• 한국식물생명공학회:학술대회논문집
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• 한국식물생명공학회 2003년도 식물바이오벤처 페스티발
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• pp.65-71
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• 2003

Bax, a mammalian pro-apoptotic member of the Bcl-2 family, induces cell death when expressed in yeast. To investigate whether Bax expression can induce cell death in plant, we produced transgenic Arabidopsis plants that contained murine Bax cDNA under control of a glucocorticoid-inducible promoter. Transgenic plants treated with dexamethasone, a strong synthetic glucocorticoid, induced Bax accumulation and cell death, suggesting that some elements of cell death mechanism by Bax may be conserved among various organisms. Therefore, we developed novel yeast genetic system, and cloned several Plant Bax Inhibitors (PBIs). Here, we report the function of two PBIs in detail. PBI1 is ascorbate peroxidase (sAPX). Fluorescence method of dihydrorho-damine 123 oxidation revealed that expression of Bax in yeast cells generated reactive oxygen species (ROS), and which was greatly reduced by co-expression with sAPX. These results suggest that sAPX inhibits the generation of ROS by Bax, which in turn suppresses Baxinduced cell death in yeast. PBI2 encodes nucleoside diphosphate kinase (NDPK). ROS stress strongly induces the expression of the NDPK2 gene in Arabidopsis thaliana (AtNDPK2). Transgenic plants overexpressing AtNDPK2 have lower levels of ROS than wildtype plants. Mutants lacking AtNDPK2 had higher levels of ROS than wildtype. $H_2O_2$ treatment induced the phosphorylation of two endogenous proteins whose molecular weights suggested they are AtMPK3 and AtMPK6. In the absence of $H_2O_2$ treatment, phosphorylation of these proteins was slightly elevated in plants overexpressing AtNDPK2 but markedly decreased in the AtNDPK2 deletion mutant. Yeast two-hybrid and in vitro protein pull-down assays revealed that AtNDPK2 specifically interacts with AtMPK3 and AtMPK6. Furthermore, AtNDPK2 also enhances the MBP phosphorylation activity of AtMPK3 in vitro. Finally, constitutive overexpression of AtNDPK2 in Arabidopsis plants conferred an enhanced tolerance to multiple environmental stresses that elicit ROS accumulation in situ. Thus, AtNDPK2 appears to play a novel regulatory role in $H_2O_2$-mediated MAPK signaling in plants.

• Journal of Microbiology and Biotechnology
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• 제33권8호
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• pp.1023-1029
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• 2023

Biosurfactants reduce surface and interfacial tension due to their amphiphilic properties and are an eco-friendly alternative for chemical surfactants. In this study, a new yeast strain JAF-11 that produces a biosurfactant was selected using drop collapse method, and the properties of the extracts were investigated. The nucleotide sequences of the strain were compared with closely related strains and identified based on the D1/D2 domain of the large subunit ribosomal DNA (LSU) and internal transcribed spacer (ITS) regions. Neodothiora populina CPC 39399^T, the closest species with strain JAF-11, showed a sequence similarity of 97.75% for LSU and 94.27% for ITS, respectively. The result suggests that the strain JAF-11 represents a distinct species that cannot be assigned to any existing genus or species in the family Dothideaceae. Strain JAF-11 produced a biosurfactant reducing the surface tension of water from 72 mN/m to 34.5 mN/m on the sixth day of culture and the result of measuring the critical micelle concentration (CMC) by extracting the crude biosurfactant was found to be 24 mg/l. The molecular weight 502 of the purified biosurfactant was confirmed by measuring the fast atom bombardment mass spectrum. The chemical structure was analyzed by measuring ¹H nuclear magnetic resonance (NMR), ¹³C NMR, and two-dimensional NMRs of the compound. The molecular formula was C₂₆H₄₆O₉, and it was composed of one octanoyl group and two hexanoyl groups to myo-inositol moiety. The new biosurfactant is the first report of a compound produced by a new yeast strain, JAF-11.

• Mycobiology
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• 제51권6호
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• pp.379-387
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• 2023

In this study, twenty-five yeast strains were isolated from soil samples collected in the gold mining ore in Gia Lai, Vietnam. Among them, one isolate named GL1^T could highly tolerate Cu²⁺ up to 10 mM, and the isolates could also grow in a wide range of pH (3-7), and temperature (10-40 ℃). Dried biomass of GL1 was able to remove Cu²⁺ effectively up to 90.49% with a maximal biosorption capacity of 18.1 mg/g at pH 6, temperature 30 ℃, and incubation time 60 min. Sequence analysis of rDNA indicated this strain was closely related to Rhodotorula mucilaginosa but with 1.53 and 3.46% nucleotide differences in the D1/D2 domain of the 28S rRNA gene and the ITS1-5.8S rRNA gene-ITS2 region sequence, respectively. Based on phylogenetic tree analysis and the biochemical characteristics, the strain appears to be a novel Rhodotorula species, and the name Rhodotorula aurum sp. nov. is proposed. This study provides us with more information about heavy metal-tolerant yeasts and it may produce a new tool for environmental control and metal recovery operations.

