• Title/Summary/Keyword: Gram positive

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Identification and Biochemical Characterization of Xylanase-producing Streptomyces glaucescens subsp. WJ-1 Isolated from Soil in Jeju Island, Korea (제주도 토양에서 분리한 xylanase 생산균주 Streptomyces glaucescens subsp. WJ-1의 동정 및 효소의 생화학적 특성 연구)

  • Kim, Da Som;Jung, Sung Cheol;Bae, Chang Hwan;Chi, Won-Jae
    • Microbiology and Biotechnology Letters
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    • v.45 no.1
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    • pp.43-50
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    • 2017
  • A xylan-degrading bacterium (strain WJ-1) was isolated from soil collected from Jeju Island, Republic of Korea. Strain WJ-1 was characterized as a gram-positive, aerobic, and spore-forming bacterium. The predominant fatty acid in this bacterium was anteiso-$C_{15:0}$ (42.99%). A similarity search based on 16S rRNA gene sequences suggested that the strain belonged to the genus Streptomyces. Further, strain WJ-1 shared the highest sequence similarity with the type strains Streptomyces spinoveruucosus NBRC 14228, S. minutiscleroticus NBRC 13000, and S. glaucescens NBRC 12774. Together, they formed a coherent cluster in a phylogenetic tree based on the neighbor-joining algorithm. The DNA G+C content of strain WJ-1 was 74.7 mol%. The level of DNA-DNA relatedness between strain WJ-1 and the closest related species S. glaucescens NBRC 12774 was 85.7%. DNA-DNA hybridization, 16S rRNA gene sequence similarity, and the phenotypic and chemotaxonomic characteristics suggest that strain WJ-1 constitutes a novel subspecies of S. glaucescens. Thus, the strain was designated as S. glaucescens subsp. WJ-1 (Korean Agricultural Culture Collection [KACC] accession number 92086). Additionally, strain WJ-1 secreted thermostable endo-type xylanases that converted xylan to xylooligosaccharides such as xylotriose and xylotetraose. The enzymes exhibited optimal activity at pH 7.0 and $55^{\circ}C$.

Generation of Transgenic Plant (Nicotiana tabacum var. Petit Havana SR1) harboring Bacillus thuringiensis Insecticidal Crystal Protein Gene, cry II A (Bacillus thuringiensis 살충성 결정단백질 유전자(cry II A)의 형질전환 식물 제작)

  • 이정민;류종석;권무식
    • Korean Journal of Plant Tissue Culture
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    • v.24 no.5
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    • pp.305-311
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    • 1997
  • Bacillus thuringiensis, a gram-positive soil bacterium, is characterized by its ability to produce crystalline inclusions during sporulation. The crystal proteins exhibit a highly specific insecticidal activity. An insecticidal crystal protein (ICP), Cry II A, is specifically toxic to both lepidopteran and dipteran insects. In this study, tobacco plants transformed by the cry II A gene have been generated. The Cry II A crystal protein was purified from E. coli JM103 harboring cry II A gene by differential solubility. The activated Cry II A was prepared by tryptic digestion. The purified protoxin (70 kDa) and the activated toxin (50 kDa) were analyzed by SDS-PAGE. To generate the transgenic tobacco having cry II A gene, the cry II A gene was subcloned to a plant expression vector, pSRL2, having two CaMV 35S promoters. The recombinant plasmid was transformed into tobacco (N. tabacum var. Petit Havana SR1) by Agrobacterium-mediated leaf disc transformation. Through the regeneration, six putative transgenic tobacco plants were obtained and three transformants were confirmed by Southern blot analysis. It has been found that one plant had single copy of cry II A gene, another had two copies of the gene, and the third had a truncated gene. After the immunochemical confirmation of cry II A expression in plants, the transgenic tobacco plants will be used to study the genetics of future generation with the insecticidal crystal protein gene cry II A.

