• Title/Summary/Keyword: r-DNA

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A Study on the Sensory Characteristic of Yogurt and Antimicrobial Activity of Lactobacillus plantarum LHC52 Isolated from Kimchi (김치에서 분리한 Lactobacillus plantarum LHC52의 항균활성과 요구르트의 관능성 연구)

  • Lee, Seung-Gyu;Han, Ki-Sung;Jeong, Seok-Geun;Oh, Mi-Hwa;Jang, Ae-Ra;Kim, Dong-Hun;Bae, In-Hyu;Ham, Jun-Sang
    • Food Science of Animal Resources
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    • v.30 no.2
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    • pp.328-335
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    • 2010
  • The aim of our study was to develop a new starter culture for fermented milk. Polymerase chain reaction screening of 103 acid-producing isolates from Kimchi identified 72 Lactobacillus strains. The ability of the strains to inhibit the growth of the food-borne human pathogens (Escherichia coli, Salmonella Enteritidis, Staphylococcus aureus) was measured, using a conventional paper disk method. Among the 72 strains, strain LHC52 displayed potent antagonistic activity. Use of 16S rDNA sequencing and the API 50CHL system identified the strain as Lactobacillus plantarum and it was designated L. plantarum LHC52. Biochemical analyses revealed especially high antibacterial activity against E. coli. Yogurt produced using L. plantarum LHC52 did not show different microbiological and physicochemical properties compared to conventionally-prepared yogurt, implicating L. plantarum LHC52 as a useful, potently antibacterial starter culture for yogurt preparation.

Development of Cleaved Amplified Polymorphic Sequence (CAPS) Marker for Selecting Powdery Mildew-Resistance Line in Strawberry (Fragaria×ananassa Duchesne) (딸기 흰가루병 저항성 계통 선발을 위한 분자마커 개발)

  • Je, Hee-Jeong;Ahn, Jae-Wook;Yoon, Hae-Suk;Kim, Min-Keun;Ryu, Jae-San;Hong, Kwang-Pyo;Lee, Sang-Dae;Park, Young-Hoon
    • Horticultural Science & Technology
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    • v.33 no.5
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    • pp.722-729
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    • 2015
  • Powdery mildew (PM) caused by Podosphaera aphanis is a major disease that can result in significant yield losses in strawberry (Fragaria ${\times}$ ananassa Duchesne). For preventing PM, pesticides are usually applied in strawberry. In this study, molecular markers were developed to increase breeding efficiency of PM-resistance cultivars by marker-assisted selection (MAS). An $F_2$ population derived from a cross between PM-resistance 'Seolhyang' and PM-susceptibility 'Akihime' was evaluated for disease resistance to PM and RAPD (random amplification of polymorphic DNA)-BSA (bulked segregant analysis). Among 200 RAPD primers tested, OPE10 primer amplified a 311bp-band present in with 331bp. Sequence alignment performed for searching polymorphisms and six single nucleotide polymorphism (SNP) were found in amplified regions. To develop polymorphic marker for distinguishing between resistant and susceptible, RAPD was converted to cleaved amplified polymorphic sequence (CAPS) marker. Among restriction enzymes associated with six SNPs, Eae I (Y/GGCCR) was successfully digested to 231bp in susceptible. The results suggest that the selected CAPS marker could be used for increasing efficiency of selecting powdery mildew resistant strawberry in breeding system.

Diversity and physiological properties of soil actinobacteria in Ulleung Island (울릉도 유래 토양 방선균의 다양성과 생리활성)

  • Yun, Bo-Ram;Roh, Su Gwon;Kim, Seung Bum
    • Korean Journal of Microbiology
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    • v.53 no.4
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    • pp.242-250
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    • 2017
  • Actinobacteria tolerating extreme conditions can be a rich source of bioactive compounds and enzymes. In this study filamentous actinobacteria were isolated from soils of Ulleung Island, and their physiological properties were examined. Soil samples were collected, serially diluted and spread on various agar media. The average viable counts of total bacteria were $1.28{\times}10^7CFU/g$ for soil sample 1 (ULS1) and $2.05{\times}10^7CFU/g$ for soil sample 2 (ULS2). As a result, 34 strains of actinobacteria were isolated and assigned to the genera Streptomyces (16 strains), Isoptericola (5 strains), Rhodococcus (4 strains), Agromyces (3 strains), Micrococcus (2 strains), Arthrobacter (1 strain), Williamsia (1 strain), Microbacterium (1 strain), and Oerskovia (1 strain) based on 16S rRNA gene sequence analysis. Enzyme activity and plant growth promoting potential were tested for representative isolates. Multiple strains of Streptomyces degraded starch, casein and Tween 80. As for plant growth promoting potential, strains of Oerskovia, Williamsia, Isoptericola, and Streptomyces solubilized phosphate, and those of Agromyces, Oerskovia, Micrococcus, Rhodococcus, Streptomyces, and Isoptericola produced 3-indole-acetic acid (IAA), respectively. Selected strains of Streptomyces exhibited strong antagonistic activity against Staphylococcus aureus and Bacillus subtilis as well as Candida albicans. This study confirms that actinobacteria from Ulleung Island can be a good source of novel bioactive compounds.

