• 미생물학회지
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• 제47권2호
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• pp.110-116
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• 2011

ScKns1는 Saccharomyces cerevisiae에서 발견되는 이중 특이성 인산화 효소이다. S288c 계열에서 $Sckns1{\Delta}$ 균주는 야생형과 차이를 보이지 않아, ScKns1의 세포 내 기능에 대해 밝혀지지 않았다. 그러나 분열 효모에서 LAMMER kinase는 세포 엉김과 균사형 생장 및 산화 스트레스와 관련된 것으로 밝혀졌다. 따라서 형태학적 변화를 관찰할 수 있는 S. cerevisiae ${\Sigma}1278b$ 균주에서 ScKNS1 결손균주($Sckns1{\Delta}$)를 제조하고 표현형 변화를 통하여 기능을 분석하였다. $Sckns1{\Delta}$ 균주는 균사형 생장을 유도하는 질소고갈 조건과 butanol 첨가 조건에서 균사형 생장의 결함을 보였으며, agar 표면의 부착생장도 감소하였다. 또한 $Sckns1{\Delta}$ 균주에 균사형 생장을 조절하는 MAPK 경로 유전자를 도입한 결과 RAS2와 STE20에 의해서는 균사형 생장 결함이 회복되지 않았으나, STE11, STE12와 TEC1에 의해서 결합이 회복되었다. 이러한 결과들은 S. cerevisiae에서도 LAMMER kinase가 MAPK 경로를 통하여 균사형 생장을 조절함을 시사한다.

• 한국동물학회:학술대회논문집
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• 한국동물학회 1998년도 한국생물과학협회 학술발표대회
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• pp.113-113
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• 1998

In Saccharomyces cerevisiae UV irradiation and a variety of chemical DNA -damaging agents induce the transcription of specific genes, including several involved in DNA repair. One of the best characterized of DNA -damage inducible genes is PHRI, which encodes the apoenzyme for DNA photolyase. Basal-level and damage-induced expression of PHRI require an upstream activation sequence, UASPHRI. Here we report the identification of the UlvIE6 gene of S. cerevisiae as a regulator of UASPHRl activity. Surprisingly, the effect of deletion of UME6 is growth phase dependent. In wild-type cells PHRI is induced in late exponential phase, concomitant with the initiation of glycogen accumulation that precedes the diauxic shift. Deletion of UNIE6 abolishes this induction, decreases the steady-state concentration of photolyase molecules and PHRI mRNA, and increases the UV sensitivity of a rad2 mutant. The results suggest that UM E6 contributes to the regulated expression of a subset of damage-responsive genes in yeast. Furthermore, the upstream repression sequence, URSPHRI, is required for repression and damage-induced expression of PHRl. Here we show identification of YER169W and YDR096W as putative regulators acting through $URS_{PHRI}$. These open reading frames were designated as RPHI (YERl69W) and RPH2 (YDR096W) indicating regulator of PHRI. Simultaneous disruption of both genes showed a synergistic effect, producing a four-fold increase in basal level expression and a similar decrease m the induction ratio following treatment of methyl methanesulfonate(MMS). Mutation of the sequence ($AG_4$) bound by Rphlp rendered the promoter of PHRI insensitive to changes in RPHI or RPH2 status. The data suggest that RPHI and RPH2 act as damage-responsive negative regulators of PHRI. Surprisingly, the sequence bound by Rphlp in vitro is found to be $AG_4$ which is identical to the consensus binding site for the regulators Msn2p and Msn4p involved in stress-induced expression. Deletion of MSN2 and MSN4 has little effect on the induction$.$ ratio following DNA damage. However, all deletions led to a significant decrease in basal-level and induced expression of PHRI. These results imply that MSN2 and MSN4 are positive regulators of P HRI but are not required for DNA damage repression. [Supported by grant from NIH]om NIH]

• 환경생물
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• 제20권2호
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• pp.165-172
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• 2002

