• Title/Summary/Keyword: 효모배양

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Characterization of a heat-resistant antimicrobial peptide secreted by Bacillus subtilis A405 (Bacillus subtilis A405 균주가 생성하는 내열성 항균 peptide의 특성 검정)

  • Koo, Bon-Sung;Lee, Seung-Bum;Yoon, Sang-Hong;Song, Gae-Kyung;Chung, Dae-Sung;Byun, Myung-Ok;Ryu, Jin-Chang
    • The Korean Journal of Pesticide Science
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    • v.2 no.3
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    • pp.28-35
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    • 1998
  • An antimicrobial peptide producing bacterium, Bacillus subtilis A405, was screened and identified among 700 of antagonistic bacteria. The heat-resistant antimicrobial peptide, AMP-405, was purified from the broth culture of B. subtilis A405 through $20{\sim}40%$ ammonium sulfate precipitation and ultrafiltration. The AMP-405 exhibited strong antimicrobial activities against Botrytis cinerea, Cercospora sp., Fusarium oxysporum, Penicillium digitatum, Celletotrichum gloeosporioides, Rhizoctonia solani, Pythium ultimum, Pyricularia oryzae, Escherichia coli, Pseudomonas spp. and Candida albicans. The molecular weight of the peptide was about 3.0 kDa determined by SDS-PAGE, Native-PAGE and Tris-Tricine gradient electrophoresis, and composed of 9 kinds of amino acid such as aspartic acid, glycine, serine, glutamine, valine, leucine, isoleucine, proline, tyrocine. To determine the efficiency of AMP-405 as a potential maintenance of fruits freshness, cherry tomato was srored at $25^{\circ}C$ for 2 weeks after treatment with $50{\mu}g/ml$ of AMP-405 and $10^{5}$ spores/ml of Botrytis cinerea simultaneously. Treatment with AMP-405 resulted in significantly less infection by Botrytis cinerea, than the treatment with tap water as a control.

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Isolation and In Vitro Antimicrobial Activity of Low Molecular Phenolic Compounds from Burkholderia sp. MP-1 (Brukholderia sp. MP-1 에서의 페놀화합물의 분리와 항균활성의 측정)

  • Mao, Sopheareth;Jin, Rong-De;Lee, Seung-Je;Kim, Yong-Woong;Kim, In-Seon;Shim, Jae-Han;Park, Ro-Dong;Kim, Kil-Yong
    • Korean Journal of Soil Science and Fertilizer
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    • v.39 no.4
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    • pp.195-203
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    • 2006
  • An antagonistic strain, Burkholderia MP-1, showed antimicrobial activity against various filamentous plant pathogenic fungi, yeasts and food borne bacteria (Gram-positive and Gram-negative). The nucleotide sequence of the 16S rRNA gene (1491 pb) of strain MP-1 exhibited close similarity (99-100%) with other Burkholderia 16S rRNA genes. Isolation of the antibiotic substances from culture broth was fractionated by ethyl acetate (EtOAc) solvent and EtOAc-soluble acidic fraction. The antibiotic substances were purified through a silica gel, Sephadex LH-20, ODS column chromatography, and high performance liquid chromatography, respectively. Four active substances were identified as phenylacetic acid, hydrocinnamic acid, 4-hydroxyphenylacetic acid and 4-hydroxyphenylacetate methyl ester by gas chromatographic-mass spectrum analysis. The minimum inhibition of concentration (MIC) of each active compound inhibited the growth of the microorganisms tested at 250 to $2500{\mu}g\;ml^{-1}$. The antimicrobial activity of crude acidic fraction at 1 mg of dry weight per 6 mm paper disc was more effective than authentic standard mixture (four active substances were mixed with the same ratio as acidic fraction) over a wide range of bacterial test.