Characterization of Cellulase from Bacillus subtilis NSC Isolated from Soil (토양으로부터 단리한 Bacillus subtilis NSC 유래 Cellulase의 특성 규명)

  • Kim, Sang Jin;Park, Chang-Su
    • Journal of Chitin and Chitosan
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    • v.23 no.4
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    • pp.228-233
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    • 2018
  • We isolated microorganisms from soil, which is sampled at forest, Kyeonbuk, Korea, as cellulolytic microorganisms. The isolated strains were identified by analysis of 16S rRNA gene from the starins. The result, four kinds of Bacillus subtilis, one kind of Bacillus amyloliquefaciens, and one kind of Bacillus cereus were identified. Among these strains, Bacillus subtilis was selected due to its high cellulase activity and this strain was named as Bacillus subtilis CNS. The optimum pH and temperature of the cellulase from Bacillus subtilis CNS was pH 5.0 and $40^{\circ}C$, respectively. In the investigation of pH and temperature stability, the cellulase from Bacillus subtilis NSC stabled pH 4.0~6.0 range and until $40^{\circ}C$ for 30 min perfectly. In the enzyme activity for various cellulosic substrate, cellulase from Bacillus subtilis CNS showed the highest activity for CM-cellulose. And, the enzyme activities for alkali swollen cellulose, Alpha-cellulose, Sigmacell-cellulose, and Avicel were approximately 31%, 8%, 8% and 4% of activity for CM-cellulose, respectively. In the degradation of CM-cellulose, the 0.26 U/ml and 0.52 U/ml of cellulase showed 0.43 and 0.76 U/ml activity for CM-cellulose after the reaction of 120 min, respectively.

First Record of the Smoothtail Mobula, Mobula thurstoni (Myliobatiformes: Myliobatidae) in Southern Korea (한국 여수에서 채집된 매가오리과 (Myliobatidae) 어류 첫기록종, Mobula thurstoni)

  • Myoung, Se Hun;Song, Young Sun;Kang, Chung-Bae;Choi, Hong-In;Kim, Jong-Gwan;Yoon, Moongeun;Im, Jaebok;Han, Dong-Jin
    • Korean Journal of Ichthyology
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    • v.33 no.2
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    • pp.148-154
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    • 2021
  • Two specimens (1770~1850 mm disc width) of Mobula thurstoni, belonging to the family Myliobatidae, order Myliobatiformes, were first collected from the central coast of the Southern Sea of Korea in September 2018. This species is characterized by an anterior margin of disc with double curvature, a white-tipped dorsal fin, and the absence of a caudal spine. This species is morphologically similar to Mobula kuhlii, but has an anterior margin of pectoral fins with a double curvature and the dorsal coloration is bluish black rather than white. In addition, M. thurstoni was well distinguished from M. kuhlii as determined by mitochondrial DNA 16S rRNA sequences with genetic distances ranging from 0.030 to 0.069. The Korean name 'Mae-kkeun-kko-li-jwi-ga-o-li' is proposed for the species M. thurstoni.

