• Title/Summary/Keyword: rDNA sequencing

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Probiotic Properties of Lactobacillus plantarum NK181 Isolated from Jeotgal, a Korean Fermented Food

  • Lee, Na-Kyoung;Kim, Hyoun-Wook;Chang, Hyo-Ihl;Yun, Cheol-Won;Kim, Seung-Wook;Kang, Chang-Won;Paik, Hyun-Dong
    • Food Science and Biotechnology
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    • v.15 no.2
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    • pp.227-231
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    • 2006
  • Strain NK181 was isolated for probiotic use from jeotkal and based on results of API 50 CHL kit and 16S rDNA sequencing was tentatively named Lactobacillus plantarum NK181. L. plantarum NK181 was highly resistant to artificial gastric juice (pH 2.5) and bile acid and demonstrated strong adherence to Caco-2 cells. In test using API ZYM kit, eight enzymes were produced. Supernatant of L. plantarum NK181 exhibited about 30% 1,1-diphenyl-2-picyryl hedrazyl (DPPH) radical-scavenging activity and reduced cholesterol by 70%. These results demonstrate potential use of L. plantarum NK181 as health-promoting probiotic.

Identification of Non-mutans Streptococci Organisms in Dental Plaques Recovering on Mitis-Salivarius Bacitracin Agar Medium

  • Yoo So Young;Kim Pyung Sik;Hwan Ho Keel;Lim Seong Hoon;Kim Kwang Won;Choe Son Jin;Min Byung Moo;Kook Joong Ki
    • Journal of Microbiology
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    • v.43 no.2
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    • pp.204-208
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    • 2005
  • The objective of this study was to both isolate and identify non-mutans streptococci organisms (non­MSO) from dental plaques recovered on mitis-salivarius sucrose bacitracin agar (MSB) plates. The dental plaque samples, which had been collected from 63 human subjects, were diluted and plated on MSB. The bacteria growing on the MSB plates were then identified with biochemical tests, as well as with 16S rDNA cloning and sequencing techniques. Our data indicated that bacteria from 30 subjects had been recovered on the MSB plates. Among the 21 typical colonies selected from the 30 subjects, 12 colonies, derived from 10 subjects, were identified as non-MSO. These 12 colonies were determined to be Streptococcus anginosus (8 colonies), S. sanguinis (1 colony), and Pantoea agglomerans (3 colonies). These results strongly suggest that a new selective medium will be required for the reliable isolation of mutans streptococci.

Purification and Physiochemical Characterization of Melanin Pigment from Klebsiella sp. GSK

  • Sajjan, Shrishailnath;Kulkarni, Guruprasad;Yaligara, Veeranagouda;Lee, Kyoung;Karegoudar, T.B.
    • Journal of Microbiology and Biotechnology
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    • v.20 no.11
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    • pp.1513-1520
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    • 2010
  • A bacterium capable of producing melanin pigment in the presence of L-tyrosine was isolated from a crop field soil sample and identified as Klebsiella sp. GSK based on morphological, biochemical, and 16S rDNA sequencing. The polymerization of this pigment occurs outside the cell wall, which has a granular structure as melanin ghosts. Chemical characterization of the pigment particles showed then to be acid resistant, alkali soluble, and insoluble in most of the organic solvents and water. The pigment got bleached when subjected to the action of oxidants as well as reductants. This pigment was precipitated with $FeCl_3$, ammoniacal silver nitrate, and potassium ferricynide. The pigment showed high absorbance in the UV region and decreased absorbance when shifted towards the visible region. The melanin pigment was further charecterized by FT-IR and EPR spectroscopies. A key enzyme, 4-hydroxyphenylacetic acid hydroxylase, that catalyzes the formation of melanin pigment by hydroxylation of L-tyrosine was detected in this bacterium. Inhibition studies with specific inhibitors, kojic acid and KCN, proved that melanin is synthesized by the DOPA-melanin pathway.

Studies on the Development of a Microbial Cryoprotectant Formulation Using a W/O/W Multiple Emulsion System

  • Bae, Eun-Kyung;Cho, Young-Hee;Park, Ji-Yong
    • Journal of Microbiology and Biotechnology
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    • v.14 no.4
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    • pp.673-679
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    • 2004
  • A microbial cryoprotectant formulation using a W/O/W multiple emulsion system was developed. The psychrotolerant microorganism, B4, isolated from soil in South Korea, was observed by the drop freezing method, in which the microorganism sample inhibited ice nucleation activity. The antifreeze activity was eliminated when the microorganism sample was treated with protease, indicating that the antifreeze activity was due to the presence of antifreeze protein. The result of the l6S rDNA sequencing indicated the B4 strain was most closely related to a species of the genus Bacillus. Culture broth of B4 strain (Bacillus sp.) and rapeseed oil containing 1 % polyglycerine polyricinolate (PGPR) were used as core and wall material, respectively. The most stable W/O emulsion was prepared at a core/oil ratio of 1:2. The highest W/O/W emulsion stability was achieved when the primary emulsion to external aqueous phase containing 0.5% caster oil polyoxyethylene ether $(COG25^{TM})$ ratio was 1:1. Microcrystalline cellulose showed better W/O/W emulsion stability than other polymer types. The viability of cells in a W/O/W emulsion was higher than free cells during storage at $37^\circ{C}$. An acidic pH and UV exposure decreased the viability of free cells, but cells in W/O/W emulsion were more stable under these conditions.

