• KSBB Journal
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• 제6권2호
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• pp.189-194
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• 1991

The effect of wtlitc light on unaeraten growth of Baker's yeast and the accompanying ethanol production has been studied in a batch process at 27$^{\circ}C$. Over the 80-hour period of the Champagne yeast process without pH control, the cull growth was inhibited by the fluorescent light. Another observed difference between the runs is that the drop and subsequent rise in redox potential occurred much sooner in the fermentation with light than in the fermentation without light. This preliminary study indicated that ethanol production could be enhanced by light as the cell concentration is repressed. The possible pathway, shift of the sugar substrate toward ethanol and away from cells was manifested by another difference as well. As observed under the microscope, many of the yeast cells grown under light budded without dividing by the normal fission process as they did in the dark. Furthermore, the undivided and branched (light grown) cell did not agglutinate at the end of the fermentation process as did the distinct spherical (dark grown) cells.

• 분석과학
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• 제35권4호
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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.

• Biotechnology and Bioprocess Engineering:BBE
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• 제10권1호
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• pp.85-90
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• 2005

A downstream process was developed for the production of yeast extract from brewer's yeast cells. Various downstream processing conditions including clarification, debittering, and the Maillard reaction were considered in the development of the process. This simple and economic clarification process used flocculating agents, specifically calcium chloride ($1\%$). After the clarification step, a Maillard reaction is initiated as a flavor-enhancing step. By investigating the effects of several operation parameters, including the type of sugar added, sugar dosage, glycine addition, and temperature, on the degree of browning (DB), giucose addition and reaction temperature were found to have significant effects on DB. A synthetic adsorption resin (HP20) was used for the debittering process, which induced a compositional change of the hydrophobic amino acids in the yeast hydrolysate, thereby reducing the bitter taste. The overall dry matter yield and protein yield for the entire process, including the downstream process proposed for the production of brewer's yeast extract were 50 and $50\%$, respectively.

• Journal of Applied Biological Chemistry
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• 제60권2호
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• pp.113-117
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• 2017

생물학적정화는 미생물을 이용하여 중금속을 포함한 오염물질을 정화하는 방법이다. 효모는 다양한 중금속을 흡착하여 정화하는 능력이 우수하다고 잘 알려졌다. 따라서 본 연구는 여러 중금속 중 하나인 아연에 저항성인 큰 효모를 인위적인 돌연변이를 통해서 얻는 것에 초점을 두었다. 대조구인 효모를 1 mM 농도의 아연이 포함된 배지에서 키운 후 점차 아연의 농도를 높여 최종적으로 80 mM이 되는 지점까지 키웠으며 이 농도는 효모가 점진적 적응을 거쳤어도 성장이 불가능한 한계점 근처이며 이렇게 유도 분리된 효모를 ZnR이라 명명하였다. ZnR은 대조구 효모보다 아연에 대한 저항성이 현저히 증가되었으며 이 외에 카드뮴과 니켈에 대한 저항성도 증가되었지만 구리에 대한 저항성은 오히려 감소 되었다. ZnR이 보여준 아연에 대한 저항성 증가는 유전자 돌연변이에 의한 영구적인 획득 결과이다. 단 기간 내에 아연에 대한 저항성 큰 효모의 인위적인 유도 및 분리는 생물학적정화 연구에 유용하게 쓰여질 정보이다.

• 미생물학회지
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• 제6권4호
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• pp.122-130
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• 1968

Action of various wavelengths of visible light on ultraviolet-radiation damage to haploid yeast cells, Saccharomyces cerevisiae 23971, was studied. The results were obtained on the basis of the survival and respiration rates by pre- and post-illuminations of various wavelengths before and after U.V.-irradiations on the yeast cells. Among the wavelengths tested, 635 $m{\mu}$, 429 $m{\mu}$ and white light which caused increase of respiration in pre-treatment alone, induced less resistance to the U. V.-damage than in the control, in both pre- and U.V.-treatment. On the contrary, such wavelengths as 574 $m{\mu}$and 530 $m{\mu}$, showing a weak effect on respiration in pre-treatment increased the susceptability to U.V.-radiation. Photoinactivation was generally obtained by both pre- and post- illuminations along with U.V.-treatment. At 635 $m{\mu}$ the PI rate was the lowest and also a low PI rate was shown at 429 $m{\mu}$. But 429 $m{\mu}$, in the post-treatment of the yeast cells pre-treated by the white light and the darkness respectively, showed the highest PI rate. In both pre- and post- treatment of 574, 530 and 473 $m{\mu}$,the PI rates were high to the same degree. Post-treatments of the wavelengths on U.V.-treated yeasts incubated rather under the white light than the darkness induced lower PI rate. It is assumed that there are great differences in action even of the same wavelength, depending upon the various combination of pre- and post-treatments, and that, moreover, the action of various wavelengths of visible light on U.V.-damage on the cells are concerned with the doses and dose rates of U.V. and visible lights. These observations led to an interpretation that each wavelength of visible light might exert distinctively different effects oil U. V.-damage, mainly causing the inhibition or stimulation of enzymes in the yeast cells.

• Journal of Microbiology and Biotechnology
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• 제30권9호
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• pp.1290-1296
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• 2020

Recently, it was reported that entire mammalian mtDNA genomes could be transplanted into the mitochondrial networks of yeast, where they were accurately and stably maintained without rearrangement as intact genomes. Here, it was found that engineered mtDNA genomes could be readily transferred to and steadily maintained in the mitochondria of genetically modified yeast expressing the mouse mitochondrial transcription factor A (Tfam), one of the mitochondrial nucleoid proteins. The transferred mtDNA genomes were stably retained in the Tfam-expressing yeast cells for many generations. These results indicated that the engineered mouse mtDNA genomes introduced in yeast mitochondria could be relocated into the mitochondria of other cells and that the transferred genomes could be maintained within a mitochondrial environment that is highly amenable to mimicry of the biological conditions in mammalian mitochondria.

