• Title/Summary/Keyword: l6S rDNA

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DNA Profiling of Leuconostoc citreum Strains in Fermented Foods by Repetitive Element Polymerase Chain Reaction

  • Kaur, Jasmine;Sharma, Anshul;Lee, Sulhee;Park, Young-Seo
    • Journal of Microbiology and Biotechnology
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    • v.27 no.10
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    • pp.1778-1782
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    • 2017
  • To identify and discriminate the bacterial species at the subspecific level, rep-PCR is a reliable genomic fingerprinting tool. Fourteen strains of bacteria were isolated from different food sources, identified as Leuconostoc citreum using 16S rRNA gene sequencing, and amplified using rep-primers (REP, ERIC, and $(GTG)_5$). Fingerprinting patterns generated bands in the range of 300-6,000 bp with REP, 150-6,000 bp with ERIC, and 200-1,700 bp with $(GTG)_5$ primers. In UPGMA dendrogram analysis, 14 strains were clustered into three clades (I, II, and III) with all the primers, thus differentiating them at the molecular level. The present study revealed the differentiation of L. citreum strains using rep-PCR.

Effects of Feeding Fermented Colostrum Feed on the Growth to Piglets (발효초유사료 급여가 자돈의 성장에 미치는 영향)

  • Na, Seuk-Han;Choi, Seong-Hyun;Renchinthand, Gereltuya;Bae, Hyoung-Churl;Nam, Myoung-Soo
    • Food Science of Animal Resources
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    • v.28 no.3
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    • pp.355-362
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    • 2008
  • This study was carried out to assess the fermentation properties of lactic acid bacteria (LAB) isolated from bovine colostrum and effects of feeding fermented colostrum feed on the growth to piglet. A total of 427 colonies were isolated from bovine colostrum on the BCP plate count agar. These LAB isolated were subcultured in 10% reconstituted skim milk, and seven strain thereof were selected for their highest acid productions, and two strain thereof were finally selected for their excellent sugar utilization. These strains were identified as Streptococcus thermophilus and Streptococcus macedonicus based on l6S rDNA sequencing data, named S. thermophilus CNB-11 and S. macedonicus CNB-11 respectively. For fermentation profiles, sugar utilization, acid production and viable cell counts were excellent in S. thermophilus CNB-11 as compared with S. macedoniclts CNB-11 after 48 hour. The effect of feeding fermented colostrum feed 0.5% using S. thermophilus CNB-11 was investigated for growth rate, analysis of blood and incidence of diarrhea. 24 heads of piglets were divided into two groups: the experimental and the control of 12 animals each. The average growth rate in the pigs fed fermented colostrum feed was higher 16.73% compared with control diet (p<0.05). There were no differences in the concentrations of blood glucose, cholesterol, albumin and globulin in pigs fed fermented colostrum feed as compared with control piglets. Incidence of diarrhea was no in pigs fed fermented colostrum feed as compared with control piglets.

Isolation of Tetracycline-resistant Lactic Acid Bacteria from Kimchi (김치에서 tetracycline 내성 유산균의 분리)

  • Kang, Hyo-Jin;Kim, Byung-Chun;Park, Wan
    • Korean Journal of Microbiology
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    • v.40 no.1
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    • pp.1-6
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    • 2004
  • Tetracycline resistant bacterial strains were isolated from 10 batches of Kimchi among 50 batches collected in Taegu restrict. The MIC of tetracycline ranged between 25 and> 100 ㎖/l. Total genomic DNA preparation from all 10 tetracycline resistant lactic acid bacterial isolates were subjected to PCR amplification with class-specific primers for tet(M) and tet(O). In only one isolate, HJ9, tet(M) was detected. By Southern blotting and hybridization with a tet(M)-specific probe, the tet(M) gene of HJ9 isolate could be localized on a plasmid. The partial nucleotide sequence and deduced amino acid sequence of tet(M) of HJ9 showed 90-99% and 94-100% homology to those of Gram positive bacteria, respectively. With sequencing of 16S rRNA, HJ9 isolate from Kimchi was identified as Lactobacillus sakei. From these results, Kimchi can be considered potential vehicle for the spread of antibiotic-resistant lactic acid bacteria along the food chain to the consumer.