Characterization of a Nitrogen Fixing Bacteria Mycobacterium hominis sp. AKC-10 Isolated from the Wetland (습지에서 분리한 질소고정 세균인 Mycobacterium hominis sp. AKC-10의 특성)

  • Hong, Sun-Hwa;Shin, Ki-Chul;Lee, Eun-Young
    • Microbiology and Biotechnology Letters
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    • v.38 no.3
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    • pp.302-307
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    • 2010
  • Nitrogen is an element need to grow plants growth. Plants take up nitrogen in the form of nitrate or ammonium. Most of plants absorb nitrogen source as fertilizers. But from 50 to 70% of fertilizers applied were washed away. This study was conducted to isolate free-living nitrogen fixing bacteria from reed and to examine its beneficial traits for developing sustainable biofertilizers. Enriched consortium obtained from a reed in Ansan was developed for the fixing of nitrogen. Nitrogen fixing bacteria isolated from an enriched culture in Congo Red Medium was analyzed by 16s rDNA sequencing. AKC-10 was isolated and shown to have excellent nitrogen fixing ability. The optimum conditions of nitrogen fixing ability were $25^{\circ}C$ ($237.50{\pm}39.65\;nmole{\cdot}mg-protein^{-1}{\cdot}h^{-1}$ and pH 7 ($168.335{\pm}12.84$ nmole/hr mg-protein). It was identified as Microbacterium hominis [(AKC-10 (similarity : 99%)]. This strain was had to IAA (indole-3-acetic acid) productivity and ACC(1-aminocyclopropane-1-carboxylic acid) deaminase activity. Therefore, Microbacterium hominis AKC-10 stimulated plant development in the soil, enhancing the efficiency of remediation.

Biocontrol of Late Blight and Plant Growth Promotion in Tomato Using Rhizobacterial Isolates

  • Lamsal, Kabir;Kim, Sang Woo;Kim, Yun Seok;Lee, Youn Su
    • Journal of Microbiology and Biotechnology
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    • v.23 no.7
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    • pp.897-904
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    • 2013
  • Seven bacterial isolates (viz., AB05, AB10, AB11, AB12, AB14, AB15, and AB17) were derived from the rhizosphere and evaluated in terms of plant growth-promoting activities and the inhibition of Phytophthora infestans affecting tomatoes in Korea. According to 16S rDNA sequencing, a majority of the isolates are members of Bacillus, and a single isolate belongs to Paenibacillus. All seven isolates inhibited P. infestans by more than 60% in vitro. However, AB15 was the most effective, inhibiting mycelial growth of the pathogen by more than 80% in vitro and suppressing disease by 74% compared with control plants under greenhouse conditions. In a PGPR assay, all of the bacterial isolates were capable of enhancing different growth parameters (shoot/root length, fresh biomass, dry matter, and chlorophyll content) in comparison with non-inoculated control plants. AB17-treated plants in particular showed the highest enhancement in fresh biomass with 18% and 26% increments in the root and shoot biomass, respectively. However, isolate AB10 showed the highest shoot and root growth with 18% and 26% increments, respectively. Moreover, the total chlorophyll content was 14%~19% higher in treated plants.

Food waste treatment using Bacillus species isolated from food wastes and production of air-dried Bacillus cell starters

  • An, Byungryul;Park, Mi-Kyung;Oh, Jun-Hyun
    • Environmental Engineering Research
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    • v.23 no.3
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    • pp.258-264
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    • 2018
  • The objectives of this research were to 1) isolate and identify thermophilic bacteria for food waste treatment; 2) investigate the capability of food waste treatment using Bacillus species; and 3) develop air-dried Bacillus starters for food waste treatment. Five Bacillus species were isolated from food wastes and identified as Bacillus licheniformis (B. licheniformis) G1, Bacillus circulans C2, Bacillus subtilis (B. subtilis) E1, Bacillus vanillea F1, and Bacillus atrophaeus G2 based on 16S rDNA sequencing. Each identified Bacillus and the mixture of Bacillus species were cultivated in the standard food waste at $45^{\circ}C$ for 8 d. Changes in cell count, solid contents, and pH of the food waste were monitored during cultivation. Air-dried Bacillus cell powders were prepared using wheat flour and lactomil as excipients, and the cell count and survival rate were determined. The cell count of B. licheniformis G1 exhibited the highest number among the tested Bacillus (${\sim}10^8CFU/mL$). The greatest reduction in solid contents of food waste was achieved by B. subtilis E1 (22.6%). The mixture of B. licheniformis G1 and B. subtilis E1 exhibited a synergistic effect on the reduction of solid contents. Lactomil was determined as better excipient than wheat flour based on the greatest survival rate of 95%.

