• Title/Summary/Keyword: Microbial isolation

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Bioprospecting Endophytic Fungi and Their Metabolites from Medicinal Tree Aegle marmelos in Western Ghats, India

  • Mani, Vellingiri Manon;Soundari, Arockiamjeyasundar Parimala Gnana;Karthiyaini, Damodharan;Preethi, Kathirvel
    • Mycobiology
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    • v.43 no.3
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    • pp.303-310
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    • 2015
  • The increasing emergence of lead drugs for the resistance produced by the pathogenic strains and arrival of new diseases have initiated the need for searching novel metabolites with best anticancer and antimicrobial properties than the existing one. With this view, the investigation was conducted for the isolation, identification, and biological evaluation of potential endophytic fungi of Aegle marmelos, a medicinal tree used for more than three decades, for curing various disorders. A total of 169 endophytic fungal strains obtained from sampling and among those 67 were pigmented strains. Upon antagonistic screening, five endophytic fungal strains exhibited antagonistic potentiality by inhibiting the pathogens. These five potent strains were characterized at molecular level by sequencing the amplified internal transcribed spacer (ITS) 1 and ITS 4 regions of rDNA and they were grouped under order Pleosporales, Eurotiales, and Capnodiales. The metabolites from the respective strains were produced in fungal culturing media and extracted using polar solvents. Further, the extracts of five endophytes manifested antimicrobial activity against tested clinical pathogens and Alternaria alternata (FC39BY), Al. citrimacularis (FC8ABr), and Curvularia australiensis (FC2AP) exhibited significant antimicrobial profile against 9 of 12 tested pathogens, showing broad spectrum activity. The antioxidant levels of all the five endophytes revealed the highest activity at least concentrations, and major activity was unveiled by the members of order Pleosporales FC2AP and FC8ABr. This research explains the value of endophytic fungal extracts and its significance of antimicrobial and antioxidant properties.

Isolation of Indole-3-acetic acid (IAA) producing Arthrobacter sp. and plant growth promotion effect (Indole-3-acetic acid (IAA) 생성 Arthrobacter sp.의 분리 및 식물 생육촉진 효과)

  • Da Som Kim;Ho-Young Shin;Song-Ih Han
    • Journal of the Korean Applied Science and Technology
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    • v.39 no.6
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    • pp.831-838
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    • 2022
  • An auxin-producing bacteria, KSD16, KSD33, and KSD36 were isolated from agricultural soil. The strain KSD16, KSD33, and KSD36 was classified as a strain of Arthrobacter sp. based on phylogenetic analysis of 16S rRNA gene. The isolated KDS16, KDS33, and KSD36 was confirmed to produce indole-3-acetic acid (IAA), which is one of the auxin hormones. When the concentration of IAA was assessed the maximum concentration of IAA, 206.62 mg L-1, was detected from the culture broth incubated in R2A medium containing 0.1% L-tryptophan for 48 h at 28 ℃. To study the effect of IAA producing bacteria on germination rate, seeds of Mung bean were prepared for each treatment. KSD16, KSD33, and KSD36 showed significant increase in root length and number of adventitious roots than the controls. To investigate the growth-promoting effects on the crops, Arthrobacter species were placed in water cultures and seed pots of mung beans. In consequence, the seed germination of mung beans was 73.4% higher than the control.

Isolation of cellulosic biomass degrading microorganisms from different sources for low cost biofuel production