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Effect of Gamma Irradiation on the Sterilization of Red Pepper Powder (고추가루의 살균(殺菌)을 위한 ${\gamma}$-선(線) 조사효과)

  • Kwon, Joong-Ho;Byun, Myung-Woo;Cho, Han-Ok
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.13 no.2
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    • pp.188-192
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    • 1984
  • In order to investigate the hygienic status of commercial red pepper powder, one sample was directly prepared from raw red pepper which was purchased at market and the other one was a commercial red pepper powder. They were used for the investigation upon the status of microbial contamination, effect of irradiation for sterilization and physicochemical changes during 3 months of storage. Total bacterial counts of commercial red pepper powders were 3.83 to $6.68{\times}10^6$ per gram and coliform group was shown to be positive in some products. Total bacteria and coliform group were sterilized by the irradiation of 9 kGy and 3 kGy, respectively and no microorganisms were grown up until 3 months of storage at room temperature. The $D_{10}$ values of total viable bacteria in red pepper powders were 1.52 to 1.58 kGy. Chemical components such as moisture, total and reducing sugars were slightly decreased during the storage period. The contents of capsanthin and capsaicin were partly affected by the high dose irradiation but the difference between unirradiated and irradiated groups was diminished with the elapse of storage period.

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Xylanase-producing Bacillus subtilis isolated from spent mushroom (Pleurotus eryngii) substrates (큰느타리버섯 폐배지에서 분리한 Xylanase 생성 Bacillus subtilis CS9)

  • Cho, Ji Jong;Hong, Su Young;Ha, Jun;Cho, Young Un;Kim, Hong Chul;Gal, Sang Wan;Yun, Han Dae;Cho, Soo Jeong
    • Journal of Mushroom
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    • v.6 no.3_4
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    • pp.138-145
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    • 2008
  • A Gram-positive, endospore-forming, rod-shaped bacterial strain was isolated from spent mushroom (Pleurotus eryngii) substrates taken from the Hoamfarm located in Keyollgnam, Korea. The isolate, designated CS9, was facultatively anaerobic, motile rod and produced xylanase. The strain grew optimally at $40^{\circ}C$ and pH 6.0. The major cellular fatty acids were anteiso-$C_{15:0}$, anteiso-$C_{17:0}$, and iso-$C_{17:0}$. The genomic DNA G+C content was 45 mol%. Comparative 16S-rDNA sequence analysis showed that the isolate CS9 formed a distinct phyletic line within the genus Bacillus and was most closely related to Bacillus subtilis YB1, with 16S DNA sequence similarity of 96.8%. Sequence similarities to other type strains were 92-94%. On the based of physiological and molecular properties, the isolate CS9 was classified within the genus Bacillus as Bacillus subtilis CS9.

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Antibacterial, Antioxidative and Anti-proliferative Activity against Human Colorectal Cell of Pimpinella brachycarpa (참나물의 항균, 항산화 활성 및 대장암세포 성장억제 활성 평가)

  • Ahn, Seon-Mi;Kim, Mi-Sun;Jung, In-Chang;Sohn, Ho-Yong
    • Food Science and Preservation
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    • v.18 no.4
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    • pp.590-596
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    • 2011
  • Pimpinella brachycarpa, called as cham-na-mul in Korea, is an edible popular herb. However, the study of biological activity of P. brachycarpa is still rudimentary in worldwide. In this study, from the cultivated P. brachycarpa, we prepared the methanol extract and its subsequent solvent fractions, and their antimicrobial, antioxidation, and anti-proliferative activities were evaluated. The fraction yields of n-hexane (H), methylene chloride (EC), ethylacetate (EA), butanol (B), and water residue (W) from the methanol extract were 18.71, 0.7, 0.56, 4.57, and 71.51%, respectively. Analysis of total flavonoid and total polyphenol showed that the EA fraction contained the highest contents (89.23 and 200 mg/g), and the W residue has the lowest contents (19.6 and 2.27 mg/g) among the factions. In antimicrobial activity assay, the EA fraction showed a broad-range antibacterial activity, while the H fraction is effective against gram positive bacteria. In antioxidation activity assay, EA and B fraction showed strong DPPH anion and ABTS cation scavenging activities including reducing power, and Hand MC fraction showed effective nitrite scavenging activity (71.43~83.82 ${\mu}g$/mL of $IC_{50}$). In a while, only B fraction showed strong anti-proliferative activity against human colorectal cancer HCT-116 (166 ${\mu}g$/mL of $IC_{50}$) as a dose-dependent manner up to 200 ${\mu}g$/mL. These results suggest that the EA and B fraction of P. brachycarpa could be developed as functional food ingredients.