두 가지 유해인자가 동시에 작용할 때 생물체에 나타나는 영향은 길항작용에서 상승작용에 이르기까지 폭넓게 나타난다. 상승작용의 생물학적 중요성에도 불구하고 유해인자간의 복합작용을 설명할 수 있는 구체적 연구결과들은 별로 없는 실정이다. 본 연구는 고온과 기타 환경 유해인자의 복합작용에 의해 생물체에 나타내는 반응에 있어서의 규칙성을 도출해 내기 위하여 수행되었다. 고온과 이온화방사선, 자외선, 초음파 등의 동시작용에 관한 효모세포 실험결과를 분석하였으며 다른 연구자들에 의하여 수행된 바이러스, 세균포자, 동식물세포 및 포유류 배양세포에 관한 실험결과를 이용하여 도출된 규칙성을 검증하였다. 본 연구를 통하여 검증된 상승 작용에 관한 규칙성은 다음과 같이 요약될 수 있다. 방사선량률이 일정할 경우 특정 온도범위 내에서만 상승작용을 관찰할 수 있다. 방사선량률을 증가시키면 최대 상승작용을 나타내는 온도 값도 증가하게 되며, 선량률을 낮추면 온도값도 따라서 낮아지게 된다. 방사선 조사시의 온도가 일정할 경우 특정범위의 선량률에서만 상승작용이 관찰되는데, 이 범위 내에 최대상승작용 유발선량률 값이 존재한다. 방사선 조사시의 온도를 낮추면 선량률의 값 또한 낮아지며 그 역도 마찬가지이다. 본 연구에서 밝혀진 규칙성은 상승작용의 신개념 적립에 실마리를 제공할 것이며 특히 생물체에 대한 유해 환경 요인의 복합영향 해석과 환경재해 예방을 위한 영향평가 및 위해 평가에 유용하게 활용될 수 있을 것이다

• Journal of Applied Biological Chemistry
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• 제54권4호
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• pp.303-308
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• 2011

전통고추장의 품질을 개선하고 기호성을 높이기 위하여 단감을 분말화 하여 고추장에 0, 3, 4, 5% 첨가하여 단감 분말을 첨가한 고추장을 제조하고 $20^{\circ}C$에서 90일 동안 숙성시키면서 미생물학적 특성 및 관능적 특성을 조사하였다. 고추장의 숙성 중 총균수는 숙성 초기부터 숙성 마지막 단계까지 서서히 증가하여 $10^8$ CFU/g 수준을 유지하였고 모든 숙성 과정에서 대조구와 단감 분말 첨가한 것 사이에 유의적 차이는 없었다(p <0.05). 효모 및 곰팡이 수는 숙성기간 동안 $10^4-10^5$ CFU/g을 유지하였다. 고추장의 관능검사를 숙성 30, 60, 90일 째 실시한 결과, 관능적 특성에 있어서 단감 분말의 첨가는 고추장의 색에 긍정적인 영향을 미치지 않았으나, 단맛의 경우에 단감 분말 첨가량에 비례적으로 높아지는 경향을 보였고, 감칠맛의 경우에도 비슷한 경향을 보였다. 전체적 기호도 측면에서 보았을 때 숙성 30일 경에는 유의적 차이가 없었으나(p <0.05), 숙성이 진행됨에 따라 4% 단감분말 첨가구가 관능적으로 높은 값을 나타냈으며 숙성 최종 단계에서 단맛과 감칠맛 그리고 전체적 기호도에서 높은 선호도를 보여 단감 분말을 첨가한 고추장을 제조 시 단감 분말을 4% 첨가하는 것이 적절할 것으로 생각된다.

• 한국미생물·생명공학회지
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• 제40권4호
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• pp.348-355
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• 2012