Mapping of Quantitative Trait Loci for Yield and Grade Related Traits in Peanut (Arachis hypogaea L.) Using High-Resolution SNP Markers

  • Liang, Yuya;Baring, Michael R.;Septiningsih, Endang M.
    • Plant Breeding and Biotechnology
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    • v.6 no.4
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    • pp.454-462
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    • 2018
  • Yield and grade are the key factors that affect production value of peanut. The objective of this study was to identify QTLs for pod yield, hundred-seed weight, and total sound mature kernel (TSMK). A total of 90 recombinant inbred lines, derived from Tamrun OL07 and a breeding line Tx964117, were used as a mapping population and planted in Brownfield and Stephenville, Texas. A genetic map was developed using 1,211 SNP markers based on double digest restriction-site associated DNA sequencing (ddRAD-seq). A total of 10 QTLs were identified above the permutation threshold, three for yield, three for hundred-seed weight and four for TSMK, with LOD score values of 3.7 - 6.9 and phenotypic variance explained of 12.2% - 35.9%. Among those, there were several QTLs that were detected in more than one field experiment. The commonly detected QTLs in this study may be used as potential targets for future breeding program to incorporate yield and grade related traits through molecular breeding.

Analysis of the oxidized low density lipoprotein receptor 1 gene as a potential marker for carcass quality traits in Qinchuan cattle

  • Gui, Lin-sheng;Raza, Sayed Haidar Abbas;Jia, Jianlei
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.1
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    • pp.58-62
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    • 2019
  • Objective: The oxidized low density lipoprotein receptor 1 (OLR1) gene plays an important role in the degradation of oxidized low-density lipoprotein and adipocyte proliferation in mammals. For this reason, we aimed at investigating the association of OLR1 gene polymorphisms with carcass quality traits in Chinese Qinchuan cattle. Methods: The single nucleotide polymorphism (SNP) was identified in the 3' untranslated region of bovine OLR1 gene by DNA sequencing. In addition, the haplotype frequency and linkage disequilibrium estimates of three SNPs were evaluated in 520 individuals. Results: Results indicated that the studied three SNPs were within the range of moderate genetic diversity (0.25< polymorphism information content<0.5). Haplotype analysis of three SNPs showed that ten different haplotypes were identified, but only five haplotypes were listed as those with a frequency of <0.05 were excluded. The Hap3 ($-G_1T_2C_3-$) had the highest haplotype frequency (42.10%). Linkage disequilibrium analysis showed that the three SNPs had a low linkage ($r^2<0.001$). The T10588C and C10647T were significantly associated with backfat thickness and intramuscular fat content in Qinchuan cattle. Conclusion: Based on our results, we believe that the OLR1 gene could be a strong candidate gene for influencing carcass quality traits in Qinchuan cattle.

Surface Film Formation in Static-Fermented Rice Vinegar: A Case Study

  • Yun, Jeong Hyun;Kim, Jae Ho;Lee, Jang-Eun
    • Mycobiology
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    • v.47 no.2
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    • pp.250-255
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    • 2019
  • In the present study, we aimed to determine the cause of surface film formation in three rice vinegars fermented using the traditional static fermentation method. The pH and total acidity of vinegar were 3.0-3.3 and 3.0-8.7%, respectively, and acetic acid was the predominant organic acid present. Colonies showing a clear halo on GYC medium were isolated from the surface film of all vinegars. Via 16S rDNA sequencing, all of the isolates were identified as Acetobacter pasteurianus. Furthermore, field-emission scanning electron microscopy analysis showed that the bacterial cells had a rough surface, were rod-shaped, and were ${\sim}1{\times}2{\mu}m$ in size. Interestingly, cells of the isolate from one of the vinegars were surrounded with an extremely fine threadlike structure. Thus, our results suggest that formation of the surface film in rice vinegar was attributable not to external contamination, to the production of bacterial cellulose by A. pasteurianus to withstand the high concentrations of acetic acid generated during fermentation. However, because of the formation of a surface film in vinegar is undesirable from an industrial perspective, further studies should focus on devising a modified fermentation process to prevent surface film formation and consequent quality degradation.