• Microbiology and Biotechnology Letters
• /
• v.36 no.3
• /
• pp.221-226
• /
• 2008

To produce human ferritins in yeast, human H-chain and L-chain ferritins were amplified from previously cloned vectors. Each amplified ferritin gene was inserted into the pYES2.1/V5-His-TOPO yeast expression vector under the control of the GAL1promoter. Western blot analysis of the recombinant yeast cells revealed that H-and L-chain subunits of human ferritin were expressed in Saccharomyces cerevisiae. Atomic absorption spectrometry (AAS) analysis demonstrated that the intracellular content of iron in the ferritin transformant was 1.6 to 1.8-fold higher than that of the control strain. Ferritin transformants could potentially supply iron-fortified nutrients for food and feed.

• Korean Journal of Microbiology
• /
• v.28 no.1
• /
• pp.1-5
• /
• 1990

The coding sequence of hepatitis B viral core antigen (HBcAg) (subtype adr) contains two in-phase initiation codons, one for precore and the other for core antigen gene. To study the expression of core antigen and the role of precore region, the coding sequence of HBcAg with or without precore (pre-C) region were subcloned into yeast expression vector containing phosphoglycerate kinase (PGK) promoter. To study the role of upstream region in the expression of the core antigen, a series of 5' deletion mutants were also subcloned into the vector. After transformation into various host strains, the expression of HBcAg were analysed by radio-immunoassat. Under optimal condition of core antigen gene expression in yeast, the highest amount of antigen was detected in the cell line SHY4 containing pGKHBc plasmid composed of the yeast PGK gene promoter, terminator and C-gene. Regardless of the presence of precore region, core antigen was not detected in the medium but in cell extract. These results suggest that precore region cannot affect the secretion of core antigen in Saccharomyces cerevisiae.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.33 no.7
• /
• pp.1180-1185
• /
• 2004

This study was conducted to investigate the optimum conditions for the medium composition after isolating and identifying yeast and lactic acid bacteria from sourdough. It was found that the best quality lactic acid bacterium with acid product and flavor was identified as Leuconostoc species among isolated 115 lactic acid bacteria, the best Quality yeast with good fermentation and flavor was identified as Saccharomyces species among isolated 8 yeast. While the microbial growth with glucose or sucrose as sugar source was good, it was, selected that sucrose which is using commercially is better than glucose. The growth of lactic acid bacterium ; was good with 1％ added sucrose whereas yeast needed more growth. Additionally, the medium for the optimum 1 growth of the yeast was composed of 0.5％ wheat flour, 0.5％ peptone and 3％ sucrose, whereas lactic acid bacterium was composed of 0.5％ wheat flour and 1％ sucrose without peptone.

• Microbiology and Biotechnology Letters
• /
• v.4 no.4
• /
• pp.152-158
• /
• 1976

Cultivation condition of yeast on the utilization of fermentable substrate from the cellulosic wastes such as rice hull, rice straw and corn starch cake was investigated. The results obtained were summarized as follows；1. Corn starch cake was respectively added to rice hull and rice straw in order to increase sugar concentration in the hydrolyzate, and then hydrolyzed. As the result, concentration of sugar in hydrolyzed solution of rice hull was 9.12％, in that of rice straw was 7.98％. 2. It was found that calcium carbonate as a neutralizer was the most effective to prepare the culture broth of yeast. 3. An optimal growth of Hansenula subpelliculosa GFY-2 was observed in the medium prepared by adding 0.3％ of ammonium sulfate, 0.4％ of potassium phosphate dibasic, 0.02％ of magnesium sulfate, sodium chloride and calcium chloride to hydrolyaed sugar solution, respectively. 4. Hansenula subpellicuiosa GFY-2 cultured in the substrate solution which of rice hull and rice straw added to corn starch cake was assimilated more than 90％ of sugar in the hydrolyzate within 48 hours. The yeast cells yielded in rice hull was 46.5％, and that of rice straw 45.4％ to utilized sugars.

