The White-Rot Fungus<i>Pleurotus ostreatus</i>Transformant Overproduced Intracellular cAMP and Laccase

Yao, Yuki; Sakamoto, Takako; Honda, Yoshio; Kagotani, Yasuyuki; Izumitsu, Kousuke; Suzuki, Kaoru; Irie, Toshikazu

doi:10.1271/bbb.130470

Cited by 8 publications

(16 citation statements)

References 30 publications

(41 reference statements)

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“…A cAMP-overproducing strain (65367Q218R2), which was previously made by introducing a mutated heterotrimeric G protein α subunit (Gα) isoform gene into strain PC9, was used for quantitative determination of CaM transcript (Yao et al 2013). Fungal strains were grown and maintained in potato-dextrose agar (PDA; Becton, Dickinson and Company, USA) medium.…”

Section: Strain and Culture Conditionsmentioning

confidence: 99%

“…1a), by inserting the CaM ORF downstream of the β-tubulin promoter of the pB-TUB_DEST_GFP vector using the Gateway system (Life Technologies). The pB-TUB_DEST vector was constructed by replacing the sdi1 promoter in pIp-DEST-GFP (Yao et al 2013) with a β-tubulin promoter that is highly expressed in strain PC9 (Salame et al 2012). To construct a repression vector (pB-TUB_PoCaM_shRNA vector, Fig.…”

Section: Cloning Of Cam Construction Of Vectors and Transformationmentioning

confidence: 99%

“…To investigate whether the CaM gene is also transcriptionally regulated by intracellular cAMP in P. ostreatus, we measured the absolute quantity of gene transcript after 7 days of culture of a cAMP-overproducing strain, which was previously made by introducing a mutated Gα isoform gene (Yao et al 2013). The absolute quantity of CaM transcript in the cAMP-overproducing strain was twofold higher than that in the wild-type PC9 strain (Fig.…”

Section: Cam Gene Transcription In a Camp-overproducing Strainmentioning

confidence: 99%

“…Whole genome sequencing also revealed one copy of a gene encoding CaM. The release of the whole genome sequence of P. ostreatus, reproducible transformation systems (Amore et al 2012;Honda et al 2000;Irie et al 2001a, b;Yao et al 2013), a gene disruption system using the Δku80 strain (Salame et al 2012(Salame et al , 2013(Salame et al , 2014 and RNAi systems (Salame et al 2010) make the fungus a good model strain for analysis of physiological processes in white-rot basidiomycetes.…”

Section: Introductionmentioning

confidence: 99%

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Effects of calmodulin on expression of lignin-modifying enzymes in Pleurotus ostreatus

et al. 2014

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Previously, we suppressed the expression of genes encoding isozymes of lignin peroxidase (LiP) and manganese peroxidase (MnP) using a calmodulin (CaM) inhibitor, W7, in the white-rot fungus Phanerochaete chrysosporium; this suggested that CaM positively regulates their expression. Here, we studied the role of CaM in another white-rot fungus, Pleurotus ostreatus, which produces MnP and versatile peroxidase (VP), but not LiP. W7 upregulated Mn(2+)-dependent oxidation of guaiacol, suggesting that CaM negatively regulates the production of the enzymes. Suppression of CaM in P. ostreatus using RNAi also led to upregulation of enzyme activity, whereas overexpression of CaM in P. ostreatus caused downregulation. Real-time RT-PCR showed that MnP1-6 and VP3 levels in the CaM-knockdown strain were higher than those in the wild-type strain, while MnP-5 and -6 and VP1 and 2 levels in the CaM-overexpressing strain were lower than in the wild type. Moreover, we also found that another ligninolytic enzyme, laccase, which is not produced by P. chrysosporium, was negatively regulated by CaM in P. ostreatus similar to MnP and VP. Although overexpression of CaM did not reduce the ability of P. ostreatus to digest beech wood powder, the percentage of lignin remaining in the digest was slightly higher than in the wild-type strain digest.

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Section: Strain and Culture Conditionsmentioning

confidence: 99%

Section: Cloning Of Cam Construction Of Vectors and Transformationmentioning

confidence: 99%

Section: Cam Gene Transcription In a Camp-overproducing Strainmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Effects of calmodulin on expression of lignin-modifying enzymes in Pleurotus ostreatus

et al. 2014

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“…These enzymes are induced, and the transcription gene of laccases is regulated by different carbon and nitrogen sources (Bertrand et al 2013). In strategies to increase production, inducers have been added to the culture medium of the fungus, such as 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) (Hou et al 2004), ferulic acid (Yao et al 2013), plant residues (Quevedo-Hidalgo et al 2015), and azo dyes (Camacho et al 2015). However, production times are still very long and costly.…”

Section: Introductionmentioning

confidence: 99%

Heterologous Expression of Laccase (LACP83) of Pleurotus ostreatus

et al. 2017

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The heterologous expression of the gene LACP83 (encoding a laccase) from Pleurotus ostreatus in Escherichia coli was characterized. The laccase enzyme activity and kinetics of bacterial growth with an inducer (IPTG) and without inducer were determined. The maximum enzymatic activity was observed at 7 h post induction with a value of 3740 ± 342 U/L, which was similar to that reported for the native strain of P. ostreatus at 144 h of culture. Furthermore, the induction of laccase with IPTG reduced the specific growth rate of recombinant E. coli BL21 by approximately 50%. These results support the use this system for the recombinant production of the enzyme on an industrial scale.

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The ligninolytic peroxidases in the genus Pleurotus: divergence in activities, expression, and potential applications

Knop

Yarden

Hadar

2014

Appl Microbiol Biotechnol

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Mushrooms of the genus Pleurotus are comprised of cultivated edible ligninolytic fungi with medicinal properties and a wide array of biotechnological and environmental applications. Like other white-rot fungi (WRF), they are able to grow on a variety of lignocellulosic biomass substrates and degrade both natural and anthropogenic aromatic compounds. This is due to the presence of the non-specific oxidative enzymatic systems, which are mainly consisted of lacasses, versatile peroxidases (VPs), and short manganese peroxidases (short-MnPs). Additional, less studied, peroxidase are dye-decolorizing peroxidases (DyPs) and heme-thiolate peroxidases (HTPs). During the past two decades, substantial information has accumulated concerning the biochemistry, structure and function of the Pleurotus ligninolytic peroxidases, which are considered to play a key role in many biodegradation processes. The production of these enzymes is dependent on growth media composition, pH, and temperature as well as the growth phase of the fungus. Mn(2+) concentration differentially affects the expression of the different genes. It also severs as a preferred substrate for these preoxidases. Recently, sequencing of the Pleurotus ostreatus genome was completed, and a comprehensive picture of the ligninolytic peroxidase gene family, consisting of three VPs and six short-MnPs, has been established. Similar enzymes were also discovered and studied in other Pleurotus species. In addition, progress has been made in the development of molecular tools for targeted gene replacement, RNAi-based gene silencing and overexpression of genes of interest. These advances increase the fundamental understanding of the ligninolytic system and provide the opportunity for harnessing the unique attributes of these WRF for applied purposes.

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The White-Rot FungusPleurotus ostreatusTransformant Overproduced Intracellular cAMP and Laccase

Cited by 8 publications

References 30 publications

Effects of calmodulin on expression of lignin-modifying enzymes in Pleurotus ostreatus

Effects of calmodulin on expression of lignin-modifying enzymes in Pleurotus ostreatus

Heterologous Expression of Laccase (LACP83) of Pleurotus ostreatus

The ligninolytic peroxidases in the genus Pleurotus: divergence in activities, expression, and potential applications

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