  • Sheikh, M. Mominul Islam;Kim, Chul-Hwan;Lee, Ji-Yong;Yeasmin, Shabina;Park, Hyeon-Jin;Kim, Gyeong-Chul;Kim, Sung-Ho;Kim, Jae-Won
    • Proceedings of the Korea Technical Association of the Pulp and Paper Industry Conference
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    • 2011.04a
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    • pp.81-91
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    • 2011
  • Current fuel ethanol research and development deals with process engineering trends for improving biotechnological production of ethanol. Recently, a large amount of studies regarding the utilization of lignocellulosic biomass as a good feedstock for producing fuel ethanol is being carried out worldwide. The plant biomass is mainly composed of cellulose, hemicellulose and lignin. The main challenge in the conversion of biomass into ethanol is the complex, rigid and harsh structures which require efficient process and cost effective to break down. The isolation of microorganisms is one of the means for obtaining enzymes with properties suitable for industrial applications. For these reasons, crude cultures containing cellulosic biomass degrading microorganisms were isolated from rice field soil, cow farm soil and rotten rice straw from cow farm. Carboxymethyl cellulose (CMC), xylan and Avicel (microcrystalline cellulose) degradation zone of clearance on agar platefrom rice field soil resulted approximately at 25 mm, 24 mm and 22 mm respectively. As for cow farm soil, CMC, xylan and Avicel degradation clearancezone on agar plate resulted around at 24mm, 23mm and 21 mm respectively. Rotten rice straw from cow farm also resulted for CMC, xylan and Avicel degradation zone almost at 24 mm, 23 mm and 22 mm respectively. The objective of this study is to isolatebiomass degrading microbial strains having good efficiency in cellulose hydrolysis and observed the effects of different substrates (CMC, xylan and Avicel) on the production of cellulase enzymes (endo-glucanase, exo-glucanase, cellobiase, xylanase and avicelase) for producing low cost biofuel from cellulosic materials.

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Microbial and Pathogenic Contamination of Ready-to-eat Fresh Vegetables in Korea (한국에 유통중인 신선편이 채소류의 미생물 품질 및 병원성 세균의 오염도 조사)

  • Bae, Young-Min;Hong, Yu-Jin;Kang, Dong-Hyun;Heu, Sung-Gi;Lee, Sun-Young
    • Korean Journal of Food Science and Technology
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    • v.43 no.2
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    • pp.161-168
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    • 2011
  • The purpose of this study was to evaluate microbiological contamination of fresh vegetables in Korea. Twenty types of vegetables were tested for total aerobic bacteria, coliforms, Escherichia coli, yeast and mold, and pathogenic bacteria such as Staphylococcus aureus, Listeria monocytogenes, Bacillus cereus, Salmonella, E. coli O157:H7, Cronobacter sakazakii, Shigella, and Campylobacter. Levels of total aerobic bacteria and coliform on 20 vegetables were between 3.74 and 8.04 log CFU/g, and 0.16 and 5.02 log CFU/g, respectively. The highest contamination levels of total aerobic bacteria were observed on water dropwort, sprouts, mungbean sprout, and ballflower root. There was no significant difference (p>0.05) in microbial contamination levels of total aerobic count, coliform, E. coli, yeast and mold between organic and nonorganic vegetables. When isolation methods using selective agars were applied, L. monocytogenes, B. cereus, Salmonella and Campylobacter were isolated from some fresh vegetable samples. Results of API kit tests showed that L. monocytogenes was identified on Chinese cabbage, cucumber, soybean sprouts, and iceberg lettuce while Salmonella was identified on Korean leek. Furthermore, Campylobacter jejuni was also identified in more than 50 of the 100 samples. However, when positive samples from API kit were tested for real-time PCR or 16S rRNA sequencing method, only B. cereus from perilla leaf, carrot, water dropwort, and sprouts showed positive results. These results indicate that selective agar and API kit detection methods might result in false positive results for some pathogens. Therefore, studies need to improve isolation or confirmation methods for such pathogens.

Microbial Hygienic Status of Poultry Meats and Eggs Collected at the Public Markets in Seoul and Kyung-gi Regions in 1996 (1996년도 서울${\cdot}$경기지역에서의 시판계육과 계란에 대한 미생물학적 위생실태)

  • Woo Yong-Ku
    • Korean Journal of Microbiology
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    • v.41 no.1
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    • pp.38-46
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    • 2005
  • To determine the actual hygienic status of domestic chicken meats sold in public markets (conventional markets and department stores), microbial contamination levels (Total cells, Coliforms and Staphylococcal cells) and zoonotic pathogens (Salmonella species, Campylobacter species, Listeria species, and Staphylococcus aureus) isolation tests were conducted. Chicken meats and eggs tested were collected from the conventional markets (Si-Jang) and department-stores located in Seoul and Kyung-gi regions in 1996. In total cells and coliforms contamination tests, chicken meats sold in department stores were much lesser contamination status than those of Si-Jang, but staphylococcal cells level was much more higher than that of conventional markets. Salmonella isolation frequency was investigated as $68.8\%$, but Campylobacter jejuni and Listeria monocytogenes isolation frequency were appeared both $64.0\%\;and\;63.3\%$. In case of eggs sold in public markets, one of S. gallinarum strain $(0.7\%)$ was isolated only on the egg-shell part among the four-hundred and fourty-six. In comparison with foreign imported chicken meats, there were no big differences in microbial contamination status. On the other hand, both Salmonella and L. monocytogenes were isolated only in the chicken wings from Korea and China, but not from U.S.A. This data suggest that more hygienic control system in order to produce the safe and hygienic chicken meats and eggs is need in our country as soon as possible.

