• Title/Summary/Keyword: Microbial culture

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Phylogenetic diversity of bacterial communities in a gray solar saltern and isolation of extremely halophilic bacteria using culturomics (토판염전 결정지 내 세균군집의 계통학적 다양성 및 Culturomics법을 이용한 고도 호염균의 분리)

  • Cho, Geon-Yeong;Han, Song-Ih;Whang, Kyung-Sook
    • Korean Journal of Microbiology
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    • v.53 no.1
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    • pp.29-38
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    • 2017
  • In this study, we investigated the phylogenetic diversity of the bacterial community and isolation of extremely halophilic bacteria using culturomics in a gray solar saltern. The number of bacterial living cells, enumerated in a gray solar saltern by direct fluorescence microscopy was three to four orders of magnitude greater than those enumerated by plate counts, suggesting the distribution of 'viable but non-culturable bacteria'. The biodiversity of bacterial communities in a gray solar saltern was investigated by pyrosequencing, 1,778 OTUs of bacteria were comprised of 18 phyla 46 classes 85 orders 140 families 243 genera with 6.16 diversity index. Archaea communities were composed of 3 phyla 6 classes 7 orders 7 families 38 genera with 4.95 diversity index from 643 OTUs. Totally 137 isolates were isolated by 59 different cultural methods based on culturomics considering culture media and conditions suitable for the growth of extremely halophilic bacteria. Phylogenetic analyses of extremely halophilic isolates based on 16S rRNA gene sequences, extremely halophilic isolates were composed of 4 phyla and 11 genera. Haloterrigena and Haloferax can be successfully isolated from culturomics. These culturomics were effective methods for collection of diversity of extremely halophilic bacteria.

Characteristics of Fermented Fruit and Vegetable Mixed Broth Using by Bacteriocin-producing Lactic Acid Bacteria and Yeast (박테리오신 생성 젖산균과 효모를 이용한 과채발효액의 특성)

  • Jung, Dong-Sun;Lee, Young-Kyung;Lim, Kyung-Wha
    • Korean Journal of Food Science and Technology
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    • v.32 no.6
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    • pp.1358-1364
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    • 2000
  • A mixed culture of bacteriocin-producing lactic acid bacteria and yeast was used to prepare unique fermented fruit and vegetable beverage which contains bacteriocin. Fruit and vegetable broth fermented by Lactococcus lactis 11454 showed inhibitory activity against foodborne pathogens such as Listeria monocytogens and Staphylococcus aureus, and Streptococcus mutans which is a major causative agent for dental caries. Bacteriocin was detected at the end of fermentation of fruit and vegetables and its antimicrobial activity was stable for 5 weeks during storage at $4^{\circ}C$. These results suggest that the presence of bacteriocin in beverages during fermentation and storage will provide a safeguard against foodborne pathogens and spoilage bacteria. To improve flavor of the fermented broth, post-fermentation of the fermented broth was carried out at $4^{\circ}C$ by using yeast isolated from raw material mixture. Total acidity of the post-fermented broth was slightly decreased, but significant increase in the concentration of succinic acid was observed in the post-fermented broth. It was also observed that bitter and disagreeable taste compounds such as phthalates were decreased, and mild acidic and fruity flavor esters and alcohols were increased by the post-fermentation of the fermented broth with yeast.

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Biochemical and cultural characteristics of mineral-solubilizing Acinetobacter sp. DDP346 (미네랄 가용화능을 갖는 Acinetobacter sp. DDP346의 생화학적 및 배양학적 특성)

  • Kim, Hee Sook;Lee, Song Min;Oh, Ka-Yoon;Kim, Ji-Youn;Lee, Kwang Hui;Lee, Sang-Hyeon;Jang, Jeong Su
    • Journal of Applied Biological Chemistry
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    • v.64 no.4
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    • pp.333-341
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    • 2021
  • In this study, to select strains suitable as microbial agent from among rhizosphere microorganisms present in rhizosphere soil and roots, the mineral solubilization ability, antifungal activity against 10 types of plant pathogenic fungi, and plant growth-promoting activity of rhizosphere microorganisms were evaluated. As a result, DDP346 was selected because it has solubilization ability of phosphoric acid, calcium carbonate, silicon, and zinc; nitrogen fixing ability; production ability of siderophore, indole-3-acetic acid, and aminocyclopropane-1-carboxylate deaminase; and antifungal activity against seven types of plant pathogenic fungi. DDP346 showed a 99.9% homology with Acinetobacter pittii DSM 21653 (NR_117621.1); phylogenetic analysis also revealed a close relationship with Acinetobacter pittii based on the 16S rRNA base sequence. The growth conditions of DDP346 were identified as temperatures in the range of 10-40 ℃, pH in the range of 5-11, and salt concentrations in the range of 0-5%. In addition, a negative correlation coefficient (r2 = -0.913, p <0.01) was shown between pH change and the solubilized phosphoric acid content of Acinetobacter sp. DDP346, and this is assumed to be due to the organic acid generated during culture. Consequently, through the evaluation of its mineral solubilization ability, antifungal activity against plant pathogenic fungi, and plant growth-promoting activity, the potential for the utilization of Acinetobacter sp. DDP346 as a multi-purpose microbial agent is presented.

