• Korean Journal of Microbiology
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• v.29 no.4
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• pp.238-242
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• 1991

Extracellular laccase secreted from Pleurotus ostreatus was activated by $Cu^{2+}$ and $Cu^{+}$ . The enzyme was strongly inactivated by 8-hydroxyquinoline, potassium cyanide, sodium azide, sodium bisulfite and 2-mercaptoethanol. The two ionogenic groups, which have pKa values of 5.60-5.70 and 6.70-6.85 respectively, were found to relate with the active site of this enzyme. The oxidation reactions were brought about by initial single electron transfer process on the active site. The enzyme was found to be a metalloprotein which had about 3.9 cupric ions per molecule of protein as a prosthetic group. The enzyme showed a strong peak at 605 nm and a weak shoulder at 330 nm in UV-Visible absorption spectrum. Both signals disappeated upon treatment of the enzyme with 4 electron equivalent ascorbate. These results indicate that type I Cu peak and type III Cu shoulder are present in laccase.

• Bulletin of the Korean Chemical Society
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• v.23 no.3
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• pp.385-390
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• 2002

Laccase could be successfully assembled on an amine-derivatized platinum electrode by glutaraldehyde coupling. The enzyme layer formed on the surface does not communicate electron directly with the electrode, but the enzymatic activity of the surf ace could be followed by electrochemical detection of enzymatically oxidized products. The well-known laccase substrates, ABTS (2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid)) and PPD (p-phenylenediamine) were used. ABTS can be detected down to 0.5 ${\mu}M$ with linear response up to 15 ${\mu}M$ and current sensitivity of 75 nA/ ${\mu}M.$ PPD showed better response with detection limit of 0.05 ${\mu}M$, linear response up to 20 ${\mu}M$, and current sensitivity of 340 nA/ ${\mu}M$ with the same electrode. The sensor responses fit well to the Michaelis-Menten equation and apparent $K_M$ values are 0.16 mM for ABTS and 0.055 mM for PPD, which show the enzymatic reaction is the rate-determining step. The laccase electrode we developed is very stable and more than 80% of initial activity was still maintained after 2 months of uses.

• Journal of Mushroom
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• v.20 no.1
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• pp.29-33
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• 2022

In this study, five different Lentinula edodes cultivar (Chamaram, Sanbaekhyang, Sanjo 713ho, Sanjo 715ho, Sanjo 718ho) were evaluated for their ability to decolorize Remazol Brilliant Blue R (RBBR) in MEB medium, respectively. Chamaram and Sanjo 713ho decolorized RBBR rapidly in MEB medium within 3 and 5 days. The activities of manganese peroxidase (MnP) and laccase were determined on the MEB medium with and without lignin. Sanjo 713ho resulted the highest ligninolytic enzyme activities on incubation day 1, indicating of 1,213 U/mg of MnP activity and 1,421 U/mg of laccase activity.

• Journal of Korea Technical Association of The Pulp and Paper Industry
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• v.46 no.1
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• pp.1-10
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• 2014

Xylanase (X) derived from Aurreobasidium pullulans and laccase-mediator system (LM) using Trichophyton sp. LKY-7 laccase (TrL) and N-hydroxy-2-pyridone analogue (NHP) as a mediator were applied in hardwood kraft pulp (HwKP) bleaching. The individual and the synergistic effects of X and LM stage were investigated in the enzymatic bleaching of HwKP. Also, the effects of subsequent alkaline extraction (E) and alkaline/hydrogen peroxide treatment (P) were examined. In X or LM treatment alone, an appreciable bleaching effect of HwKP was not observed, whereas subsequent E or P stage enhanced the increase of brightness and the decrease of kappa number. Especially, P stage significantly enhanced the bleaching effect of pulp. Bleaching of HwKP with XLM sequentially gave significantly higher pulp brightness and lower kappa number than that obtained after the treatment of HwKP with X+LM simultaneously. When HwKP was sequentially treated with XLM followed by P stage, the brightness increased by about 11% ISO and the kappa number decreased by about 3.6 in comparison with the initial pulp. Xylanase and laccase were strongly inactivated by NHP both in the absence and the presence of pulp.

• Journal of Plant Biotechnology
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• v.35 no.1
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• pp.41-45
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• 2008

Laccase (EC 1.10.3.2) is a small group of enzymes that catalyze the oxidation of a broad range of phenolic compounds including hazardous and recalcitrant pollutants in the environment. This study attempted to develop an efficient system for production of a recombinant laccase by chloroplast genetic transformation of tobacco. Chloroplast transformation vector was constructed and introduced into the tobacco chloroplast genome using particle bombardment. Chloroplast-transformed plants were subsequently regenerated. PCR and southern blot analyses confirmed stable integration of the laccase gene into the chloroplast genome. Northern blot analysis revealed that mRNA of the laccase gene was highly expressed in chloroplast-transformed plants.

