• Mycobiology
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• 제40권4호
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• pp.258-262
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• 2012

cDNA of the glx1 gene encoding glyoxal oxidase (GLX) from Phanerochaete chrysosporium was isolated and expressed in Pichia pastoris. The recombinant GLX (rGLX) produces $H_2O_2$ over 7.0 nmol/min/mL using methyl glyoxal as a substrate. Use of rGLX as a generator of $H_2O_2$ improved the coupled reaction with recombinant manganese peroxidase resulting in decolorization of malachite green up to $150{\mu}M$ within 90 min.

• 펄프종이기술
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• 제31권2호
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• pp.8-14
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• 1999

A screening has been performed to find hyper-ligninolytic fungi, which degtrade beech and pine lignin extensively in order to broaden the understanding of the ligninolytic enzymes elaborated by various white-rot fungi. One hundred and twenty two ligninolytic strains were selected from decayed woods with a selective medium for screening ligninolytic wood-rotting fungi. Two of them, Phanerochaete sordida YK-624 and YK-472, showed much higher ligninolytic activity and selectivity in beech-wood degradation than typical lignin-degrading fungi, phanerochaete chrysosporium and Coriolus versicolor. They also degraded birch dioxane lignin and residual lignin in unbleached kraft pulp(UKP) much more extensively than P. chrysosporium and C. versicolor. During fungal treatment of beech wood-powder, the fungus strain P. sordida YK-624 showed higher activity of extracellular manganese peroxidase (MnP) in the medium than P. chrysosporium. It also showed MnP activity, which would not be lignin peroxidast during treatment of oxygen-bleached kraft pulp(OKP) and under enzyme-inducing conditin.

• Mycobiology
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• 제34권4호
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• pp.159-165
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• 2006

One of the most economically viable processes for the bioconversion of many lignocellulosic waste is represented by white rot fungi. Phanerochaete chrysosporium is one of the important commercially cultivated fungi which exhibit varying abilities to utilize different lignocellulosic as growth substrate. Examination of the lignocellulolytic enzyme profiles of the two organisms Phanerochaete chrysosporium and Rhizopus stolonifer show this diversity to be reflected in qualitative variation in the major enzymatic determinants (ie cellulase, xylanase, ligninase and etc) required for substrate bioconversion. For example P. chrysosporium which is cultivated on highly lignified substrates such as wood (or) sawdust, produces two extracellular enzymes which have associated with lignin deploymerization. (Mn peroxidase and lignin peroxidase). Conversely Rhizopus stolonifer which prefers high cellulose and low lignin containg substrates produce a family of cellulolytic enzymes including at least cellobiohydrolases and ${\beta}-glucosidases$, but very low level of recognized lignin degrading enzymes.

• Journal of the Korean Wood Science and Technology
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• 제33권4호통권132호
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• pp.45-52
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• 2005

백색부후균 Phanerochaete chrysosporium으로부터 유래한 Manganese peroxidase (mnp5)를 methylotrophic yeast인 Pichia pastoris에서 이종 발현을 하였다. 이종발현으로부터 얻어진 단백질은 클로닝으로부터 예상되어지는 분자량보다 높은 분자량인 45 kDa으로 나타났다. 이것은 mnp5가 가지고 있는 glycosylation site에 의한 것이며, N-linked hyperglycosylation이 효소 활성에 영향을 미치는지를 site direct mutation에 의해 확인하였다. Sodium dodesyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE)와 Coomassie Brilliant Blue (CBB) 염색에 의해 분자량을 확인한 결과 약 37 kDa으로 나타났으며, 효소활성을 측정한 결과 glycosylation이 효소 활성에 영향을 미치지 않는 것으로 나타났다. 따라서 본 연구로부터 P. pastoris에서 mnp5의 이종발현이 성공적으로 이루어졌으며 이러한 결과로부터 heme을 포함하고 있는 단백질의 이종발현 생산의 가능성을 보여주었다.

