• Title/Summary/Keyword: Activity Sequencing

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Purification and Characterization of an Antibacterial Substance from Aerococcus urinaeequi Strain HS36

  • Sung, Ho Sun;Jo, Youl-Lae
    • Journal of Microbiology and Biotechnology
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    • v.30 no.1
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    • pp.93-100
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    • 2020
  • A bacterial strain inhibiting the growth of Vibrio anguillarum, the causative agent of vibriosis, was isolated from fish intestines. The isolated strain HS36 was identified as Aerococcus urinaeequi based on the characteristics of the genus according to Bergey's Manual of Systematic Bacteriology and by 16S rRNA sequencing. The growth rate and antibacterial activity of strain HS36 in shaking culture were higher than those in static culture, while the optimal pH and temperature for antibacterial activity were 7.0 and 30℃, respectively. The active antibacterial substance was purified from a culture broth of A. urinaeequi HS36 by Sephadex G-75 gel chromatography, Sephadex G-25 gel chromatography, and reverse-phase high-performance liquid chromatography. Its molecular weight, as estimated by Tricine SDS-polyacrylamide gel electrophoresis, was approximately 1,000 Da. The antibacterial substance produced by strain HS36 was stable after incubation for 1 h at 100℃. Although its antibacterial activity was optimal at pH 6-8, activity was retained at a pH range from 2 to 11. The purified antibacterial substance was inactivated by proteinase K, papain, and β-amylase treatment. The newly purified antibacterial substance, classified as a class II bacteriocin, inhibited the growth of Klebsiella pneumoniae, Salmonella enterica, and Vibrio alginolyticus.

Characterization and Antifungal Activity against Candida albicans of Vaginal Lactobacillus spp. Isolated from Korean Women (질 내 유산균의 Candida albicans에 대한 항진균 효과 연구)

  • Jung, Yeojung;Kang, Chang-Ho;Shin, YuJin;So, Jae-Seong
    • KSBB Journal
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    • v.32 no.2
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    • pp.146-152
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    • 2017
  • Vulvovaginal candidiasis (VVC) is one of the urogenital infections occurring in women worldwide. Candida albicans is generally observed among various types of microorganisms causing VVC. Antibiotic therapy is typical, and the use of Lactobacilli probiotics is to be recognized as a promising alternative. The aim of this study was to select vaginal lactobacilli with probiotic properties against C. albicans. In a previous study, we isolated 38 lactobacilli from vagina of Korean women and 20 isolates were shown to inhibit C. albicans. We further selected 10 isolates which were able to inhibit C. albicans less than $10^5CFU/mL$. Among these selected strains, Lactobacillus salivarius MG242 (identified by 16s rRNA sequencing) was finally selected based on its strong anti-candidal activity, acid/bile salt resistance and adhesion property. Indirect adhesion activity of MG242 measured by auto-aggregation assay showed more than 60% auto-aggregation after 5 h standing. Taken these results together, the selected strain MG242 may have potential for application in vagina health related products.

Antibacterial Activity and Probiotic Properties of Lactic Acid Bacteria Isolated from Traditional Fermented Foods (전통발효식품에서 분리한 유산균의 항균활성 및 프로바이오틱스 기능성 연구)

  • Kang, Chang-Ho;Han, Seul Hwa;Kim, Yonggyeong;Jeong, Yulah;Paek, Nam-Soo
    • KSBB Journal
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    • v.32 no.3
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    • pp.199-205
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    • 2017
  • The aim of this study was to investigate probiotic characteristics and fermentation profile of selected lactic acid bacteria (LAB) isolated from traditional fermented foods. Antibacterial activity against various pathogens, acid and bile salt tolerance, cell hydrophobicity, and antibiotic resistance were examined. 16S rRNA sequencing was carried out to identify eight presumptive LAB isolates. In general, all identified LAB (Enterococcus faecium MG89-2, Lactobacillus plantarum MG207, L. paracasei MG310, L. casei MG311, Streptococcus thermophilus MG510, L. bulgaricus MG515, L. helveticus MG585, and L. fermentum MG590) showed strong antimicrobial activity. Also, the selected strains were resistant to bile acid up to 3% and their autoaggregation rates were as high as 60%. All selected strains tested were sensitive to chloramphenicol, tetracycline, and ampicillin, whereas resistant to nalidixic acid and kanamycin.

