• Title/Summary/Keyword: novel yeast species

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Functional Screening for Cell Death Suppressors and Development of Multiple Stress-Tolerant Plants

  • Moon, Hae-Jeong;Baek, Dong-Won;Lee, Ji-Young;Nam, Jae-Sung;Yun, Dae-Jin
    • Proceedings of the Korean Society of Plant Biotechnology Conference
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    • 2003.04a
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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.

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Production of a New Biosurfactant by a New Yeast Species Isolated from Prunus mume Sieb. et Zucc.

  • Jeong-Seon Kim;Miran Lee;Dae-Won Ki;Soon-Wo Kwon;Young-Joon Ko;Jong-Shik Kim;Bong-Sik Yun;Soo-Jin Kim
    • Journal of Microbiology and Biotechnology
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    • v.33 no.8
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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 39399T, 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.

Copper Tolerance of Novel Rhodotorula sp. Yeast Isolated from Gold Mining Ore in Gia Lai, Vietnam

  • Kim Cuc Thi Nguyen;Phuc Hung Truong;Cuong Tu Ho;Cong Tuan Le;Khoa Dang Tran;Tien Long Nguyen;Manh Tuan Nguyen;Phu Van Nguyen
    • Mycobiology
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    • v.51 no.6
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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 GL1T 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.

Isolation of Wild Yeasts Obtained from Waters and Soils of Riversides in Sangjubo of Nakdong River and Daechung Dam of Geumgang Midstream, Korea and Characterization of Unrecorded Wild Yeasts (낙동강 상주보와 금강중류 대청댐 주변으로부터 야생효모의 분리 및 국내 미기록 효모들의 균학적 특성)

  • Han, Sang-Min;Park, Dong-Jae;Kim, Ji-Yoon;Lim, Hyo Jin;Lee, Hyang Burm;Lee, Jong-Soo
    • The Korean Journal of Mycology
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    • v.48 no.3
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    • pp.237-249
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    • 2020
  • The goal of this study was to isolate wild yeasts from waters and soils in Sangjubo of Nakdong river and Daechung dam in Daejeon city, Korea and investigate characteristics of previously unrecorded novel wild yeasts. In total, 94 strains from 39 different species of wild yeasts were isolated from 47 soils and waters samples obtained from Sangjubo in Nakdong river. Among these, 3 strains, including Rhodosporidium azoricum JSL-GRU-009, represented novel yeast strains which were previous recorded. Two hundred nine strains from 105 different species of wild yeasts were isolated from 130 soils and water samples obtained from Daechungdaegyo and Daechung dam of Shintanjin in Daejeon city, Korea. Ten strains including Bullera pseudoalba JSL-GRU-005 also represented newly recorded strains in Korea. All the 13 previously unrecorded yeasts were oval or global in shape, and nine strains, including R. azoricum JSL-GRU-009, formed ascospores. Ten strains grew well in vitamin-free medium. R. azoricum JSL-GRU-009 grew well in 20% glucose-containing yeast extract-peptone-dextrose (YPD) medium and 3 strains, including Rhodotorula oryzicola JSL-GRU-003, grew well in 5% NaCl-containing YPD medium.

Isolation of Wild Yeasts from the Water and Riverside Soil of Geumgang Midstream in Sejong City, Korea, and Characterization of Unrecorded Wild Yeasts (세종특별자치시 주변의 금강 중류 물과 토양에서 야생 효모의 분리 및 국내 미기록 효모의 특성)