Antioxidant and antimicrobial activities of ginger with aging and fermentation (발효숙성생강의 항산화 및 항균 활성)

  • Seo, Young-Ho
    • Food Science and Preservation
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    • v.24 no.8
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    • pp.1180-1187
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    • 2017
  • Zingiber officinale Roscoe, commonly known as ginger, has long been used as a powerful health-promoting antioxidant that supports cellular health of the human body. The objective of this study was to compare the antioxidant and antimicrobial activities of the samples with aging and fermentation. Antioxidant activities of the samples were compared using total phenol, flavonoid contents, ABTS cation radical scavenging activity and DPPH radical scavenging activity. Antimicrobial activities were also examined using the paper disc method and minimum inhibitory concentration (MIC). Acidity of the fermented ginger (FG) with lactic acid bacteria showed a significantly higher value than that of the ginger (GG). The content of 6-gingerol, a bioactive component in ginger, decreased in all fermented gingers but 6-shogaol which is also one of the main valuable ingredients showed the increased content at ginger fermented with Streptococcus thermophilus and Lactobacillus acidphilus. Flavonoid contents of the FG and GG did not show significant differences. However, ABTS cation radical scavenging activity and DPPH radical scavenging activity were 10-30% increased in the samples with fermentation (p<0.05), respectively. The samples of the disc showed an inhibitory effect on growth of gram positive Staphylococcus aureus and Listeria monocytogenes. Zinger with fermentation showed higher antioxidant and antimicrobial activities. Thus, we conclude that aging and fermentation can be a helpful process to increase the functional effects of ginger.

Study on the Anti-oxidative, Anti-microbial and Anti-cancer effect of Bibangtalmyungsan. (비방탈명산(秘方奪命散)의 항산화 ${\cdot}$ 항암 ${\cdot}$ 항균 효과 연구)

  • Han, Hong-Jun;Choi, Jeong-Hwa;Park, Su-Yeon;Kim, Jong-Han
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.20 no.1 s.32
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    • pp.115-129
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    • 2007
  • Objectives : This study was carried out to evaluate anti-oxidative, anti-microbial and anti-cancer effect for clinical application of Bibangtalmyungsan (BTS) Results : 1. The oxidative effects were measured by polyphenol, DPPH radical scavenging activity. BTS water extract was showed more effective than ethanol extract and also various solvent fractions from BTS water extract showed effective in the following order : ethyl acetate fraction > butanol fraction > hexane fraction ${\fallingdotseq}$ chloroform fraction ${\fallingdotseq}$ aqueous fraction 2. The results of anti-microbial effects were as follows. 1) Antibacterial activities of BTS extracts against Gram's negative and positive bacteria were ineffective 2) Antifungal activities of the BTS extracts against Aspergillus spp. , Trichohyton mentagrophyte KTCC 1077 were not effective. 3. The result of anti-cancer effects were as followings: 1) BTS ethanol extract was more effective than water extract against Caco-2, Calu-6, but it had cytotoxic effect against NIH3T3. 2) On the Caco-2, effective only in ethyl acetate $fraction(IC_{50}:$ 35.81 ${\mu}g/ml)$. 3) On the Calu-6, the most effective in ethyl acetate $fraction(IC_{50}:$ 189.65 ${\mu}g/ml)$ and effective In butanol $fraction(IC_{50}:$ 299.74 ${\mu}g/ml)$ and hexane $fraction(IC_{50}:$ 345.13 ${\mu}g/ml)$ 4) On the SUN-601, $IC_{50}$ value was within 80 ${\mu}g/ml$ in ethyl acetate fraction and hexane fraction. 5) On the HCT-1l6, the most effective in ethyl acetate $fraction(IC_{50}:$ 82.94 ${\mu}g/ml)$ and effective in hexane $fraction(IC_{50}:$ 374.56 ${\mu}g/ml)$. 6) On the AML-2/WT, the most effective in ethyl acetate $fraction(IC_{50}:$ 41.44 ${\mu}g/ml)$ and effective In hexane $fraction(IC_{50}:$ 303.01 ${\mu}g/ml)$. 7) On the NIH3T3, effective only in ethyl acetate $fraction(IC_{50}:$ 203.42 ${\mu}g/ml)$, but it was more ineffective than other cancer cells. Conclusion : These result suggest that BTS has antioxidative, antifungal activities and cytotoxic effects against Caco-2, Calu-6, SUN-601, HCT-116, AML-2/WT and NIH3T3, especially ethyl acetate fraction from water extract has more effective in antioxidative and anticancer effects.