• Microbiology and Biotechnology Letters
• /
• v.6 no.4
• /
• pp.155-159
• /
• 1978

Phenol utilizing yeast No. 558 isolated from soil sewage sediment was able to use substantial amount of phenol as the sole carbon source, and the biomass productivity by this organism was very excellent. This organism could grow well in 1000 ppm of phenol concentration, the maxim-um specific growth rate obtainable at pH 5.0, 3$0^{\circ}C$ was 0.27/hr., and the biomass yield coefficient Y vs. consumed phenol was 3.2. Maximum production rate of biomass was observed at 35$^{\circ}C$, pH 3.5 to pH 4.5, and the addition of the 0.005~0. 01% yeast extract was the most effective. Addition of HgCl$_2$ and phenyl hydrazine, inhibitors of oxide-reductase, in the phenol containing cultural liquid caused this organism no-growth at the concentration of 10$^{-5}$ M, 10$^{-3}$ M respectively. This organism could utilize not only phenol but catechol, resorcinol and benzidine.

• Microbiology and Biotechnology Letters
• /
• v.36 no.4
• /
• pp.307-313
• /
• 2008

The yeast surface expression system, pCTXYN (6.8 kb), of Bacillus endoxylanase gene (xynB, 642 bp) was constructed and introduced into Saccharomyces cerevisiae EBY100 cell. The transformed yeast cell showing the highest endoxylanase activity was selected through the active staining of colonies grown on YPDG medium containing xylan. With the yeast transformant, EBY100/pCTXYN, grown on galactose containing medium, it was found that the endoxylanase was successfully displayed on the yeast cell surface and the xylooligosaccharides were efficiently produced from xylan. The most of endoxylanase activity was detected in the cell fraction and reached about 1.9 unit/mL after 48 h cultivation. The optimized conditions for xylooligosaccharides production from xylan were determined as follows: substrate and its concentration, oat spelt xylan 6%; concentration of yeast whole-cell, 5 unit/mL; temperature, $50^{\circ}C$, and reaction time $2{\sim}4\;h$. When the oat spelts xylan and corncob xylan were hydrolyzed by treatment with cell surface-displayed endoxylanase, xylotriose was formed as a main product.

• Journal of the Korea Organic Resources Recycling Association
• /
• v.9 no.4
• /
• pp.99-104
• /
• 2001

The influence of agitation speed on the yeast growth was investigated in the production of probiotic feed from pulverized liquified food wastes by aerobic fermentation. A yeast Kluyvermyces marxianus was selected through a preliminary screening. The yeast was cultured by 2liter jar fermenter. in 10% solid(w/v) substrate of liquified food waste at $35^{\circ}C$ with each different agitation speed of 500, 900 and 1200 rpm. For the acceleration of enzyme excretion mixed culture with Aspergillus oryzae was also attempted and the results were compared to those of single culture. As results the viable cell number was increased by increasing agitation speed. But it showed highest value in 900rpm and then decreased in 1200rpm. The mixed culture increased amylase activity and growth rate, but did not seem to enhance the highest viable cell count in the final fermentation stage.

