Laccase Gene Expression and Vinasse Biodegradation by Trametes hirsuta Strain Bm-2

ABSTRACTLaccase is a copper-containing polyphenol oxidase that has great potential in industrial and biotechnological applications. Previous research has suggested that fungal laccase may be involved in the defense against oxidative stress, but there is little direct evidence supporting this hypothesis, and the mechanism by which laccase protects cells from oxidative stress also remains unclear. Here, we report that the expression of the laccase gene from white rot fungus inPichia pastoriscan significantly enhance the resistance of yeast to H2O2-mediated oxidative stress. The expression of laccase in yeast was found to confer a strong ability to scavenge intracellular H2O2and to protect cells from lipid oxidative damage. The mechanism by which laccase gene expression increases resistance to oxidative stress was then investigated further. We found that laccase gene expression inPichia pastoriscould increase the level of glutathione-based antioxidative activity, including the intracellular glutathione levels and the enzymatic activity of glutathione peroxidase, glutathione reductase, and γ-glutamylcysteine synthetase. The transcription of the laccase gene inPichia pastoriswas found to be enhanced by the oxidative stress caused by exogenous H2O2. The stimulation of laccase gene expression in response to exogenous H2O2stress further contributed to the transcriptional induction of the genes involved in the glutathione-dependent antioxidative system, includingPpYAP1,PpGPX1,PpPMP20,PpGLR1, andPpGSH1. Taken together, these results suggest that the expression of the laccase gene inPichia pastoriscan enhance the resistance of yeast to H2O2-mediated oxidative stress by stimulating the glutathione-based antioxidative system to protect the cell from oxidative damage.

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Use of Multiplex Reverse Transcription-PCR To Study the Expression of a Laccase Gene Family in a Basidiomycetous Fungus

Applied and Environmental Microbiology ◽

10.1128/aem.69.12.7083-7090.2003 ◽

2003 ◽

Vol 69 (12) ◽

pp. 7083-7090 ◽

Cited By ~ 17

Author(s):

Tania González ◽

María C. Terrón ◽

Ernesto J. Zapico ◽

Alejandro Téllez ◽

Susana Yagüe ◽

...

Keyword(s):

Gene Expression ◽

Reverse Transcription ◽

White Rot Fungi ◽

Gene Families ◽

Veratryl Alcohol ◽

White Rot ◽

Laccase Gene ◽

Reverse Transcription Pcr ◽

Basidiomycetous Fungus ◽

Laccase Genes

ABSTRACT Laccases produced by white rot fungi are involved in the degradation of lignin and a broad diversity of other natural and synthetic molecules, having a great potential for biotechnological applications. They are frequently encoded by gene families, as in the basidiomycete Trametes sp. strain I-62, from which the lcc1, lcc2, and lcc3 laccase genes have been cloned and sequenced. A multiplex reverse transcription-PCR method to simultaneously study the expression of these genes was developed in this study. The assay proved to be quick, simple, highly sensitive, and reproducible and is particularly valuable when numerous samples are to be analyzed and/or if the amount of initial mRNA is limited. It was used to analyze the effect of 3,4-dimethoxybenzyl alcohol (veratryl alcohol) and two of its isomers (2,5-dimethoxybenzyl alcohol and 3,5-dimethoxybenzyl alcohol) on differential laccase gene expression in Trametes sp. strain I-62. These aromatic compounds produced different induction patterns despite their chemical similarity. We found 2,5-dimethoxybenzyl alcohol to be the best inducer of laccase activity while also producing the highest increase in gene expression; 3,5-dimethoxybenzyl alcohol was the next best inducer. Transcript amounts of each gene fluctuated dramatically in the presence of these three inducers, while the total amounts of laccase mRNAs seemed to be modulated by a coordinated regulation of the different genes.

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Melanoidin-containing wastewaters induce selective laccase gene expression in the white-rot fungus Trametes sp. I-62

Research in Microbiology ◽

10.1016/j.resmic.2007.10.005 ◽

2008 ◽

Vol 159 (2) ◽

pp. 103-109 ◽

Cited By ~ 23

Author(s):

Tania González ◽

María Carmen Terrón ◽

Susana Yagüe ◽

Howard Junca ◽

José María Carbajo ◽

...

