• The Korean Journal of Pesticide Science
• /
• v.2 no.3
• /
• pp.70-78
• /
• 1998

Dissipation, adsorption and desorption of oxadiazon were examined in two soils containing different amounts of soil organic matter. In addition, reactivity of oxadiazon with humic monomers was searched to clarify binding mechanism of oxadiazon to soil organic matter in the presence of a laccase of Myceliophthera thermophila. Half lives of oxadiazon were 38 days in Soil I and 45 days in Soil II. Freundlich constant, k values of fresh soils were higher than those of oxidized soils. Adsorption rates of oxadiazon were increased 17.1% in Soil I and 9.3% in Soil II in the presence of a laccase but no significant increase was observed in oxidized soils. Desorption rates of oxadiazon in fresh soils were lower than those in oxidized soils. Desorption rates of adsorbed oxadiazon in soils addes with the enzyme were not changed in oxidized soils but decreased in fresh soils. The herbicide oxadiazon alone underwent no transformation by a laccase but in the presence of catechol, guaiacol and gallic acid as humic monomer, transformation rates of it were from 20% to 24%.

• Mycobiology
• /
• v.41 no.1
• /
• pp.37-41
• /
• 2013

Laccases (EC 1.10.3.2) are copper-containing polyphenol oxidases found in white-rot fungi. Here, we report the cloning and analysis of the nucleotide sequence of a new laccase gene, fvlac7, based on the genomic sequence of Flammulina velutipes. A primer set was designed from the putative mRNA that was aligned to the genomic DNA of F. velutipes. A cDNA fragment approximately 1.6-kb long was then amplified by reverse transcriptase-PCR using total RNA, which was subsequently cloned and sequenced. The cDNA sequence of fvlac7 was then compared to that of the genomic DNA, and 16 introns were found in the genomic DNA sequence. The fvlac7 protein, which consists of 538 amino acids, showed only 42~51% identity with 12 different mushroom species containing two laccases of F. velutipes, suggesting the fvlac7 is a novel laccase gene. The first 25 amino acids of Fvlac7 correspond to a predicted signal sequence, four copper-binding sites, and four N-glycosylation sites. Fvlac7 cDNA was heterologously overexpressed in an Escherichia coli system with an approximate expected molecular weight of 60 kDa.

• Journal of Mushroom
• /
• v.17 no.4
• /
• pp.247-254
• /
• 2019

Trametes hirsuta, a white rot fungus, exhibits the ability to degrade synthetic aromatic dyes such as congo red (CR), methylene blue (MB), crystal violet (CV), and remazol brilliant blue R (RBBR). The mycelia of T. hirsuta degraded RBBR and CR more efficiently than CV and MB in the PDB liquid medium (supplemented with 0.01% 4 aromatic dyes). In these mycelia the activities of three ligninolytic enzymes-laccase, manganese peroxidase (MnP), and lignin peroxidase (LiP)-were observed. Among these, laccase was identified to be the major enzyme responsible for the degradation of the four aromatic dyes. The degradation of bisphenol A was also investigated by culturing the mycelia of T. hirsuta in YMG medium supplemented with 100 ppm bisphenol A. The mycelia of T. hirsuta were found to degrade bisphenol A by 71.3, 95.3, and 100 % within incubation periods of 12, 24, and 36 hr, respectively. These mycelia also showed ligninolytic enzyme-like activities including those similar to laccase, MnP, and LiP. Therefore, these results indicate that T. hirsuta could emerge as a potential tool for the remediation of environmental contamination by aromatic dyes and bisphenol A.

• Biotechnology and Bioprocess Engineering:BBE
• /
• v.5 no.3
• /
• pp.211-214
• /
• 2000

The roles of lignin peroxidase, manganese peroxidase, and laccase were inverstigated in the biodegration of pentachlorphenol (PCP) by several which rot fungi. The disappearance of pentachlorophenol from cultures of wild type strains, P. chrysosporium, Trametes sp. and of pentachlorophenol from cultures of wild type strains, P. cheysocporium, Trametes sp. and Pleurotus ap., was observed. The activities of mangnese peroxidase and laccase was detected in Trametes sp. and pleurotus sp. cultures. However, the activities showed that PCP was degraded under ligninolytic as well as nonligninoytic condicationg that lignin peroxidase, manganese peroxidase, and laccase are not essential in the biodegradation of PCP by white rot fungi.