Isolation of the Protease-producing Yeast Pichia anomala CO-1 and Characterization of Its Extracellular Neutral Protease (세포 외 중성 단백질분해효소를 생산하는 Pichia anomala CO-1의 분리 동정 및 효소 특성)

  • Kim, Ji Yeon
    • Journal of Life Science
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    • v.29 no.10
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    • pp.1126-1135
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    • 2019
  • From a sample of bamboo byproduct, the protease-producing yeast strain CO-1 was newly isolated. Strain CO-1 is spherical to ovoid in shape and measures $3.1-4.0{\times}3.8-4.4{\mu}m$. For the growth of strain CO-1, the optimal temperature and initial pH were $30^{\circ}C$ and 4.0, respectively. The strain was able to grow in 0.0-15.0%(w/v) NaCl and 0.0-9.0%(v/v) ethanol. Based on a phylogenetic analysis of its 18S rDNA sequences, strain CO-1 was identified as Pichia anomala. The extracellular protease produced by P. anomala CO-1 was partially purified by ammonium sulfate precipitation, which resulted in a 14.6-fold purification and a yield of 7.2%. The molecular mass of the protease was recorded as approximately 30 kDa via zymogram. The protease activity reached its maximum when 1.0%(w/v) CMC was used as the carbon source, 1.0%(w/v) yeast extract was used as the nitrogen source, and 0.3%(w/v) $MnSO_4$ was used as the mineral source. The protease revealed the highest activity at pH 7.0 and $30^{\circ}C$. This enzyme maintained more than 75% of its stability at a pH range of 4.0-10.0. After heating at $65^{\circ}C$ for 1 hr, the neutral protease registered at 60% of its original activity. The protease production coincided with growth and attained a maximal level during the post-exponential phase.

A Study of the Diversity and Profile for Extracellular Enzyme Production of Aerobically Cultured Bacteria in the Gut of Muraenesox cinereus (갯장어(Muraenesox cinereus) 장으로부터 호기적 조건에서 분리된 미생물의 다양성 및 세포외 효소 생산능 분석에 관한 연구)

  • Lee, Yong-Jik;Oh, Do-Kyoung;Kim, Hye Won;Nam, Gae-Won;Sohn, Jae Hak;Lee, Han-Seung;Shin, Kee-Sun;Lee, Sang-Jae
    • Journal of Life Science
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    • v.29 no.2
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    • pp.248-255
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    • 2019
  • This research confirmed the diversity and characterization of gut microorganisms isolated from the intestinal organs of Muraenesox cinereus, collected on the Samcheonpo Coast and Seocheon Coast in South Korea. To isolate strains, Marine agar medium was basically used and cultivated at $37^{\circ}C$ and pH7 for several days aerobically. After single colony isolation, totally 49 pure single-colonies were isolated and phylogenetic analysis was carried out based on the result of 16S rRNA gene DNA sequencing, indicating that isolated strains were divided into 3 phyla, 13 families, 15 genera, 34 species and 49 strains. Proteobacteria phylum, the main phyletic group, comprised 83.7% with 8 families, 8 genera and 26 species of Aeromonadaceae, Pseudoalteromonadaceae, Shewanellaceae, Enterobacteriaceae, Morganellaceae, Moraxellaceae, Pseudomonadaceae, and Vibrionaceae. To confirm whether isolated strain can produce industrially useful enzyme or not, amylase, lipase, and protease enzyme assays were performed individually, showing that 39 strains possessed at least one enzyme activity. Especially the Aeromonas sp. strains showed all enzyme activity tested. This result indicated that isolated strains have shown the possibility of the industrial application. Therefore, this study has contributed for securing domestic genetic resources and the expansion of scientific knowledge of the gut microbial community in Muraenesox cinereus of South Korea.

Analysis of Efficiency of Recombinant pOPINEneo-3C-GFP Vector with p53 Tumor Suppression Gene Inserted (p53 암억제 유전자가 삽입된 재조합 pOPINEneo-3C-GFP 벡터의 효율 분석)

  • Sa, Young-Hee;Choi, Chang-Shik;Lee, Ki Hwan;Hong, Seong-Karp
    • Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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    • 2019.05a
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    • pp.533-536
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    • 2019
  • Recombinant baculoviruses are widely used to express heterologous genes in cultured insect cells. Recombinant baculoviruses can serve as gene-transfer vectors for expression of recombinant proteins in a wide range of mammalian cell types. Baculovirus system has significant benefits in view of safety, large-scale, and high level of gene expression. In this study, baculoviral vectors which were reconstructed from pOPINEneo-3C-GFP vector, were recombined with cytomegalovirus (CMV) promoter, green fluorescent protein (GFP), and p53 with NcoI and XhoI. These recombinant vectors were infected with various cells and cell lines. The baculovirus vector thus developed was analyzed by comparing the metastasis and expression of the recombinant genes with conventional vectors. These results suggest that the baculovirus vector has higher efficiency in metastasis and expression than the control vector. This work was supported by a grant from Mid-Career Researcher Program(NRF-2016R1A2B4016552) through the National Research Foundation of Korea(NRF) funded by the Ministry of Science, ICT & Future Planning(MSIP).

