• Journal of Microbiology and Biotechnology
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• v.16 no.2
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• pp.226-231
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• 2006

Several white-rot fungi are able to produce extracellular lignin-degrading enzymes such as manganese peroxidase (MnP), lignin peroxidase (LiP), and laccase. In order to enhance the production of laccase and MnP using Trametes versicolor KCTC 16781 in suspension culture, the effects of major medium ingredients, such as carbon and nitrogen sources, on the production of the enzymes were investigated. The decolorization mechanism in terms of biodegradation and biosorption was also investigated. Among the carbon sources used, glucose showed the highest potential for the production of laccase and MnP. Ammonium tartrate was a good nitrogen source for the enzyme production. No significant difference in the laccase production was observed, when glucose concentration was varied between 5 g/l and 30 g/l. As the concentration of nitrogen source increased, a lower MnP activity was observed. The optimal C/N ratio was 25 for the production of laccase and MnP. When the concentrations of glucose and ammonium tartrate were simultaneously increased, the laccase and MnP activities increased dramatically. The maximum laccase and MnP activities were 33.7 U/ml at 72 h and 475 U/ml at 96 h, respectively, in the optimal condition. In this condition, over 90% decolorization efficiency was observed.

• Proceedings of the Korea Technical Association of the Pulp and Paper Industry Conference
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• 2001.04a
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• pp.18-25
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• 2001

A new wood-degrading fungus Trichophyton rubrum LKY-7 secretes a high level of laccase in a glucose-peptone liquid medium. The production of laccase by the fungus was barely induced by 2,5-xylidine. The laccase has been purified to homogeneity through three chromatography steps in an overall yield of 40%. The molecular mass of the purified laccase was about 65 kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The purified laccase had the distinct blue color and had basic spectroscopic features of a typical blue laccase: two absorption maxima at 278 and 610 nm and a shoulder at 338 nm. The N-terminus of the laccase has been sequenced, revealing high homology to laccases from wood-degrading white-rot fungi such as Ceriporiopsis subvermispora. The enzyme had a "low" redox potential (0.5 V vs normal hydrogen electrode), yet it was one of the most active laccases in oxidizing a series of representative substrates/mediators. Compared with other fungal laccases, the laccase has a very low Km value with ABTS [2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid] as a substrate and a very high Km value with violuric acid as a substrate. The laccase has the isoelectric point of 4.0. The laccase had very acidic optimal pH values (pH 3-4) while it was more stable at neutral pH than at acidic pH. The laccase oxidized hydroquinone faster than catechol and pyrogallol. The oxidation of tyrosine by the laccase was not detectable under the reaction conditions. The laccase was strongly inhibited by sodium azide and sodium fluoride. fluoride.

• Korean Journal of Microbiology
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• v.48 no.3
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• pp.225-227
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• 2012

Lignin degrading enzymes from white rot fungi show broad substrate specificities, and therefore they can degrade variety of recalcitrant compounds. We have used three different protocols for the generation of immobilized laccase and manganese peroxidase crude enzymes from the genetically transformed strains of Merulius tremellosus Fr. These immobilized enzymes were used in the decolorization of Remazol Brilliant Blue R (RBBR), and they showed about 75% decolorization rates during the 48 h reactions. Although the decolorization efficiency decreased by 10-15% after a repeated use of the immobilized enzymes, these can be reused in various degrading reactions.