Isolation of N-Iauroyl Tyrosine Antibiotic in E. coli Carrying N-acyl Amino Acid Synthase Gene from Environmental DNA in Korean Soils (한국 토양 환경유래의 N-acyl amino acid synthase 유전자에 의한 대장균 내 항생제 N-lauroyl tyrosine 생산)

  • Yeo, Yun-Soo;Lim, Yoon-Ho;Kim, Jeong-Bong;Yang, Jung-Mo;Lee, Chang-Muk;Kim, Soo-Jin;Park, Min-Seon;Koo, Bon-Sung;Yoon, Sang-Hong
    • Applied Biological Chemistry
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    • v.50 no.4
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    • pp.262-267
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    • 2007
  • To access the natural product antibiotics produced by uncultured microorganisms, six cosmid libraries of DNA extracted directly from soil samples (environmental DNA, eDNA) were constructed and screened for the production of antibacterial active molecules. Of the approximately 60,000 clones screened, one antibacterial clone (YS92B) was detected. Ethyl acetate extracts of clone YS92B showed antibacterial activity against various pathogenic bacteria (Listeria monocytogenes, Bacillus subtilis, Pseudomonas syringae, Xanthomonas campestris pv. oryzae, Staphylococcus epidemis). Active constituents from cultures of YS92B were isolated and purified using a bioassay-guided fractionation against B. subtilis through a series of procedures (ethyl acetate extraction, Sephadex LH20 column chromatography, High Performance Liquid Chromatography). NMR (Nuclear Magnetic Resonance) spectral analysis of a major antibacterial active YS92B-VII indicated that it is a lauric acid linked to tyrosine. This report describes the characterization of antibacterially active long chain N-acyl derivatives of tyrosine that are produced by eDNA clones hosted in Escherichia coli from Korean soils.

Isolation and Characterization of Bio-active Materials from Prickly Lettuce (Lactuca serriola) (가시상추 유래 생리활성물질의 분리 및 특성 규명)

  • Moon, Sung-Il;Kim, Sung-Whan;Huh, Wan;Kim, Soon-Young;Kim, Jong-Sik;Lee, Kon-Joo
    • Journal of Life Science
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    • v.19 no.2
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    • pp.206-212
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    • 2009
  • The bio-active materials were isolated from prickly lettuce (Lactuca serriola) by using several extraction solvents. The contents of general compounds, vitamins, polyphenols and flavonoids of prickly lettuce were analysed. In addition, nitrite scavenging activity, anti-oxidative activity and anti-microbial activity of methanol extracts were measured. The ethylacetate (EtOAc) fraction of methanol extracts of prickly lettuce showed the best on the nitrite scavenging activity, anti-oxidant activity and anti-microbial activity. And also, the contents of polyphenols and flavonoids were the highest among all fractions. The subfraction 2 (EtOAc:MeOH=3:1) of EtOAc fraction showed the strongest anti-oxidative activity among 5 subfractions. The subfraction 2 was identified as ${\rho}$-coumaric acid and caffeic acid by GC-MS. The anti-microbial activity of each solvent fraction from prickly lettuce was measured. The EtOAc fraction showed the strongest anti-microbial activity. Overall, these results may provide the basic data to understand the biological activities of bio-active materials derived from prickly lettuce.

Seasonal Changed of Microbial Population in the Field Soil (계절에 따른 토양중 미생물의 밀도 변화)