• Proceedings of the KSAR Conference
• /
• 2001.03a
• /
• pp.16-16
• /
• 2001

Apoptosis로 일컬어지는 예정된 세포사멸(programmed cell death)은 개별 세포의 입장에서는 곧바로 사멸을 의미하지만, 정상적인 고등 생물의 입장에서는 개체의 발생과 분화하는데 프로그램된 과정이다. 자발적 세포사멸은 다른 조직에 비해 생식 조직인 난소나 정소에서 복잡한 apoptosis 기작들을 가지리라 사료된다. 본 연구는 Bcl-2 family중 apoptotic protein인 Bax에 대해 suppression하는 유전자를 yeast system을 활용하여 돼지 정소와 난소로부터 각각 cDNA library를 구축한 후 탐색하였다. 탐색에 활용된 cDNA library는 돼지의 정소와 난소로부터 mRNA를 분리하여 yeast vector인 pAD-GAL4-2.1에 구축하였고, 마우스 bax 유전자는 gal 1 promoter의 조절 하에 glucose 배지에서는 유도되지 않고, galactose 배지에서만 선택적으로 Bax를 발현할 수 있는 효모 vector(pL19-bax)를 구축하였다. Bax에 의한 apoptosis suppressor를 탐색하기 위해 우선 효모 W303에 pL19-bax를 transform하여 glucose 배지에서 Bax의 발현을 억제하였다. pL19-bax를 가진 효모에 정소와 난소로부터 구축된 cDNA library를 transform 시키고, transform된 효모는 각각 Bax에 의한 toxicity를 저해하는 유전자를 찾기 위해 스크린되었다. 이러한 방법으로 정소 cDNA library 탐색에서는 5 $\times$ $10^{6}$ transformant중 39개, 난소cDNA library 탐색에서는 2 $\times$ $10^{6}$ transformant중 26개의 콜로니가 생존하였다. 이들 콜로니로부터 유전자를 분리하여 분석해 본 결과 여러 그룹으로 분류할 수 있었다. 각 그룹의 관련 유전자는 protein synthesis/degradation 12종, oxidation/reductation 5종, detoxin/ cell cycle promoter 3종, signal transduction/growth factor 5종, 그리고 알려지지 않은 유전자 9종이었다. 그 중, bax-toxicity inhibition에 강력한 survival phenotype을 가지는 유전자(pSEDL)를 동정하였다. 이것은 T3-4-1 콜로니로부터 분리하였는데 140개 아미노산으로 이루어진 인간 SEDL(GenBank, XM_013096) 유전자와 매우 유사한 homology를 가지며, bax와 관련된 기능은 밝혀져 있지 않다. 이외에도 분리된 유전자에는 NADH, thioreduction, 그리고 cytochrome oxidase와 같은 positive 유전자 군이 크로닝되어, Bax를 이용한 효모에서 apoptosis suppressor에 관련된 유전자를 손쉽게 스크린하는 것이 가능하고, 분리된 유전자의 기능을 예측할 수 있어 지금까지 보고된 유전자 크로닝법 보다는 강력한 수단으로 활용될 수 있다는 사실을 시사하였다. 그러나, ORF에 관계없이 Bax 발현에 저항하는 유전자군이 선발된다든지 하는 문제점은 금후 검토가 필요하리라 사료된다.

• Microbiology and Biotechnology Letters
• /
• v.17 no.2
• /
• pp.88-93
• /
• 1989

To improve a new yeast strain capable of converting xylan to ethanol directly, we tried protoplast fusion between xylose fermenting yeast (Candida sp. X-6-41) and xylan assimilating yeast (Crypto-coccus sp. XB-33), finally selected the most promising two fusants (XFU-1 and XFU-2). As the optimum conditions for protoplast formation, the yeast cells were cultured to exponential phase in YPD and YPX containing 0.6M KCI, respectively, and then treated with zymolyase (0.25mg/$m\ell$), cellulase(4mg/$m\ell$) and 100mM 2-mercaptoethanol at pH 8 and 3$0^{\circ}C$. The protoplasts of parental auxotrophs were fused in the presence of 20mM CaCl$_2$and 40% polyethylene glycol(M.W.4000). The physiological and morphological characteristics of the fusants, such as assimilation of carbon sources, cell size, growth rate, xylanase activity and xylan fermentation ability were investigated. Xylanase activity of fusants that cultured in chemically minimal medium was higher than that of fusants that cultured in completed medium, because xylanase producing activity of xylose fermenting yeast(X-6-41) was inhibited by isoleucine.

• KSBB Journal
• /
• v.10 no.3
• /
• pp.249-256
• /
• 1995

${\gamma}$-Glutamylcysteine production by the immobilized microbial system of recombinant Escherichia coli and yeast was investigated. ${\gamma}$-Glutamylcysteine was synthesized from L-glutamic acid and L-cysteine in the presence of ATP by the reaction catalyzed by ${\gamma}$-glutamylcysteine synthetase. An immobilized microbial cell system was developed for the efficient ${\gamma}$-glutamylcysteine production. Recombinant Escherichia coli and yeast were immobilized by alginate. Production of ${\gamma}$-glutamylcysteine was better with the recombinant Escherichia coli for both the synthesis of ${\gamma}$-glutamylcysteine and the ATP regeneration than the mixed system of recombinant Escherichia coli and yeast. The proper radio of recombinant Escherichia coli to yeast was experimentary observed to be 1:4 in the mixed system. Although the immobi1ized system had the slower reaction rate, its reaction stability was increased by 10%.