• Mycobiology
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• v.30 no.2
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• pp.82-87
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• 2002

Species of Phellinus were known to harmful fungi causing white pocket rot and severe plant disease such as canker or heartrot in living trees in the West, but some species have been used to traditional medicines in the Orient for a long time. In this study the partial D1-D2 nucleotide sequences of 28S ribosomal DNA from 13 Phellinus strains were determined and compared with the sequences of 21 strains obtained from GenBank database. According to the neighbor-joining(NJ) method comparing the sequence data the phylogenetic tree was constructed. The phylogenetic tree displayed the presence of four groups. Group I includes P. ferreus, P. gilvus and P. johnsonianus, Group II contains P. laevigatus, P. conchatus and P. tremulae, Group III possesses P. linteus, P. weirianus, P. baumii, P. rhabarbarinus and P. igniarius, and Group IV comprises P. pini, P. chrysoloma. P. linteus and P. baumii, which were used mainly in traditional medicine, belong to the same group, but exactly speaking both were split into two different subgroups. To detect P. linteus only, we developed the PCR primer, D12HR. The primer showed the specific amplification of P linteus, which is permitted to medicinal mushroom in the East. The results make a potential to be incorporated in a PCR identification system that could be used for the rapid identification of this species from its related species, P. linteus especially.

• The Korean Journal of Mycology
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• v.14 no.1
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• pp.43-47
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• 1986

Among the representative phenolic compounds in relation to lignin derivatives and protein synthesis inhibitors, the most effective inducer for the extracellular polyphenol oxidase (PO) of Lentinus edodes JA01 was gallic acid and ferulic acid for Pleurotus ostreatus. Optimum concentration of these inducers was 2.0 mM and 1.0 mM, respectively. Addition of gallic acid after two days culture had the best effect on production of PO enzyme of L. edodes JA01 and for P. ostreatus, and addition of ferulic acid after three days culture had the best effect. Also, in case of L. edodes JA01, polyphenol oxidase activity was parallel to growth curve, whereas the maximum enzyme activity of P. ostreatus was shown at exponential growth phase and declined thereafter.

• Journal of Korean Society of Environmental Engineers
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• v.27 no.7
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• pp.712-718
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• 2005

Stepwise reductions of glucose and Orange II concentration were observed from the experiment of both white-rot fungi such as T. versicolor and P. chrysosporium. As a result, typical removal patterns in those dual substrate system were categorized through several distinctive steps: initial lag period, primary and secondary carbon consumption periods. Also, based on the total removal amounts of Orange II, COD and Color during the experimental period, similar removal extent were observed from both species experiments, within the maximal error range of 5%. However, it was refereed that the internal steps of Orange II removal on enzymatic level should be different between two species: Enzyme Lac showed good affinity for Orange II removal in T. versicolor, however in P. chrysosporium enzyme LiP represented more close affinity to the similar experimental condition. Thus, even though the superficial removal amount of calcitrant Orange II at different fungal species was merely similar, removal pathway of enzymatic levels and intermediates produced during the fungal decomposition would be different.

• Plant Resources
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• v.3 no.2
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• pp.110-117
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• 2000

The recycling method of enokitake cultural waste and the potentiality of second flush for enokitake were determined, because this fungus is not as prolific as the more commonly cultivated white rot fungi in the conversion of sawdust to mycelial mass. The mycelial growth of F. velutipes on several substrates, variously treated with rice bran was promoted at ratios of 10-20% (w/w) on all substrates, but suppressed at above ratios, although some difference was there. The mycelial densities generally increased correlated to the supplementation contents of rice bran. It could be concluded that F. velutipes preferred mild acidic to acidic conditions for mycelial growth, considering that the mycelial growth rate was highest on waste of pH 6.01, treated with 0.1 % Ca(OH)$_2$ and on populus mixed waste of pH 6.02, non treated. The ranges of substrate bulk densities, which was pertinent for mycelial linear growth were from B.D. (g/cc) 0.17 to 0.23 on waste and populus mixed waste all. The pertinent contents of rice bran supplementation in bottle cultivation was from 20 to 30% on waste and 20% on populus mixed waste, considering the requried duration for pinheading and fruiting yields. Standard bulk density for filling and utilizing the waste and populus mixed waste for commercial f. velutipes cultivation were B.D.(g/cc) 0.19 ~ 0.23, and 0.23~ 0.25, which could be conversed to 510~ 540g/900m1 and 520~ 570g/900m1, respectively, The second flush of F. velutipes was tried and the re-inoculation by sawdust and liquid spawn showed somewhat good results, indicating the potentiality of second crop and suggesting further research for it.