본 연구에서는 S. cerevisiae와 P. pastoris에서 bovine pancreatic (bp-) DNase I의 과발현과 재조합 DNase I의 특성을 조사하였다. bp-DNase I 유전자는 GAL10 promoter, $MF{\alpha}$, GAL7 terminator 사이에 삽입하여 재조합 plasmid인 pGAL-$MF{\alpha}$-DNaseI (6.4 kb)를 구축하였다. 그리고 bp-DNase I 유전자를 AOX1 promoter, $MF{\alpha}$, AOX1 terminator 에 삽입하여 재조합 plasmid인 pPEXI (8.8 kb)를 구축하였다. 재조합 plasmid인 pGAL-$MF{\alpha}$-DNaseI과 pPEXI를 각각 S. cerevisiae와 P. pastoris 숙주세포에 형질전환시켰다. 형질전환된 효모세포들을 galactose와 methanol 배지에서 $30^{\circ}C$, 48시간 배양하면 bp-DNase I은 대부분이 배양 상등액으로 과발현되었다. P. pastoris 형질전환체는 배양 상등액에서 45.5 unit/mL의 DNase I 활성을 보였으며, 반면에 S. cerevisiae 형질전환체는 37.7 unit/mL의 DNase I 활성을 보였다. 또한 DNA 분해 특성을 조사한 결과, P. pastoris 재조합 DNase I으로 기질 DNA(calf thymus)를 처리하였을 때 1분 이내 DNA가 분해되는 것을 확인할 수 있었으며 이는 상업용 bp-DNase I과 S. cerevisiae 재조합 DNase I으로 처리했을 때보다 빠른 분해 패턴을 보였다.

• 한국식품영양과학회지
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• 제33권4호
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• pp.711-715
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• 2004

홍국균이 생산하는 색소를 상업적으로 이용하기 위한 기초연구로서 Monascus purpureus CBS 281.34 80% ethanol 가용성 색소의 열 pH, 유기산 존재 시의 안정성을 알아보았다. 또한 ethanol 가용성 색소 추출물에 증류수를 가해 수분 함량이 0∼80%가 되도록 하고 수용성 색소와 함께 저장하면서 이들의 안정성도 검토하였다. 80% ethanol 가용성 홍국색소는 1$0^{\circ}C$의 저온에서 4주간 저장 시 9.74%의 분해율을 보였으나, 60∼8$0^{\circ}C$의 고온에서는 24시간과 12시간 저장 시 각각 23.06%와 30.36%나 분해되어 색소의 안정성은 단기간 내 크게 저하하였다. pH 2와 pH 10에서의 80% ethanol 가용성 색소 분해율은 pH 4∼8보다 높았고, 0.6% citric acid, 6% acetic acid 및 1.5% lactic acid를 가해 pH 4로 조정하여 4주간 저장 시에는 대조구와 대조구와의 차이가 가장 큰 acetic acid 하에서의 분해율과의 차이도 6.08%로 거의 나타나지 않아 80% ethanol 가용성 홍국 색소는 유기산하에서 비교적 안정한 것으로 사료되었다. 또한 ethanol 가용성 색소에 증류수를 가해 저장하면 색소 안정성은 저하하였는데, 색소 분해율은 수분함량 20%에서 가장 높았고 80%에서 가장 낮았다. 한편 수중에서의 ethanol 가용성 색소는 2$0^{\circ}C$보다 1$0^{\circ}C$에서 현저하게 안정하였다.

• Asian-Australasian Journal of Animal Sciences
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• 제19권10호
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• pp.1429-1436
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• 2006

