• Journal of Life Science
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• v.13 no.4
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• pp.522-528
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• 2003

The RAD4 gene of Saccharomyces cerevisiae is essential for the incision step of UV-induced excision repair. A yeast RAD4 gene has been previously isolated by functional complementation. In order to identify the RAD4 homologous gene from fungus Coprinus cinereus, we have constructed cosmid libraries from electrophoretically separated chromosomes of the C. cinereus. The 13 C. cinereus chromosomes were resolved by pulse-field gel electrophoresis, hybridized with S. cerevisiae RAD4 DNA, and then isolated homologous C. cinereus chromosome. The insert DNA of the RAD4 homolog was contained 3.2 kb. Here, we report the characterization of fungus C. cinereus homolog of yeast RAD4 gene. Southern blot analysis confirmed that C. cinereus contains the RAD4 homolog gene and this gene exists as a single copy in C. cinereus genome. When total RNA isolated from C. cinereus cells was hybridized with the 1.2 kb PvuII DNA fragment of the S. cerevisiae RAD4 gene, a 2.5 kb of transcript was detected. In order to investigation whether the increase of transcripts by DNA damaging agent, transcripts levels were examined after treating the cells. The level of transcript did not increase by untraviolet light (UV). This result indicated that the RAD4 homologous gene is not UV inducible gene. Gene deletion experiments indicate that the RAD4 homologous gene is essential for cell viability.

• The Korean Journal of Mycology
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• v.22 no.1
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• pp.31-35
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• 1994

Coprinus cinereus resistant strain against validamycin was successfully isolated. Germination rate of oidiospores, branching pattern, and localization of actin protein of the resistant strain were compared with normal strain. The resistant strain showed better germiantion rate of oidia (about 20 fold), more frequent branching, and even actin localization on validamycin plate where actin content was severely reduced in case of normal strain.

• Mycobiology
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• v.32 no.4
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• pp.164-169
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• 2004

Development of oidia, a type of thallic spores from monokaryotic mycelium of Coprinus cinereus was examined with electron microscope and light microscopes. Oidia formation in this fungus is unique in its mode of formation compared with other types of asexual sporogenesis. Oidiogenesis in C. cinereus is carried out in three steps: 1) Formation of oidiophore from the parent mycelium, 2) Formation of initials of oidial cells from swollen oidiophore, 3) Segmentation and detachment of mature oidial cell. Oidiophores appear to spring out singly as a swollen hyphal branches from the normal foot hyphae or sometimes coiled hypha. From the oidiophore, oidial branches sprout out forming a group of $2{\sim}6$, most often 4 oidial cells and each oidial cell undergoes a single mitosis resulting in 2 oidia. One of the sibling oidial cells in a group is frequently transformed into a new oidiophore, thus oidiogenic structures are tandemly produced at the several different levels.

• KSBB Journal
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• v.23 no.4
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• pp.340-344
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• 2008

Coprinus cinereus peroxidase (CiP) was often used as a catalyst for oxidative polymerization of a variety of phenol derivatives to produce a new class of polyphenols. Economical point of view, to know the mechanism of enzyme deactivation is significantly important because cost of enzyme is critically high. Hydrogen peroxide being used as oxidizing agent induced deactivation of peroxidase by destruction of heme structure. In the presence of hydrogen peroxide the stability of peroxidase was unexpectedly improved by adding organic solvent. Especially 2-propanol significantly improved enzyme stability among tested solvents. Radical scavenging by organic solvents may play a major role in protecting peroxidase from the oxidation of oxidizing radicals.

• Journal of Microbiology and Biotechnology
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• v.19 no.9
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• pp.966-971
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• 2009

Peroxidase from Coprinus cinereus (CiP) has attracted attention for its high specific activity and broad substrate spectrum compared with other peroxidases. In this study, the functional expression of this peroxidase was successfully achieved in the methylotrophic yeast Pichia pastoris. The expression level of CiP was increased by varying the microbial hosts and the expression promoters. Since a signal sequence, such as the alpha mating factor of Saccharomyces cerevisiae, was placed preceding the cDNA of the CiP coding gene, expressed recombinant CiP (rCiP) was secreted into the culture broth. The Mut Pichia pastoris host showed a 3-fold higher peroxidase activity, as well as 2-fold higher growth rate, compared with the $Mut^s $ Pichia pastoris host. Furthermore, the AOX1 promoter facilitated a 5-fold higher expression of rCiP than did the GAP promoter.