Improving Biomass Productivity of Freshwater microalga, Parachlorella sp. by Controlling Gas Supply Rate and Light Intensity in a Bubble Column Photobioreactor (가스공급속도 및 광도조절을 이용한 담수미세조류 Parachlorella sp.의 바이오매스 생산성 향상)

  • Z-Hun Kim;Kyung Jun Yim;Seong-Joo Hong;Huisoo Jang;Hyun-Jin Jang;Suk Min Yun;Seung Hwan Lee;Choul-Gyun Lee;Chang Soo Lee
    • Journal of Marine Bioscience and Biotechnology
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    • v.15 no.2
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    • pp.41-48
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    • 2023
  • The objective of the present study was to improve the biomass productivity of newly isolated freshwater green microalga Parachlorella sp. This was accomplished by culture conditions optimization, including CO2 concentration, superficial gas velocity, and light intensity, in 0.5 L bubble column photobioreactors. The supplied CO2 concentration and gas velocity varied from 0.032% (air) to 10% and 0.02 m/s - 0.11 m/s, respectively, to evaluate their effects on growth kinetics. Next, to maximize the production rate of Parachlorella sp., a lumostatic operation based on a specific light uptake rate (qe) was applied. From these results, the optimal CO2 concentration in the supplied gas and the gas velocity were determined to be 5% and 0.064 m/s, respectively. For the lumostatic operation at 10.2 µmol/g/s, biomass productivity and photon yield showed significant increases of 83% and 66%, respectively, relative to cultures under constant light intensity. These results indicate that the biomass productivity of Parachlorella sp. can be improved by optimizing gas properties and light control as cell concentrations vary over time.

Two-year Quaternary Isolation of Gram-positive Bacilli Using MALDI-TOF MS in Positive Blood Culture of a University Hospital (일개 대학병원의 혈액배양에서 MALDI-TOF MS를 이용한 Gram-positive Bacilli의 2년간 분기별 분리율)

  • Choi, Jin-Un;Yu, Young-Bin;Kim, Sang-Ha;Won, Seungho;Kim, Young-Kwon
    • Korean Journal of Clinical Laboratory Science
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    • v.50 no.4
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    • pp.414-421
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    • 2018
  • In this study, Gram-positive bacilli (GPB) were identified by MALDI-TOF MS and analyzed according to the quaternary and microbial strains in the blood culture medium over a two year period at a university hospital. The results were as follows. The overall positive rate of blood culture was 9.97%. In 713 isolated GPB, 410 strains (57.5%) were identified using a microflex MALDI Biotyper. The positive rate of GPB among the blood culture positive bacteria was 8.2%, and the quarterly isolation rate was 9.8% in the third quarter of 2015, 8.7% in the second quarter of 2016, 8.1% in the third quarter of 2016, 8.1% in the first quarter of 2015, 7.9% in the first quarter of 2015, 7.9% in the second quarter of 2015, 6.8% in the first quarter of 2016, and 6.7% in the fourth quarter of 2015. The isolates were Corynebacterium striatum 89 (12.4%), Bacillus cereus 60 (8.4%), Bacillus subtilis 30 (4.2%), Paenibacillus urinalis 29 (4.1%), and Listeria monocytogenes 25 (3.5%). The results of 16S rRNA sequencing of 43 isolates (86.0%) were consistent with those of the other 50 isolates. Five out of the seven unmatched weeks were not identified by MALDI-TOF MS.

