• Title/Summary/Keyword: E-Isolation

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Isolation of Bacillus licheniformis Producing Antimicrobial Agents against Bacillus cereus and Its Properties (Bacillus cereus 증식 억제능을 가지는 Bacillus licheniformis SCK 121057의 분리 및 특징)

  • Kim, Yong-Sang;Yun, Suk-Hyun;Jeong, Do-Yeon;Hahn, Kum-Su;Uhm, Tai-Boong
    • Korean Journal of Microbiology
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    • v.46 no.3
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    • pp.270-277
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    • 2010
  • In order to manufacture Bacillus cereus-free fermented soybean products, an antimicrobial agentproducing isolate against B. cereus was obtained from 150 traditionally fermented soybean products. The morphological and biochemical tests and the phylogenetic relationship among 16S rRNA gene sequences indicated that the isolate named as the strain SCK 121057 was most closely related to Bacillus licheniformis. The B. licheniformis isolate began to produce the antimicrobial agent after 48 h of incubation. The agent was nonproteinaceous and insensitive to heat, long term storage and protease K. Electron microscopic observation indicated that the agent attacked the membrane of B. cereus, leaving the ghost cell. The isolate inhibited growth of B. subtilis, Lactobacillus brevis and various types of pathogenic strains including Escherichia coli, E. faecalis, Micrococcus luteus, Staphylococcus aureus, Aspergillus flavus, A. ochraceus, and A. parasiticus as well as B. cereus. After coinoculation of B. licheniformis SCK 121057 and B. cereus in the ratio (as the basis of CFU/g sample) of 10 to 1 on the surface of cooked soybeans, cell numbers of B. cereus had been dramatically reduced after 31 days of incubation compared to those of single inoculation of B. cereus.

Isolation of Alliin in Garlic and Its Quantitative Determination by High Performance Liquid Chromatography and Studies on the Antimicrobial Efforts of Alliin and Ethanol Extracts from Korean Garlic(Alliium sativum L.) (마늘 중 고속 액체 크로마토그래피에 의한 알린의 분리 및 정량과 Alliin과 에탄올 추출물의 항균효과에 관한 연구)

  • 위성언
    • The Korean Journal of Food And Nutrition
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    • v.16 no.4
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    • pp.296-302
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    • 2003
  • First. the purification and analysis of alliin in garlic from different origins by alliin-HPLC determination method were studied. Allinase in garlic was inactivated by heating in boiling water followed by extraction of alliin in garlic with 80% methanol. To remove free amino acids and alliin homologs in garlic, garlic extract was separated by cation exchange column which was packed with amberlite CG-120 resin using 40L d-water as eluent. Alliin in garlic extract was crystallized in a mixture of acetone (50$^{\circ}C$):H$_2$O:acetic acid=70:29:1 and then recrystallized in a mixture of acetone (50$^{\circ}C$):H$_2$O:acetic acid=75:24:1. Obtained alliin was identified by melting point. TLC, microscope observation and mass spectrometry. High performance liquid chromatography (HPLC) following pre-column derivatization of cystein derivatives with o-phthaldialdehyde/2-mercaptoethanol has succeessfully been applied to the analysis of various garlics. Each alliic of standard solution and garlic extract was derivatized to isoindole derivative by o-phthaldialdehyde /2-mercaptoethanol and then analyzed by HPLC. Six point calibration was done by using alliin peak area. Lineality was observed at 0 ∼ 1.0mg/ml of alliin concentration. Weighted regression line function was Y=6254X - 256077. By this function, alliin contents in various garlics were 0.34 ∼ 0.73% fresh weight. Second study was designed to evaluate the effects of garlic extracts of various concentrations on the growth of various pathogenes (Eubacterium limonsum, Bacteroides fragilis, Salmonella typhimurium, Salmonella typhi, Shigella sonnei, Kiebsiella pneumoniae, Enterobacter cloacae, Pserdomonas aeruginosa, Escherichia coli). For antimicrobial effects against microorganism, totally minimal inhibition concentrations (MIC) of alliin were from 5,000 to 20,000ppm. MIC of ethanol extract were 1,250 to 10,000ppm.

