• 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.

• Biotechnology and Bioprocess Engineering:BBE
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• v.11 no.2
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• pp.96-99
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• 2006

In this study, we have attempted to determine the optimum concentration of inducers responsible for efficient laccase production by the white-rot fungus, Trametes sp. Variations in laccase activity were investigated with changing concentrations of 2,5-xylidine, syringaldazine, ABTS, and guaiacol. Enhancement of peak laccase activity was achieved via the combination of 2,5-xylidine with ABTS, syringaldazine, or guaiacol, resulting in increases of up to 359, 313, and 340%, respectively, as compared to control values. Among the tested inducers, the addition of 0.1mM of ABTS coupled with 1.0mM of 2,5-xylidine in the medium after 24 h of cultivation proved optimal with regard to laccase enzyme production.

• Microbiology and Biotechnology Letters
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• v.42 no.3
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• pp.225-231
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• 2014

In this study about the optimum conditions for the production of laccase, a polyphenol oxidase involved in lignin degradation, from Marasmius scorodonius, a white-rot fungus garlic mushroom, were determined. Amongst the tested media used for the enzyme's production, YM medium (1% dextrose, 0.5% malt extract, 0.3% yeast extract) allowed for the highest activity of the enzyme. Then, to optimize the culture conditions for laccase activity, the influence of various carbon and nitrogen sources was investigated in YM medium. Among various carbon and nitrogen sources, 1% galactose and 0.4% yeast extract resulted in the highest production of the enzyme, respectively. Enzyme production attained its highest level after cultivation for 15 days at $25^{\circ}C$. Zymogram analysis of the culture supernatant showed two isoenzymatic bands with molecular masses of 60-70 kDa. The optimum pH and temperature for enzyme activity were 3.4 and $75^{\circ}C$, respectively.

• Journal of Microbiology and Biotechnology
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• v.26 no.9
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• pp.1570-1578
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• 2016

Commercial application of laccase is often hampered by insufficient enzyme stocks, with very low yields obtained from natural sources. This study aimed to improve laccase production by mutation of a Coriolopsis gallica strain and to determine the biological properties of the mutant. The high-yield laccase strain C. gallica TCK was treated with N-methyl-N-nitro-N-nitrosoguanidine and ultraviolet light. Among the mutants isolated, T906 was found to be a high-production strain of laccases. The mutant strain T906 was stabilized via dozens of passages, and the selected ones were further processed for optimization of metallic ion, inducers, and nutritional requirements, which resulted in the optimized liquid fermentation medium MF9. The incubation temperature and pH were optimized to be 30℃ and 4.5, respectively. The mutant strain T906 showed 3-times higher laccase activity than the original strain TCK under optimized conditions, and the maximum laccase production (303 U/ml) was accomplished after 13 days. The extracellular laccase isoenzyme 1 was purified and characterized from the two strains, respectively, and their cDNA sequence was determined. Of note, the laccase isoenzyme 1 transcription levels were overtly increased in T906 mycelia compared with values obtained for strain TCK. These findings provide a basis for C. gallica modification for the production of high laccase amounts.

• The Korean Journal of Mycology
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• v.31 no.3
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• pp.181-186
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• 2003

The production of laccase by Fomitella fraxinea was studied. The addition of minerals were necessary far laccase production by Fomitella fraxinea. Jar fermentor and Air-sparging fermentor performed high productivity In laccase activity by F. fraxinea. Laccase activity reached 3,540 in 8 days (Jar fermentor) and 3,100 in 6 days (Air-sparging fermentor) respectively.

• Journal of Microbiology
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• v.43 no.3
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• pp.301-307
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• 2005

The feasibility of laccase production by immobilization of Pleurotus ostreatus 1804 on polyurethane foam (PUF) cubes with respect to media composition was studied in both batch and reactor systems. Enhanced laccase yield was evidenced due to immobilization. A relatively high maximum laccase activity of 312.6 U was observed with immobilized mycelia in shake flasks compared to the maximum laccase activity of free mycelia (272.2 U). It is evident from this study that the culture conditions studied, i.e. biomass level, pH, substrate concentration, yeast extract concentration, $Cu^{2+}$ concentration, and alcohol nature, showed significant influence on the laccase yield. Gel electrophoretic analysis showed the molecular weight of the laccase produced by immobilized P. ostreatus to be 66 kDa. The laccase yield was significantly higher and more rapid in the packed bed reactor than in the shake flask experiments. A maximum laccase yield of 392.9 U was observed within 144 h of the fermentation period with complete glucose depletion.

