• Journal of Microbiology and Biotechnology
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• v.25 no.12
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• pp.1989-1996
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• 2015

Ice-binding proteins (IBPs) can inhibit ice recrystallization (IR), a major cause of cell death during cryopreservation. IBPs are hypothesized to improve cell viability after cryopreservation by alleviating the cryoinjury caused by IR. In our previous studies, we showed that supplementation of the freezing medium with the recombinant IBP of the Arctic yeast Glaciozyma sp. (designated as LeIBP) could reduce post-thaw hemolysis of human red blood cells and increase the survival of cryopreserved diatoms. Here, we showed that LeIBP could improve the viability of cryopreserved mammalian cells. Human cervical cancer cells (HeLa), mouse fibroblasts (NIH/3T3), human preosteoblasts (MC3T3-E1), Chinese hamster ovary cells (CHO-K1), and human keratinocytes (HaCaT) were evaluated. These mammalian cells were frozen in dimethyl sulfoxide (DMSO)/fetal bovine serum (FBS) solution with or without 0.1 mg/ml LeIBP at a cooling rate of -1℃/min in a -80℃ freezer overnight. The minimum effective concentration (0.1 mg/ml) of LeIBP was determined, based on the viability of HeLa cells after treatment with LeIBP during cryopreservation and the IR inhibition assay results. The post-thaw viability of mammalian cells was examined. In all cases, cell viability was significantly enhanced by more than 10% by LeIBP supplementation in 5% DMSO/5% FBS: viability increased by 20% for HeLa cells, 28% for NIH/3T3 cells, 21% for MC3T3-E1, 10% for CHO-K1, and 20% for HaCaT. Furthermore, addition of LeIBP reduced the concentrations of toxic DMSO and FBS down to 5%. Therefore, we demonstrated that LeIBP can increase the viability of cryopreserved mammalian cells by inhibiting IR.

• Korean Journal of Microbiology
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• v.35 no.4
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• pp.302-306
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• 1999

Production of biomass by fed-batch culture of Saccharomyces cerevisiae Hansen CBS5926, which is used to treat intestinal disorders, was investigated using ethanol as the sole carbon source. Ethanol was a better carbon source than glucose for high cell density culture of the st-rain since it could decrease the frequency of contamination while increasing the efficiency and final productivity of the fermentation process. Under optimal conditions, 38 g/ℓ of dry cell weight with $2.2{\times}10^{9}$ cfu/㎖ of maximum viable cell count was achieved after 72h cultivation. Freeze-drying of the cultured yeast cells resulted in severe reduction of viability. Of the freeze-drying protectants tested, 20% sucrose and 30% lactose were most effective for the preservation of yeast cells with a viability level of 16.3%. A combination of skim milk and lactose with 20% sucrose(w/v) exerted no synergistic influence upo the viability of the cells during cryopreservation by freeze-drying.

• 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.

• Proceedings of the Microbiological Society of Korea Conference
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• 2005.05a
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• pp.105-109
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• 2005

Heat Shock Transcription Factor (HSF), and the promoter heat Shock Element (HSE), are among the most highly conserved transcriptional regulatory elements in nature. HSF mediates the transcriptional response of eukaryotic cells to heat, infection and inflammation, pharmacological agents, and other stresses. While HSF is essential for cell viability in yeast, oogenesis and early development in Drosophila, extended life-span in C. elegans, and extra-embryonic development and stress resistance in mammals, little is known about its full range of biological target genes. We used whole genome analyses to identify virtually all of the direct transcriptional targets of yeast HSF, representing nearly three percent of the genomic loci. The majority of the identified loci are heat-inducibly bound by yeast HSF, and the target genes encode proteins that have a broad range of biological functions including protein folding and degradation, energy generation, protein secretion, maintenance of cell integrity, small molecule transport, cell signaling, and transcription. Approximately 30% of the HSF direct target genes are also induced by the diauxic shift, in which glucose levels begin to be depleted. We demonstrate that phosphorylation of HSF by Snf1 kinase is responsible for expression of a subset of HSF targets upon glucose starvation.

• Journal of Microbiology and Biotechnology
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• v.15 no.4
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• pp.694-700
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• 2005

Defects in the mitotic spindle or in the attachment of chromosomes to the spindle are believed to release an activated form of spindle checkpoint complex that inhibits APC-dependent ubiquitination and subsequently arrests the cell cycle at metaphase. When the spindle assembly is disrupted, the fission yeast mitotic arrest deficient (mad) mutants fail to arrest and rapidly lose viability. To enhance our understanding of the molecular mechanisms for the pathway of checkpoint function, the functional characterizations of Mad 1 p from Schizosaccharomyces pombe involved in this process have been carried out. Yeast two-hybrid and various deletion analyses of S. pombe Mad1 p reveal that the C terminus of Mad1p is critical for the binding of Mad2p and maintenance of Mad 1 p-Mad2p interaction. In addition, it was found. that the Mad1p region (residues 206-356) is essential for Mad1p-other checkpoint components. Mad1p truncating this region is sufficient to bind Mad2p but abolishes the checkpoint function, indicating that the checkpoint function is necessary for interaction of Mad 1 p-other checkpoint components. The possible functions of S. pombe Mad1p at the cell cycle checkpoint are discussed.

