseII 유전자를 E. coli에서 발현시켜 29kDa의 재조합 단백질인 XYNI과 29.5 kDa의 XYNII를 생한하여 정제하였고, Rumicoccus albus로부터 cloning되어 변형된 50kDa의 EGIV-CBDII를 동일한 방법으로 cellulase를 생산 정제하였다. 정제된 효소들을 각각 xylan과 CMC를 기질로하여 효소 활성도를 측정한 결과 XYNI이 XYNII보다 효소활성이 더 높았으며, EFIV-CBDII 또한 높은 활성을 나타내었다. 이렇게 생산 정제된 xylanase와 cellulase를 이용하여 침엽수 Kraft 펄프를 대상으로 연속처리에 의한 biobleaching test를 실시하였다. XYNI을 처리하였을 때 백색도는 29.9% 증가하였고, XYNI를 1차 처리한 후 2차로 EGIV-CBDII를 처리하였을 때 백색도는 1차 처리구에 비해 9.1% 증가하였다. 그 후 3차 XYNI를 처리 하였을 때 2차에 비해 7.3%의 bleaching의 효과를 볼 수 있었다. 또한 Kappa No.는 처리단계에 따라 8.1, 4.6 그리고 3.2% 각각 지속적으로 감소하였다. 이 실험결과를 통해 재조합 효소 XYNI과 EGIV-CBDII를 함께 이용하여 Karft 펄프에 대한 biobleaching의 상승효과를 확인하였다.

• Journal of Microbiology and Biotechnology
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• 제12권1호
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• pp.153-156
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• 2002

The characterization of a partially purified, cellulase-free, thermostable alkaline xylanase from thermoalkalophilic Bacillus sp. JB-99 was investigated. The xylanase production was the highest when birchwood xylan was added to a medium containing finely powdered rice bran, showing 4,826 IU$ml^-1$ of activity for 15 h of incubation. The partially purified xylanase exhibited an optimum temperature and pH at $70^C{\circ}$ and 10, respectively. The enzyme was stable at pH 5-11 at $50^C{\circ}$. The xylanase activity was strongly inhibited by $Hg^2+$, while dithiothreitol, cysteine, and ${\beta}$-mercaptoethanol enhanced the activity.

• Journal of Microbiology and Biotechnology
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• 제22권12호
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• pp.1636-1643
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• 2012

Enzymatic pre-bleaching by modification of pulp fibers with xylanases is an attractive approach to reduce the consumption of toxic bleaching chemicals in the paper industry. In this study, an alkaliphilic endoxylanase gene was isolated from metagenomic DNA of a structurally stable thermophilic lignocellulose-degrading microbial consortium using amplification with conserved glycosyl hydrolase family 10 primers and subsequent genome walking. The full-length xylanase showed 78% sequence identity to an endo-${\beta}$-1,4-xylanase of Clostridium phytofermentans and was expressed in a mature form with an N-terminal His6 tag fusion in Escherichia coli. The recombinant xylanase Xyn3F was thermotolerant and alkaliphilic, working optimally at $65-70^{\circ}C$ with an optimal pH at 9-10 and retaining >80% activity at pH 9, $60^{\circ}C$ for 1 h. Xyn3F showed a $V_{max}$ of 2,327 IU/mg and $K_m$ of 3.5 mg/ml on birchwood xylan. Pre-bleaching of industrial eucalyptus pulp with no prior pH adjustment (pH 9) using Xyn3F at 50 IU/g dried pulp led to 4.5-5.1% increase in final pulp brightness and 90.4-102.4% increase in whiteness after a single-step hypochlorite bleaching over the untreated pulp, which allowed at least 20% decrease in hypochlorite consumption to achieve the same final bleaching indices. The alkaliphilic xylanase is promising for application in an environmentally friendly bleaching step of kraft and soda pulps with no requirement for pH adjustment, leading to improved economic feasibility of the process.

• Journal of Microbiology
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• 제44권3호
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• pp.263-268
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• 2006

Pleurotus ostreatus is a widely cultivated white-rot fungus. Owing to its considerable enzymatic versatility p. ostreatus has become the focus of increasing attention for its possible utility in biobleaching and bioremediation applications. Interactions between microorganisms can be an important factor in those processes. In this study, we describe the presence of a bacterial species associated with P. ostreatus strain G2. This bacterial species grew slowly (approximately 30 days) in the liquid and semi-solid media tested. When p. ostreatus was inoculated in solid media containing Tween 80 or Tween 20, bacterial microcolonies were detected proximal to the fungal colonies, and the relevant bacterium was identified via the analysis of a partial 16S rDNA sequence; it was determined to belong to the Burkholderia cepacia complex, but was not closely related to other fungus-isolated Burkholderiaceae. New specific primers were designed, and confirmed the presence of in vitro P. ostreatus cultures. This is the first time that a bacterial species belonging to the B. cepacia complex has been found associated with P. ostreatus.

