• Journal of Microbiology and Biotechnology
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• v.19 no.4
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• pp.391-396
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• 2009

The chemical compositions, and antibacterial and antifungal effects of essential oils extracted from three coniferous species, Pinus densiflora, Cryptomeria japonica, and Chamaecyparis obtusa, were investigated. Gas chromatography mass analysis of the essential oils revealed that the major components and the percentage of each essential oil were 16.66% $\beta$-phellandrene and 14.85% $\alpha$-pinene in P. densiflora; 31.45% kaur-16-ene and 11.06% sabinene in C. japonica; and 18.75% bicyclo [2,2,1] heptan-2-ol and 17.41% 2-carene in Ch. obtusa. The antimicrobial assay by agar disc diffusion method showed that $2.2{\mu}g$ of Ch. obtusa oil inhibited most effectively the growth of Escherichia coli ATCC 33312 and Klebsiella oxytoca ATCC 10031, whereas the C. japonica oil gave weak antimicrobial activity. The minimal inhibitory concentration(MIC) values for bacterial strains were in the range of 5.45-21.8 mg/ml depending on essential oils, but most Gram-negative bacteria were resistant even at 21.8 mg oil/ml. P. densiflora oil showed the most effective antifungal activity and the MIC values for Cryptococcus neoformans B42419 and Candida glabrata YFCC 062CCM 11658 were as low as 0.545 and 2.18 mg/ml, respectively. Cryp. neoformans B42419 was the most sensitive to all essential oils in the range of 0.545-2.18 mg/ml. Our data clearly showed that the essential oils from the three conifers had effective antimicrobial activity, especially against fungi.

• Korean Journal of Veterinary Research
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• v.22 no.2
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• pp.139-147
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• 1982

A total of 133 isolates of yeast-like fungi was tested for sensitivity to seven different antifungal agents. The yeast-like fungi tested were isolated from the milk from normal or mastitic bovine quaters or from bovine feces. They were 5 Candida albicans (C. albicans) isolates, 63 C. krusei, 27 C. tropicalis, 5 C. parapsilosis, 10 Torulopsis glabrata, 6 Rhodotorula sp., 6 Hansenula sp. and 1 Pichia sp. isolate. The antifungal agents tested were nystatin, griseofulvin, cycloheximide, 5-fluorocytosine, miconazol, clotrimazole and tolnaftate. In general, clotrimazole, miconazol and 5-fluorocytosine were more effective in antifungal activity in vitro against the test organisms than the rest of the agents tested. However, some of the isolates showed higher resistance to certain antifungal agents compared to the other isolates of the some species. They were: 1 C. albicans isolate to 5-fluorocytosine; 1 C. albicans to 5-fluorocytosine, miconazol and clotrimazole; 1 C. krusei to 5-fluorocytosine and cycloheximide; and 11 C. tropicalis isolates to cycloheximide. The minimum inhibitory concentrations(MIC) of clotrimazole were $12.5{\mu}g/ml$ or lower for all isolates tested except one C. albicans isolate, for which MIC of the drug was $100{\mu}g/ml$. On the other hand, the MIC's of cycloheximide were $6.5{\mu}g/ml$ or lower for all isolates except the following; all isolates of C. albicans ($100{\mu}g/ml$), C. pseudotropicalis ($200{\mu}g/ml$) and Rhodotorula sp. ($25-50{\mu}g/ml$), 11 C. tropicalis isolates ($100{\mu}g/ml$) and 1 C. krusei isolate ($200{\mu}g/ml$).

• Journal of Life Science
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• v.28 no.11
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• pp.1277-1284
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• 2018

In this study, a repeated yeast integrative plasmid (R-YIp) harboring Cre/loxP system was constructed to integrate various gene expression cassettes into the yeast chromosome. The R-YIp system contains a reusable selective marker (CgTRP1), loxP sequence, and target sequence for integration. Therefore, many gene expression cassettes can be integrated into the same position of the same yeast chromosome. In the present study, several model enzymes involving xylan/xylose metabolism were examined, including endoxylanase (XYLP), ${\beta}$-xylosidase (XYLB), xylose reductase (GRE3) and xylitol dehydrogenase (XYL2). Efficient expression of these genes was obtained using two promoters (GAL10p and ADH1p) and various plasmids (pGMF-GENE and pAMF-GENE plasmids) were constructed. The XYLP, XYLB, GRE3, and XYL2 genes were efficiently expressed under the control of the GAL10 promoter. Subsequently, R-YIps containing the GAL10p-GENE-GAL7t cassette were constructed, resulting in pRS-XylP, pRS-XylB, pRS-Gre3, and pRS-Xyl2 plasmids. These plasmids were sequentially integrated into chromosome VII of a Saccharomyces cerevisiae strain by repeated gene integration and selective marker rescue. These genes were integrated by the R-YIp system and were stably expressed in the yeast transformants to produce active recombinant enzymes. Therefore, we expect that the R-YIp system will be able to overcome current limitations of the host cells and allow selective marker selection for the integration of various genes into the yeast chromosome.