• Title/Summary/Keyword: bacterial isolation

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Characteristics on the Incubation of Sulfur Compound-Oxidizing Strains Separated for the Removal of Malodor (악취제거를 위하여 분리한 황화합물 산화균주의 배양특성)

  • Lim, Dong Joon;Lim, Kwang-Hee
    • Korean Chemical Engineering Research
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    • v.47 no.6
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    • pp.788-794
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    • 2009
  • Both strains of KD-1212 and DAH-1056 were isolated and identified from animal manure-contaminated soil by screening bacterial strains for the removal of sulfur compound-malodor with such substrate as sodium thiosulfate or free sulfur. Then the characteristics on the incubation of these microbes were observed under various incubating-condition such as pH, temperature, aerobic or anaerobic, substrate(sulfur compound) concentration, nitrogen and carbon source and rotating speed for mixing, and the optimum incubating condition was established. The optimum pHs of KD-1212 and DAH-1056 were 7.0 and 4.0, respectively, and their optimum temperatures were in the range of $30{\sim}35^{\circ}C$. Another autotrophic strain, ED-1138, was isolated from contaminated soil. The strain DAH-1056 excelled a strain Thiobacillus sp. IW in eliminating hydrogen sulfide during the process of malodor-biofiltration with a fixed strain. The characteristics on the incubation of strain KD-1212 were observed under various substrate-concentrations, nitrogen and carbon sources. KD-1212 favored glucose and maltose, and yeast extract as carbon sources and nitrogen source, respectively. The optimum concentrations of substrate and nitrogen source were 25 mM of sodium thiosulfate and 0.5% yeast extract, respectively for the growth of strain KD-1212.

Isolation and Characterization of Extracellular Polymeric Substances (EPS)-producing bacteria for restoration of burnt forest soils (산불토양복원을 위한 Extracellular Polymeric Substances (EPS) 생성세균의 분리, 동정 및 특성에 관한 연구)

  • Lee, Gun-Young;Song, In-Geun;Chung, Jae-Chun;Kim, Young-Jun
    • Journal of the Korea Organic Resources Recycling Association
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    • v.12 no.4
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    • pp.139-147
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    • 2004
  • We have isolated two bacterial strains, FM-02 and AL-02, which produced EPS from forest soil for the restoration of forest fire by promoting soil aggregation. FM-02 was found to be Gram negative rod and belong to Beta Proteobacterium sp. through 16s-rDNA sequence analysis, and AL-02 was Gram positive rod and showed 81% of similarity to Zoogloea sp. through the analysis of 16s-rDNA sequence. FM-02 and AL-02 produced about 1.8g and 8.3g of EPS, respectively, per 1L of culture as dry weight. Flocculation activity (FA) was also measured in two strains. FM-02 showed 2.31 FA against active carbon, and AL-02 showed 6.21 FA against kaolin clay. From these results, we expect that AL-02 strain will be applied as a good biological material for the reduction of forest soil erosion by wild and rain after fire through promoting coagulation of soil particles.

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Isolation and identification of protease-producing bacteria from the intertidal zone in Jeju Island, Korea (제주 조간대로부터 단백질 가수분해효소를 생산하는 세균의 분리 및 동정)

  • Moon, Young-Gun;Dharaneedharan, Subramanian;Kim, Dong-Hwi;Park, So-Hyun;Heo, Moon Soo
    • Korean Journal of Microbiology
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    • v.51 no.4
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    • pp.382-388
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    • 2015
  • Eleven protease-producing bacteria were isolated from the organisms' external shells and the inorganic materials collected from intertidal zone of Jeju Island, Republic of Korea. The samples were diluted serially, inoculated on Zobell agar plates with 1% skim milk and incubated at $20^{\circ}C$. Clear zone forming colonies were selected as protease-producing bacteria and each strain was identified based on the phylogenetic analysis with their 16S rDNA sequences. Strains JJM125, JJM129, YG47 and YG49 belong to the marine bacterial genus Pseudoalteromonas; strain JJM122 belong to the genus Microbulbifer; strains YG51, YG52, YG62 and YG63 belong to the genus Vibrio; and strain YG65 belong to genus Bacillus. Hence, the present study suggests that these protease producing bacteria could be further used to develop new varieties of protease with various biotechnological applications.