Isolation and Characterization of Xylanolytic Bacteria from Horse Manure (말똥으로부터 xylan 분해 균주의 분리 및 특성)

  • Kim, Jung-Kon;Kim, Tae-Hyun
    • KSBB Journal
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    • v.26 no.5
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    • pp.465-470
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    • 2011
  • Twenty six microorganisms were isolated from soil and horse manure samples from in Iowa, U.S. Microorganisms were cultivated and screened by using plate count agar (PCA) at $35^{\circ}C$ containing 1% (w/v) oat spelt xylan instead of glucose. The xylanase activities of bacterial strains were analyzed by measuring the concentration of reducing sugar by DNS method. All isolated strains were characterized as the rod form and gram positive strains. Among the isolated strains, the HM6 strains gave the highest xylanase activity. This strain was identified as Bacillus pumilus HM6 by 16S rDNA sequence, morphological and biochemical analysis. Optimal culture temperature and initial medium pH for B. pumilus HM6 were $30-35^{\circ}C$ and pH 6-7, respectively. The maximum xylanase activity of 6879 IU/mL was obtained after growth of HM6 with 1% (w/v) oat spelt xylan at $35^{\circ}C$ for 6 days. Studies on enzymatic properties showed that the optimum conditions for the highest xylanase activity were $60^{\circ}C$ and pH 8.0. In addition, xylanase activity was stable over 2 hours at $50^{\circ}C$, whereas activity decreased after 30 min at $70^{\circ}C$.

Effect on the Concentration of Glucose and Sucrose on the Hydrogen Production using by the Facultative Anaerobic Hydrogen Producing Bacterium Rhodopseudomonas sp. MeL 6-2 (통성혐기성 수소생산균주 Rhodopseudomonas sp. MeL 6-2를 이용한 수소생산효율에 미치는 포도당 및 자당 농도의 영향)

  • Lee, Eun-Young
    • Microbiology and Biotechnology Letters
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    • v.37 no.2
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    • pp.176-182
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    • 2009
  • Hydrogen producing bacterium, strain MeL 6-2 was isolated from the sludge of the factory areas in Anyang through the acclimation in basal salt medium (BSM) supplemented with 10 g/L of sucrose. Isolated strain MeL 6-2 was a facultative anaerobe which could grow in both aerobic and anaerobic environments. An aerobically grown pure culture isolated from enriched culture was analyzed by 16S rDNA sequencing and identified as Rhodopseudomonas sp. MeL 6-2. Effects of the concentrations of glucose and sucrose on the hydrogen production rate and the hydrogen production yield were investigated. When glucose in the range of 1~12 g/L was supplemented to the BSM, strain MeL 6-2 could grow without lag phase. An increased glucose concentration increased the specific hydrogen production rate linearly to $4.2\;mmol-H_2{\cdot}L^{-1}{\cdot}h^{-1}$ at 10 g/L, and $60\;mmol-H_2{\cdot}mg-DCW^{-1}{\cdot}h^{-1}$, but decreased slightly as the concentration increased to 12 g/L. The hydrogen production yield was maintained over a range from 2.6 to $3.1\;mol-H_2{\cdot}mol-glucose^{-1}$. When sucrose in the range of 1~12 g/L was supplemented to the BSM, strain MeL 6-2 could grow after ten hours. An increased sucrose concentration increased the specific hydrogen production rate and the hydrogen production yield to $163\;mmol-H_2{\cdot}mg-DCW^{-1}{\cdot}h^{-1}$ and to $4.5\;mol-H_2{\cdot}mol-sucrose^{-1}$, respectively.

Enhanced Production of Carboxymethylcellulase by a Newly Isolated Marine Microorganism Bacillus atrophaeus LBH-18 Using Rice Bran, a Byproduct from the Rice Processing Industry (미강을 이용한 해양미생물 Bacillus atrophaeus LBH-18 유래의 carboxymethylcellulase 생산의 최적화)