Drug Resistance Profiles of Salmonella sp. and Shigella sp. Isolated from Diarrheal Patients in Pusan, Korea (부산지역의 설사환자로부터 분리한 Salmonella sp. 및 Shigella sp.의 약제내성 유형에 관한 연구)

  • 차인호;김용환;빈재훈;김경숙
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.23 no.6
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    • pp.927-932
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    • 1994
  • A total of 34 Salmonella sp. an d25 Shigella sp. were isolated from 311 patients with diarrhea. The isolation rates of Salmonella sp. ad Shigella sp. were 10.9% and 8%, respectively. The serogroups of 34 Salmonella sp. were in order of group D(50%), group B(38.25), group E(8.8%) and group C 92, 9%0. the serogroups of 25 Shigella sp. were group D(96%) and group B(4%). Seasonal distribution of isolated Salmonella sp. and Shigella sp. were shown the most high at July, 17.65% and 64%, respectively. Age group distribution of isolated Salmonella sp. and Shigella sp. were shown the most high at twenties and thirties (23.5%), and teens(76%), respectively. The Salmonella isolates were resistant in order of prevalence use of streptomycin(SM) (100%), erythromycin (EM) and movobiocin (NB)(90.6%), penicillin G(PG) (65.6%) and cephalexin (CPX)(46.9%). the isolates of Shigella sp. were resistant in order of prevalence use of EM (95.8%), NB(91.7%), SM(87.5%). Eighteen kinds of resistant patterns of Salmonella ioslates were detected. The multiple resistance patterns of Shigella isolates were mostly SM, EM, NB type (79.2%). The minimum inhibitory concentration of Salmonella sp. and Shigella sp. and Shigella sp. isolated from patients with diarrhea were tabulated.

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Sequence Analysis of a Cryptic Plasmid pKW2124 from Weissella cibaria KLC140 and Construction of a Surface Display Vector

  • Kim, Soo Young;Oh, Chang Geun;Lee, Young Joo;Choi, Kyu Ha;Shin, Doo Sik;Lee, Si Kyung;Park, Kab Joo;Shin, Hakdong;Park, Myeong Soo;Lee, Ju-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.23 no.4
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    • pp.545-554
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    • 2013
  • Plasmid isolation of kimchi-derived Weissella cibaria KLC140 revealed six different plasmids. The smallest plasmid, pKW2124, was DNA sequenced and characterized, showing 2,126 bp with a GC content of 36.39% and five putative open reading frames (ORFs). In silico analysis of these ORFs showed ORF1 encodes a putative replication protein similar to rolling circular replication proteins from other lactic acid bacteria. However, a single-stranded intermediate was not detected when S1 nuclease was treated, suggesting it may follow theta replication. Interestingly, the replication initiation site of this plasmid is 100% identical to other plasmids from lactic acid bacteria, suggesting it may function for replication initiation. To construct a surface layer expression vector, pTSLGFP, slpA encoding the surface layer protein from Lactobacillus acidophilus was PCR amplified and fused with the gfp gene, forming a SLGFP fused gene. The plasmid pKW2124 was cloned into the XbaI site of pUC19, forming an Weissella-E. coli shuttle vector pKUW22. NheI-linearized pTSLGFP was ligated into pKUWCAT containing pKUW22 and the chloramphenicol acetyltransferase gene from pEK104, resulting in an 8.6 kb pKWCSLGFP surface layer expression vector. After transformation of this vector into W. cibaria KLC140, a GFP fluorescence signal was detected on the surface of the transformant, substantiating production of SLGFP fused protein and its secretion. This is the first report for construction of a Weissella surface layer expression vector, which may be useful for surface layer production of beneficial proteins in Weissella.