Isolation and characterization of lactic acid bacteria for use as silage additives (사일리지 제조를 위한 유산균 탐색 및 특성연구)

  • Ro, Yu-Mi;Lee, Gwan-Hyeong;Park, InCheol;Kim, Wan-Gyu;Han, Byeong-Hak;You, Jaehong;Ahn, Jae-Hyung
    • Korean Journal of Microbiology
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    • v.52 no.4
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    • pp.444-454
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    • 2016
  • Sixteen lactic acid bacterial strains were isolated from silage and cow dung samples, and characterized to identify their potential as silage additives. They were identified as the members of the genera Lactobacillus, Enterococcus, and Weissella, and clustered into nine groups based on the sequences of the genes for 16S rRNA, RNA polymerase alpha subunit, 60-kDa heat shock protein, and phenylalanyl-tRNA synthase alpha subunit. Among them, the three strains which were genetically similar to L. plantarum showed the fastest growth and pH decrease in MRS and rye extract media, the highest numbers of available carbohydrates, and the widest ranges of pH, temperature, and salinity for growth. In addition, they showed no amplified DNA products in the PCR examination targeting the genes for the production of biogenic amines, and the MRS media where they had been cultured showed relatively high inhibition effect against the growth of silage-spoiling microorganisms, including fungi, yeast, and clostridia. The results suggest that these strains are good candidates for silage additives. However, the rye extract media where the lactic acid bacteria had been cultured had no effect on or stimulated the growth of the silage-spoiling microorganisms, and the causes must be established for the practical use of the lactic acid bacteria as silage additives.

Early Results of Mitral Valve Reconstruction in Mitral Regurgitation (승모판막 폐쇄부전에 있어 승모판막 성형술의 단기성적)

  • Kim, Kyung-Hwan;Won, Tae-hee;Kim, Ki-Bong;Ahn, Hyuk
    • Journal of Chest Surgery
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    • v.33 no.1
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    • pp.32-37
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    • 2000
  • Background: Reconstruction surgery of mitral valve regurgitation is now considered as an effective operative technique and has shown good long-term results. Although reconstructive surgery of mitral valve has been performed since 1970s, we have started only in early 1990s in full scale because of small number of the mitral regurgitation compared to mitral stenosis and lack of knowledge from the viewpoint of patients and physicians. Material and Method: From January 1992 to December 1996, 100 patients underwent repair of the mitral valve for mitral regurgitation with or without mitral stenosis in Seoul National University Hospital. 45(45%) of the patients were men and 55(55%) were women. The mean age was 39.9$\pm$14.4 years. The causes of the mitral regurgitation were rheumatic in 61, degenerative in 28 and others in 11. According to the Carpentier's pathological classification of mitral regurgitation 5 patients were type I. 55 patients were type II and 40 patients were type III. 7 patients underwent concomitant aortic valvuloplasty and 8 patients underwent aortic valve replacement. 7 patients underwent Maze operation or pulmonary vein isolation. Result: There were no operative death but 3 major operative complications: 2patients were postoperative low cardiac output syndrome(needed intra-aortic ballon pump support) and 1 patient was postoperative bleeding. There was one late death(1.0%) The cause of death was sepsis secondary to acute bacterial endocarditis. 3 patients required reoperation for recurred mitral regurgitation. There were no statistically significant risk factors for reoperation. The other 96 patients showed no or mild degree of mitral regurgitation 99 survivors were in NYHA functional class I or II. There were two throumboembolisms but no anticoagulation-related complications. Conclusion: We concluded that mitral valve repair could be performed successfully in most cases of mitral regurgitation even in the rheumatic and combined lesions with very low operative mortality and morbidity. The early results are very promising.

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Anaerobic Bacterial Degradation for the Effective Utilization of Biomass