• Microbiology and Biotechnology Letters
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• v.45 no.4
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• pp.330-342
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• 2017

Statistical design of experiments was employed to optimize the media composition for the production of laccase from Bacillus sp. PK4. In order to find the key ingredients for the best yield of enzyme production from the selected eleven variables viz yeast extract, glucose, zinc sulphate, copper sulphate, potassium chloride, magnesium sulphate, calcium chloride, ferrous sulphate, sodium chloride, potassium dihydrogen phosphate ($KH_2PO_4$) and dipotassium hydrogen phosphate ($K_2HPO_4$), Plackett-Burman design was applied. The $MgSO_4$, $FeSO_4$, and $CuSO_4$ showed positive estimate, and their concentration optimized further. The steepest ascent method and Box-Behnken method revealed that 1.5 mM $MgSO_4$, 0.33 g/l $FeSO_4$ and 1.41 mM $CuSO_4$ were optimal for the laccase production by Bacillus sp. PK4. This optimization strategy leads to enhancement of laccase production from 2.13 U/ml to 40.79 U/ml. Agro-wastes residues replace the carbon source glucose in the optimized media namely sugarcane bagasse, wheat bran, rice husk, and groundnut shell, among these groundnut shells (117 U/ml) was found to enhance the laccase production significantly. The laccase produced by Bacillus sp. PK4 was found to have the potential to degrade persistent organic pollutant benzo[a]pyrene.

• Microbiology and Biotechnology Letters
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• v.37 no.1
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• pp.62-68
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• 2009

The culture conditions to maximize the production of laccase (EC 1.10.3.2) from Fomitopsis pinicola mycelia were investigated. Among the tested media for the enzyme production, mushroom complete medium (MCM ; 2% dextrose, 0.2% peptone, 0.2% yeast extract, 0.05% $KH_2PO_4$, and 0.05% $MgSO_4{\cdot}7H_2O$) showed the highest activity of the enzyme. To optimize the culture condition for the laccase activity, influence of various carbon and nitrogen sources was investigated in MCM. Among various carbon and nitrogen sources, 2% glucose and 0.4% peptone showed the highest production of the enzyme, respectively. For the phosphorus and inorganic source, 0.05% $NaH_2PO_4$ and 0.05% $CaCl_2$ were best for the enzyme activity. The enzyme production was reached to highest level after the cultivation for 8 days at $25^{\circ}C$. Native polyacrylamide gel electrophoresis (PAGE) followed by the laccase activity staining using 2, 2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) as the substrate was performed to identify the laccase under culture conditions studied. Zymogram analysis of the culture supernatant showed a laccase band with molecular mass of 52 kDa. The optimum pH and temperature for the enzyme activity were $80^{\circ}C$ and pH 3.0.

• The Korean Journal of Mycology
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• v.13 no.2
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• pp.111-114
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• 1985

The production of lac case by the funguson various media was studied. The characteristics of the enzyme were also studied regarding to the optimum pH, stability, Km value, and inactivation. The maximum activity of laccase reached the 40 days of incubation and the barley straw extract appeared to be a strong inducer for laccase. The enzyme showed stability at wide range of pH with optimum pH of 6.6. Temperature stability of the enzyme was high. Laccase was not inactivated by the organic solvents used for the precipitation. The enzyme, how­ever, was completely inactivated by trichloroacetic acid and sodium azide.

• Journal of the Korean Wood Science and Technology
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• v.24 no.4
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• pp.74-81
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• 1996

리그닌분해능(分解能)이 높은 균주(菌株)로 선발(選拔)된 Coriolus versicolor-13 (CV-13), LKY-7 및 LKY-12세 균주(菌株)에 대하여 균체외(菌體外) laccase 생산(生産)을 검토(檢討)하였다. Glucose-peptone broth에서 균체외(菌體外) laccase활성(活性)은 CV-13의 경우 3일 이상배양후(以上培養後)에 나타났고 LKY-7과 LKY-12균주(菌株)의 laccase 활성(活性)은 배양(培養) 2일째에 검출(檢出)되었다. 탄소원(炭素源)으로서는 maltose가 glucose와 비슷한 laccase 생산효과(生産效果)를 나타냈고 질소원(窒素源)으로서는 유기태질소(有機態窒素)가 무기태(無機態) 질소(窒素)보다 효과적(效果的)이었다. Laccase 유도물질(誘導物質)로서는 2,5-Xylidine이 가장 우수하였으며 1mM 이하(以下)의 농도(濃度)에서는 유도효과(誘導效果)가 크게 나타났으나 1.5mM 이상(以上)의 농도(濃度)에서는 laccase생산(生産)이 억제(抑制)되었고, 균사생장(菌絲生長) 초기(初期)에 첨가(添加)하는 것이 효과적(效果的)으로 나타났다. SDS-PAGE 후, CV-13 균주(菌株)의 균체외(菌體外) 단백질(蛋白質)에서는 약 69, 66, 25, 23, 19kDa 크기의 laccase band가 5개 나타났고 LKY-7 균주(菌株)에서는 27kDa과 19kDa 크기의 2개 band가, LKY-12 균주(菌株)에서는 22, 20, 17kDa 크기의 laccase band가 3개 나타났다.