• Korean Journal of Microbiology
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• v.30 no.2
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• pp.108-112
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• 1992

To investigate the expression and the secretion of heterologous proteins in yeast, we constructed an yeast secretion vector and produced a human secretory protein, .alpha.-1-antitrypsin (.alpha.-1-AT), from yeast cells. The secretion vector pGAT8 was constructed by inserting the signal sequence of yeast acid phosphatase gene (PH05) into the .alpha.1-AT expression vector pGAT6 which contained .alpha.-1-AT cDNA fused to GAL10-CYC1 promotor. The .alpha.-1-AT was produced efficiently in the yeast cells transformed with plasmid pGAT8, which was onfirmed both by the .alpha.-1-AT activity assay and by the immunoblot method using .alpha.-1-AT antibody. We also showed the secretion of .alpha.-1-AT into the culture media and into the periplasmic space by immunoblot.

• Environmental Analysis Health and Toxicology
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• v.17 no.3
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• pp.253-259
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• 2002

The phototoxicity inhibitory activity of 15 natural products having antiinflammatory effect was screened by three in vitro methods: yeast growth inhibition test with Candida albicans, RBC photohemolysis and MTT assay. We induced phototoxic reaction by irradiating UVA (365 nm) on chlorpromazine (CPZ) that has been widely documented as phototoxic agent in clinical and experimental studies and then observed the effects of the natural products after treating them with CPZ. In yeast growth inhibition test, X. stramonium showed the inhibitory effect on the UVA phototoxicity and E. officinalis, Yeast, P. suffruticosa showed phototoxicity inhibitory effect in that their ％ hemolysis compared with control were 36.14${\pm}$ 2.69, 42.82${\pm}$1.35, 36.41${\pm}$0.48 on UVA. In MTT assay, all tested natural products increased cell viability compared with the control.

• Journal of Microbiology
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• v.40 no.3
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• pp.245-247
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• 2002

A genomic DNA library of the fungus Trametes versicolor was constructed in a yeast integration vector which contains the URA3 gene of the budding yeast Saccharomyces cerevisiae and the gene responsible for hygromycin B resistance, and fragments acting as autonomously replicating sequences (ARSes) in the budding yeast were identified from the genomic DNA library. Sixteen recombinant plasmids from the library transformed the budding yeast Saccharomyces cerevisiae to Ura+ at high frequencies. They were maintained stably under selective conditions, but were gradually lost from yeast cells at different rates under nonselective conditions, indicating that they contain eukaryotic origins of DNA replication and exist as extrachromosomal plasmids. Base sequences of four ARS DNAs among the 16 cloned fragments revealed that all or the four contain at least one 11 bp ［(A/T)TTTA(T/C)(A/G)TTT(A/T)］consensus sequence of the budding yeast ARS.

• Proceedings of the Korean Institute of Resources Recycling Conference
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• 2001.05b
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• pp.121-125
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• 2001

For the probiotic feed production, aerobic liquid fermentation of pulverized food wastes was attempted with a yeast Kluyveromyces marxianus. After grinding finely, optimal fermentation conditions of the substrate was investigated by shaking culture. The most active growth of the yeast was shown at solid content of 10%. The proper addition of urea(0.5g/l), o-phosphate(0.4g/l), molasses(4g/l), and yeast extract (1g/1) increased cell growth rate and viable cell count. For optimizing, the nutrients were all added to substrate and fermentation was carried in 2 litre jar fermenter. For the stimulation of hydrolyzing enzyme excretion, mixed culture with Aspersillus oryzae was also conducted. In 12 hours of fermentation, viable cell count of the yeast Kluyveromyces marxianus amounted to the number of 1.4 $\times$10$^{10}$ /1 in the culture medium.