• BMB Reports
• /
• v.33 no.1
• /
• pp.37-42
• /
• 2000

Based on the study by Leatham and Stabmann concerned with the rates (v) of amines and phenolic compounds oxidation catalyzed by laccase of basidiomycete Lentinus edodes (Berk.) Sing., as well as on the results of semiempirical quantum chemical computations using the PM3 method, the linear correlations of v and lnv values with first vertical ionization potentials of the substrates molecules and radicals derived from them, spin densities on N and O atoms of the above radicals, and with the radicals reorganization energies have been found.

• Journal of Microbiology and Biotechnology
• /
• v.29 no.9
• /
• pp.1383-1390
• /
• 2019

In this study, we expressed cotA laccase from Bacillus subtilis on the surface of B. subtilis spores for efficient decolorization of synthetic dyes. The cotE, cotG, and cotY genes were used as anchoring motifs for efficient spore surface display of cotA laccase. Moreover, a $His_6$ tag was inserted at the C-terminal end of cotA for the immunological detection of the expressed fusion protein. Appropriate expression of the CotE-CotA (74 kDa), CotG-CotA (76 kDa), and CotY-CotA (73 kDa) fusion proteins was confirmed by western blot. We verified the surface expression of each fusion protein on B. subtilis spore by flow cytometry. The decoloration rates of Acid Green 25 (anthraquinone dye) for the recombinant DB104 (pSDJH-EA), DB104 (pSDJH-GA), DB104 (pSDJH-YA), and the control DB104 spores were 48.75%, 16.12%, 21.10%, and 9.96%, respectively. DB104 (pSDJH-EA) showed the highest decolorization of Acid Green 25 and was subsequently tested on other synthetic dyes with different structures. The decolorization rates of the DB104 (pSDJH-EA) spore for Acid Red 18 (azo dye) and indigo carmine (indigo dye) were 18.58% and 43.20%, respectively. The optimum temperature for the decolorization of Acid Green 25 by the DB104 (pSDJH-EA) spore was found to be $50^{\circ}C$. Upon treatment with known laccase inhibitors, including EDTA, SDS, and $NaN_3$, the decolorization rate of Acid Green 25 by the DB104 (pSDJH-EA) spore decreased by 23%, 80%, and 36%, respectively.

• Korean Journal of Clinical Laboratory Science
• /
• v.38 no.2
• /
• pp.87-93
• /
• 2006

This study was done to know a kind and change (transition) of enzymes produceed by Streptomyces halstedii ssp. scabies SA1-27 and Streptomyces violaceusinger C1-6 which showed good lignolytic activity and a good decolorization ratio of remazol brilliant blue R(RBBR) dye. These strains were isolated from soil and identified by the author. The basal medium containg 0.2% glucose was used to measure enzyme activity, Lignin peroxidase 1 (Lip 1) was measured by the methods of Choi, and Bourbonnais and Paice. Lignin peroxidase 2 (Lip 2) was measured by the methods of Ishida et al and Ramachandra et al using 2.4-dichlorophenol(2.4 DCP), manganese peroxidase(Mnp), veratryl alcohol oxidase (VAO), and laccase. They were measured by each of the methods of Choi and Paszczynski et al, and Bourbonnais and Paice, and De Jong et al. In the results, the kind of enzymes produced by Streptomyces halstedii ssp. scabies SA1-27 were Lip 1, Lip 2, VAO, and laccase, and their activities indicated the highest value as each 4.95 nmol/mg protein, $8.45({\times}100^{-3})unit$, 10.25 nmol/mg protein, 9.20 nmol/mg protein on the sixth day of the culture and decreased gradually over time. The kind of enzymes produced by Streptomyces violaceusinger C1-6 were Lip 1, Lip 2, Mnp, VAO, and laccase, and their activities indicated the highest value as each 4.90 nmol/mg protein, $13.85({\times}100^{-3})unit$, 3.10 nmol/mg protein, 11.30 nmol/mg protein, 4.45 nmol/mg protein on the sixth day of the culture and decreased gradually over time. Consequently, the author knew the fact that there were few differences in the kind and quantity of enzymes produced by the two Streptomyces strains, but all enzyme activities indicated the highest value on the sixth day of the culture and decreased gradually over time.