Characterization of an Amylase-sensitive Bacteriocin DF01 Produced by Lactobacillus brevis DF01 Isolated from Dongchimi, Korean Fermented Vegetable

  • Kang, Tae-Kyu;Kim, Wang-June
    • Food Science of Animal Resources
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    • v.30 no.5
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    • pp.795-803
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    • 2010
  • A DF01 strain that inhibits tyramine-producing Lactobacillus curvatus KFRI 166 was isolated from Dongchimi, a traditional Korean fermented vegetable, and identified as Lactobacillus brevis by biochemical analysis and reverse transcriptase sequencing of 16S rRNA. The antimicrobial compound produced by L. brevis DF01 was secreted at a maximum level of 640 AU/mL in late exponential phase in MRS broth, and its activity remained constant during stationary phase. The activity of bacteriocin DF01 was totally inactivated by $\alpha$-chymotrypsin, pronase E, proteinase K, trypsin, and $\alpha$-amylase, but not by catalase, which indicates the compound was glycoprotein in nature. The activity was not affected by pH changes ranging from 2 to 12 or heat treatment (60, 80, and $100^{\circ}C$ for 30 min), but was reduced after autoclaving. Bacteriocin DF01 had bacteriolytic activity and a molecular weight of approximately 8.2 kDa, as shown by tricine-SDS-PAGE analysis. Therefore, bacteriocin DF01 can be used in the manufacture of fermented meat products due to its inhibition of tyramine-producing L. curvatus and non-inhibition of L. sake, which is used as a starter culture for meat fermentation.

Purification and Characterization of Fibrinolytic Enzyme from Armillariella mellea (뽕나무버섯으로부터 Fibrinolytic enzyme의 정제 및 특성 연구)

  • Kim, Jun-Ho;Kim, Yang-Sun
    • The Korean Journal of Mycology
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    • v.26 no.4 s.87
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    • pp.583-588
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    • 1998
  • A fibrinolytic enzyme has been isolated from the edible honey mushroom, Armillariella mellea and purified. The apparent molecular mass of purified enzyme was estimated to be 19800Da by SDS polyacryl amide gel electrophoresis and 19900Da by gel filtration, indicating that it was a monomer. The enzyme was optimal at pH 7, suggesting that the purified enzyme was a neutral proteinase. It shows the maximum fibrinolytic activity at $55^{\circ}C$, is completely inactivated above $65^{\circ}C$, and still indicates 40% of activity at $37^{\circ}C$. The fibrinolytic activity has been decreased by the addition of EDTA. Fifteen amino acid sequence was determined by protein sequencing techniques.

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Cloning and Characterization of a Multidomain GH10 Xylanase from Paenibacillus sp. DG-22

  • Lee, Sun Hwa;Lee, Yong-Eok
    • Journal of Microbiology and Biotechnology
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    • v.24 no.11
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    • pp.1525-1535
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    • 2014
  • The xynC gene, which encodes high molecular weight xylanase from Paenibacillus sp. DG-22, was cloned and expressed in Escherichia coli, and its nucleotide sequence was determined. The xynC gene comprised a 4,419bp open reading frame encoding 1,472 amino acid residues, including a 27 amino acid signal sequence. Sequence analysis indicated that XynC is a multidomain enzyme composed of two family 4_9 carbohydrate-binding modules (CBMs), a catalytic domain of family 10 glycosyl hydrolases, a family 9 CBM, and three S-layer homologous domains. Recombinant XynC was purified to homogeneity by heat treatment, followed by Avicel affinity chromatography. SDS-PAGE and zymogram analysis of the purified enzyme identified three active truncated xylanase species. Protein sequencing of these truncated proteins showed that all had identical N-terminal sequences. In the protein characterization, recombinant XynC exhibited optimal activity at pH 6.5 and $65^{\circ}C$ and remained stable at neutral to alkaline pH (pH 6.0-10.0). The xylanase activity of recombinant XynC was strongly inhibited by 1 mM $Cu^{2+}$ and $Hg^{2+}$, whereas it was noticeably enhanced by 10 mM dithiothreitol. The enzyme exhibited strong activity towards xylans, including beechwood xylan and arabinoxylan, whereas it showed no cellulase activity. The hydrolyzed product patterns of birchwood xylan and xylooligosaccharides by thin-layer chromatography confirmed XynC as an endoxylanase.

Phenolic Compounds Production, Enhancement and Its Antioxidant Activity of Blue Berry Powder with Bacillus subtilis Light Mediated Fermentation Compounds

  • Elumalai, Punniyakotti;Lim, Jeong-Muk;Mohan, Harshavardhan;Lee, Jeong-Ho;Oh, Byung-Taek
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2018.04a
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    • pp.66-66
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    • 2018
  • Light fermentation has been conducted under different light conditions such as normal dark light, white light, and light emitting diodes (LEDs) various color (blue, green, red, white on blueberry powder with fermenting bacteria Bacillus subtilis (B2). The bacteria B2 was isolated and identified by 16S rRNA sequencing method. RYRP biologically converted to secondary metabolites through light fermentation in the presence of Bacillus subtilis, the bacteria actively involved in bioconversion process. LEDs fermentation to enhance the production of phenolic content while comparing to normal dark and white light. Among the different color LEDs, blue LEDs mediated fermentation showed higher amount of total phenolic and flavonoid content. Then blue LEDs mediated fermented compound were characterized by FTIR and GC-MS, subsequently the compound was analyzed antioxidant activity tests and the antioxidant activity exhibited higher. This is the first study to demonstrate that B. subtilis-LEDs mediated fermentation is useful for facilitating phenolic compound production and enhancing antioxidant activity, which may have greater application fermentation fields.