Fermentation characteristics, nutrient retention and aerobic stability of barley silages prepared using 6 commercial inoculants were evaluated using 126 mini-silos (3-L) in a completely randomized design. Whole barley forage was chopped, wilted to 39% DM and treated with water (control, S) or one of six inoculants: A (containing Lactobacillus plantarum); B (L. plantarum and Enterococcus faecium); C (L. plantarum and Pediococcus cerevisiae); D (L. plantarum, Pediococcus pentosaceus and Propionibacterium freudenreichii, plus hydrolytic enzymes); E (Lactobacillus buchneri plus hydrolytic enzymes); F (L. buchneri and P. pentosaceus plus hydrolytic enzymes). Samples of treated forage were collected for analysis at the time of ensiling, and then 18 silos of each treatment were filled, capped and weighed. Triplicate silos were weighed and opened after 1, 3, 5, 7, 33, and 61 d. On d 61, $400{\pm}5g$ of material from each silo was placed in 1-L styrofoam containers, covered with cheesecloth and held at room temperature. Silage temperature was recorded hourly for 14 d via implanted thermocouple probes. Chemical composition of the forage at ensiling was consistent with previously reported values. At d 61, pH was lowest (p<0.01) in silage S. Ammonia-N was lower (p<0.05) in silage A than in silages S, B, E, or F. Compared to pre-ensiling values, water soluble carbohydrate concentrations were elevated in silages S, A, B, C and D, and decreased in E and F. Lactic acid concentrations were similar (p>0.10) across treatments. Acetic acid levels were highest (p<0.01) in silage E and lowest (p<0.01) in silage D. Recovery of DM was lower (p<0.01) in silage F than in silages S, A, B, C, or D. On d 61, yeasts were most numerous (p<0.01) in silage D, which was the only silage in which temperature rose more than $2^{\circ}C$ above ambient during aerobic exposure. Silage D also had the highest (p<0.01) pH and ADIN content after aerobic exposure. Lactic acid and WSC content of silage D decreased dramatically during the 14-d aerobic exposure period. Yeast counts (at d 14 of exposure) were lowest (p<0.01) in silages E and F. In general, the commercial inoculants did not appear to enhance the fermentation of barley silage to any appreciable extent in laboratory silos.

• Asian-Australasian Journal of Animal Sciences
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• 제20권3호
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• pp.378-384
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• 2007

Three microorganisms and one chemical preservative were tested for their effects on the fermentation and aerobic stability of whole-crop wheat, sorghum and maize silages. Wheat at the early dough stage, sorghum at the late milk stage and maize at the one-third milk line stage were harvested and ensiled in 1.5-l anaerobic jars untreated or after the following treatments: control (no additives); Lactobacillus plantarum (LP) at $1.0{\times}10^6$ colony-forming units (CFU)/g of fresh forage; L. buchneri (LB) at $1.0{\times}10^6$ CFU/g; Propionibacterium acidipropionici (PA) at $1.0{\times}10^6$ CFU/g; and a formic acid-based preservative (FAP) at 3 ml/kg of fresh forage weight. Three jars per treatment were sampled on d 90 after ensiling, for chemical and microbiological analysis. At the end of the ensiling period, 90 d, the silages were subjected to an aerobic stability test lasting 5 d. In this test, $CO_2$ produced during aerobic exposure was measured along with chemical and microbiological parameters which serve as spoilage indicators. The silages inoculated with LP had higher concentration of lactic acid compared with the controls and the other treated silages (p<0.05). The controls and LP-inoculated silages spoiled upon aerobic exposure faster than LB, PA and FAP-treated silages. The controls and LP-inoculated silages spoiled upon aerobic exposure faster than LB, PA and FAP-treated silages due to more $CO_2$ production (p<0.05) in these two groups and development of yeasts unlike the other groups. In the experiment, the silages treated with LB, PA and FAP were stable under aerobic conditions. However, the numbers of yeasts was higher in the LP-inoculated wheat, sorghum and maize silages compared with the LB, PA and FAP-treated silages. The LB, PA and FAP improved the aerobic stability of the silages by causing more extensive heterolactic fermentation that resulted in the silages with high levels of acetic and propionic acid. The use of LB, PA and FAP as silage additives can improve the aerobic stability of whole-crop wheat, sorghum and maize silages by inhibition of yeast activity.

• 한국막학회:학술대회논문집
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• 한국막학회 1998년도 제6회 하계 Workshop (98 한국막학회, 국립환경연구원 국제 Workshop, 수자원 보전과 막분리 공정)
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• pp.135-153
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• 1998