  • Kim, Yi-Joon;Cao, Wa;Lee, Yu-Jeong;Lee, Sang-Un;Jeong, Jeong-Han;Lee, Jin-Woo
    • Journal of Life Science
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    • v.22 no.10
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    • pp.1295-1306
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    • 2012
  • A microorganism producing carboxymethylcellulase (CMCase) was isolated from seawater and identified as Bacillus atrophaeus. This species was designated as B. atrophaeus LBH-18 based on its evolutionary distance and the phylogenetic tree resulting from 16S rDNA sequencing and the neighbor-joining method. The optimal conditions for rice bran (68.1 g/l), peptone (9.1 g/l), and initial pH (7.0) of the medium for cell growth was determined by Design Expert Software based on the response surface method; conditions for production of CMCase were 55.2 g/l, 6.6 g/l, and 7.1, respectively. The optimal temperature for cell growth and the production of CMCase by B. atrophaeus LBH-18 was $30^{\circ}C$. The optimal conditions of agitation speed and aeration rate for cell growth in a 7-l bioreactor were 324 rpm and 0.9 vvm, respectively, whereas those for production of CMCase were 343 rpm and 0.6 vvm, respectively. The optimal inner pressure for cell growth and production of CMCase in a 100-l bioreactor was 0.06 MPa. Maximal production of CMCase under optimal conditions in a 100-l bioreactor was 127.5 U/ml, which was 1.32 times higher than that without an inner pressure. In this study, rice bran was developed as a carbon source for industrial scale production of CMCase by B. atrophaeus LBH-18. Reduced time for the production of CMCase from 7 to 10 days to 3 days by using a bacterial strain with submerged fermentation also resulted in increased productivity of CMCase and a decrease in its production cost.

Effect of Chlorella vulgaris CHK0008 Fertilization on Enhancement of Storage and Freshness in Organic Strawberry and Leaf Vegetables (Chlorella vulgaris CHK0008 시비가 유기농 딸기와 엽채소의 저장성과 신선도 향상에 미치는 영향)

  • Kim, Min-Jeong;Shim, Chang-Ki;Kim, Yong-Ki;Park, Jong-Ho;Hong, Sung-Jun;Ji, Hyeong-Jin;Han, Eun-Jung;Yoon, Jung-Chul
    • Horticultural Science & Technology
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    • v.32 no.6
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    • pp.872-878
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    • 2014
  • This study aimed to enhance storage and freshness of strawberry fruits and foliage vegetables by spray treatment with Chlorella vulgaris as a bio-fertilizer. The tested strain, C. vulgaris CHK0008, was isolated from an organically cultivated rice paddy and identified as C. vulgaris by its morphology and 18S rDNA and 23S rDNA sequence homology. We successfully cultured C. vulgaris CHK0008 in BG11 modified medium (BG11MM) and adjusted $2.15{\times}10^6cell/mL$ C. vulgaris CHK0008 to one OD value by measuring the optical density at 680 nm using a UV-vis spectrophotometer. The soluble solid content of 'Seolhyang' and 'Yukbo' strawberry fruits treated by spray application with C. vulgaris CHK0008 was enhanced by 22.2% and 11.5% respectively, compared to untreated controls. Additionally, the decay rates of treated 'Seolhyang' and 'Yukbo' strawberry fruits decreased 63.8% and 74.4% respectively, compared to untreated control. Surface color changes and chlorosis of leaves in leaf vegetables such as lettuce, kale, red ornamental kale, white ornamental kale and beet were observed in samples treated with water spray for 10 days after cold storage. However, the decay rate of leafy vegetables treated with foliar application of 25% C. vulgaris CHK0008 liquid culture was significantly decreased compared to that of the untreated control during storage at $4^{\circ}C$.

Molecular and Cultivation-Based Characterization of Bacterial Community Structure in Rice Field Soil