Full-Wave Analysis, Design and Fabrication of Duplexer by Mode Matching Method for Ka-Band Transponder (모드정합법에 의한 Ka-밴드 위성중계기용 듀플렉서의 Full-Wave 분석 및 설계${\cdot}$제작에 관한 연구)

  • Lee, Yong-Min;Ra, Keuk-Hwan
    • Journal of the Korean Institute of Telematics and Electronics D
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    • v.36D no.8
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    • pp.36-44
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    • 1999
  • This paper presents the design and fabrication of the duplexer for a Ka-band satellite transponder which is analyzed transmission characteristics by calculating the generalized scattering matrix using mode matching method. It is composed of 2 bandpass filters, coupled with H-plane T-junction having symmetrical inductive iris and E-plane metal insert structures. Compared with the size and weight of the Rx and Tx filter loaded with a transponders respectively, those of the duplexer can be effectively reduced. In a high power transmission, the variation of the filter characteristics is minimized by the scheme that irises are extended to the exterior of H-plane of the waveguide. This scheme needs no extra heat sinks for dissipating high power. The duplexer is designed to improve the simplification, durability and reliability by eliminating tuning screws, which have been used to compensate for the characteristics of fabricated filters. The bandpass filters of the duplexer show the insertion loss of less than 1.2 dB and the return loss in excess of 15 dB. The isolations of more than 65 dB have been achieved between Rx and Tx filter.

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Fabrication, Estimation and Trypsin Digestion Experiment of the Thermally Isolated Micro Teactor for Bio-chemical Reaction

  • Sim, Tae-Seok;Kim, Dae-Weon;Kim, Eun-Mi;Joo, Hwang-Soo;Lee, Kook-Nyung;Kim, Byung-Gee;Kim, Yong-Hyup;Kim, Yong-Kweon
    • JSTS:Journal of Semiconductor Technology and Science
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    • v.5 no.3
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    • pp.149-158
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    • 2005
  • This paper describes design, fabrication, and application of the silicon based temperature controllable micro reactor. In order to achieve fast temperature variation and low energy consumption, reaction chamber of the micro reactor was thermally isolated by etching the highly conductive silicon around the reaction chamber. Compared with the model not having thermally isolated structure, the thermally isolated micro reactor showed enhanced thermal performances such as fast temperature variation and low energy consumption. The performance enhancements of the micro reactor due to etched holes were verified by thermal experiment and numerical analysis. Regarding to 42 percents reduction of the thermal mass achieved by the etched holes, approximately 4 times faster thermal variation and 5 times smaller energy consumption were acquired. The total size of the fabricated micro reactor was $37{\times}30{\times}1mm^{3}$. Microchannel and reaction chamber were formed on the silicon substrate. The openings of channel and chamber were covered by the glass substrate. The Pt electrodes for heater and sensor are fabricated on the backside of silicon substrate below the reaction chamber. The dimension of channel cross section was $200{\times}100{\mu}m^{2}$. The volume of reaction chamber was $4{\mu}l$. The temperature of the micro reactor was controlled and measured simultaneously with NI DAQ PCI-MIO-16E-l board and LabVIEW program. Finally, the fabricated micro reactor and the temperature control system were applied to the thermal denaturation and the trypsin digestion of protein. BSA(bovine serum albumin) was chosen for the test sample. It was successfully shown that BSA was successfully denatured at $75^{\circ}C$ for 1 min and digested by trypsin at $37^{\circ}C$ for 10 min.