  • Park, Dong-Jin;Lee, Sang-Hwa;Kim, Chang-Jin
    • Korean Journal of Microbiology
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    • v.34 no.3
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    • pp.144-148
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    • 1998
  • Soil microorganisms including bacteria, fungi, and actinomycetes were seasonally isolated at depths (0.5~2, $10{\pm}1$, $50{\pm}1cm$) of field. The frequency of microbial isolates was employed for the determination of microbial population (CFU/g dry soil) and distribution ratio (%) in soil. Both bacteria (24-fold) and actinomycetes (7-fold) exhibited the biggest change at the depth of $50{\pm}1cm$, whereas fungi showed the maximum (13-fold) at $10{\pm}1cm$. On the whole, the bacterial population was high in spring soil, fungi in winter, and actinomycetes in autumn. Soil microorganisms also exhibited the seasonal variation on their distribution ratio (%). The maximum distribution ratio (85.7%) of bacteria was observed at the depth of $50{\pm}1cm$ in spring, whereas bacteria showed the minimum (35.2%) at the depth of $10{\pm}1cm$ in spring. The maximum distribution ratio (23.0%) of fungi was found at the depth of $50{\pm}1cm$ in spring, whereas its minimum (0.5%) at the depth of $10{\pm}1cm$ in spring. Actinomycetes exhibited the maximum distribution ratio (45.2%) at the depth of $10{\pm}1cm$ in spring, whereas its minimum (12.2%) was showed at the depth of $50{\pm}1cm$ in spring.

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Isolation and Characterization of Endophytic Bacteria from Rice Root Cultivated in Korea (한반도 중부지방의 벼 뿌리로부터 내생 세균의 분리와 특성 분석)

  • Park, Soo-Young;Yang, Sung-Hyun;Choi, Soo-Keun;Kim, Ji-Hyeon;Kim, Jong-Guk;Park, Seung-Hwan
    • Microbiology and Biotechnology Letters
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    • v.35 no.1
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    • pp.1-10
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    • 2007
  • The 44 endophytic bacterial strains were isolated from surface-sterilized root of rice cultivated in seven different locations of Chungcheong province, Korea. Each isolate was introduced into rice seedlings grown gnotobiotically by inoculating scissor-cut first true leaf with cell suspensions, and the colonization capacity of each isolate in root tissue was analyzed at 7 days after inoculation. Sixteen out of 44 isolates were re-isolated from root successfully with the frequency of $10^{3-5}$ CFU/g tissue. Interestingly, seven out of 16 isolates were identified as Burkholderia species. The identity between inoculated strains and re-isolates was confirmed by genomic finger-printing and 16S rDNA sequence analysis. By a confocal laser scanning microscopic observation it was revealed that KJ001 strain, one of the sixteen isolates tagged with gfp colonized in root tissue especially around xylem. Six out of seven Burkholderia strains obtained in this study showed antagonizing activities against seven different fungal pathogens, contain nifH gene, and five of them enhanced growth of cucumber over 30%. The isolates showed no hypersensitive response on tobacco leaves and no pathogenecity in rice. From these results it was found that the endophytic Burkholderia strains will be useful in agriculture to develop a biocontrol agent or a bio-fertilizer.

Isolation of Biogenic Amine Non-producing Lactobacillus brevis SBB07 and Its Potential Probiotic Properties (바이오제닉 아민 비생성 Lactobacillus brevis SBB07의 분리 및 잠재적 프로바이오틱스 특성 분석)

  • Yang, Hee-Jong;Jeong, Su-Ji;Jeong, Seong-Yeop;Ryu, Myeong Seon;Jeong, Do-Youn
    • Journal of Life Science
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    • v.28 no.1
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    • pp.68-77
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    • 2018
  • The purpose of this study was to isolate the probiotic lactic acid bacteria, and verify the possibility of the final selection strain as probiotic material. For screening of biogenic amines non-producing microorganisms, 42 lactic acid bacteria were isolated from various berries, extract and vinegar grown in Sunchang. Isolates were investigated for various physiological activities such as extracellular enzyme, antimicrobial and antioxidant activities, and 5 isolates were firstly screened. SBB07 was finally selected by analyzing the biogenic amine, and named Lactobacillus brevis SBB07 by 16S rRNA sequencing analysis. Next, SBB07 was assayed for their survival ability when exposed to acidic and bile conditions as well as heat and antibiotic resistance. As a result, SBB07 showed more than 86% and 54% higher survival rate in acidic condition at pH 2.0 and bile resistance with 0.5% oxgall. In addition, SBB07 showed a survival rate of more than 113% in $60^{\circ}C$, and also confirmed that it has resistant to various antibiotics. As a result of confirming the possibility of prebiotics, SBB07 showed the best utilization of GOS as a prebiotic substrate, and utilization of FOS and inulin were also high. These results suggest that SBB07 have good potential for application as probiotic lactic acid bacteria.