• 한국균학회지
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• 제48권3호
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• pp.237-249
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• 2020

경상북도 낙동강의 상주보와 금강중류인 대전시 신탄진의 대청댐주변의 야생효모들의 종 분포특성을 알아보기 위해 먼저 이들지역의 토양과 물들로부터 야생효모들을 분리, 동정하였다. 낙동강의 상주보에서 채취한 시료 47점에서 모두 39종 94 균주의 야생효모들이 분리, 동정되었고 Cryptococcus속 균들이 가장 많았다. 이들중Rhodosporidium azoricum JSL-GRU-009를 포함한 3균주들이 아직까지 국내에 보고되지 않은 미기록 효모들로 최종 선별되었다. 또한, 금강중류지역인 대전시 신탄진의 대청대교와 대청댐주변 토양과 물등 130종의 시료들로부터 105종 209균주들이 분리, 동정되었고 이들 야생효모 균주들로부터 Bullera pseudoalba JSL-GRU-005를 포함한 10종의 균들이 국내 미기록 효모들로 최종 선별되었다. 이들 낙동강 상주보와 대청댐등에서 선별된 13균주들의 균학적 특성을 조사한 결과 이들 미기록 균주들은 구형~타원형으로 출아, 분열법으로 영양 증식하였다. R. azoricum JSL-GRU를 포함한 9균주들이 포자를 형성하였고 10균주들이 비타민을 첨가하지 않은 YPD 배지에서 생육하였다.R. azoricum JSL-GR균주는 20% 포도당을 함유한 YPD 배지에서 잘 생육하였고, R. oryzicola JSL-GRU-003를 포함한 3 균주는 5% NaCl을 함유한 YPD 배지에서 생육하였다.

• 한국균학회지
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• 제47권1호
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• pp.51-61
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• 2019

우리나라 금강의 물과 주변 토양에 분포하는 야생 효모들의 종 다양성을 확립하고자 세종특별자치시 불티교와 햇무리교를 흐르는 금강 중류의 물과 주변 토양에서 야생 효모들을 분리, 동정하였고 이들 중 국내 미기록 효모들을 선별하여 이들의 균학적 특성을 조사하였다. 세종시 불티교 밑의 물과 주변 토양들로부터 32종, 37균주의 야생 효모들을 분리, 동정하였다. 이들 중 Holtermanniella takashimae SW048 (NNIBRFG9314), Cystofilobasidium infirmominiatum SW013 (NNIBRFG9310), Mrakia cryoconite SW015(NNIBRFG9316), Pichia sporocuriosa SW085(NNIBRFG9326)와 Cryptococcus aspenensis SW008 (NNIBRFG9309) 5균주들을 국내 미기록 효모 균주들로 최종 선별하였고, 이들은 구형-난형으로 출아에 의해 영양증식을 하였다. M. cryoconite SW015 외 모두 자낭포자를 형성하였고, P. sporocuriosa SW085 와 C. aspenensis SW008 균주는 20% 포도당을 함유한 YPD 배지에서도 생육하는 내당성 야생 효모들이었다. 또한, 세종시 햇무리교의 물과 주변 토양시료에서 모두 8종 26균주의 야생 효모들을 분리, 동정하였고 이들 야생 효모들 중 Occultifur kilbournensis HB060 (NNIBRFG9317), Sampaiozyma vanillica HB014 (NNIBRFG9332), Xenoramularia neerlandica HB039 (NNIBRFG9335), Candida norvegica HB315 (NNIBRFG9306), C. melibiosicaHB316 (NNIBRFG9305), C. quercuum GB014 (NNIBRFG9307)와 C. succiphila GB015 (NNIBRFG9308) 7균주 국내 미기록 효모들로 분류되었다. O. kilbournensis HB060와 X. neerlandica HB039는 자낭포자와 의균사를 형성하지 못하였고 C. norvegica HB315는 5 % NaCl를 함유한 YPD 배지에서 생육하는 내염성을 보였다. 본 연구결과는 앞으로 금강의 야생 효모 종 다양성 확립에 귀중한 자료로 활용될 것이다.

• 생약학회지
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• 제37권1호통권144호
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• pp.21-27
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• 2006

Yeast based estrogenicity assay is the simplest and useful for the assay and the discovery of novel estrogenic substances in natural specimens, The estrogen receptor(ER) modulation activity of 50% EtOH extracts of 101 traditional medicinal herbs was assessed using a recombinant yeast assay system with both a human estrogen receptor expression plasmid and a receptor plasmid. Among them, 14 species proved to be active. Pureariae Flos (flower of Puerraria thunbergiana BENTH.) had the highest estrogenic relative potency$(7.75{\times}10^{-3})$ $(EC_{50}=9.39\;{\mu}g/ml)$. The $EC_{50}$ value of $17{\beta}-estradiol$ used as the positive control was $0.073\;{\mu}g/ml)$ (Relative Potency=1.00). There results demonstrated that some of the traditional medical herb may be useful in the therapy of estrogen replacement.

