• Animal cells and systems
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• 제6권4호
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• pp.317-322
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• 2002

To improve conventional yeast one-hybrid screening, we have developed an efficient mammalian one-hybrid system that allows rapid isolation of com-plementary DNAs which are able to induce human p14$^{ARF}$. tumor suppressor gene. A 1.5 kb promoter region of p14$^{ARF}$ was fused to EGFP to generate ARF promoter-EGFP reporter vector. This reporter plasmid was stably trans-fected into NIH3T3 cells for generation of reporter cell line. When the reporter cell line was infected with E2F-1 together with excess amounts of empty vector, the cells that received the positive modulator were readily identifiable by green fluorescence using FACS. The GFP-positive cells were cloned directly from the cultured cells and expanded in bulk culture. The genomic DNAs from GFP-positive cells were prepared and the CDNA insert in integrated retroviral genome was recovered by PCR using primers annealing to the retroviral vector sequences flanking the insert-cloning site. This system should be useful for efficient screening of expression CDNA libraries in mammalian cells to identify novel upstream regulators for spe-cific genes by one-hybrid interaction.ion.

• Food Science and Biotechnology
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• 제16권1호
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• pp.133-137
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• 2007

The Strain TFM-7, Which has an antitumor effect, was isolated from Kefir and identified based on analysis using the API 50 CHL kit and 265 rDNA sequencing. Strain TFM-7 was confirmed to belong to the genus Kluyveromyces. Analysis of the 265 rDNA nucleotide sequences found strain TFM-7 to be related to Kluyveromyces marxianus. NRRL Y-828IT. K. marxianus. TFM-7 was cultured with potato dektrose broth medium at $27^{\circ}C$ for 72 hr, and its inhibition effects on the proliferation of seven tumor cell lines and a normal cell line were assessed using the MTT assay. The antitumor effects and growth characteristics of K. marxianus TFM-7 were investigated during a culture period of 7 days. By the $3^{rd}\;day$, K. marxianus TFM-7 showed a dry cell weight 2.39 g/L, a pH of 4.39, an ethanol content of 0.89%, and an inhibition effect on the proliferation of seven tumor cell lines above 50%, except for A-549 tumor cell line. K. marxianus TFM-7 was the most effective at inhibiting the growth of Hep-2 cell line among all tumor cell lines tested. Growth inhibition of a normal cell line, NIH/3T3, was less than 35%, suggesting a decreased level of cytotoxicity toward normal cells. These results indicate that K. marxianus TFM-7 may have used as a yeast strain with antitumor activity.

• 한국미생물·생명공학회지
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• 제30권1호
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• pp.57-62
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• 2002

Acetobacter xylinum GS11 균주를 이용하여 다양한 배양조건에서 미생물 셀룰로오스의 생산성을 검토하였다. 탄소원으로 glucose 이외에 첨가한 기질은 영향이 없었으며, 2% glucose 농도 범위에서 셀룰로오스 생산량이 2.8 g/L으로 가장 양호한 것으로 나타났다. 질소원은 Y.E와 soytone등의 유기질소원 첨가가 효과적이었으나, 무기질소원의 효과는 나타나지 않았다. 무기염류 및 아미노산의 첨가는 대부분 효과적이었으며, 이 중 무기염류는$ MgSO_4$가 1.5배, 아미노산은 phenylalanine이 1.4배의 셀룰로오스 생산성을 나타냈다. 비타민은 $\alpha$-tocopherol의 첨가 시 1.4배의 생산성을 보였다. 또한 기본배지에 Coconut milk와 0.5%의 lignosulfonate의 첨가가 대조구와 비교하여 각각 2.2, 2.1배의 셀룰로오스의 생산성을 나타내어 실험 처리 중 가장 효과적이었다.

• 한국식품과학회지
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• 제30권6호
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• pp.1394-1398
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• 1998

L. bulgaricus LBS 47과 S. serevisiae SCS 5를 접종하여 전통안동식혜를 발효 후 저장하는 동안에 안정제에 의한 침전억제 효과는 Na-Alginate가 가장 우수하였다. 침전 안정제를 넣어서 관능 검사를 한 결과 균질화 하지 않은 식혜의 경우는 Na-alginate는 0.1에서 0.15% 처리했을 때 Carrageenan은 0.05% 첨가했을 때 가장 맛이 우수하였으며 식품 안정제의 종류 및 첨가 농도간의 상호 효과는 침전 안정제의 종류간에는 통계적 유의성이 없으나 첨가 농도간에는 유의성이 있었다. 식혜의 현탁안정성을 증진시킬 목적으로 안정제를 첨가하여 숙성시켰을 때 CMC, Na-alginate은 2일째 carrageenan은 비균질화 식혜는 2일에서 4일째 균질화한 식혜는 1일째 최고의 점도를 나타내었다.