Nanofiltration (NF) is a recently introduced term in membrane separation. In 1988, Eriksson was one of the first authors using the word 'nanofiltration' explicitly. Some years before, FilmTech started to use this term for their NF50 membrane which was supposed to be a very loose reverse osmosis membrane or a very tight ultrafiltration membrane. Since then, this term has been introduced to indicate a specific boundary of membrane technology in between ultrafiltration and reverse osmosis. The application fields of the NF membranes are very broad as follows: Demeneralizing water, Cleaning up contaminated groundwater, Ultrapure water production, Treatment of effleunts containing heavy metals, Offshore oil platforms, Yeast production, Pulp and paper mills, Textile production, Electroless copper plating, Cheese whey production, Cyclodextrin production, Lactose production. The earliest NF membrane was made by Cadotte et al, using piperazine and trimesoyl chloride as monomers for the formation of polyamide active layer of the composite type membrane. They coated very thin interfacially potymerized polyamide on the surface of the microporous polysulfone supports. The NF membrane exhibited low rejections for monovalent anions (chloride) and high rejections for bivalent anions (sulphate). This membrane was called NS300. Some of the earliest NF membranes, like the NF40 membrane of FilmTech, the NTR7250 of Nitto-Denko and the UTC20 and UTC60 of Toray, are formed by a comparable synthesis route as the NS300 membrane. Commercially available NF membranes nowadays are as follows: ASP35 (Advanced Membrane Technology), MPF21; MPF32 (Kiryat Weizmann), UTC20; UTC60; UTC70; UTC90 (Toray), CTA-LP; TFCS (Fluid Systems), NF45; NF70 (FilmTec), BQ01; MX07; HG01; HG19; SX01; SX10 (Osmonics), 8040-LSY-PVDI (Hydranautics), NF CA30; NF PES 10 (Hoechst), WFN0505 (Stork Friesland). The typical ones among the commercially available NF membranes are polyamide composite membrane consisting of interfacially polymerized polyamide active layer and microporous support. While showing high water fluxes and high rejections of multivalent ions and small organic molecules, these membranes have relatively low chemical stability. These membranes have low chlorine tolerance and are unstable in acid or base solution. This chemical instability is appearing to be a big obstacle for their applications. To improve the chemical stability, we have tried, in this study, to prepare chemically stable NF membranes from PVA. The ionomers and interfacially polymerized polyamide were used for the modification of'the PVA membranes. For the detail study of the active layer, homogeneous NF membranes made only from active layer materials were prepared and for the high performance, composite type NF membranes were prepared by coating the active layer materials on microporous polysulfone supports.

• Asian-Australasian Journal of Animal Sciences
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• 제15권2호
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• pp.218-221
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• 2002

Whole forage sorghum (saccharatum) cultivar FS5 was harvested at the soft dough ($303{\pm}9g\;kg^{-1}$ DM) stage of maturity. The sorghum was chopped into approximately 20 mm pieces and ensiled under laboratory conditions in 1.5 L Weck glass jars. At ensiling, it was treated with two commercial silage inoculants: Pioneer 1188 (Inoculant A) and Eco-corn (Inoculant B). The inoculant A and B was applied at ca $2{\times}10^5$ or $2{\times}10^4$ colony forming units $g^{-1}$ DM., respectively. Silage with no additives served as a control. Three jars per treatment were opened on days 2, 4, 8, 15 and 60 post-ensiling to study fermentation dynamics. After 60 days of ensiling the silages were analyzed and subjected to an aerobic stability test lasting 5 days. Results showed that both inoculants caused a more rapid rate of pH decrease and a higher amount of lactic acid production. All the silages were well preserved and were stable upon exposure to air. Inoculants did not influence (p>0.05) the ash and total N contents, but tended to reduce acetic acid (p<0.05), butyric acid (p<0.01) and propionic acid (p<0.01) contents, and to increase the lactic acid content (p<0.01). The lower DM content of silages treated with Inoculant A agrees with the greater gas loss resulting from the DM loss, which was in good agreement with the higher yeast counts upon aerobic exposure. Silage treated with inoculant B had the highest DM (p<0.05) and lactic acid contents (p<0.01), and the lowest acetic acid content (p<0.05), which agrees with the rapid reduction of pH and smaller gas loss. Inoculant B reduced the ADF (p<0.01), ADL and NDF (p<0.05) contents, which also indicates smaller losses of organic soluble material. The control silages contained the highest levels of volatile fatty acids but no lactic acid, indicating secondary fermentation. It was concluded that both inoculants may improve the fermentation process, since silages from all treatments were stable upon aerobic exposure, noadvantage could be attributed to any of the inoculants used.