• Archives of Pharmacal Research
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• v.29 no.8
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• pp.707-711
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• 2006

Bifidobacteria-loaded alginate poly-l-lysine microparticles (bap microparticles) were prepared using an air atomization method and then freeze-dried. The viability of the bap microparticles was investigated as a function of the amount of the bifidobacteria cultures, and the addition of a yeast extract, cryoprotectants, antioxidants and neutralizer. The size of the bap microparticles with and without the bifidobacteria was $84.8{\pm}28.5\;{\mu}m$ ($mean{\pm}standard$ deviation) and $113.1{\pm}38.5\;{\mu}m$, respectively. The surface morphology was slightly ellipsoid and wrinkled regardless of the incorporating bifidobacteria. The viability gradually decreased with increasing freeze-drying time. Free-flowing powdered bap microparticles were obtained at least 12 h after freeze-drying the wetted slurry of bap microparticles. However, the particles tended to aggregate when either lactose or ascorbic acid was added. The addition of a yeast extract, cryoprotectants (glycerol and lactose), antioxidants ($NaHSO_3$ and ascorbic acid) and neutralizer $(Mg_3(PO_4)_2)$ resulted in a significantly higher viability of the bifidobacteria in the bap microparticles after freeze-drying (0.34-1.84 log) compared with the culture alone.

• Journal of Microbiology
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• v.40 no.1
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• pp.82-85
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• 2002

The aerobic batch growth of Issatchenkia orientalis DY252 with glucose and fructose medium was investigated at 32$\^{C}$ and pH 5.0. Aerobic ethanol production was evident with yeast I, orientalis. A diauxic lag of about 1 h between growth on glucose and growth on ethanol during batch culture was observed. However, no diauxic growth occurred with fructose. As the incubation temperature was increased from 32 to 39$\^{C}$, viability at the end of each batch culture declined significantly, from 93 to 43%, Unlike the effect of temperature, viability was not greatly affected by incubation pH, and cell yield values in a range of 0.45-0.48 were obtained. In order to overcome overflow metabolism, a fedbatch culture under glucose limitation was carried out. Compared with aerobic batch culture, about 10% improvement in cell yield was achieved with a fed-batch culture in optimal conditions.

• Microbiology and Biotechnology Letters
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• v.21 no.4
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• pp.373-380
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• 1993

When the cells of yeast K35 were immobilized in Ca-alginate gel, cell concentration and viability decreased as alginate concentration increased. Considering the results, 2% (w/v) Ca-alginate concentration would be suitable. Among various concentrations of additives and cross-lin-king agent, the addition of 1.67% (w/v) of bentonite together with 0.33% (v/v) of glutaraldehyde (ABG bead) resulted in the highest ethanol production of 1.8%(w/v), using YPD medium containing 2% glucose. ABG bead seemed to be more resistant to phosphate ion than Ca-alginate bead. 0.33%(w/v) of phosphate was a proper concentration for the ethanol production by ABG bead. Scanning electron microscopic observation depicted that the immobilized cells on the bead surface were coated by alginate gel and that the cells in the internal bead were cross-linked with alginate matrix. When repeated-batch culture was performed with ABG bead for 40 days in a packed-bed reactor, ethanol concentration of about 90~110 g/l-gel was maintained. Cell viability was maintained around 70%, and outgrowing cell concentration was below 6.3% of total cell concentration. Consequently, the results showed that ABG head was a potential carrier for continuous production of ethanol compared to conventional Ca-alginate bead.

• Microbiology and Biotechnology Letters
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• v.48 no.2
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• pp.113-120
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• 2020

The objective of the study was to assess the survival of microencapsulated Lactobacillus plantarum ATCC8014 produced using the emulsion technique in alginate gel combined with pectin and maltodextrin components. The microcapsules were then added to cupcake dough that was further baked at 200℃ for 12 min. The viability of L. plantarum was assessed during baking and the 10 days of storage at 4℃ as well as in simulated gastrointestinal conditions. In addition, yeast-mold and water activity were investigated. After baking, the samples with microencapsulated L. plantarum contained more than 5 log CFU/g, which was higher compared to the bacterial concentration of the control samples. The concentration of L. plantarum was more than 6 logs CFU/g after the end of the storage; therefore, the probiotic functioned as a biopreservative in the cake. The prebiotic component strengthened the microcapsules network and helped protect the viability of L. plantarum in simulated gastric fluid (SGF) and simulated intestinal fluid (SIF) media. The results show that the addition of L. plantarum microencapsules did not affect the sensory scores of the cupcake while ensuring the viability of the probiotic during baking and storing.

• Environmental Mutagens and Carcinogens
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• v.16 no.2
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• pp.77-82
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• 1996

The RAD3 gene of Saccharomyces cerevisiae is required for excision repair and is essential for cell viability. RAD3 encoded protein possesses a single stranded DNA-dependent ATPase and DNA-RNA helicase activies. To examine the extent of conservation of structure and function of RAD3 during eukaryotic evolution, we have cloned the RAD3 homolog, HRD3, from the distantly related yeast Schizosaccharomyces pombe. Here, we report the partial cloning and characterization of HRD3 gene (Homologous of RAD3 gene) which was isolated by PCR amplification using conserved domain of Saccharomyces cerevisiae RAD3 gene. Chromosomal DNA isolated from S. pombe had similar restriction patterns to those from S. cerevisiae, as determined by Southern blot analysis. The 2. 8 kb transcript of mRNA was identified by Northern hybridization. The level of transcript did not increase upon UV-irradiation, suggesting that the HRD3 gene in S. pombe is not UV-inducible.