Changes in Non-saponin Fatty Acid Content and Increases in Inhibitory Activities of Collagenase and Elastase by Treatment with Saccharomyces cerevisiae of the Supercritical Fluid Extracted Oil of the Adventitious Roots Culture of Wild Mountain Ginseng (초임계 유체로 추출된 산삼 부정 배양근 오일의 효모균 처리에 의한 비사포닌계 지방산 함량 변화 및 Collagenase 및 Elastase 저해 활성 증대)

  • Kim, Chul Joong;Shim, Jae Kwon;Kwon, Kyung Cheol;Lim, Jung Dae;Choi, Seon Kang;Yu, Chang Yeon;Lee, Jae Geun
    • Korean Journal of Medicinal Crop Science
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    • v.26 no.2
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    • pp.170-180
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    • 2018
  • Background: To obtain useful cosmetic resources, this study aimed to determine the non-saponin fatty acid and inhibitory activities of collagenase and elastase by treatment of Saccharomyces cerevisiae in supercritical fluid extracted oil of the adventitious root culture of wild mountain ginseng. Methods and Results: We performed supercritical fluid extraction at various conditions such as pressure, temperature, time, and use of co-solvents, unlike the n-hexane extraction for the adventitious roots culture of wild mountain ginseng. The non-saponin-fatty acid obtained from the oil of the adventitious roots culture was incresed by treatment with S. cerevisiae. The supercritical fluid extraction was conducted using gas chromatography. Non-saponin-fatty acid content, in the oil of adventitious roots culture of wild mountain ginseng treated with S. cerevisiae for 2 days were three times higher than that in the control. In addition, the oil of the adventitious roots culture treated with S. cerevisiae was investigated for the anti-wrinkle effect by using collagenase and elastase. The oil of adventitious roots culture treated with S. cerevisiae exhibited higher collagenase and elastase inhibitory activities than those in the control. Conclusions: Supercritical fluid extracted oil of the adventitious roots culture of wild mountain ginseng treated with S. cerevisiae was found to have decreased ratio of saturated fatty acids and incresed ratio and content of unsaturated fatty acids increased. Furthermore, it showed anti-wrinkle effects in vitro.

FACTORS INFLUENCING THE FORMATION OF INSOLUBLE GLUCAN BY STREPTOCOCCUS SOBRINUS (Streptococcus sobrinus의 비수용성 글루캔 합성에 영향을 미치는 인자)

  • Chung, Jin;Kim, Shin
    • Journal of the korean academy of Pediatric Dentistry
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    • v.27 no.1
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    • pp.90-97
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    • 2000
  • There are various kinds of factors associated with the formation of dental plaque in oral cavity such as nutrient molecules and chemical agents. The factors influencing the formation of insoluble glucan by Streptococcus sobrinus and its replication were examined on orthodontic wires. The results were as follows: 1. Insoluble glucan was well produced in the media initially adjusted at pH 7.0 than pH 5.5 or pH 8.5 like bacterial replication. 2. The synthesis of insoluble glucan and bacterial replication were significantly increased in the media containing 2.5% yeast extract. The formation of insoluble glucan was inhibited by 10 folds in the media containing 20% of sucrose than 1.25%, but the replication of bacteria was increased by 20 folds. 3. Insoluble glucan was significantly formed at a concentration of 1.0mM of calcium chloride, 40mM of potassium chloride, 0.1mM of magnesium chloride, while the replication of bacteria was little influenced by them regardless their concentration. 4. The formation of insoluble glucan and bacterial replication were significant in the media containing 10mM of sodium bicarbonate, but both were completely inhibited at 100mM or above. The production of insoluble glucan and the bacterial replication were largely decreased at 10mM of Tris while insoluble glucan was formed in abundance at 100mM of Tris. 5. The synthesis of insoluble glucan and the bacterial replication were inhibited at 10mM or above of sodium phosphate and potassium phosphate.

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Verification of Mold Determination Method using Slide Culture by Monitoring (모니터링을 이용한 Slide Culture 곰팡이 시험법 검증)

  • Lee, Hee-Sook;Park, Kun-Sang;Sin, Yeong-Min;Lee, Myung-Ja;Lim, Jong-Mi;You, Hyun-Jeong;Kim, Ki-Hyun;Cho, Dae-Hyun;Kim, Dai-Byung;Kim, Ok-Hee
    • Journal of Food Hygiene and Safety
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    • v.22 no.3
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    • pp.137-144
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    • 2007
  • Koji is steamed rice that has had koji-kin, or koji mold spores, cultivated onto it. The isolation, culture, and microscopic examination of molds in the koji require the use of the selective media and special microscopic slide techniques. If simple wet mount slides of molds were attempted, it became apparent that wet mount slides made from mold colonies usually don't reveal the arrangement of spores that is so necessary in identification. The process of merely transferring hyphae to a slide breaks up the hyphae and sporangiophores in such a way that identification becomes very difficult. The slide culture method is superior to wet mounts in that the hyphae, sporangiophores, and spores remain more or less intact when transferred. The procedure that will be used to produce a mold culture on a slide that can be observed directly on the slide. We investigated the contamination rate of penicillium spp. on the 21 kinds of koji distributed at Korea. The contamination rate of Penicillium spp. were not detected at 21 products by slide culture method. These results will be used to reestablish a mold determination of koji and food in Food Additives Code.

