Production of laccase from Trametes trogii TEM H2: a newly isolated white-rot fungus by air sampling
ABSTRACT This work represents the first report of isolation of potential laccase producers by air sampling using media supplemented with 2,2-azinobis (3-ethylbenzothiazoline-6-sulfonate) and guaiacol for laccase production and secretion indicators. Nine fungal isolates showed positive reactions with 2,2-azinobis (3-ethylbenzothiazoline-6-sulfonate) and guaiacol. The isolate named TEM H2 exhibited the largest and intensive oxidation zones with 2,2-azinobis (3-ethylbenzothiazoline-6-sulfonate) (85 mm) and guaiacol (66 mm) and therefore it was selected for detailed investigations. The strain was identified as Trametes trogii TEM H2 due to the morphological characteristics and the comparison of internal transcribed spacer ribosomal DNA gene sequences. The laccase production was screened in different liquid cultures. The best laccase production medium was determined as soluble starch yeast extract medium in which laccase production was reached to a maximum level (989.6 U l(-1) ) on the 8(th) day of cultivation. Effects of different initial pH values on laccase production were tested. Optimum pH value for laccase production in soluble starch yeast extract medium was determined as pH 3.0 with 15425.0 U l(-1) laccase production at 12(th) day of cultivation. In addition, effects of eight inducers (veratryl alcohol, ferulic acid, 1-Hydroxybenzotriazole, syringic acid, 2,2-azinobis (3-ethylbenzothiazoline-6-sulfonate), 1 mmol l(-1) CuSO(4) , 3% ethanol, guaiacol) were examined. Only cultures with 2,5-xylidine exhibited 1.9 fold increase in laccase activity reaching to 28890.0 U l(-1) . (© 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim).
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ABSTRACT: The capability of the fungi Nigrospora sp. CBMAI 1328 and Arthopyrenia sp. CBMAI 1330 isolated from marine sponge to synthesise laccases (Lcc) in the presence of the inducer copper (1–10 μM) was assessed. In a liquid culture medium supplemented with 5 μM of copper sulphate after 5 days of incubation, Nigrospora sp. presented the highest Lcc activity (25.2 U·L−1). The effect of copper on Lcc gene expression was evaluated by reverse transcriptase polymerase chain reaction. Nigrospora sp. showed the highest gene expression of Lcc under the same conditions of Lcc synthesis. The highest Lcc expression by the Arthopyrenia sp. was detected at 96 h of incubation in absence of copper. Molecular approaches allowed the detection of Lcc isozymes and suggest the presence of at least two undescribed putative genes. Additionally, Lcc sequences from the both fungal strains clustered with other Lcc sequences from other fungi that inhabit marine environments.03/2015; 5(1):19. DOI:10.1186/s13568-015-0106-7
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ABSTRACT: A basidiomycete strain MT showing lignin degradation capability was isolated and identified as a Trametes trogii strain, based on the morphological characteristics and the ITS 5.8S sequence. Enzyme production by the organism under liquid fermentation and during the decay process of a 120-day incubation period on Populus wood was studied. The results showed that the strain can produce high activities of laccase and manganese peroxidases, but no lignin peroxidases, and has the ability to simultaneously degrade lignin, cellulose, as well as hemicellulose of poplar wood. The decay pattern and process were explored using scanning electron microscopy and infrared spectroscopy. The results showed that the poplar wood is decomposed from the lumen to the middle lamellae, all the wood cell wall components are degraded, and the lignin and carbohydrate decay with no obvious differences in the levels of degradation. The decay patterns of MT in poplar wood showed the simultaneous type characteristics. The secretome of MT was surveyed using a shotgun strategy, and 65 proteins were assigned unambiguously, including the proteins with the functions of carbohydrate metabolism, cell wall and lignin degradation, fatty acid metabolism, protein metabolism and other functions. The secretome data showed that the fungus might have a complex enzymatic system implicated in lignocellulose degradation. The information presented in this paper is helpful to better understand the lignocellulose degradation mechanisms of T. trogii strains.International Biodeterioration & Biodegradation 11/2012; 75:55–62. DOI:10.1016/j.ibiod.2012.09.001 · 2.24 Impact Factor
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ABSTRACT: Laccase production by a temperature and pH tolerant fungal strain (GBPI-CDF-03) isolated from a glacial site in Indian Himalayan Region (IHR) has been investigated. The fungus developed white cottony mass on potato dextrose agar and revealed thread-like mycelium under microscope. ITS region analysis of fungus showed its 100% similarity with Trametes hirsuta. The fungus tolerated temperature from 4 to 48°C ± 2 (25°C opt.) and pH 3-13 (5-7 opt.). Molecular weight of laccase was determined approximately 45 kDa by native PAGE. Amplification of laccase gene fragment (corresponding to the copper-binding conserved domain) contained 200 bp. The optimum pH for laccase production, at optimum growth temperature, was determined between 5.5 and 7.5. In optimization experiments, fructose and ammonium sulfate were found to be the best carbon and nitrogen sources, respectively, for enhancing the laccase production. Production of laccase was favored by high carbon/nitrogen ratio. Addition of CuSO4 (up to 1.0 mM) induced laccase production up to 2-fold, in case of 0.4 mM concentration. Addition of organic solvents also induced the production of laccase; acetone showed the highest (2-fold) induction. The study has implications in bioprospecting of ecologically resilient microbial strains.04/2013; 2013:869062. DOI:10.1155/2013/869062