Fusaric Acid Production in Fusarium oxysporum Transformants Generated by Restriction Enzyme-Mediated Integration Procedure (Restriction Enzyme-Mediated Integration 방법으로 확보한 Fusarium oxysporum 형질전환체의 후자리산 생성능 분석)

  • Lee, Theresa;Shin, Jean Young;Son, Seung Wan;Lee, Soohyung;Ryu, Jae-Gee
    • Research in Plant Disease
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    • v.19 no.4
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    • pp.254-258
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    • 2013
  • Fusaric acid (FA) is a mycotoxin produced by Fusarium species. Its toxicity is relatively low but often associated with other mycotoxins, thus enhancing total toxicity. To date, biosynthetic genes or enzymes for FA have not been identified in F. oxysporum. In order to explore the genetic element(s) for FA biosynthesis, restriction enzyme mediated integration (REMI) procedure as an insertional mutagenesis was employed using FA producing-F. oxysporum strains. Genetic transformation of two F. oxysporum strains by REMI yielded more than 7,100 transformants with efficiency of average 3.2 transformants/${\mu}g$ DNA. To develop a screening system using phytotoxicity of FA, eleven various grains and vegetable seeds were tested for germination in cultures containing FA: Kimchi cabbage seed was selected as the most sensitive host. Screening for FA non-producer of F. oxysporum was done by growing each fungal REMI transformant in Czapek-Dox broth for 3 weeks at $25^{\circ}C$ then observing if the Kimchi cabbage seeds germinated in the culture filtrate. Of more than 5,000 REMI transformants screened, fifty-three made the seeds germinated, indicating that they produced little or fewer FA. Among them, twenty-six were analyzed for FA production by HPLC and two turned out to produce less than 1% of FA produced by a wild type strain. Sequencing of genomic DNA regions (252 bp) flanking the vector insertion site revealed an uncharacterized genomic region homologous (93%) to the F. fujikuroi genome. Further study is necessary to determine if the vector insertion sites in FA-deficient mutants are associated with FA production.

Effect of Microbial Additives on Metabolic Characteristics in Sheep and Milking Performance of Lactating Dairy Cows (미생물제제의 첨가가 면양의 반추대사 및 젖소의 유생산성에 미치는 영향)

  • Kim, G.L.;Choi, S.K.;Choi, S.H.;Song, M.K.
    • Journal of Animal Science and Technology
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    • v.49 no.6
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    • pp.819-828
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    • 2007
  • Two experiments were conducted to observe the effects of direct fed microbials on metabolic characteristics in sheep and milking performance in dairy cows. A metabolic trial with four ruminally cannulated sheep(60±6kg) was conducted in a 4×4 Latin square design to investigate the supplementation effects of Saccharomyces cerevisiae, Clostridium butyricum or mixed microbes of S. cerevisiae and C. butyricum on ruminal fermentation characteristics and whole tract digestibility. Sheep were fed 1.25 kg of total mixed ration(TMR, DM basis) supplemented with S. cerevisiae (2.5g/day), C. butyricum (1.0g/day) or its mixture(S. cerevisiae 1.25g/day+C. butyricum 1g/day), twice daily in an equal volume. But control sheep were fed only TMR. A feeding trial with 28 lactating Holstein cattle was also conducted for 12 weeks to investigate the effects of the same microbial supplements as for the metabolic trial on milking performance. The cows were fed the TMR(control), and fed S. cerevisiae(50g/day), C. butyricum(15g/day) or its mixture (S. cerevisiae 25g/day + C. butyricum 7.5g/day) with upper layer dressing method. Total VFA concentration and the digestibility of whole digestive tract in the sheep increased by supplementation of S. cerevisiae, C. butyricum or their combined microbials compare to control group. The proportion of propionic acid at 1h(P<0.039) and 3h(P<0.022) decreased by supplementation of S. cerevisiae while tended to increase acetic acid proportion at the same times. Daily dry matter intake(DMI) was not influenced by the microbial treatments, but milk yield(P<0.031) and feed efficiency(milk yield/DMI, P<0.043) were higher for the cow received C. butyricum than those for other treatments. The milk fat content was higher (P<0.085) when cows fed S. cerevisiae(4.11%) than that fed the control (4.08%), the diets with C. butyricum (3.85%) and the microbial mixture. Based on the results obtained from the current experiments, supplementation of C. butyricum or mixture with S. cerevisiae might be increased milk fat content and milk productivity of lactating daily cows. (Key words:Saccharomyces cerevisiae, Clostridium butyricum, Fermentation characteristics,

Effects on the Rumen Microbial Fermentation Characteristics of Lignosulfonate Treated Soybean Meal (Lignosulfonate처리 대두박의 반추위 내 미생물 발효특성에 미치는 영향)