• Journal of Life Science
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• v.14 no.6 s.67
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• pp.1023-1027
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• 2004

The RAD3 gene of Saccharomyces cerevisiae is essential for the incision step of UV-induced excision repair. An yeast RAD3 gene has been previously isolated by functional complementation. In order to identify the RAD3 homologous gene from fungus Coprinus cinereus, we have constructed cosmid libraries from electrophoretically separated chromosomes of the C. cinereus. The 13 C. cinereus chromosomes were resolved by pulse-field gel electrophoresis, hybridized with S. cerevisiae RAD3 DNA, and then isolated RAD3 homologous DNA from C. cinereus chromosome. The RAD3 homolog DNA was contained in 3.2 kb DNA fragment. Here, we report the results of characterization of a fungus C. cinereus homolog to the yeast RAD3 gene. Southern blot analysis confirmed that the C. cinereus chromosome contains the RAD3 homolog gene and this gene exists as a single copy in C. cinereus genome. When total RNA isolated from the C. cinereus cells were hybridized with the 3.4 kb PvuII DNA fragment of the S. cerevisiae RAD3 gene, transcripts size of 2.8 kb were detected. In order to investigate whether the increase of the amount of transcripts by DNA damaging agent, transcript levels were examined after treating agents to the cells. The level of transcripts were not increased by untraviolet light (UV). This result indicated that the RAD3 homologous gene is not UV inducible gene. Gene deletion experiments indicate that the HRD3 gene is essential for viability of the cells and DNA repair function. These observations suggest an evolutionary conservation of other protein components with which HRD3 interacts in mediating its DNA repair and viability functions.

• Journal of Korea Technical Association of The Pulp and Paper Industry
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• v.31 no.5
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• pp.12-17
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• 1999

This study was to evaluate applications of the alkalophilic enzymes from Coprinus cinereus 2249 with old newsprint(ONP). A enzymatic deinking process based on alkalopholic enzymes was investigated . It was found that alkalophilic enzymes could effectively deink old newsprint. When applied on deinking of the old newsprint, it increases the freeness and brightness due to effect of hydrolysis at 0.1% enzyme concentration . Also, the physical properties of deinked pulp were improved.

• Journal of Life Science
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• v.10 no.2
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• pp.50-54
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• 2000

The RAD4 gene of Saccharomyces cerevisiae is essential for the incision step of UV-induced excision repair. A yeast RAD4 gene has been previously isolated by functional complementation. In order to identify the RAD4 homologous gene from fungus Coprinus cinereus, we have constructed cosmid libraries from electrophoretically separated chromosomes of the C. cinereus. The 13 C. cinereus chromosomes were resolved by pulse-field gel electrophoresis, hybridized with S. cerevisiae RAD4 DNA, and then isolated homologous C. cinereus chromosome. Here, we report the cloning and characterization of fungus C. cinereus homolog of yeast RAD4 gene. Southern blot analysis confirmed that C. cinereus contains the sequence homologous DNA to RAD4 gene and this gene exists as a single copy in C. cinereus genome. When total RNA isolated from C. cinereus cells was hybridized with the 3.4 kb BglII DNA fragment of the S. cerevisiae RAD4 gene, a 2.5 kb of transcript was detected. The isolated gene encodes a protein of 810 amino acids.

• Environmental Mutagens and Carcinogens
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• v.17 no.1
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• pp.1-6
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• 1997

The RAD4 gene of Saccharomyces cerevisiae is essential for the incision step of UV-induced excision repair. A yeast RAD4 gene has been previously isolated by functional complementation. In order to identify the RAD4 homologous gene from fungus Coprinus cinereus, we have constructed cosmid libraries from electrophoretically separated chromosomes of the C. cinereus. The 13 C. cinereus chromosomes were resolved by pulse-field gel electrophoresis, hybridized with S. cerevisiae RAD4 DNA, and then isolated homologous C. cinereus chromosome. The insert DNA of the RAD4 homolog was contained 3.2 kb. Here, we report the partial cloning and characterization of fungus C. cinereus homolog of yeast RAD4 gene. Southern blot analysis confirmed that C. cinereus contains the sequence homologous DNA to RAD4 gene and this gene exists as a single copy in C. cinereus genome. When total RNA isolated from C. cinereus cells was hybridized with the 1.2 kb PvuII DNA fragment of the S. cerevisiae RAD4 gene, a 2.5 kb of transcript was detected. The level of the transcript did not increase upon UV-irradiation, suggesting that the RAD4 homologous gene in C. cinereus is not UV-inducible.

• Journal of Korea Technical Association of The Pulp and Paper Industry
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• v.36 no.1
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• pp.24-29
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• 2004

Alkaline cellulolytic enzymes from cultured medium of Coprinus cinereus 2249 were purified with gel and ion-exchange chromatography and characteristics of those enzyme proteins were investigated. A fiber length distribution and a crystallinity of cellulose and sugar composition of enzyme treated Mixed Office Wastepaper(MOW) and Unbleached Kraft Pulp(UKP) were analysed. The conclusion could summarized as follows; \circled1 Alkaline and acidic, endo- and exo-glucanases were purified from cultured medium of Coprinus cinereus 2249. \circled2 The approximate molecular weight of alkaline endo-glucanase was 42 kDa, and also that of alkaline exo-glucanase was 50 kDa. A fiber length distribution and a crystallization of cellulose and sugar composition of enzyme treated MOW and UKP were not so much changed with original paper and pulp.