• 펄프종이기술
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• 제29권1호
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• pp.43-51
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• 1997

The various culture conditions of Trichoderma viride(ATCC 3454) and Phanerochaete chrysosporium(ATCC 26921) with glucose-pepton medium, Mandels medium, YMG medium for wood degradable enzyme were examined. Mycellium of the two species grew profusely on glucose-pepton medium. Maximum fungal growth was observed about 10days. But CMCase, Fpase, laccase activity in the culture medium with glucose-pepton was not detected. When grown in fermenter culture using Mandels medium, Trichoderma viride produced CMCase and Fpase. Its CMCase activity was 0.15 lU/ml and Fpase activity was 0.3 IU/ml within about 4-6days. Phanerochaete chrysosporium grown in a YMG medium gave the best enzyme activity when they were grown under stationary culture with an atmosphere of 100% oxygen. Levels of laccase activity of 3.0 mull were achieved in stationary culture under 100% oxygen. The enzyme condensation by ultrafiltration method caused a 2-fold(cellulase) and 6-fold(laccase) as compared to control activity.

• Mycobiology
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• 제39권2호
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• pp.121-124
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• 2011

The cDNA of endo-1,$4-{\beta}-xylanaseA$, isolated from Phaenerocheate chrysosporium was expressed in Pichia pastoris. Using either the intrinsic leader peptide of XynA or the ${\alpha}$-factor signal peptide of Saccharomyces cerevisiae, xylanaseA is efficiently secreted into the medium at maximum concentrations of 1,946 U/L and 2,496 U/L, respectively.

• Journal of the Korean Wood Science and Technology
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• 제26권3호
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• pp.53-62
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• 1998

In this research, 7 species of white rot fungi were used for determining the resistance against pentachlorophenol (PCP). Three fungi with good PCP resistance were selected for evaluating the biodegradability, and biodegradation mechanism by HPLC and GC/MS spectrometry. Among 7 fungi, there were significant differences on PCP resistance on 4 different PCP concentrations. In the concentrations of 50 and 100ppm ($\mu$g of PCP per g of 2% malt extract agar), most fungi were easily able to grow, and well suited to newly PCP-added condition, but in that of more than 250ppm, the mycelia growths of Ganoderma lucidum 20435, G. lucidum 20432, Pleurotus ostreatus, and Daldinia concentrica were significantly inhibited or even stopped by the addition of PCP to the culture. However, Trametes versicolor, Phanerochaete chrysosporium, and Inonotus cuticularis still kept growing at 250ppm, indicating the potential utilization of wood rot fungi to high concentrated PCP biodegradation. Particularly, P. chrysosporium even showed very rapid growth rate at more than 500ppm of PCP concentration. Three selected fungi based on the above results showed an excellent biodegradability against PCP. P. chrysosporium degraded PCP up to 84% on the first day of incubation, and during 7 days, most of added PCP were degraded. T. versicolor also showed more than 90% of biodegradability at 7th day, and even though the initial stage of degradation was very slow, I. cuticularis has been approached to 90% at 21 st day after incubation with dense growing pattern of mycelia. Therefore, the PCP biodegradability was definitely dependent on the rapid suitability of fungi to newly PCP-added condition. In addition, the PCP biodegradation by filtrates of P. chrysosporium, T. versicolor, and I. cuticularis was very minimal or limited, suggesting that the extracellular enzyme system may be not so significantly related to the PCP biodegradation. Among the biodegradation metabolites of PCP, the most abundant one was pentachloroanisole which resulted in a little weaker toxicity than PCP, and others were tetrachlorophenol, tetrachloro-hydroquinone, benzoic acid, and salicylic acid, suggesting that PCP may be biodegraded by several sequential reactions such as methylation, radical-induced oxidation, dechlorination, and hydroxylation.