• Mycobiology
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• v.44 no.4
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• pp.260-268
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• 2016

Regulation of alkaline-resistant laccase from Perenniporia tephropora KU-Alk4 was proved to be controlled by several factors. One important factor was the initial pH, which drove the fungus to produce different kinds of ligninolytic enzymes. P. tephropora KU-Alk4 could grow at pH 4.5, 7.0, and 8.0. The fungus produced laccase and MnP at pH 7.0, but only laccase at pH 8.0. The specific activity of laccase in the pH 8.0 culture was higher than that in the pH 7.0 culture. At pH 8.0, glucose was the best carbon source for laccase production but growth was better with lactose. Low concentrations of glucose at 0.1% to 1.0% enhanced laccase production, while concentrations over 1% gave contradictory results. Veratryl alcohol induced the production of laccase. A trace concentration of copper ions was required for laccase production. Biomass increased with an increasing rate of aeration of shaking flasks from 100 to 140 rpm; however, shaking at over 120 rpm decreased laccase quantity. Highest amount of laccase produced by KU-Alk4, 360 U/mL, was at pH 8.0 with 1% glucose and 0.2 mM copper sulfate, unshaken for the first 3 days, followed by addition of 0.85 mM veratryl alcohol and shaking at 120 rpm. The crude enzyme was significantly stable in alkaline pH 8.0~10.0 for 24 hr. After treating the pulp mill effluent with the KU-Alk4 system for 3 days, pH decreased from 9.6 to 6.8, with reduction of color and chemical oxygen demand at 83.2% and 81%, respectively. Laccase was detectable during the biotreatment process.

• Journal of the Korean Wood Science and Technology
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• v.35 no.1
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• pp.64-72
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• 2007

The biodeterioration behaviors of square post to support the wooden printing blocks Shelves in the Janggeongpanjeon were investigated according to the positions and parts of square post, and environmental conditions. It was found that a high differences of deterioration in the progressing levels of wood decay, according to the positions and parts of square post, and environmental conditions. The decay levels were very high and still progressing in the contacted areas with stone foundation which are about up to 50 cm above it. In the decay type, white rot fungi was mainly affected in the inside of building which mainly made of softwood. The decay in the square posts to support the wooden printing block shelves inside of building was worse in the rear side, compared to front side. The insects was not found in most of square posts excluding the post which was neighboring at the infected round column by insect.

• Journal of Microbiology and Biotechnology
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• v.34 no.4
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• pp.930-939
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• 2024

Mushroom laccases play a crucial role in lignin depolymerization, one of the most critical challenges in lignin utilization. Importantly, laccases can utilize a wide range of substrates, such as toxicants and antibiotics. This study isolated a novel laccase, named HeLac4c, from endophytic white-rot fungi Hericium erinaceus mushrooms. The cDNAs for this enzyme were 1569 bp in length and encoded a protein of 523 amino acids, including a 20 amino-acid signal peptide. Active extracellular production of glycosylated laccases from Saccharomyces cerevisiae was successfully achieved by selecting an optimal translational fusion partner. We observed that 5 and 10 mM Ca²⁺, Zn²⁺, and K⁺ increased laccase activity, whereas 5 mM Fe²⁺ and Al³⁺ inhibited laccase activity. The laccase activity was inhibited by the addition of low concentrations of sodium azide and ⳑ-cysteine. The optimal pH for the 2,2'-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt was 4.4. Guaiacylglycerol-β-guaiacyl ether, a lignin model compound, was polymerized by the HeLac4c enzyme. These results indicated that HeLac4c is a novel oxidase biocatalyst for the bioconversion of lignin into value-added products for environmental biotechnological applications.