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The Establishing Test Method of Bactericidal Activity and the Evaluating of Korean Disinfectants/Sanitizers Efficacy (살균소독력 시험법 확립 및 살균소독력 평가)

  • Kim, Hyung-Il;Lee, Kwang-Ho;Kwak, In-Shin;Eom, Mi-Ok;Jeon, Dae-Hoon;Sung, Jun-Hyun;Choi, Jung-Mi;Kang, Han-Saem;Kim, Yong-Soo;Kang, Kil-Jin
    • Korean Journal of Food Science and Technology
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    • v.37 no.5
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    • pp.838-843
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    • 2005
  • Bacterial suspension test was used to establish Standardization Test Method to investigate bactericidal activity of disinfectant/sanitizer product. Using acceptable verification methodology, test substance showing 5 log or higher reduction in viable count against Escherichia coli ATCC 10536 and Staphylococcus aureus ATCC 6538, representing Gram-negative and -positive bacteria, respectively, under test conditions for $5\;min{\pm}10\;sec\;at\;20{\pm}1^{\circ}C$ was considered to have sanitizing capability. All disinfectant/sanitizer products tested under manufacturer's recommended in-use condition gave good reduction values against major food-poisoning bacteria. This standardized method was valuable for evaluating efficacy of disinfectants/sanitizers and could be used as Standardization Test Method for assessing bactericidal activity

Isolation and Identification of a Marine Bacterium, Pseudomonas sp. BK1 Producing Extracellular Enzymes Capable of Decomposing Multiple Complex Polysaccharides (복합 다당류 분해 효소들을 생산하는 해양미생물 Pseudomonas sp. BK1의 분리 및 특성)

  • Kim, Beom-Kyu;Jeon, Beong-Sam;Cha, Jae-Young;Park, Jeong-Won;Kim, Sam-Woong;Kim, Ji-Yoon;Park, Yong-Lark;Cho, Young-Su;Song, Jae-Young
    • Journal of Life Science
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    • v.13 no.6
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    • pp.871-878
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    • 2003
  • A marine bacterium (strain BKl) that produces extracellular enzymes capable of decomposing complex polysac-charides, such as agar, chitin, carboxymethylcellulose, xylan and mannan, was isolated from the marine red alga Porphyra dentata. Strain BKl was gram-negative, aerobic, catalase- and oxidase-positive, polarly flagellated bacilli that produce gelatinase and urease, but not decarboxylases. The G+C content of the DNA was 51.6 mol%. The major isoprenoid quinone component was identified as an ubiquinone-8, and the major cellular fatty acids were C16:0, C16:1 w6c and C18:1 w7c. Comparative 16S rRNA sequence analysis placed strain BK1 with members of the genus Pseudomonas. On the basis of phenotypic and genotypic data, the strain BK1 was shown to be a member of the subgroup of Pseudomonas, and named as Pseudomonas sp. BK1.

Screening of Antimicrobial Activity Compounds from Korea Ginseng Fine Root (고려인삼의 세근을 이용한 항균성 물질 탐색)

  • Kim, Ah-Reum;Lee, Myung-Suk
    • Journal of Life Science
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    • v.21 no.9
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    • pp.1244-1250
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    • 2011
  • The study was performed to evaluate the antibacterial and antiviral activities of ginseng fine root in order to search for antibacterial substances. Among 8 kinds of fermentation strains, Lactobacillus plantarum was selected based on viable cell count and antibacterial activities during incubation. Optimum conditions of ginseng fine root fermentation for L. plantarum were incubation at $35^{\circ}C$ for 48 hr in 5% ginseng fine root broth. That methanolic extract of fermented ginseng fine root broth was observed to be antibacterial and have antiviral activities. The results of paper disc method of non-fermented extract and fermented extract measured against E. coli was 11 mm and 20 mm, S. aureus was 15 mm and 22 mm, respectively. Shaking flask method was observed to inhibit the growth E. coli and S. aureus in fermented extract by 99.9%. However, antiviral activity of Feline calicivirus (FCV) was mostly activated. Fermented extract was used to investigate the compositional changes of ginsenosides on HPLC analysis. By fermentation, ginsenoside Rg1, Re and Rd were increased, with Rd showing a significant increase of 50 ${\mu}g/g$. These results suggest that ginseng fine root extract is a useful resource.