Keyword(s):

Gene Expression ◽

White Rot ◽

White Rot Fungus ◽

Laccase Gene

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Differential regulation of laccase gene expression in Coriolopsis rigida LPSC No. 232

Fungal Biology ◽

10.1016/j.funbio.2010.09.010 ◽

2010 ◽

Vol 114 (11-12) ◽

pp. 999-1006 ◽

Cited By ~ 29

Author(s):

Mario Saparrat ◽

Pedro A. Balatti ◽

María Jesús Martínez ◽

Miguel Jurado

Keyword(s):

Gene Expression ◽

Differential Regulation ◽

Laccase Gene

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The genotype-specific laccase gene expression and lignin deposition patterns in apple root during Pythium ultimum infection

Fruit Research ◽

10.48130/frures-2021-0012 ◽

2021 ◽

Vol 1 (0) ◽

pp. 1-9

Author(s):

Yanmin Zhu ◽

◽

Zhe Zhou ◽

Keyword(s):

Gene Expression ◽

Pythium Ultimum ◽

Laccase Gene ◽

Deposition Patterns ◽

Lignin Deposition

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Enhancing the Laccase Production and Laccase Gene Expression in the White-Rot Fungus Trametes velutina 5930 with Great Potential for Biotechnological Applications by Different Metal Ions and Aromatic Compounds

PLoS ONE ◽

10.1371/journal.pone.0079307 ◽

2013 ◽

Vol 8 (11) ◽

pp. e79307 ◽

Cited By ~ 30

Author(s):

Yang Yang ◽

Fuxiang Wei ◽

Rui Zhuo ◽

Fangfang Fan ◽

Huahua Liu ◽

...

Keyword(s):

Gene Expression ◽

Metal Ions ◽

Aromatic Compounds ◽

White Rot ◽

White Rot Fungus ◽

Laccase Production ◽

Laccase Gene ◽

Biotechnological Applications

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Highly Efficient Production of Laccase by the Basidiomycete Pycnoporus cinnabarinus

Applied and Environmental Microbiology ◽

10.1128/aem.70.11.6379-6384.2004 ◽

2004 ◽

Vol 70 (11) ◽

pp. 6379-6384 ◽

Cited By ~ 68

Author(s):

Alexandra M. C. R. Alves ◽

Eric Record ◽

Anne Lomascolo ◽

Karin Scholtmeijer ◽

Marcel Asther ◽

...

Keyword(s):

Gene Expression ◽

Laccase Activity ◽

Expression System ◽

Schizophyllum Commune ◽

Laccase Production ◽

Efficient Production ◽

Laccase Gene ◽

Mrna Accumulation ◽

Pycnoporus Cinnabarinus ◽

Monokaryotic Strain

ABSTRACT An efficient transformation and expression system was developed for the industrially relevant basidiomycete Pycnoporus cinnabarinus. This was used to transform a laccase-deficient monokaryotic strain with the homologous lac1 laccase gene placed under the regulation of its own promoter or that of the SC3 hydrophobin gene or the glyceraldehyde-3-phosphate dehydrogenase (GPD) gene of Schizophyllum commune. SC3-driven expression resulted in a maximal laccase activity of 107 nkat ml−1 in liquid shaken cultures. This value was about 1.4 and 1.6 times higher in the cases of the GPD and lac1 promoters, respectively. lac1-driven expression strongly increased when 25 g of ethanol liter−1 was added to the medium. Accordingly, laccase activity increased to 1,223 nkat ml−1. These findings agree with the fact that ethanol induces laccase gene expression in some fungi. Remarkably, lac1 mRNA accumulation and laccase activity also strongly increased in the presence of 25 g of ethanol liter−1 when lac1 was expressed behind the SC3 or GPD promoter. In the latter case, a maximal laccase activity of 1,393 nkat ml−1 (i.e., 360 mg liter−1) was obtained. Laccase production was further increased in transformants expressing lac1 behind its own promoter or that of GPD by growth in the presence of 40 g of ethanol liter−1. In this case, maximal activities were 3,900 and 4,660 nkat ml−1, respectively, corresponding to 1 and 1.2 g of laccase per liter and thus representing the highest laccase activities reported for recombinant fungal strains. These results suggest that P. cinnabarinus may be a host of choice for the production of other proteins as well.

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