• Mycobiology
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• 제44권4호
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• pp.283-290
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• 2016

A double-stranded RNA (dsRNA) mycovirus was detected in malformed fruiting bodies of Pleurotus ostreatus strain ASI2792, one of bottle cultivated commercial strains of the edible oyster mushroom. The partial RNA-dependent RNA polymerase (RdRp) gene of the P. ostreatus ASI2792 mycovirus (PoV-ASI2792) was cloned, and a cDNA sequences alignment revealed that the sequence was identical to the RdRp gene of a known PoSV found in the P. ostreatus strain. To investigate the symptoms of PoV-ASI2792 infection by comparing the isogenic virus-free P. ostreatus strains with a virus-infected strain, isogenic virus-cured P. ostreatus strains were obtained by the mycelial fragmentation method for virus curing. The absence of virus was verified with gel electrophoresis after dsRNA-specific virus purification and Northern blot analysis using a partial RdRp cDNA of PoV-ASI2792. The growth rate and mycelial dry weight of virus-infected P. ostreatus strain with PoV-ASI2792 mycovirus were compared to those of three virus-free isogenic strains on 10 different media. The virus-cured strains showed distinctly higher mycelial growth rates and dry weights on all kinds of experimental culture media, with at least a 2.2-fold higher mycelial growth rate on mushroom complete media (MCM) and Hamada media, and a 2.7-fold higher mycelial dry weight on MCM and yeast-malt-glucose agar media than those of the virus-infected strain. These results suggest that the infection of PoV mycovirus has a deleterious effect on the vegetative growth of P. ostreatus.

• 미생물학회지
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• 제54권3호
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• pp.244-253
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• 2018

이 연구는 분리된 토양 세균에 의해 생성된 항균물질의 효과를 평가하기 위해 수행되었다. 2000여개의 세균 분리주 중 Paenibacillus elgii DS381이 여러 인간 피부 상재균과 병원성 세균에 대해 높은 항균활성을 나타내었다. DS381 균주는 agar well diffusion test에서 모든 대상 세균과 효모에 대해 15.3~26.0 mm 직경의 저해대를 형성하였다. DS381이 생성한 항균 펩티드는 모든 대상 미생물에 낮은 최소저해농도 (0.039-5.000 mg/ml)를 나타내었다. DS381 균주는 lipopeptide 같은 생물계면활성제 생산을 나타내었는데, 배양 상등액의 표면장력을 60.0에서 40.3 mN/m으로 낮추었다. DS381은 또한 $1.56{\pm}0.13U/ml$의 chitinase 활성도 나타내었다. 이 결과들은 P. elgii DS381이 일부 중요한 인간 피부 상재균과 병원성 세균에 대한 효율적인 생물제어제로 사용될 수 있음을 가리킨다.

• 한국식품과학회지
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• 제19권4호
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• pp.324-330
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• 1987

토양, 호수, 개천, 연못, 부때우유, 단무지로부터 메탄올을 자화하는 효모와 세균을 분리하였다. 효모 중 가장 생육이 빠른 균주를 선발하여 동정하고 메탄올 배양에서의 생육특성을 조사하였다. 그 결과 분리된 효모 YF-3은 Candida boidinii로 동정되었다. 이 균주의 특성은 biotin을 요구하며 $C_1$화합물 외에 다른 화합물도 이용할 수 있는 facultative methylotroph로서 최적 pH는 6.0, 최적 온도는 $30^{\circ}C$, 최적 매탄올 농도는 배지의 1%(v/v)였다. 균체 생산량은 건조균체로 2.72g/l 균체 수율은 $0.39\;g\;g^{-1}$로 측정되었다. 균체의 총핵산 함량은 5.9%, 조단백질 함량은 45.5%, 총아미노산 함량은 35.22%도 분석 되었다.

