• Journal of Mushroom
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• v.2 no.1
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• pp.33-37
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• 2004

To develop neohaplonts for Ganoderma breeding, protoplasts were isolated from dikaryotic mycelium and regenerated. Selection rate of neohaplonts varied between ASI7074, ASI7091, ASI7094, ASI7100 and ASI7115, showing 5.24% on the average. Auxotropic mutants from Ganoderma monokarions were recovered by UV irradiation on protoplasts. Protoplast survival rates were 1.9% ASI 7074, 0.17% ASI 7091, and zero percent ASI 7100 using 300 second irradiation. Four auxotrophic strains were recovered from 1,536 colonies screened that will be further utilized for protoplast fusion and transformation.

• Microbiology and Biotechnology Letters
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• v.27 no.2
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• pp.145-152
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• 1999

Brettanomyces custersii CBS 5512 which has reported as a thermotolerant glucose-cellobiose co-fermentable yeast strain was mutated with UV and NTG to improve ethanol yield at higher than 4$0^{\circ}C$ B. custersii H1-23, H1-39, H1-55 and H1062 were finally selected for hyper-fermentable strains at higher than 4$0^{\circ}C$ from thermotolerant 7510 colonies through 5th selection. Among the selected strains, H1-39 mutant had better fermentability at 4$0^{\circ}C$ and 43$^{\circ}C$ from different concentrations of glucose. H1-39 and H1-23 mutants yielded more than 70% of the theoretical ethanol yield in 4 and 8% mixed sugars at above 4$0^{\circ}C$, which was 5-11% higher than those by original strain. Especially, H1-39 mutant had better fermentability in 4% mixed sugar. It showed 78.5% of the theoretical yield at 4$0^{\circ}C$ and 72.2% of the theoretical yield at 43$^{\circ}C$. On the other hand, theoretical yield of ethanol by H1-39 mutant in 8% mixed sugar at 4$0^{\circ}C$ and 43$^{\circ}C$ were 75.2% and 70.2%, respectively. Theses values increased up to 7-11% as compared to those by orginal strain. By the simultaneous saccharification and fermentation, ethanol production by H1-39 mutant increased up to more than 23% as compared to that by original strain.

• Journal of Animal Science and Technology
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• v.51 no.6
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• pp.527-536
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• 2009

A potent protein degrading bacterium was isolated from soil samples of different environments. Polyphasic taxonomic studies and phylogenetic 16S rRNA sequence analyses led to identify the isolate IB No. 11 as a strain of Bacillus subtilis. The isolated strain was recognized to produce protease constitutively, and the maximum production (1.64 units/ml) was attained in a shake flask culture when the isolate was grown at $40^{\circ}C$, for 32 h in basal medium supplemented with starch (0.25%) and gelatin (1.25%) as sole carbon and nitrogen source, respectively. The optimum pH and temperature for the protease activity were determined to be pH 7.0 and $50^{\circ}C$, respectively. $Ca^{2+}$ and $Mn^{2+}$ enhanced remarkably the protease activity but neither showed positive effect on the protease's thermal stability. In addition, it was observed that the protease was fairly stable in the pH range of 6.5-8.0 and at temperatures below $50^{\circ}C$, and it could be a good candidate for an animal feed additive. The inhibition profile of the protease by various inhibitors indicated that the enzyme is a member of serine-proteases. A combination of UV irradiation and NTG mutagenesis allowed to develop a protease hyper-producing mutant strain coded as IB No. 11-4. This mutant strain produced approximately 3.23-fold higher protease activity (6.74 units/mg) than the parent strain IB No. 11 when grown at $40^{\circ}C$ for 32h in the production medium. The protease production profile of the selected mutants was also confirmed by the zymography analysis.

• Microbiology and Biotechnology Letters
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• v.37 no.2
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• pp.140-146
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• 2009

Cyclosporin, an immunosuppressant, is a representative group of biologically active secondary metabolites produced by the fungus Tolypocladium inflatum. The amount and ratio of cyclosporin derivatives in the culture broth are an important factors for the production of cyclosporin A and the purification in the industrial process. Therefore, we studied the effect of amino acids and complex organic nitrogen sources using Tolypocladium inflatum mutants on the productivity of cyclosporin A and the ratio of cyclosporin derivatives. Overproducing mutant YHC-004 having seven times higher productivity than mother strain's could be obtained through the artificial mutation by UV irradiation. The concentration and kind of organic nitrogens and amino acids shows the profound effect on the productivity of cyclosporin A and ratio of cyclosporin derivatives. As a result, it was possible to raise the productivity and the ratio of cyclosporin A up to 3,430 mg/L and 93% respectively, but on the other hand the other cyclosporin derivatives decreased less than 2% in the culture broth.