Characterization of ${\alpha}$-amylase Producing Hybrid Constructed between Saccharomycopsis and Saccharomyces (Saccharomycopsis속과 Saccharomyces속의 잡종형성 균주에서 생산하는 ${\alpha}$-amylase의 특성)

  • Yang, Young-Ki;Moon, Myeng-Nim;Lim, Chae-Young;Rhee, Young-Ha;Kim, Jeong-Ho
    • Korean Journal of Microbiology
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    • v.35 no.4
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    • pp.315-321
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    • 1999
  • This study has been performed to deveope a yeast strain having high ${\alpha}$-amylase production ability using nuclear transfer method. Hybrids formed between the strains of Saccharomyces fiburigera KCTC 7393 and Saccharomyces cerevisiae KCTC 7049 (tyr-, ura-)were obtained by nuclear transfer technique. Nuclei isolated from the wild type S. fiburigera strain were transfered into auxotrophic mutants S. cerevisiae and selected the hybrids showing an increased starch degrading capability were selected (MN-16). This transformant grew best and produced maximal ${\alpha}$-amylase activity on the medium containing 2% (V/V) soluble starch. ${\alpha}$-Amylase from MN-16 was purified electrophoretically homogenety and its properties were investigated. The enzyme was purified about 10.6 fold with an overall yield 9.7% from the culture medium by ammonium sulfate fractionation. DEAE-Sephacel column chromatography, and Sephacryl S-200 column chromatography. The purified enzyme showed a single band on SDS-polyacrylamide gel electrophoresis. The molecular weight of the ${\alpha}$-amylase was estimated to be 53,000 daltons by SDS-PAGE and by gel permeation chromatography on Sephacryl S-200. The purified enzyme showed the maximum activity at pH 5.5 and 40${\circ}C$. The km value for soluble starch was 2.5㎎/㎖. The enzyme activity increased in the presence of $Ca^{2+}, Co^{2+}, EDTA, Mg^{2+}, Mn^{2+}, Zn^{2+}$, but inhibited by $Cu^{2+}, Fe^{2+}$, and $Ni^{2+}$

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Expression of Tkermomonoepora fusea Exoglucanase in Saccharomyces cerevisiae and Its Application to Cellulose Hydrolysis (Saccharomyces cerevisiae에서 Tkermomonospora fusca Exoglucanase의 발현 및 Cellulose분해에의 응용)

  • Park Hyun-Soon;Kim Hyun-Chul;Shin Dong-Ha;Kim Joong-Kyun;Nam Soo-Wan
    • Microbiology and Biotechnology Letters
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    • v.33 no.4
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    • pp.267-273
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    • 2005
  • To develop effective and powerful probiotic, Saccharomyces cerevisiae strains producing cellulolytic enzymes were genetically brooded. For the production of exoglucanase, the plasmid pVT-TExo (8.8 kb) was constructed, in which Thermomonosporafusca exoglucanase gene (E3) was under the control of ADHl promoter, and introduced into S. cerevisiae SEY2102. When the transformant, S. cerevisiae SEY2102/pVT-TExo, was cultivated on YPD medium, the total expression level of avicelase reached about 190 unit/l. The secretion efficiency and plasmid stability were about $50\%\;and\;91\%$, respectively. Recombination exoglucanase enzyme bound to avicel better than Clostridium endoglucanase (CelA) and Trichoderma endoglucanase (C4) enzymes. The mixing ratio of E3 and CelA displaying the best synergistic hydrolysis for avicel was observed at 4:1. The mixture of endoglucanase (CelA) and exoglucanase (E3) resulted in 3.2-fold increase of avicelase activity and 2.5-fold enhanced production of sugar production from avicel, compared to the single enzyme treatment.

