Induction of laccase activity in the white rot fungus Pleurotus ostreatus using water polluted with wheat straw extracts

Parenti, Alejandra; Muguerza, Elaia; Iroz, Amaia Redin; Omarini, Alejandra; Conde, Enma; Alfaro, Manuel; Castanera, Raúl; Santoyo, Francisco; Ramı́rez, Lucı́a; Pisabarro, Antonio G.

doi:10.1016/j.biortech.2013.01.072

Cited by 54 publications

(26 citation statements)

References 31 publications

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“…In contrast, mnp3 showed a more constant expression throughout the cycle. Specifically, the lacc10 expression profile in SmF medium was correlated with the typical growth curve of P. ostreatus in rich media (24), as its expression rose in concordance with the biomass increment and sugar consumption. Nevertheless, the profiles in the s-SSF and SSF media were exactly the opposite, with expression decreasing along the time course.…”

Section: Resultsmentioning

confidence: 84%

“…Laccase production by lignin-degrading fungi in culture media is regulated by aromatic compounds related to lignin (24,39), metals (41,42), and nitrogen and carbon sources (43,44). According to the findings of previous studies, laccase production is highly induced by lignin-related compounds, such as ferulic acid or veratryl alcohol (45), which are present in aqueous wheat straw extracts (24). These compounds can be released into the excess water in s-SSF medium, leading to the induction of laccase transcription.…”

Section: Resultsmentioning

confidence: 99%

“…The solid-state condition of the SSF medium would make these compounds less accessible to the hyphae, which could be the reason for the lower level of expression. In the case of SmF medium, the initial repression of lacc10 likely originated from the glucose concentration in the fresh medium compared with that in the inoculum (which is consumed after 6 days of growth [24]). In other studies, we observed that glucose represses lacc10 expression while it induces the expression of other laccase genes, such as lacc6.…”

Section: Resultsmentioning

confidence: 99%

“…Experimental conditions. To include different growth stages in our analyses, the sampling times were based on the findings of previous studies analyzing the variations observed during P. ostreatus growth (24). Three independent biological replicates were sampled for each culture medium.…”

Section: Methodsmentioning

confidence: 99%

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Validation of Reference Genes for Transcriptional Analyses in Pleurotus ostreatus by Using Reverse Transcription-Quantitative PCR

Castanera

López-Varas

Pisabarro

et al. 2015

Appl Environ Microbiol

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Recently, the lignin-degrading basidiomycete Pleurotus ostreatus has become a widely used model organism for fungal genomic and transcriptomic analyses. The increasing interest in this species has led to an increasing number of studies analyzing the transcriptional regulation of multigene families that encode extracellular enzymes. Reverse transcription (RT) followed by real-time PCR is the most suitable technique for analyzing the expression of gene sets under multiple culture conditions. In this work, we tested the suitability of 13 candidate genes for their use as reference genes in P. ostreatus time course cultures for enzyme production. We applied three different statistical algorithms and obtained a combination of stable reference genes for optimal normalization of RT-quantitative PCR assays. This reference index can be used for future transcriptomic analyses and validation of transcriptome sequencing or microarray data. Moreover, we analyzed the expression patterns of a laccase and a manganese peroxidase (lacc10 and mnp3, respectively) in lignocellulose and glucose-based media using submerged, semisolid, and solid-state fermentation. By testing different normalization strategies, we demonstrate that the use of nonvalidated reference genes as internal controls leads to biased results and misinterpretations of the biological responses underlying expression changes.T he basidiomycete Pleurotus ostreatus is an efficient producer of extracellular enzymes, such as laccases (Lacs; EC 1.10.3.2) and manganese peroxidases (MnPs; EC 1.11.1.13), which are notable for their application in multiple industrial and biotechnological processes (1, 2). It has a special relevance in agroindustry due to its worldwide cultivation for mushroom production, and it has also been studied for its nutritional and medicinal value (3). Currently, two genomic sequences of P. ostreatus are publicly available, and it has become a very popular model organism for the study of fungal genetics and comparative genomics (4-6). Additionally, many studies on the production of ligninolytic enzymes, with a focus on culture optimization for enzyme production, have recently been published. Most of these enzymes are encoded by genes organized in gene families, which are occasionally arranged into clusters as a result of gene duplications.Despite the information uncovered by sequencing efforts, substantial work on the functional characterization of gene family members remains to be performed. In this sense, transcriptomic analyses are powerful tools for providing an understanding of gene regulation and the presumptive function of genes. Despite the high productivity and low cost of transcriptome sequencing (RNA-seq), reverse transcription (RT) followed by real-time PCR (RT-quantitative PCR [RT-qPCR]) is likely the best option for analyzing the expression patterns of certain genes under multiple conditions. Additionally, it is the most reliable technique for validating RNA-seq and microarray data because of its specificity, reproducibility, and capacity...

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Section: Resultsmentioning

confidence: 84%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

Validation of Reference Genes for Transcriptional Analyses in Pleurotus ostreatus by Using Reverse Transcription-Quantitative PCR

Castanera

López-Varas

Pisabarro

et al. 2015

Appl Environ Microbiol

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“…10,11,12,13 However, the laccase production by basidiomycetes has been reported in solid state fermentation (SSF) 14,15 as well as in submerged liquid fermentation (SmF) sys-tem. 16,17 SSF is generally defined as the growth of microorganisms on solid materials in the absence or near absence of free water. It is also treated as an adequate method for filamentous fungi cultivation and lignocellulolytic enzyme production.…”

Section: Introductionmentioning

confidence: 99%

Effect of Solid State Fermentation Medium Optimization on Pleurotus ostreatus Laccase Production

Belšak-Šel

Gregori

Leitgeb

et al. 2015

ACSi

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The objective of this work was to increase laccase production by Pleurotus ostreatus PLAB through culture medium optimization using solid-state culture conditions. Increased laccase activity was obtained through the design of experiments (DOE) using the Taguchi orthogonal array (OA). Seven factors, viz. lignocellulose, glucose, yeast extract, peptone, KH 2 PO 4 , MgSO 4 · 7H 2 O and MnSO 4 · H 2 O at three levels and pH at two levels with OA layout of L18 (2 1 × 3 7 ) were selected for the proposed experimental design using Minitab 17 software. Data analysis showed that lignocellulose (20%) and glucose (10 g L were the optimal conditions to maximize laccase production. The model predicted a 30.37 U g -1 dry wt., which agreed with the experimentally obtained laccase activity 29.15 U g -1 dry wt. at optimal conditions.

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Pleurotus ostreatus: A Biofactory for Lignin-Degrading Enzymes of Diverse Industrial Applications

Enshasy

Agouillal

Mat

et al. 2019

Recent Advancement in White Biotechnology Through Fungi

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Induction of laccase activity in the white rot fungus Pleurotus ostreatus using water polluted with wheat straw extracts

Cited by 54 publications

References 31 publications

Validation of Reference Genes for Transcriptional Analyses in Pleurotus ostreatus by Using Reverse Transcription-Quantitative PCR

Validation of Reference Genes for Transcriptional Analyses in Pleurotus ostreatus by Using Reverse Transcription-Quantitative PCR

Effect of Solid State Fermentation Medium Optimization on Pleurotus ostreatus Laccase Production

Pleurotus ostreatus: A Biofactory for Lignin-Degrading Enzymes of Diverse Industrial Applications

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