Serotypes and genotypes of Salmonella isolates from slaughtered pigs (도축돈에서 분리된 살모넬라의 혈청형 및 유전형)

  • Choi, Won-Zong;Jung, Ji-Hun;Won, Ho-Keun;Kang, Zheng-Wu;Hahn, Tae-Wook
    • Korean Journal of Veterinary Service
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    • v.31 no.1
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    • pp.1-16
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    • 2008
  • Salmonella infections cause the disease in pigs but also some zoonotic Salmonella serotypes can be transmitted to human through swine products, resulting in food poisoning. The objective of this study was to investigate the bacteriological prevalence and detection of invA gene using Salmonella specific polymerase chain reaction (PCR), the epidemiological characteristics related to Salmonella strains cultured from pig samples in Gangwon areas using serotyping, random amplified polymorphic DNA (RAPD) and pulsed field gel electrophoresis (PFGE) methods. During the period of November 2001 through April 2002, 1,174 ileocecal lymph node were collected from the slaughtered pigs raised in 38 farms located in Gangwon province. The samples were submerged in boiling water and macerated in saline and lymph node homogenates were inoculated into Tetrathionate broth with iodine (TTB, Difco, 0.5% iodine was added) for enrichment growth. Then additional tests were performed using several mediums, and suspects were identified by API 20E kit (BioMerieux) and PCR. Of total 1,174 samples from 38 farms, 44 (3.7%) were isolated as Salmonella spp from 13 farms (34.2%). Of 44 isolates, 31 were in Yangyang region, followed by 9 in Goseong, 2 in both Gangneung and Sokcho. However, there was no difference in regional isolation frequency. All isolates have a 521bp amplified product in Salmonella specific PCR with primer invA which encodes in proteins for invasion of epithelial cells. Of 44 recovered serotypes, 23 (52.3%) were S Eingedi, 10 (22.7%) S Schwarzengrund, 9 (20.5%) S Typhimurium, and 2 (4.5%) S Mbandaka. In RAPD analysis, there appeared to be unique bands distinguishing each serotype, although similarities exist between the different serotypes. Four serotypes of 44 Salmonella isolates appeared to fall into 14 different RAPD types. In PFGE analysis, 9 S Typhimurium were tested with XbaI enzyme and SpeI enzyme. The combination of results obtained with two enzymes subdivided the 9 S Typhimurium into 4 PFGE types.

Studies on the Distribution of Streptomyces spp. in Soil in Korea (한국토양(韓國土壤)의 방선균(放線菌) 분포(分布)에 관(關)한 연구(硏究))

  • Park, J.E.;Choi, Y.C.;Sin, Y.H;Lee, K.H.
    • Korean journal of applied entomology
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    • v.25 no.1 s.66
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    • pp.47-51
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    • 1986
  • A study was undertaken to investigate the distribution of Streptomyces spp. in soil in Korea. Among the different types of soil surveyed, the highest population of Streptomyces spp. recording $5.6{\times}10^5\;CFU/g$ was observed in upland soil. With reference to the soil depth, most of their population was distributed from soil surface to 5cm depth and the highest value was found in $0{\sim}2cm$ soil depth. Comparing the population of Streptomyces spp. with different soil color (by Munsell soil color chart), the highest value of $9.2{\times}10^\;CFU/g$ was showed in Oliver yellow soil (2.5Y, 6/4). On the basis of the acidity of soil samples subjected to Streptomyces spp. isolation, it is considered that the optimum pH range for Streptomyces spp. in soil lies between 6.0 to 7.5, showing the highest value of $1.05{\times}10^6CFU/g$ at pH 7.5. Among the colors of isolated colonies, gray and white colonies occupied 60% and 26% of the total isolates respectively.

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A STUDY OF NI-RESISTANT BACTERIA ISOLATED FROM GINGIVAL CREVICULAR FLUID ON THE PATIENTS WEARING NI-CR ALLOY PROSTHESIS (IN TERMS OF MOLECULAR BIOLOGICAL ASPECTS) (니켈-크롬 합금 보철물 주위 치은 열구내에서 발견된 니켈 내성 균주에 관한 분자생물학적 연구)