• Journal of Mushroom
• /
• v.2 no.4
• /
• pp.184-191
• /
• 2004

Mutual growth limitation was observation when the two antagonistic fungi was come in contact with each other. Brown line was formed 2day after contact with Trichoderma spp., and then, green spores formed overnight. The laccase activity of L. edodes was stimulated when this fungus wsa co-incubated with Trichoderma spp. for a few days in liquid media. In sawdust-rice bran nixtures, outstanding broun line developed when the two antagonistic fungi co-cultured. The pH of the substrates changed from 5.5 to 4.5 after overgrowth, suggesting a difference in the degradation ability and the preference of the two fungi for the lignocellulose material.

• Journal of Microbiology and Biotechnology
• /
• v.20 no.7
• /
• pp.1069-1076
• /
• 2010

A novel laccase from Tricholoma mongolicum was purified by using a procedure that entailed ion-exchange chromatographies on DEAE-cellulose, CM-cellulose, and Q-Sepharose, and FPLC-gel filtration on Superdex 75. The purified enzyme was obtained with a specific activity of 1,480 U/mg-protein and a final yield of 15%. It was found to be a monomeric protein with a molecular mass of 66 kDa as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Its N-terminal amino acid sequence was GIGPVADLYVGNRIL, similar to some but also different to other mushroom laccases. The optimum pH and temperature for the purified enzyme were pH 2 to pH 3 and $30^{\circ}C$, respectively. It displayed a low $K_m$ toward 2,7-azinobis (3-ethylbenzothiazolone-6-sulfonic acid) diammonium salt (ABTS) and high $k_{cat}/K_m$ values. The purified laccase oxidized a wide range of lignin-related phenols, but exerted maximal activity on ABTS. It was significantly inhibited by $Hg^{2+}$ ions, and remarkably stimulated by $Cu^{2+}$ ions. It inhibited HIV-1 reverse transcriptase and proliferation of hepatoma HepG2 cells and breast cancer MCF7 cells with an $IC_{50}$ of 0.65 ${\mu}M$, 1.4 ${\mu}M$, and 4.2 ${\mu}M$, respectively, indicating that it is also an antipathogenic protein.

• Journal of Microbiology and Biotechnology
• /
• v.27 no.6
• /
• pp.1120-1127
• /
• 2017

Marasmius scorodonius secretes an extracellular laccase in potato dextrose broth, and this enzyme was purified up to 206-fold using $(NH_4)_2SO_4$ precipitation and a Hi-trap Q Sepharose column. The molecular mass of the purified laccase was estimated to be ~67 kDa by SDS-PAGE. The UV/vis spectrum of the enzyme was nontypical for laccases, and metal content analysis revealed that the enzyme contains 1 mole of Fe and Zn and 2 moles of Cu per mole of protein. The optimal pH for the enzymatic activity was 3.4, 4.0, and 4.6 with 2,2'-azino-bis(3-ethylbenzothazoline-6-sulfonate) (ABTS), guaiacol, and 2,6-dimethoxy phenol as the substrate, respectively. The optimal temperature of the enzyme was $75^{\circ}C$ with ABTS as the substrate. The enzyme was stable in the presence of some metal ions such as $Ca^{2+}$, $Cu^{2+}$, $Ni^{2+}$, $Mg^{2+}$, $Mn^{2+}$, $Ba^{2+}$, $Co^{2+}$, and $Zn^{2+}$ at a low concentration (1 mM), whereas $Fe^{2+}$ completely inhibited the enzymatic activity. The enzymatic reaction was strongly inhibited by metal chelators and thiol compounds except for EDTA. This enzyme directly decolorized Congo red, Malachite green, Crystal violet, and Methylene green dyes at various decolorization rates of 63-90%. In the presence of 1-hydroxybenzotriazole as a redox mediator, the decolorization of Reactive orange 16 and Remazol brilliant blue R was also achieved.