• Food Science and Biotechnology
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• 제17권4호
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• pp.766-771
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• 2008

The proteolytic enzyme from Saccharomyces sp. B101 was purified to homogeneity by ammonium sulfate fractionation, ultrafiltration, diethyl aminoethyl (DEAE)-Sephadex A-50 ion-exchange chromatography, and Sephadex G-100 gel filtration chromatography from the culture supernatant of Saccharomyces sp. B101. The specific activity and the purification fold of the purified enzyme were 4,688.9 unit/mg and 18, respectively. The molecular weight of the purified enzyme was estimated to be 33 kDa by sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The optimum pH and temperature for the enzyme activity were pH 8.5 and $30^{\circ}C$, respectively. The enzyme activity was relatively stable in the pH range of 6.5-8.5 at below $35^{\circ}C$. The salt-tolerance and stability for the enzyme activity were relatively stable even at NaCl concentrations of 10 and 15%. The activity of enzyme was inhibited by $Ag^{2+}$ and $Fe^{2+}$, and activated by $Mn^{2+}$. In addition, the enzyme activity was potently inhibited by ethylenediaminetetraacetic acid (EDTA) and phenylmethyl sulfonylfluoride (PMSF). Based on these findings we concluded that the purified enzyme was a serine protease. Km and Vmax values for hammastein milk casein were 1.02 mg/mL and 278.38 unit/mL, respectively.

• 한국초지조사료학회지
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• 제41권3호
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• pp.176-182
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• 2021

Ensiling is the most preferred technology to preserve the silage quality with high nutrients by the presence of lactic acid bacteria. In this study, lactic acid bacteria RJ1 and S22 were used to make the silages from different leguminous plants such as alfalfa, hairy vetch and red clover. Experimental groups were divided into control and LAB inoculated groups. LAB inoculated group; all legumes treated with a mixture of RJ1 and S22 and made an anaerobic condition for 45d. Without the addition of LAB considered the control group. The results showed that the lactic acid content was higher in all silages in response to LAB treatment and acetic acid content was slightly increased except red clover by LAB compared to control silages. A poor silage quality marker butyric acid was reduced all legume silages in response to LAB inoculation than control silages. The organic acid is closely associated with microbial population experimental silages. We noted that higher LAB and lower yeast were found in the silage in response to LAB treatment. The contents of crude protein (CP), acid detergent fiber (ADF), neutral detergent fiber (NDF), and total digestible nutrient (TDN) were not altered significantly between control and LAB treated silages. Overall data suggested that the inclusion of additional LAB potentially enhance the silage quality and preserved the nutrients for long period.

• 대한화장품학회:학술대회논문집
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• 대한화장품학회 2003년도 IFSCC Conference Proceeding Book I
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• pp.700-718
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• 2003

Fructan, a polysaccharide existing in plants or produced by microorganisms, is a sugar polymer of fructose with $\beta$-2,6 linkages. In this study, we investigated some cosmeceutical properties of Fructan such as moisturizing effect, cell proliferation effect, anti-inflammation effect and cell cytotoxicity. Zymomonas mobilis, a microorganism producing Fructan, was cultured in a medium containing 10％ sucrose and 2％ yeast extract as main components for 24 hours at 37$^{\circ}C$ and pH 7. Fructan was obtained by precipitation from the cultured medium by adding alcohol (alcohol ratio of 1:3) after removing the enzyme by centrifuging. Fructan exhibited almost same moisturizing effect as hyaluronic acid and cell proliferation effect on human fibroblast and keratinocyte as well. Moreover, on cell proliferation test on bio-artificial skin constructed by 3-dimensional(3-D) culture after inducing primary skin inflammation with 0.5％ sodium lauryl sulfate (SLS), the 3-D artificial skin treated with 0.01 mg/ml, 0.05mg/ml of Fructan exhibited higher cell proliferation than the 3-D artificial skin treated with SLS only. On anti-inflammation test on 3-D artificial skin evaluated by measuring secreted quantity of interleukin-1$\alpha$ (IL-1$\alpha$) which is a pre-inflammatory mediator induced by SLS, the quantity of IL-1$\alpha$on the 3-D artificial skin treated with 0.01 mg/ml, 0.05mg/ml of Fructan was less than the one on the 3-D artificial skin treated with SLS only. As a result of these studies, Fructan has anti-inflammation effect against inflammatory reaction by a skin irritant as well as cell proliferation effect in bio-artificial skin. Fructan was also evaluated as a safe material without any toxicity in safety tests using fibroblasts and animals.