  • Chae Young-Ah;Woo Yi-Hyung;Kwon Kung-Rock
    • The Journal of Korean Academy of Prosthodontics
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    • v.41 no.2
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    • pp.207-222
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    • 2003
  • As a material of metal-ceramic prosthesis, nickel as a form of Ni-Cr alloy has been used for many dental prostheses in many cases. However, several problems in use of the alloy have been revealed (ex ; tissue stimulation, skin allergy, hypersensitivity cytotoxicity and carcinogenecity). Little is known about nickel with respect to the relationship between Ni-prosthesis and gaining of Niresistance in oral microorganisms. The present study was undertaken to check whether use of Ni-prosthesis leads to occurrence of Ni-resistant microorganisms. So this study may suggest the possible relationships between the oral microorganisms and nickel-resistance in oral environment. Bacteria were isolated from the gingival crevicular fluid on the patients wearing Ni-Cr prosthesis. The isolated bacteria were tested fir their Ni-resistance in nickel containing media at different concentration from 3mM to 110mM. E. coli HB101 was used as control. The Ni-resistant bacteria were isolated and biochemically identified. The Ni-resistant bacteria were tested several biochemical, molecular-biological tests. Performed tests were : measuring the growth curve, antibiotic test, growth ability test in liquid media, isolation of the chromosome and plasmid, digestion of DNA by restriction enzyme, electrophoresis of chromosome and plasmid DNA, identification of Ni-resistant genes by the DNA hybridization. The results were as follows 1) The bacteria isolated from gingival crevicular fluid on the patients wearing Ni-Cr alloy prosthesis showed nickel-resistance. 2) The isolated microorganisms grew at nickel containing media of high concentrations (60mM-110mM). 3) Based on the biochemical tests, the isolated microorganisms were identified as Enterococcus faecalis(13 cases), Klebsiella pneumoniae(1 case) and Enterobacter gergoviae(1 case). 4) Enterococcus faecalis expressed not only nickel resistance but also the multi-drug resistance to several antibiotics ; chloramphenicol, kanamicin, streptomycin, lincomycin, clindamycin, However, all strain showed the sensitivity against the tetracycline. 5) DNA hybridization result suggests that there is no homology between the previously known gene of nickel resistance in Klebsiella pneumoniae and chromosomal DNA of Enterococcus faecalis.

Isolation of Wild Yeasts from Humus-rich Soil in City Park of Daejeon Metropolitan City, Korea, and Characterization of the Unrecorded Wild Yeasts (대전광역시 도시 자연공원 부엽토와 주변 산림토양들로부터 야생효모의 분리, 동정 및 국내 미기록 야생효모들의 특성)

  • Han, Sang-Min;Lee, Sang-Yeop;Lee, Jong-Soo
    • The Korean Journal of Mycology
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    • v.46 no.1
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    • pp.75-82
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    • 2018
  • Totally 91 strains of wild yeasts were isolated from the humus rich soil in the Bogyong city park of Daejeon city, Korea. Majority of the strains belonged to Cryptococcus spp., which included 11 strains of Cryptococcus aureus. Among them, Kwoniella mangroviensis JSS0500, Candida corydalis JSS0501, Candida bombi JSS0503, Candida multigemmis JSS0504, Cryptococcus dimennae JSS0506, Cryptococcus saitoi JSS0507, Cryptococcus victoriae JSS0508, Metschnikowia pulcherrima JSS0502, Papiliotrema aurea JSS0505, Debaryomyces vanrijia JSS0509, Occultifur externus JSS0510, Filobasidium floriforme JSS0511 and Yamadazyma scolyti JSS0512 represented newly recorded yeast strains in Korea, and their microbiological characteristics were investigated. All of these unrecorded yeasts showed oval shape and also formed ascospores and pseudomycelia, except for Kwoniella mangroviensis JSS0500, Candida bombi JSS0503, and Metschnikowia pulcherrima JSS0502. Seven strains including Candida corydalis JSS0501 grew in vitamin-free medium, and 4 of the wild yeasts including Cryptococcus victoriae JSS0508 were halotolerant, i.e., capable of growing in 10% NaCl-containing yeast extract-peptone-dextrose (YPD) broth. Debaryomyces vanrijia JSS0509 was found to be a thermophilic yeast that grew at $37^{\circ}C$.