• Journal of Microbiology and Biotechnology
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• 제24권3호
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• pp.408-420
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• 2014

Yeast surface-displayed antibody libraries provide an efficient and quantitative screening resource for given antigens, but suffer from typically modest library sizes owing to low yeast transformation efficiency. Yeast mating is an attractive method for overcoming the limit of yeast transformation to construct a large, combinatorial antibody library, but the optimal conditions have not been reported. Here, we report a large synthetic human Fab (antigen binding fragment) yeast surface-displayed library generated by stepwise optimization of yeast mating conditions. We first constructed HC (heavy chain) and LC (light chain) libraries, where all of the six CDRs (complementarity-determining regions) of the variable domains were diversified mimicking the human germline antibody repertoires by degenerate codons, onto single frameworks of VH3-23 and $V{\kappa}1$-16 germline sequences, in two haploid cells of opposite mating types. Yeast mating conditions were optimized in the order of cell density, media pH, and cell growth phase, yielding a mating efficiency of ~58% between the two haploid cells carrying HC and LC libraries. We constructed two combinatorial Fab libraries with CDR-H3 of 9 or 11 residues in length with colony diversities of more than $10^9$ by one round of yeast mating between the two haploid HC and LC libraries, with modest diversity sizes of ${\sim}10^7$. The synthetic human Fab yeast-displayed libraries exhibited relative amino acid compositions in each position of the six CDRs that were very similar to those of the designed repertoires, suggesting that they are a promising source for human Fab antibody screening.

• Journal of Microbiology and Biotechnology
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• 제10권3호
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• pp.293-299
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• 2000

Many methods have been developed for screening chemicals with hormonal activity. Using recombinant yeasts expressing either human estrogen receptor [Saccharomyces cerevisiae ER + LYS 8127 (YER)] or androgen receptor [S. cerevisiae AR + 8320 (YAR)], we evaluated the hormonal activities of several chemicals by induction of ${\beta}-galactosidase$ activity. The chemicals were $17{\beta}-estradiol$ (E2), testosterone (T), ${\rho}-nonylphenol$ (NP), bisphenol A (BPA), genistein (GEN), 2-bromopropane (2-BP), dibutyl phthalate (DBP), di-(2-ethylhexyl) phthalate (DEHP), 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), and butylparaben (BP). To assess the estrogenicity of NP, the result of the in vitro recombinant yeast assay was compared with an E-screen assay using MCF-7 human breast cancer cells and an uterotrophid assay using ovariectomized mice. In the YER yeast cells, E2, NP, BPA, GEN, and BP exhibited estrogenicity in a doseresponse manner, while TCDD did not. All the chemicals tested, except T, did not show androgenicity in the YAR yeast cell. The sensitivity of the yeast (YER) assay system to the estrogenic effect of NP was similar to that of the E-screen assay. NP was also estrogenic in the uterotrophic assay. However, in terms of convenience and costs, the yeast assay was superior to the E-screen assay or uterotrophic assay. These results suggest that the recombinant yeast assay can be used as a rapid tool for detecting chemicals with hormonal activities.

• Journal of Microbiology and Biotechnology
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• 제17권7호
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• pp.1139-1146
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• 2007

Cultures of Saccharomyces cerevisiae were subjected to the effect of PEF (pulse electric field) and a source of selenium. The culture period after which yeast cells were subjected to PEF treatment was optimized, as was the duration of the exposure. Optimization of the nutrient medium composition in S. cerevisiae cultures resulted in an over 1.8-fold increase in selenium accumulation with relation to cultures on the initial substrate. Optimization of the pH value and of culture duration resulted in selenium accumulation increase by approximately 78%. A significant correlation was found between the accumulation of selenium in yeast cells and its concentration in the culture substrate. The highest accumulation of selenium in the biomass of yeast, approx. $240\;{\mu}g/g$ d.m., was obtained after 15-min exposure to PEF on a 20-h culture. An approx. 50% higher content of selenium in cells was recorded, as compared with the control culture without the application of PEF.

• 미생물학회지
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• 제29권2호
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• pp.75-79
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• 1991

The yeast L-A virus (4.6 kb dsRNA genome) encodes the major coat protein and a "gag-pol" fusion minor coat protein that separately encapsidate itself and $M_{1}$, a 1.8 kb dsRNA satellite virus encoding a secreted protein toxin (the killer toxin). The teast chromosomal SKI genes prevent viral cytopathology by lowering the virus copy number. Thus, $ski^{-}$ mutants are ts and cs for growth. We transformed a ski2-2 virus-infested mutant with a yeast bank in a high copy cloning vector and selected the rare healthy transformants for analysis. One type of transformant segregated M-O L-A-O cells with high frequency. Elimination of the DNA clone from the ski2-2 strain eliminated this phinotype and introduction of the DNA clone recovered from such transformants into the parent ski2-2 strain, or into ski3 or ski6 mutants gave the same phenotype. This killer-curing phenotype was due to the curing of the helper L-A dsRNA virus. The 6.5 kb insert only had this activity when carried on a high copy vector and in $ski^{-}$ cells (not in $SKI^{+}$ cells). This 6.5 kb insert acts as a mutagen on L-A dsRNA producing a high rate of deletion mutations.mutations.