• The Korean Journal of Mycology
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• v.41 no.1
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• pp.1-8
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• 2013

COP9 signalosome (CSN), which is originally identified as the regulator of the photomorphogenic development in plant, is highly conserved protein complex in diverse eukaryotic organisms. Most eukaryotic CSN complex is composed of 8 subunits, which is structurally and functionally similar to the lid subunit of 26S proteasome and eIF3 translation initiation complex. CSN play important functions in the regulation of cell cycle and checkpoint response by controlling Cullin-Ring E3 ubiquitin ligases (CRL) activities. CSN exhibits an isopeptidase activity which cleaves the neddylated moiety of cullin components. In fission yeast, S-phase cell cycle progression was delayed and the sensitivity to g-ray or UV was increased in CSN1 and CSN2 deletion mutants, indicating that yeast CSN is also involved in the checkpoint regulation. CSN in fungal system more closely resembles that of the higher organisms in the structure and assembly of their components. Functionally, CSN is associated with the regulation of conidiation rhythms in Neurospora crassa and the sexual development in Aspsergillus nidulans. Recent studies also revealed that CSN functions as an essential cell cycle regulator, playing key roles in the regulation of DNA replication and DNA damage response in Aspergillus. Overall, CSN of microorganisms, such as fission yeast and fungi, share functionally common aspects with higher organisms, implying that they can be useful tools to study the role of CSN in the CRL-mediated diverse cellular activities.

• Microbiology and Biotechnology Letters
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• v.34 no.2
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• pp.136-142
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• 2006

Haematococcus pluvialis is a great producer of astaxanthin (3,3'-dihydroxy-$\beta$,$\beta$-carotene-4,4'-dione). The activities of astaxanthin include potential cancer prevention, immune response enhancement, antioxidant activity, and so on. Nevertheless, it tried to manipulate by mutation for overcoming low growth rate of wild type and limited production of astaxanthin. Mutated colony that is lager and more reddish one than wild type was selected by attempting to expose strains to UV irradiation and to treat chemical such as EMS and colchicines as mutagen. Selected mutants were further screened using inhibitors of the carotenoid biosynthetic pathway. Inhibitors used were nicotine and diphenylamine and both had decreased the survival rate by 40-50%. Among over 50,000 mutant colonies screened, two strains were selected. One selected mutant strain (U15-5) from UV treatment showed 1.68-fold higher total carotenoid contents per cell than that of the wild type strain. On the other hand, the other selected mutant strains (DS, M4-3) from colchicine treatment showed 20$\sim$30% faster cell growth than the wild type strain.

• The Korean Journal of Mycology
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• v.19 no.4
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• pp.266-275
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• 1991

The gene CDC70 encoding the${\alpha}-subunit$ of G protein has been known to be a component involved in mating pheromone signalling in the yeast, Saccharomyces cerevisiae. To isolate mutations of the genes involved in the signal transduction, Saccharomyces cerevisiae the strain bearing the cdc70-5 mutation was mutagenized to be forced to recover the ability of colony-formation at restrictive temperature, which means the new mutation can suppress the temperature sensitivity of the cdc70-5 phenotypes. Among these suppressors, $sir^-$ and $mat{\alpha}2^{-}$ mutations are excluded because of no relationship to signal transducer. And the selected suppressors were analyzed for the linkage relationships by the tetrad analysis. Out of fifteen suppressors isolated, twelve were classified into four linkage groups, designated as sga1, sga2, sga3, sga4 by the tetrad analysis. The other three genes were determined for the linkage.