  • Han, Sang-Min;Kim, Ji-Yoon;Lee, Jong-Soo
    • The Korean Journal of Mycology
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    • v.47 no.1
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    • pp.51-61
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    • 2019
  • The goal of this study was to elucidate wild yeast diversity of Geumgang midstream near Sejong metropolitan autonomous city, Korea. Thirty-seven strains of 32 species of wild yeasts were isolated from 43 water and soil samples under the Bulti bridge of Sejong city, Korea. Seven yeasts of each Candida spp. and Cryptococcus spp. were the predominant species isolated from samples near the Bulti bridge. Holtermanniella takashimae SW048 (NNIBRFG9314), Cystofilobasidium infirmominiatum SW013 (NNIBRFG9310), Mrakia cryoconite SW015 (NNIBRFG9316), Pichia sporocuriosa SW085 (NNIBRFG9326) and Cryptococcus aspenensis SW008 (NNIBRFG9309) represented novel yeast strains found in Korea for the first time. All of these previously unrecorded yeasts, except for Mrakia cryoconite SW015 had ascospores and grew well in yeast extract-peptone-dextrose (YPD), yeast extract-malt extract (YM) and potato-extrose (PD) media. Pichia sporocuriosa SW085 grew well in vitamin-free medium and Holtermanniella takashimae SW048, which was a halotolerant wild yeast, grew well YPD medium containing 5 % NaCl. Twenty-six strains representing eight species of wild yeast were isolated from 22 water and soil samples under the Haetmuri bridge of Sejong city, Korea. Candida pseudolambica (12 strains) and Aureobasidium pullulans (11 strains) were the predominant isolates from samples near the Haetmuri bridge. Occultifur kilbournensis HB060 (NNIBRFG9317), Sampaiozyma vanillica HB014 (NNIBRFG9332), Xenoramularia neerlandica HB039 (NNIBRFG9335), Candida norvegica HB315 (NNIBRFG9306), C. melibiosica HB316 (NNIBRFG9305), C. quercuum GB014 (NNIBRFG9307), and C. succiphila GB015 (NNIBRFG9308) represented novel yeast strains recorded in Korea for the first time. O. kilbournensis HB060 and X. neerlandica HB039 did not form ascospores or pseudo-mycelia. All of these previously unrecorded yeasts, except S. vanillica HB014 and X. neerlandica HB039, grew well in vitaminfree medium, and C. norvegica HB315 and C. succiphila GB015, which were halotolerant wild yeasts, which grew well in YPD medium containing 5 % NaCl.

In vitro Screening of Medicinal Plants with Estrogen Receptor Modulation Activity (생약의 여성호르몬 수용체 조절 활성 검색)

  • Lee, Chang-Min;Kang, Se-Chan;Oh, Joa-Sub;Choi, Han;Li, Xue-Mei;Lee, Jae-Hyun;Lee, Mi-Hyun;Choung, Eui-Su;Kawk, Joung-Hwan;Zee, Ok-Pyo
    • Korean Journal of Pharmacognosy
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    • v.37 no.1 s.144
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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.

Thermococcus onnurineus sp. nov., a Hyperthermophilic Archaeon Isolated from a Deep-Sea Hydrothermal Vent Area at the PACMANUS Field

  • Bae, Seung-Seob;Kim, Yun-Jae;Yang, Sung-Hyun;Lim, Jae-Kyu;Jeon, Jeong-Ho;Lee, Hyn-Sook;Kang, Sung-Gyun;Kim, Sang-Jin;Lee, Jung-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.16 no.11
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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).

Description of Vishniacozyma terrae sp. nov. and Dioszegia terrae sp. nov., Two Novel Basidiomycetous Yeast Species Isolated from Soil in Korea

  • Soohyun Maeng;Yuna Park;Gi-Ho Sung;Hyang Burm Lee;Myung Kyum Kim;Sathiyaraj Srinivasan
    • Mycobiology
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    • v.50 no.6
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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 YP579T 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 (KCTC27988T) and YP579T (KCTC 27998T), respectively.

Dual Role of Acidic Diacetate Sophorolipid as Biostabilizer for ZnO Nanoparticle Synthesis and Biofunctionalizing Agent Against Salmonella enterica and Candida albicans

  • Basak, Geetanjali;Das, Devlina;Das, Nilanjana
    • Journal of Microbiology and Biotechnology
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    • v.24 no.1
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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.

Molecular Cloning and Characterization of DNA Repair Related Gene in Yeast

  • Kang, Seon-Ah;Park, In-Soon
    • Journal of Life Science
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    • v.10 no.1
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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.