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Study on the promotion of inflammation and whitening of natural materials using bioconversion technology

  • Lee, Se-Won;Lim, Jeong-Muk;Lee, Seong-Hyeon;Lee, Jeong-Ho;Oh, Byung-Teak
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2018.10a
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    • pp.116-116
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    • 2018
  • Bioconversion, the enzymatic process by microorganism on organic precursor to desired products. The natural extract is converted into a form that can be easily absorbed into the skin, while scaling up of to higher quantity is possible. Selection of naturally processed raw material rather than chemically processed is preferred. Therefore, fermentation was carried out by mixing Rubus coreanus Miquel, soybean, Zanthoxylum schinifolium as bioconverting materials, the possibility of inflammation, whitening material were checked. In this study, useful microorganisms were isolated from various salted fish, and 16S rDNA sequence was analyzed to confirm their genetic characteristics. Combining the three natural materials using bioconversion technology to study their activity before and after fermentation. To evaluate the antioxidant activity and the active ingredient content the DPPH radical scavenging activity and the total polyphenol content were examined. Raw 264.7 cells were used to evaluate MTT assay, NO and $TNF-{\alpha}$ production inhibitory activity. Also, to evaluate the whitening activity, tyrosinase inhibitory activity and melanin formation inhibitory activity were measured using B16F10 cells. In total 34 strains were obtained from various salted fish. The effective strains useful for the bioconversion process, showed that DPPH radical scavenging ability and polyphenol content were increased in the two kinds of microbial treatment groups compared to the untreated group. 16S rDNA sequencing analysis of the strains showed excellent in Pediococcus pentosaceus B1 in comparison. An increase of up-to 156% in anti-oxidative activity and 141% in polyphenol content was observed after bioconversion. In addition, after mixed fermentation the toxidty of Raw 264.7 and B16F10 cells tended to decrease and a significant increase was observed in anti-inflammatory activity as well. Also, tyrosinase activity and melanin significantly. synthesis decreased significantly.

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Implementation and Application of the SCORM 2004 S&N and the Traffic-Signal-Lamp Metaphor for a Web-based Adaptive Learning Management (웹기반 적응형 학습관리를 위한 SCORM 2004 S&N과 교통신호메타포 구현 및 적용)

  • Bang, Chan-Ho;Kim, Ki-Seok
    • The Journal of Korean Association of Computer Education
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    • v.9 no.1
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    • pp.61-70
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    • 2006
  • In the area of e-learning education, SCORM2004 that is suggested by ADL and is a defacto standard allows to design and apply various interrelations among learning objects which organize learning process through consolidating IMS Simple Sequencing into S&N. In this paper, we intend to realize a web_based adaptive learning management that enable to guide experientially the learning activity through the SCORM 2004 S&N and the Traffic-Signal-Lamp Metaphor. This adaptive system allows professor to design the learning courseware realizing various learning strategies to be able to reuse same learning contents and student to be leaded a adaptive learning through being supplied immediately the state and evaluation of learning.

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Medium optimization for keratinase production by a local Streptomyces sp. NRC 13S under solid state fermentation

  • Shata, Hoda Mohamed Abdel Halim;Farid, Mohamed Abdel Fattah
    • Journal of Applied Biological Chemistry
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    • v.56 no.2
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    • pp.119-129
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    • 2013
  • Thirteen different Streptomyces isolates were evaluated for their ability to produce keratinase using chicken feather as a sole carbon and nitrogen sources under solid state fermentation (SSF). Streptomyces sp. NRC 13S produced the highest keratinase activity [1,792 U/g fermented substrate (fs)]. The phenotypic characterization and analysis of 16S rDNA sequencing of the isolate were studied. Optimization of SSF medium for keratinase production by the local isolate, Streptomyces sp. NRC13S, was carried out using the one-variable-at-a-time and the statistical approaches. In the first optimization step, the effect of incubation period, initial moisture content, initial pH value of the fermentation medium, and supplementation of some agro-industrial by-products on keratinase production were evaluated. The strain produced about 2,310 U/gfs when it grew on chicken feather with moisture content of 75% (w/w), feather: fodder yeast ratio of 70:30 (w/w), and initial pH 7 using phosphate buffer after 8 days. Based on these results, the Box-Behnken design and response surface methodology were applied to find out the optimal conditions for the enzyme production. The corresponding maximal production of keratinase was about 2,569.38 U/gfs.