• Journal of Life Science
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• v.15 no.3 s.70
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• pp.415-422
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• 2005

library of the mutants was prepared by transposon mutagenesis of the Mycobacterium bovis BCG. We screened this library for the resistance to an anti-tuberculosis antibiotic, PA-824. Most of the mutants resistant to the PA-824 were not able to synthesize the coenzyme $F_{420}$ which is normally produced by the wild type M. bovis BCG strains. HPLC analysis of the cellular extract showed that one of those mutants which lost the ability to synthesize $F_{420}$ still produced F0. The insertion site of the transposon in this mutant was determined by an inverse PCR and the transposon was found to be inserted in the Rv2435c open reading frame (ORF). Rv2435c ORF is predicted to encode an 80.3 kDa protein. Rv2435c protein appears to be bound to the cytoplasmic membrane, its N-terminal present in the periplasm and C-terminal in the cytoplasm. The C-terminal portion of this protein is highly homologous with the adenylyl cyclases of both prokaryotes and eukaryotes. There are 15 ORFs which have homology with the class III AC proteins in the genome of the M. tuberculosis and M. bovis. Two of those, Rv1625c and Rv2435c, are highly homologous with the mammalian ACs. We cloned the cytoplasmic domain of the Rv2435c ORF and expressed it with six histidine residues attached on its C-terminal in Escherichia coli to find out if this protein is a genuine AC. Production of that protein in E. coli was proved by purifying the histidine-tagged protein by using the Ni-NTA resin. This protein, however, failed to complement the cya mutation in E. coli, indicating that this protein lacks the AC activity. All of the further attempts to convert this protein to a functional AC by a mutagenesis with UV or hydroxylamine, or construction of several different fusion proteins with Rv1625c failed. It is, therefore, possible that Rv2435c protein might affect the conversion of F0 to $F_{420}$ not by synthesizing cAMP but by some other way.

• Korean Journal of Food Science and Technology
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• v.14 no.1
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• pp.72-79
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• 1982

Several mutants were isolated from the parent strain of Aspergillus niger CF: the first mutant strain CF-11 was obtained by UV irradiation, and the second mutant strain CF-21 and CF-22 were from NTG (N-methyl-N'-nitroso-N-nitroso-guanidine) treatment on the CF-11. These mutants were characterized, and their enzyme and acid production on wheat bran Koji and wheat flour Koji were studied. Asp. niger CF-22 mutant appeared to be tan type which conidial heads were discolored. It's glucoamylase activity was inreased approximately two times and its ${\alpha}-amylase$ about 50 percent as compared with that of the parent strain of Asp. niger CF, when grown on wheat bran Koji under the optimal conditions. Asp. niger CF-21 mutant showed slower growth rate and poor sporulation than the wild type, although its conidial heads were not discolored. Approximately 4-fold increment in its acid production was observed as compared with the weld type. The activities of glucoamylase and ${\alpha}-amylase$ of the Asp. niger CF-22 and CF-21 mutants were higher than those of the wild type, but their protease activity was rather lower. The maximum production of glucoamylase by the Asp. niger CF-22 mutant was obtained after 2 to 3 days incubation on wheat bran at 30 to $35^{\circ}C;$ ${\alpha}-amylase$2 days incubation at 30 to $35^{\circ}C$. The maximal levels of acid production by the mutant CF-21 was appeared after 2 days incubation on wheat bran Koji, and after 3 days on wheat flour Koji at $30^{\circ}C$. Little differences in the levels of acid production were observed between on wheat bran and flour Koji.

• Korean Journal of Veterinary Research
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• v.40 no.3
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• pp.471-478
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• 2000

The pharmacokinetic properties of KR-V compounds, recently developed as new anti-HIV agents, were studied after i.v. and p.o. administration in rats. The concentrations of the KR-V series were determined in rat plasma using an high-performance liquid chromatography (HPLC)-UV detection system. Of the 19 KR-V compounds investigated in the present study, only KR-V 3, 10, 14, 16 and 18-1 showed oral bioavailability. The plasma concentration-time data could be adequately described by an one-compartment open model. In the i.v. kinetic study (10mg/kg), the CLt of KR-V 3, 10, 14 and 16 (>4L/hr/kg) were significantly higher than that of KR-V 18-1 (1.1 L/hr/kg). The AUC of KR-V 18-1 was greater ($8.97{\mu}g{\cdot}hr/ml$) than that of the other compounds, but the Vd (0.58 L/kg) was lower. In the p.o. kinetic study (50mg/kg), although the t-1/2 of KR-V 18-1 was shorter than that of the other compounds, the AUC ($3.659{\mu}g{\cdot}hr/ml)$ and $C_{max}(1.891{\mu}g/ml$) were markedly higher. In a seperated in vitro experiment, only KR-V 18-1, of the 5 compounds with bioavailibility, exhibits potent activity against HIV-1 mutant strains. Therefore, KR-V 18-1 is expected to become a new potent anti-AIDS drug candidate/lead compound.