• Journal of the Korean Wood Science and Technology
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• v.26 no.3
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• pp.41-47
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• 1998

Some white-rot fungi, screened at the Laboratory of Forest Products Microbiological Chemistry, Chungbuk National University were cultured and added the inducer of laccase enzyme, 2,5-xylidine. The fungi named by CB-13, CB-20, CB-99, CB-100 and CB-123 strains showed positive results in the decolorization of aromatic compounds, carminic acid and Rhemazol brilliant blue R. Concerned to the inducing effect of 2,5-xylidine on laccase activity, CB-20, CB-100 and CB-123 strains showed very high activity by addition of 2,5-xylidine, whilst CB-13, CB-99 and CB-124 strains produced relatively high laccase enzymes, regardless of inducer addition. There were no any laccase activities on CB-25, CB-64 and CB-139, even in addition of inducer. It is confirmed that some screened fungi have decolorizing ability on aromatic compounds, carminic acid and Rhemazol brilliant blue R. Also, the addition of inducer, 2,5-xylidine, has increased the activity of laccase enzyme which is secreted from some white-rot fungi.

• Korean Journal of Microbiology
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• v.25 no.2
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• pp.148-156
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• 1987

Extracellular laccase (E.C. 1.10.3.2) from the culture filtrate of Pleurotus ostreatus was purified by ammonium sulfate precipctation, protamine sulfate precipitation, DEAE-Sephadex A-50 ion exchange chromatography and Sephadex G-100 gel permeation chromatography. The molecular weight of the enzyme was estimated by SDS-polyacrylamide gel electrophoresis to be 58,000 and the isoelectric point was 3.75. The optimum temperature for the enzyme was about $45^{\circ}C$ and the optimum pH was 6.5. The enzyme was found to be stable at temperature below $35^{\circ}C$ and rapidly inactivated at higher temperatures. Km values for ferulic acid, vanillic acid, dihydroxyphenylalanine (DOPA) were 48.6.$\mu$M, 0.52mM, and 2.73mM, respectively, which indicates that the enzyme has much higher affinity towards ferulic acid. The reaction products of the enzyme were separated by TLC and HPLC.

• The Korean Journal of Mycology
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• v.39 no.1
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• pp.48-52
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• 2011

Breeding of Lentinula edodes generates a number of hybrid strains that are subject to evaluation for good traits for the mushroom production. As an effort to understand biochemical properties of the hybrid strains, this study tried to develop a fast and easy method for comparison of the ability of producing extracellular laccase among hybrid strains of Lentinula edodes. For this aim, we estimated the effect of media types and chromagenic chemicals on the detection of extracellular laccase in seven hybrid strains of L. edodes. When Remazol Brilliant Blue R (RBBR) dye was used for chromagenic reaction, the detection of the enzyme activity was feasible both in the solid and liquid media containing not potato dextrose but malt extract as a nutrient component. When guaiacol was used for chromagenic reaction, the detection of the enzyme activity was feasible both in the solid and liquid media containing either potato dextrose or malt extract as a nutrient component. Malt extract-based liquid culture with RBBR or guaiacol in 2 ml microfuge tube allowed us to economically and quantitatively detect and compare the enzyme activity within 3 days among the tested hybrid strains of L. edodes.

• Mycobiology
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• v.42 no.2
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• pp.193-197
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• 2014

Laccase activity of Pleurotus ostreatus is significantly increased by the addition of apple pomace. Among various conditions, the best concentration of apple pomace and cultivation time for the production of laccase by P. ostreatus was 2.5% and 9 days, respectively. Reverse transcription polymerase chain reaction analyses of laccase isoenzyme genes, including pox1, pox3, pox4, poxc, poxa3, and poxa1b, revealed a clear effect of apple pomace on transcription induction. Our findings reveal that the use of apple pomace can be a model for the valuable addition of similar wastes and for the development of a solid-state fermenter and commercial production of oyster mushroom P. ostreatus.

• Korean Journal of Microbiology
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• v.46 no.1
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• pp.93-95
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• 2010

White rot fungi which have lignin degrading enzymes show high degrading activity to diverse recalcitrant compounds such as polycyclic aromatic compounds, dyes, explosives and endocrine disrupting chemicals. We have examined decolorizing activity of dyes by Phlebia tremellosa and two transformants which had genetically transformed using laccase or manganese peroxidase (MnP) gene. In case of methyl green, wild type strain showed 50% decolorization while laccase transformant (TF2-1) and MnP transformant (T5) showed more than 90% decolorization on day 3. Remazol brilliant blue R(RBBR) was decolorized up to 85% by two transformants while the wild type showed 67% decolorization on day 3. Transformants TF2-1 and T5 both showed increased laccase and MnP activity respectively during the whole growing phase.