• KSBB Journal
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• v.15 no.1
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• pp.76-79
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• 2000

Complex, ill-defined mixtures of natural origin are often used as nutrients in the production of biological products through microbial fermentation. Product yields are affected by variation in these natural products. Yeast extract is a typical example of these natural products. Since it is a mixture of amino acids, peptides and nucleic acids, its composition is not well characterized. In this study, we investigated the properties of thiamine hydrochloride, riboflavin and pyridoxine hydrochlride in yeast extract by using a gel filtration chromatography, ion exchange chromatography and high performance liquid chromatography. Yeast extract solution was fractionated by gel filtration chromatography and ion exchange chromatography, and then, each fraction was analyzed by using a high performance liquid chromatography.

• Journal of Species Research
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• v.13 no.1
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• pp.105-110
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• 2024

The purpose of this study was to isolate and identify wild yeasts from soil samples collected in Seoul and Daejeon, Republic of Korea. To identify wild yeast strains, pairwise sequence comparisons of D1/D2 region of the 26S rRNA gene sequence were done using Basic Local Alignment Search Tool (BLAST). The cell morphologies were observed by phase contrast microscope and carbon source assimilation test were done using API 20C AUX kit. Among the 13 isolated strains, 11 strains were previously reported, but two strains have never been reported from Republic of Korea. The 13 strains were assigned to the phylum Basidiomycota. The two unrecorded yeast strains B2UV-201 and DJ1-5-B-10C belong to the genera Rhodotorula and Rhodosporidiobolus, respectively. The two unrecorded yeast strains are oval shaped and polar budding cells. This research focuses on the morphological and biochemical properties of the two unreported yeast species that have not officially been reported in Korea.

• The Journal of Natural Sciences
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• v.9 no.1
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• pp.25-29
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• 1997

To study the mechanism of lipocortin-1, the 37 kDa protein, one of the annxin superfamily thought to be a second messenger during the Glucocorticoid dependent anti-inflammatory action, the gene for lipocortin-1 binding protein was isolated using the yeast two hybrid assay, the yeast based genetic assay recognizing the protein-protein interaction. The results showed that this gene has a weak homology to the for the human serine proteinase.

• Korean Journal of Microbiology
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• v.42 no.3
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• pp.195-198
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• 2006

Lethal toxin is a critical virulence factor of anthrax. It is composed two protein: protective antigen (PA) and lethal factor (LF). PA binds to specific cell surface receptors and, forms a membrane channel that mediates entry of LF into the cell. LF is a zinc-dependent metalloprotease, which cleaves MKKs [MAPK (mitogen-activated protein kinase) kinases] at peptide bonds very close to their N-termini. In this study, we suggest application of cell-based assays in the early phase of drug discovery, with a particular focus on the use of yeast cells. We constructed MEK1 expression system in yeast to determine LF activity and approached cell-based assay system to screen inhibitors, in which the results covering the construction of LF-substrate in yeast expression vector, expression, and LF-mediated proteolysis of substrate were described. These results could provided the basic steps in design of cell-based assay system with the high efficiency, rapidly and easy way to screening of inhibitors.

• Analytical Science and Technology
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• v.35 no.4
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• pp.181-188
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• 2022

This paper describes a comparative analysis of yeast cell viability at exponential and stationary growth phases using multiple conventional techniques and statistical tools. Overall, cellular responses to various viability assays were asynchronous. Results of optical density measurement and direct cell counting were asynchronous both at exponential and stationary phases. Proliferative capacity measurement using SP-SDS indicated that cells at the end of the stationary phase were proliferative as much as exponentially growing cells. Metabolic activity assays using two different dyes concluded that the inside of cells at stationary phase is slightly less reducing compared to that of exponentially growing cells, implying that the metabolic activity imperceptibly declined as cells were aged. These results will be helpful to understand the details of yeast cell viability at exponential and stationary growth phases.

• The Korean Journal of Mycology
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• v.26 no.3 s.86
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• pp.361-364
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• 1998

Among the organic sources of nitrogen tested, yeast extract and soytone were excellent for the mycelial growth of Tricholoma matsutake DGUM 26001. The mycelial growth was enhanced, when yeast extract at the concentration up to 1.0% was added to the starchpyridoxine medium. After 30-day cultivation of the mycelia at $24^{\circ}C$ in the medium supplemented with yeast extract, 518 mg/50 ml of dry mycelia could be harvested.