• Journal of the Korean Wood Science and Technology
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• 제29권4호
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• pp.103-110
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• 2001

목재 내 리그닌의 선택적 분해 특성을 지닌 백색부후균 중, Phanerochaete chrysosporium KCCM 34740 균주를 현사시나무 목재 칩에 전처리하여 bio-kraft pulping 적용 가능성을 실험적으로 평가하였다. 전처리 결과, 무처리 대조구에 비해 펄프의 정선수율은 전처리 10일에서 최고 약 2%의 증가를 보였으며, 전처리 기간의 증가에 따라 여수도의 감소, WRV의 증가 경향을 나타냈다. 또한 수초지의 물성 개선에도 효과가 있었으며, 주사전자현미경 관찰을 통하여 이러한 효과가 백색부후균의 생물고해 작용, 즉 펄프 섬유의 미세섬유화 및 다공질화에 기인한 것임을 확인할 수 있었다. 이러한 결과로써 향후 kraft 증해 약액 및 제지공정상의 고해 동력에너지의 소비를 절감할 수 있을 것으로 기대한다.

• 펄프종이기술
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• 제31권5호
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• pp.95-100
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• 1999

In order to find fungi having high treatment activity of kraft pulp bleaching (E1) effluent without any additional nutrietns, 124 strains of white-rot fungi were isolated from decayed wood samples. The author isolated five fungi(KS-62, MZ-400 , YK-719, YK-472 and Phanerochaete sordida YK-624) having high-decolorization activity of the E1 effluent. Particularly, the fugus KS-62 show the high effect of the decolorization and the degradation of the chlorinated lignin in the E1 , effluent compared with Coriolus versicolor and Phanerochaete chrysosporium.

• 임산에너지
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• 제17권1호
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• pp.56-70
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• 1998

This study was carried out to investigate the structural characteristics of kraft lignin and the wood degrading characteristics, the productivity of ligninolytic enzymes and the enzymatic degradation of kraft lignin by white-rot fungi. To purify kraft lignin, precipitation of kraft pulping black liquors of pitch pine meal was done by titration with lN $H_{2}SO_{4}$ reaching to pH 2, and isolation of the precipitates done by centrifugation. The isolated precipitates from pitch pine were redissloved in lN NaOH, reprecipitated by titration with lN $H_{2}SO_{4}$, washed with deionized water, and kept ofr analysis after freeze drying. Fractionation of the precipitates in solution by successive extraction with $CH_{2}Cl_{2}$ and MeOH, and the fractionates were named SwKL, SwKL I, SwKL II, and SwKL III for pitch pine kraft lignin. The more molecular weights of kraft lignin increased, the less phenolic hydroxyl groups and the more aliphatic hydroxyl groups. Because as the molecular weights increased, the ratio of etherified guaiayl/syringyl(G/S ratio) and the percentage were increased. The spectra obtained by 13C NMR and FTIR assigned by comparing the chemical shifts of various signals with shifts of signals from autherized ones reported. The optimal growth temperature and pH of white-rot fungi in medium were $28^{\circ}C$ and 4.5-5.0, respectively. Especially, in temperature and pH range, and mycelial growth, the best white-rot fungus selected was Phanerochaete chrysosporium for biodegradation. For the degradation pathways, the ligninolytic fungus jcultivated with stationary culture using medium of 1% kraft lignin as a substrate for 3 weeks at $28^{\circ}C$. The weight loss of pitch pine kraft lignin was 15.8%. The degraded products extracted successively methoanol, 90% dioxane and diethyl ether. The ether solubles were analyzed by HPLC. Kraft lignin degradation was initiated in $\beta$-O-4 bonds of lignin by the laccase from Phanerochaete chrysosporium and the degraded compounds were produced from the cleavage of $C\alpha$-$C\beta$ linkages at the side chains by oxidation process. After $C\alpha$-$C\beta$ cleavage, $C\alpha$-Carbon was oxidized and changed into aldehyde and acidic compounds such as syringic acid, syringic aldehyde and vanilline. And the other compound as quinonemethide, coumarin, was analyzed. The structural characteristics of kraft lignin were composed of guaiacyl group substituted functional OHs, methoxyl, and carbonyl at C-3, -4, and -5 and these groups were combinated with $\alpha$ aryl ether, $\beta$ aryl ether and biphenyl. Kraft lignin degradation pathways by Phanerochaete chrysosporium were initially accomplished cleavage of $C\alpha$-$C\beta$ linkages and $C\alpha$ oxidation at the propyl side chains and finally cleavage of aromatic ring and oxidation of OHs.