• Journal of Microbiology and Biotechnology
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• 제16권11호
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• pp.1826-1831
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• 2006

A novel hyperthermophilic, anaerobic, heterotrophic archaeon, designated strain $NA1^T$, was isolated from a deep-sea hydrothermal vent area (depth, 1,650 m) within the Papua New Guinea-Australia-Canada-Manus (PACMANUS) field. Cells of this strain were motile by means of polar flagella, coccoid-shaped with a diameter of approximately $0.5-1.0{\mu}m$, and occurred as single cells. Optimal temperature, pH, and NaCl concentration for growth were $80^{\circ}C$, 8.5, and 3.5%, respectively. The new isolate was an obligate heterotroph that utilized yeast extract, beef extract, tryptone, peptone, casein, and starch as carbon and energy sources. Elemental sulfur was required for growth and was reduced to hydrogen sulfide. The G+C content of the genomic DNA was 52.0 mol%. Phylogenetic analysis of the 16S rRNA gene indicated that strain $NA1^T$ belongs to the genus Thermococcus, and the organism is most closely related to T. gorgonarius, T. peptonophilus, and T. celer; however, no significant homology was observed among species by DNA-DNA hybridization. Strain $NA1^T$ therefore represents a novel species for which the name Thermococcus onnurineus sp. novo is proposed. The type strain is $NA1^T$ (=KCTC 10859, =JCM 13517).

• Mycobiology
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• 제50권6호
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• pp.439-447
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• 2022

Two strains, YP344 and YP579 were isolated from soil samples in Pocheon City, Gyeonggi Province, South Korea. The strains YP344 and YP579 belong to the genus Vishniacozyma and Dioszegia, respectively. The molecular phylogenetic analysis showed that the strain YP344 was closely related to Vishniacozyma peneaus. Strain YP344T differed by four nucleotide substitutions with no gap (0.70%) in the D1/D2 domain of the LSU rRNA gene and 16 nucleotide substitutions with 8 gaps (5.76%) in the ITS region. On the other hand, the strain YP579^T varied from the type strain of the most closely related species, Dioszegia zsoltii var. zsoltii, by 6 nucleotide substitutions with four gaps (1.64%) in the D1/D2 domain of LSU rRNA gene and 26 nucleotide substitutions with 14 gaps (8.16%) in the ITS region. Therefore, the name Vishniacozyma terrae sp. nov. and Dioszegia terrae sp. nov. are proposed, with type strains YP344T (KCTC27988^T) and YP579^T (KCTC 27998^T), respectively.

• Journal of Microbiology and Biotechnology
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• 제24권1호
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• pp.87-96
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• 2014

In the present study, a yeast species isolated from CETP, Vellore, Tamilnadu was identified as Cryptococcus sp. VITGBN2 based on molecular techniques and was found to be a potent producer of acidic diacetate sophorolipid in mineral salt media containing vegetable oil as additional carbon source. The chemical structure of the purified biosurfactant was identified as acidic diacetate sophorolipid through GC-MS analysis. This sophorolipid was used as a stabilizer for synthesis of zinc oxide nanoparticles (ZON). The formation of biofunctionalized ZON was characterized using UV-visible spectroscopy, XRD, scanning electron microscopy (SEM), and Fourier transform infrared spectroscopy. The antimicrobial activities of naked ZON and sophorolipid functionalized ZON were tested based on the diameter of inhibition zone in agar well diffusion assay, microbial growth rate determination, protein leakage analysis, and lactate dehydrogenase assay. Bacterial pathogen Salmonella enterica and fungal pathogen Candida albicans showed more sensitivity to sophorolipid biofunctionalized ZON compared with naked ZON. Among the two pathogens, S. enterica showed higher sensitivity towards sophorolipid biofunctionalized ZON. SEM analysis showed that cell damage occurred through cell elongation in the case of S. enterica, whereas cell rupture was found to occur predominantly in the case of C. albicans. This is the first report on the dual role of yeast-mediated sophorolipid used as a biostabilizer for ZON synthesis as well as a novel functionalizing agent showing antimicrobial property.

• Journal of Life Science
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• 제10권1호
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• pp.40-44
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• 2000

The SNF2/SW ATPase/helicase family comprises proteins form a variety of species with in vivo functions, such as transcriptional regulation, maintenance of chromosome stability during mitosis, and various types of DNA repair. Here, we reported the characterization of h게2+gene which was iolated by PCR amplification using the conserved domain of SNF2 motifs. Sequence analysis of PCR product showed striking evolutionary conservation among the SNF2 family of proteins. Two transcripts of 6.7 and 3.4 Lb were detected by Northern blot analysis. furthermore, the intensities of these two bands were increased by ultraviolet(UV) irradiation. These results indicate that the hrp2+ is a novel member of the SNF2 family of proteins and is one of the UV-inducible genes in S. pombe. To determine the level of transcripts of hrp2+ gene during cellular growth, Northern blot analysis were performed. This result indicates that the level of hrp2+transcript reached its maximum before cells entered the exponential growth phase. This suggests that hrp2+ gene is experssed mainly at the early stage of cell growth.