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Parvatrema duboisi (Digenea: Gymnophallidae) Life Cycle Stages in Manila Clams, Ruditapes philippinarum, from Aphae-do (Island), Shinan-gun, Korea

  • Jung, Bong-Kwang;Chang, Taehee;Shin, Hyejoo;Ryoo, Seungwan;Hong, Sooji;Lee, Jeonggyu;Song, Hyemi;Cho, Jaeeun;Kim, Deok-Gyu;Jun, Hojong;Kim, Min-Jae;Won, Eun Jeong;Han, Eun-Taek;Shin, Eun-Hee;Chai, Jong-Yil
    • Parasites, Hosts and Diseases
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    • v.59 no.1
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    • pp.83-88
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    • 2021
  • Life cycle stages, including daughter sporocysts, cercariae, and metacercariae, of Parvatrema duboisi (Dollfus, 1923) Bartoli, 1974 (Digenea: Gymnophallidae) have been found in the Manila clam Ruditapes philippinarum from Aphae-do (Island), Shinan-gun, Jeollanam-do, Korea. The daughter sporocysts were elongated sac-like and 307-570 (av. 395) ㎛ long and 101-213 (av. 157) ㎛ wide. Most of the daughter sporocysts contained 15-20 furcocercous cercariae each. The cercariae measured 112-146 (av. 134) ㎛ in total length and 35-46 (av. 40) ㎛ in width, with 69-92 (av. 85) ㎛ long body and 39-54 (av. 49) ㎛ long tail. The metacercariae were 210-250 (av. 231) ㎛ in length and 170-195 (av. 185) ㎛ in width, and characterized by having a large oral sucker, genital pore some distance anterior to the ventral sucker, no ventral pit, and 1 compact or slightly lobed vitellarium, strongly suggesting P. duboisi. The metacercariae were experimentally infected to ICR mice, and adults were recovered at day 7 post-infection. The adult flukes were morphologically similar to the metacercariae except in the presence of up to 20 eggs in the uterus. The daughter sporocysts and metacercariae were molecularly (ITS1-5.8S rDNA-ITS2) analyzed to confirm the species, and the results showed 99.8-99.9% identity with P. duboisi reported from Kyushu, Japan and Gochang, Korea. These results confirmed the presence of various life cycle stages of P. duboisi in the Manila clam, R. philippinarum, playing the role of the first as well as the second intermediate host, on Aphae-do (Island), Shinan-gun, Korea.

Isolation of Simiduia sp. SH-2 and Characterization of Its β-Agarase (한천분해세균 Simiduia sp. SH-2 균주의 분리 및 β-agarase의 특성조사)

  • Lee, Dong-Geun;Kim, Geun-Dae;Lee, Sang-Hyeon
    • Journal of Life Science
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    • v.32 no.10
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    • pp.778-783
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    • 2022
  • This study isolated a new agarase-producing bacterium and characterized its agarase. A new agar-degrading strain was isolated from the seashore of Namhae in Gyeongnam province, Korea, and was purely cultured using the Marine Agar 2216 media. The isolated bacterium was identified as Simiduia sp. SH-2 after 16S rRNA gene sequencing. The crude agarase was obtained from the culture medium of the Simiduia sp. SH-2 strain, and the agar-degrading activity was measured. The highest level of activity of the Simiduia sp. SH-2-derived agar-degrading enzyme was 625 U/l. Agar degradation activity was most significant at 40℃ and pH 7.0. Compared to the activity at 40℃, the relative activity was 31% at 20℃ and 71% at 30℃. Compared to the activity at pH 7.0, the relative activity was 94% and 89% at pH 6.0 and pH 8.0, respectively. Residual activity was greater than 96% after exposure to 20℃ and 30℃ for 2 hr and more than 49% after exposure to 40℃ for 2 hr. Simiduia sp. SH-2 was identified as a strain producing β-agarase that creates neoagarooligosaccharides, such as neoagarotetraose and neoagarohexaose. Therefore, the Simiduia sp. SH-2 strain and its β-agarase are expected to be useful functional material producers in the food, cosmetic, and pharmaceutical industries.