  • Lee, Hun-Jong;Lee, Seung-Heon;Bae, Gui-Seck;Park, Je-Hwan;Chang, Moon-Baek
    • Journal of Animal Science and Technology
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    • v.52 no.5
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    • pp.413-426
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    • 2010
  • This study was conducted to investigate the effects on fermentation characteristics of rumen microorganism by different types and levels of lignosulfonate treated soybean meal (LSBM) in in vitro test and rumen simulation continuous culture (RSCC) system in dairy cows. The experiment I was control and 12 treatments (each with 3 replications) in vitro test to demonstrate composition of different types of treatments with lignosulfonate (Desulfonate, Na, Ca and solution) and levels (2, 4 and 8%) of soybean meal in the dairy cow diet. LSBM source treatments in the dairy cow diet showed pH value, $NH_3$-N concentration and total VFA concentration lower than control at all levels and incubation times (p<0.05). Dry matter digestibility of LSBM source treatments showed lower than control (p<0.05). Gas production and rumen microbial synthesis was decreased by rumen microbial fermentation for incubation times. Undegradable protein (UDP) concentration of all LSBM treatments was decreased for incubation times, and significantly higher than control (p<0.05). In the experiment II compared diets of the control, LSBM Na 2%, LSBM Sol 2%, which are high performance to undegradable protein (UDP) concentration experiment I in vitro test, and heated treatment lignosulfonate (LSBM Heat) 2% in the dairy cow diet from four station RSCC system ($4{\times}4$ Latin square). A rumen microbial fermentation characteristic was stability during 12~15 days of experimental period in all treatments. The pH value of LSBM treatments was higher than control treatment (p<0.05). The $NH_3$-N concentration, VFA concentration and rumen microbial synthesis of LSBM treatments were lower than control (p<0.05). The undegradable protein (UDP) showed LSBM Na 2% (45.28%), LSBM Sol 2% (43.52%) and LSBM Heat 2% (43.49%) higher than control (41.55%), respectively (p<0.05). Those experiments were designed to improve by-pass protein of diet and milk protein in the dairy cows. We will conduct those experiments the in vivo test by LSBM treatments in dairy cows diet.

Effects of Acidification on the Changes of Microbial Diversity in Aquatic Microcosms

  • Young-Beom Ahn;Hong-Bum Cho;Byung Re Min;Yong-Keel Choi
    • Animal cells and systems
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    • v.3 no.2
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    • pp.153-159
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    • 1999
  • In an artificial pH-gradient batch culture system, the effects of acidification on the species composition of a heterotrophic bacterial community were analyzed. As a result of this study, it was found that total bacteria numbers were not affected by acidification and that the population of hetero-trophic bacteria decreased as pH became lower. The heterotrophic bacteria isolated from the entire pH gradient were 12 genera and 22 species. Among them, 64% were gram negative and 36% were gram positive bacteria. As pH decreased, the distribution rate of gram negative bacteria increased while that of gram positive bacteria decreased. The diversity of genera decreased from 13 to 5 as pH decreased from 7 to 3. The G+C content of all of the 202 isolated strains varied from 22.8 to 77.0%, and increased in interspecies of same genus as pH decreased. As a result of clustering analysis, the diversity index of species ranged from 1.13 to 2.37, and it had lower indices as pH decreased. In order to evaluate the diversity of numbers of sample of different size, a rarefaction method was used to analyze the expected number of species appearance according to pH. The statistical significance of species diversity was verified by the fact that the number decreased at lower pH.

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Optimization of Physical Factor for amylase Production by Arthrobacter sp. by Response Surface Methodology (반응표면분석법을 통한 Arthrobacter sp.의 amylase 생산 최적화)

  • Kim, Hyun-do;Im, Young-kum;Choi, Jong-il;Han, Se Jong
    • Korean Chemical Engineering Research
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    • v.54 no.1
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    • pp.140-144
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    • 2016
  • In this study, the physical factors for amylase production by Arthrobacter sp. were optimized using response surface methodology(RSM). Antarctic microorganism Arthrobacter sp. PAMC 27388 was obtained from the Polar and Alpine Microbial Collection(PAMC) at the Korea Polar Research Institute. This microorganism was confirmed for the excretion of amylase with Lugol's solution. The amylase activity was after flask culture was as low as 1.66 mU/L before optimization. The physical factors including the inoculum volume, the initial culture pH, and the medium volume were chosen to be optimized for the enhanced amylase production. The calculated results using RSM indicate that the optimal physical factors were 2.49 mL inoculum volume, 6.85 pH and 42.87 mL medium volume with a predicted amylase production of 2.84 mU/L. The experimentally obtained amylase activity was 2.50 mU/L, which was a 150% increase compared to the level before optimization.