  • KIM MI-SOON;AHN JAE-HYUNG;JUNG MEE-KUM;YU JI-HYEON;JOO DONGHUN;KIM MIN-CHEOL;SHIN HYE-CHUL;KIM TAESUNG;RYU TAE-HUN;KWEON SOON-JONG;KIM TAESAN;KIM DONG-HERN;KA JONG-OK
    • Journal of Microbiology and Biotechnology
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    • v.15 no.5
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    • pp.1087-1093
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    • 2005
  • The population diversity and seasonal changes of bacterial communities in rice soils were monitored using both culture-dependent approaches and molecular methods. The rice field plot consisted of twelve subplots planted with two genetically-modified (GM) rice and two non-GM rice plants in three replicates. The DGGE analysis revealed that the bacterial community structures of the twelve subplot soils were quite similar to each other in a given month, indicating that there were no significant differences in the structure of the soil microbial populations between GM rice and non-GM rice during the experiment. However, the DGGE profiles of June soil after a sudden flooding were quite different from those of the other months. The June profiles exhibited a few intense DNA bands, compared with the others, indicating that flooding of rice field stimulated selective growth of some indigenous microorganisms. Phylogenetic analysis of l6S rDNA sequences from cultivated isolates showed that, while the isolates obtained from April soil before flooding were relatively evenly distributed among diverse genera such as Arthrobacter, Streptomyces, Terrabacter, and Bacillus/Paenibacillus, those from June soil after flooding mostly belonged to the Arthrobacter species. Phylogenetic analysis of 16S rDNA sequences obtained from the soil by cloning showed that April, August, and October had more diverse microorganisms than June. The results of this study indicated that flooding of rice fields gave a significant impact on the indigenous microbial community structure; however, the initial structure was gradually recovered over time after a sudden flooding.

Analysis of Total Bacteria, Enteric Members of γ-proteobacteria and Microbial Communities in Seawater as Indirect Indicators for Quantifying Biofouling

  • Lee, Jin-Wook;Kim, Sung-Min;Jung, Ji-Yeon;Oh, Byung-Soo;Kim, In S.;Hong, Soon-Kang
    • Environmental Engineering Research
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    • v.14 no.1
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    • pp.19-25
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    • 2009
  • In this study, total bacteria, enteric members of the $\gamma$-proteobacteria, and microbial communities in seawater were analyzed as indirect indicators for quantifying biofouling. Biomass in seawater can significantly affect feed water pretreatment and membrane biofouling of reverse osmosis desalination processes. The purpose of this paper is to investigate microbiological quantity and quality of seawater at the potential intake of a desalination plant. For this analysis, the total direct cell count (TDC) using 4'-6-diamidino-2-phenylindole (DAPI)-staining and DNA-based real-time PCR were used to quantify the total bacteria and relative content of enteric members of $\gamma$-proteobacteria in seawater, respectively. In addition, microbial communities were examined using 16S rRNA gene cloning and bacterial isolation to identify the most abundant bacteria for a further biofouling study. The experimental results of this study identified about $10^6$ cells/mL of (total) bacteria, $10^5$ 16S rRNA gene copies/mL of enteric $\gamma$-proteobacteria, and the presence of more than 20 groups of bacteria.

Isolation, Identification, and Characterization of Bacillus spp. from the Traditionally fermented Cheonggukjangs in the Gyeonggi and the Gangwon Provinces (경기.강원 지역의 전통발효 청국장으로부터 Bacillus균주의 분리 동정 및 특성 분석)

  • Lee, Nam-Keun;Jeon, Eun-Hee;Lee, Hyo-Jin;Cho, Il-Jae;Hahm, Young-Tae
    • Applied Biological Chemistry
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    • v.49 no.4
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    • pp.276-280
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    • 2006
  • Traditionally fermented Cheonggukjangs were collected from Gyeonggi and Gangwon provinces and 22 strains were isolated and identified by using 165 rDNA sequences. Most of the identified strains were Bacillus subtilis and B. licheniformis, B. subtilis and B. licheniformis are dominant in the Gyeonggi area and B. licheniformis in the Gangwon area. In the growth pattern of the isolated strains, the duration of lag phase was generally 5 to 7 hours and stationary phase was reached after 23 to 40 hours of incubation. Total cell populations at the stationary phase were between $1{\times}10^6\;CFU/ml$ and $5{\times}10^7\;CFU/ml$. The fermenting ability of carbohydrates of isolates showed some differences among the regions. The isolated strains from Yong-In, Gyeonggi showed higher fermenting abilities with D-xylose, xylitol, D-tagatose and Methyl-$\alpha$-D-mannopyranoside. D-lactose, D-tagatose, D-xylose, Methyl-$\alpha$-D-mannopyranoside, amygdalin, arbutin, esculin and 2-keto-gluconate were well fermented with the An-Seong's strains; L-rhamnose, inositol, D-mannitol, D-sorbitol, celibiose and gluconate with the Kawang-Ju's stains; and D-lactose with the Odaesan's strains.