Structure, organization, and transcription of a cellobiohydrolase gene cluster from Phanerochaete chrysosporium

Covert, Sarah F.; Wymelenberg, Amber Vanden; Cullen, Dan

doi:10.1128/aem.58.7.2168-2175.1992

Cited by 88 publications

(53 citation statements)

References 40 publications

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“…However, the protein characteristics of Cel7 isozymes have not been investigated, except for two major isozymes (Cel7C and D) (Eriksson & Pettersson, 1975a;Streamer et al, 1975;Uzcategui et al, 1991;Igarashi et al, 1998), mainly because the minor Cel7 isozymes are extremely difficult to separate from each other. Transcription of these Cel7s has been examined by means of Northern analysis (Sims et al, 1988), reverse transcription (RT)-PCR Broda et al, 1995), and competitive RT-PCR (Covert et al, 1992b;Vanden Wymelenberg et al, 1993;Lamar et al, 1995;Vallim et al, 1998). Recently, expressed sequence tag analysis (Sato et al, 2009) and microarray analysis (Vanden Wymelenberg et al, 2009) were performed for the quantification of cel7 gene transcripts.…”

Section: Introductionmentioning

confidence: 99%

Quantitative transcriptional analysis of the genes encoding glycoside hydrolase family 7 cellulase isozymes in the basidiomycetePhanerochaete chrysosporium

Suzuki

Igarashi

Samejima

2009

FEMS Microbiology Letters

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Cellulolytic fungi generally secrete a cellulase mixture consisting mainly of glycoside hydrolase family 7 cellulases (Cel7s) during degradation of crystalline cellulose. Although several Cel7s have been investigated so far, the marked similarity in their amino acid and nucleotide sequences makes independent quantitative analysis difficult. Here, we present a real-time PCR method for the detection and quantification of Cel7 genes (cel7A-F/G) in the basidiomycete Phanerochaete chrysosporium using PCR primer sets designed based on the 3' untranslated region sequences. It was confirmed by agarose gel electrophoresis, sequencing, and dissociation curve analysis of the PCR products that each cel7 transcript was specifically amplified by the corresponding primers. We applied this real-time reverse-transcription PCR method using the presented primer sets to evaluate quantitatively the expression changes of cel7 genes in P. chrysosporium under conditions of carbon catabolite derepression.

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Section: Introductionmentioning

confidence: 99%

Quantitative transcriptional analysis of the genes encoding glycoside hydrolase family 7 cellulase isozymes in the basidiomycetePhanerochaete chrysosporium

Suzuki

Igarashi

Samejima

2009

FEMS Microbiology Letters

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“…Three kinds of these six genes were shown to be clustered within a 25-kbp region of the P. chrysosporium genome DNA. The transcript levels were di¡erent among the three kinds of genes in cellulose-nutrient medium in submerged culture [17]. On the other hand, no signi¢cant di¡erences of transcription among the cel1, cel2, and cel3 genes of I. lacteus were found by Northern hybridization.…”

Section: Phylogenetic Analysismentioning

confidence: 80%

Cloning and characterization of a new exo-cellulase gene, cel3, in Irpex lacteus

Hamada

1999

FEMS Microbiology Letters

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A new cellulose-inducible gene (named cel3) was isolated from a strain of the white rot basidiomycete, Irpex lacteus MC-2. The cel3 open reading frame, containing two introns, encodes a polypeptide of 526 amino acids residues with a molecular mass of 55 794 Da. Expression of the cel3 gene was induced by various insoluble celluloses and CM-cellulose. Transcription of cel3 was abolished when cells were cultivated in media containing the above cellulosic substrates, but added with glucose, fructose or lactose, while addition of glycerol or mannitol did not affect the cel3 mRNA level. The amino acid sequence of the catalytic domain of the Cel3 protein was homologous to that of fungal exo-type cellulases belonging to family 7 of the glycosyl hydrolases. A phylogenetic study showed that these exo-type cellulases can be clearly separated from family 7 endo-type cellulases. z

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“…According to the glycosyl hydrolases classification system that is based on amino acid sequence similarity of the catalytic domains, CBH have been placed in families 6 and 7 (Coutinho and Henrissat 1999). Typically, basic structures of fungal CBH are composed of three domains or regions: a cellulose-binding domain containing conserved cysteine residues, a catalytic domain containing conserved certain aspartic and glutamic acid residuces and a hinge region (linker) rich in glycine, serine and threonine residues (Gilkes et al 1991;Tomme et al 1995), such as T. reesei CBHI (Rouvinen et al 1990) and Phanerochaete chrysosporium CBHI (Covert et al 1992). However, CBH from H. grisea (Srisodsuk et al 1993;Takashima et al 1998), Tal.…”

Section: Discussionmentioning

confidence: 99%

Cloning of a gene encoding thermostable cellobiohydrolase from the thermophilic fungusChaetomium thermophilumand its expression inPichia pastoris

et al. 2009

Journal of Applied Microbiology

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Aims: A new cellobiohydrolase (CBH) gene (cbh3) from Chaetomium thermophilum was cloned, sequenced and expressed in Pichia pastoris. Methods and Results: Using RACE‐PCR, a new thermostable CBH gene (cbh3) was cloned from C. thermophilum. The cDNA of the CBH was 1607 bp and contained a 1356 bp open reading frame encoding a protein CBH precursor of 451 amino acid residues. The mature protein structure of C. thermophilum CBH3 only comprises a catalytic domain and lacks cellulose‐binding domain and a hinge region. The gene was expressed in P. pastoris. The recombinant CBH purified was a glycoprotein with a size of about 48·0 kDa, and exhibited optimum catalytic activity at pH 5·0 and 60 °C. The enzyme was more resistant to high temperature. The CBH could hydrolyse microcrystalline cellulose and filter paper. Conclusions: A new thermostable CBH gene of C. thermophilum was cloned, sequenced and overexpressed in P. pastoris. Significance and Impact of the Study: This CBH offers an interesting potential in saccharification steps in both cellulose enzymatic conversion and alcohol production.

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Structure, organization, and transcription of a cellobiohydrolase gene cluster from Phanerochaete chrysosporium

Cited by 88 publications

References 40 publications

Quantitative transcriptional analysis of the genes encoding glycoside hydrolase family 7 cellulase isozymes in the basidiomycetePhanerochaete chrysosporium

Quantitative transcriptional analysis of the genes encoding glycoside hydrolase family 7 cellulase isozymes in the basidiomycetePhanerochaete chrysosporium

Cloning and characterization of a new exo-cellulase gene, cel3, in Irpex lacteus

Cloning of a gene encoding thermostable cellobiohydrolase from the thermophilic fungusChaetomium thermophilumand its expression inPichia pastoris

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