• Journal of the Korean Wood Science and Technology
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• 제35권2호
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• pp.85-95
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• 2007

Manganese dependent peroxidase (MnP) is the most ubiquitous enzyme produced by white-rot fungi, MnP is known to be involved in lignin degradation, biobleaching and oxidation of hazardous organopollutants. Bjerkandera fumosa is a nitrogen-unregulated white-rot fungus, which produces high amounts of MnP in the excess of N-nutrients due to increased biomass yield. The objective of this study was to optimize the MnP production in N-sufficient cultures by varying different physiological factors such as Mn concentration, culture pH, and incubation temperature. The growth of fungus was optimal in pH 4.5 at $30^{\circ}C$, $N_2$-unregulated white-rot fungus produces high amounts of MnP in the excess N-nutrients. The fungus produced the highest level of MnP (up to $1000U/{\ell}$) with $0.25g/{\ell}$ asparagine and $1g/{\ell}$ $NH_4Cl$ as N source at 1.5 mM $MnCl_2$ concentration, pH value of 4.5 at $30^{\circ}C$. Purification of MnP revealed the existence of two isoforms: MnPl and MnP2. The molecular masses of the purified MnPl and MnP2 were in the same range of 42~45 kDa. These isoforms of B. fumosa strictly require Mn to oxidize phenolic substrates. Concerned to kinetic constants of B. fumosa MnPs, B. fumosa has similar Km value and Vmax compared to the other white-rot fungi.

• Journal of Microbiology and Biotechnology
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• 제22권4호
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• pp.462-469
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• 2012

A metagenomic fosmid library was constructed from genomic DNA isolated from the microbial community residing in hindguts of a wood-feeding higher termite (Microcerotermes sp.) collected in Thailand. The library was screened for clones expressing lignocellulolytic activities. Fourteen independent active clones (2 cellulases and 12 xylanases) were obtained by functional screening at pH 10.0. Analysis of shotgun-cloning and pyrosequencing data revealed six ORFs, which shared less than 59% identity and 73% similarity of their amino acid sequences with known cellulases and xylanases. Conserved domain analysis of these ORFs revealed a cellulase belonging to the glycoside hydrolase family 5, whereas the other five xylanases showed significant identity to diverse families including families 8, 10, and 11. Interestingly, one fosmid clone was isolated carrying three contiguous xylanase genes that may comprise a xylanosome operon. The enzymes with the highest activities at alkaline pH from the initial activity screening were characterized biochemically. These enzymes showed a broad range of enzyme activities from pH 5.0 to 10.0, with pH optimal of 8.0 retaining more than 70% of their respective activities at pH 9.0. The optimal temperatures of these enzymes ranged from $50^{\circ}C$ to $55^{\circ}C$. This study provides evidence for the diversity and function of lignocellulose-degrading enzymes in the termite gut microbial community, which could be of potential use for industrial processes such as pulp biobleaching and denim biostoning.

• Journal of Microbiology and Biotechnology
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• 제21권8호
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• pp.861-868
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• 2011

A xylanase gene, xyn7c, was cloned from Paenibacillus sp. 12-11, an alkalophilic strain isolated from the alkaline wastewater sludge of a paper mill, and expressed in Escherichia coli. The full-length gene consists of 1,296 bp and encodes a mature protein of 400 residues (excluding the putative signal peptide) that belongs to the glycoside hydrolase family 10. The optimal pH of the purified recombinant XYN7C was found to be 8.0, and the enzyme had good pH adaptability at 6.5-8.5 and stability over a broad pH range of 5.0-11.0. XYN7C exhibited maximum activity at $55^{\circ}C$ and was thermostable at $50^{\circ}C$ and below. Using wheat arabinoxylan as the substrate, XYN7C had a high specific activity of 1,886 U/mg, and the apparent $K_m$ and $V_{max}$ values were 1.18 mg/ml and 1,961 ${\mu}mol$/mg/min, respectively. XYN7C also had substrate specificity towards various xylans, and was highly resistant to neutral proteases. The main hydrolysis products of xylans were xylose and xylobiose. These properties make XYN7C a promising candidate to be used in biobleaching, baking, and cotton scouring processes.