Experimental Analysis on Regularities of Synergistic Interaction of Temperature with Physico-Chemical Environmental Factors (온도와 물리화학적 환경요인에 의한 상승작용의 규칙성에 관한 실험적 고찰)

  • 김진규;신해식;블라디슬라프페틴;이영엽
    • Korean Journal of Environmental Biology
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    • v.20 no.2
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    • pp.165-172
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    • 2002
  • The combined action of two factors on organisms can be either antagonistic, non-effective, additive or synergistic. Although synergism is of biological importance, the common features of synergistic interaction between harmful environmental factors are largely unknown. The purpose of this study is to establish general rules describing the response of various organisms to the combined action of heat with another inactivating agent. Synergistic interaction due to the simultaneous treatment of hyperthermia with ionizing or non-ionizing radiation has been analyzed using the experimental data mainly obtained with yeast cells. In addition, the results reported by others for viruses, bacterial spores, cultured mammalian cells, plants and animals were also analyzed to check the regularities revealed. The common rules of the synergistic interaction obtained in this study can be summarized as follows. For any constant rate of exposure, the synergy can be observed only within a certain temperature range. An increase in exposure rate resulted in an increase of this specific temperature and vice versa. For a constant temperature at which the irradiation occurs, synergy can be observed within a certain dose rate range. As the exposure temperature is reduced, the optimal intensity decreases and vice versa. A new conception taken into consideration those regularities can make a clue for environmental disaster preventive analysis of the synergy of radiation with the other factor.

Expression and Regulatory Analysis of Sporulation Gene (spo 5) in Schizosaccharomyces pombe (Schizosaccharomyces pombe 포자형성유전자 (spo 5)의 발현조절기구의 해석)

  • KIM Dong-Ju;SHIMODA Chikasi
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.30 no.1
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    • pp.46-54
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    • 1997
  • Sporulation in the fission yeast Schizosaccharomyces pombe has been regarded as an important model of cellular development and differentiation. S. pombe cells proliferate by mitosis and binary fission on growth medium. Deprivation of nutrients especially nitrogen sources, causes the cessation of mitosis and initiates sexual reproduction by malting between two sexually compatible cell types. Meiosis is then followed in a diploid cell in the absence of nitrogen source. DNA fragment complemented with the mutations of sporulation gene was isolated from the S. pombe gene library constructed in the vector, pDB 248' and designated as pDB (spo 5)1. We futher analyzed six recombinant plasmids, pDB (spo 5)2, pDB(spo 5)3, pDB(spo 5)4, pDB(spo 5)5, pDB(spo 5)6, pDB(spo 5)7, and found each plasmids is able to rescue the spo 5-2, spo 5-3, spo 5-4, spo 5-5, spo 5-6, spo 5-7, mutations, respectively. Mapping of the integrated plasmid into the homologous site of the S. pombe chromosomes demonstrated that pDB (spo 5)1, and pDB (spo 5)R1 contained the spo 5 gene. Transcipts of spo 5 gene were analyzed by Northern hybridization. Two transcripts of 3.2 kb and 25 kb were detected with 5 kb Hind III fragment containing a part of the spo 5 gene as a probe. The small mRNA (2.5 kb) appeared only when a wild-type strain was cultured in the absence of nitrogen source in which condition the large mRNA (3.2 kb) was produced constitutively. Appearance of a 2.5 kb spo 5-mRNA depends upon the function of the mei1, mei2 and mei3 genes.

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Characterization of Bacteriocin-Like Substances Produced by Bacillus subtilis MJP1 (Bacillus subtilis MJP1이 생산하는 Bacteriocin-Like Substances)

  • Yang, Eun-Ju;Chang, Hae-Choon
    • Microbiology and Biotechnology Letters
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    • v.35 no.4
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    • pp.339-346
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    • 2007
  • The MJP1 bacterial strain, which possesses antifungal activity, was isolated from meju and identified as Bacillus subtilis based on its morphological and biochemical properties, as well as its 16S rRNA sequence. Antimicrobial activity was found against various species of Gram-positive bacteria, yeasts, and molds, including food-spoilage microorganisms. The antifungal activity was found to be stable after heat and proteolytic enzyme treatment, and in the pH range of $6.0{\sim}10.0$. The antibacterial activity was stable in the pH range of $6.0{\sim}10.0$, but about 50% of the activity was lost after 24 hr at $30^{\circ}C$. The antibacterial compound was also inactivated by proteolytic enzyme treatment, indicating its proteinaceous nature. The apparent molecular masses of the partially purified antifungal and antibacterial compounds, as indicated by using the direct detection method in Tricine-SDS-PAGE, were approximately 2.4 kDa and 4.5 kDa, respectively. These studies suggest that B. subtilis MJP1 produces two bacteriocin-like substances with antifungal and antibacterial activities.