  • Ohmiya, Kunio;Sakka, Kazuo;Kimura, Tetsuya
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.6
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    • pp.482-493
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    • 2005
  • Biomass is originally photosynthesized from inorgainic compounds such as $CO_2$, minerals, water and solar energy. Recent studies have shown that anaerobic bacteria have the ability to convert recalcitrant biomass such as cellullosic or chitinoic materials to useful compounds. The biomass containing agricultural waste, unutilized wood and other garbage is expected to utilize as feed, food and fuel by microbial degradation and other metabolic functions. In this study we isolated several anaerobic, cellulolytic and chitinolytic bacteria from rumen fluid, compost and soil to study their related enzymes and genes. The anaerobic and cellulolytic bacteria, Clostridium thermocellum, Clostridium stercorarium, and Clostridium josui, were isolated from compost and the chitinolytic Clostridium paraputrificum from beach soil and Ruminococcus albus was isolated from cow rumen. After isolation, novel cellulase and xylanase genes from these anaerobes were cloned and expressed in Escherichia coli. The properties of the cloned enzymes showed that some of them were the components of the enzyme (cellulase) complex, i.e., cellulosome, which is known to form complexes by binding cohesin domains on the cellulase integrating protein (Cip: or core protein) and dockerin domains on the enzymes. Several dockerin and cohesin polypeptides were independently produced by E. coli and their binding properties were specified with BIAcore by measuring surface plasmon resonance. Three pairs of cohesin-dockerin with differing binding specificities were selected. Two of their genes encoding their respective cohesin polypeptides were combined to one gene and expressed in E. coli as a chimeric core protein, on which two dockerin-dehydrogenase chimeras, the dockerin-formaldehyde dehydrogenase and the dockerin-NADH dehydrogenase are planning to bind for catalyzing $CO_2$ reduction to formic acid by feeding NADH. This reaction may represent a novel strategy for the reduction of the green house gases. Enzymes from the anaerobes were also expressed in tobacco and rice plants. The activity of a xylanase from C. stercorarium was detected in leaves, stems, and rice grain under the control of CaMV35S promoter. The digestibility of transgenic rice leaves in goat rumen was slightly accelerated. C. paraputrificum was found to solubilize shrimp shells and chitin to generate hydrogen gas. Hydrogen productivity (1.7 mol $H_2/mol$ glucos) of the organism was improved up to 1.8 times by additional expression of the own hydrogenase gene in C. paraputrficum using a modified vector of Clostridiu, perfringens. The hydrygen producing microflora from soil, garbage and dried pelletted garbage, known as refuse derived fuel(RDF), were also found to be effective in converting biomass waste to hydrogen gas.

Isolation and Characterization of Bacillus subtilis CA105 from Spent Mushroom (Pleurotus ostreatus) Substrates (느타리버섯 수확후배지로부터 분리한 Bacillus subtilis CA105의 특성)

  • Kim, Hye Soo;Kim, Chul Hwan;Kwon, Hyun Sook;Lee, Chan-Jung;Kong, Won-Sik;Cho, Soo Jeong
    • Journal of Mushroom
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    • v.13 no.4
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    • pp.305-309
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    • 2015
  • In order to isolate compost-promoting bacteria with high activity of cellulase and xylanase, spent mushroom substrates with sawdust were collected from mushroom cultivation farm, Jinju, Gyeongnam in Korea. Among of the isolates, one strain, designated CA105 was selected by agar diffusion method. The strain CA105 was identified as members of the Bacillus subtilis by biochemical characteristics using VITEK 2 system. Comparative 16S rRNA gene sequence analysis showed that isolate CA105 formed a distinct phylogenetic tree within the genus Bacillus and was most closely related to Bacillus subtilis with 16S rRNA gene sequence similarity of 98.9%. On the basis of its physiological properties, biochemical characteristics and phylogenetic distinctiveness, isolate CA105 was classified within the genus Bacillus subtilis, for which the name Bacillus subtilis CA105 is proposed. The cellulase and xylanase activity of B. subtilis CA105 was slightly increased according to bacterial population from exponential phase to stationary phase in growth curve for Bacillus sp. CA105.

Isolation, Identification and Antagonisms of Rhizospheric Antagonists to Cucumber Wilt Pathogen, Fusarium oxysporum f. sp. cucumerinum Owen (오이 덩굴쪼김병균에 대한 오이 근권길항미생물의 분리, 동정 및 길항작용)

  • Jee Hyeong Jin;Kim Hee Kyu
    • Korean Journal Plant Pathology
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    • v.3 no.3
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    • pp.187-197
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    • 1987
  • Bacteria and fungi antagonistic to Fusarium oxysporum f. sp. cucumerinum Owen were effectively isolated with each of modified Triple Layer Agar (TLA) technique from rhizosphere soil where cucumber had been grown healthily in plastic film house. Three predominant bacterial isolates selected were identified as Pseudomonas fluorescens, and P. putida, Serratia sp. and three fungal isolates were Gliocladium sp. Trichoderma harzianum, and T. viride. Antagonistic bacteria inhibited $26-45\%$ of germination and $41-56\%$ of germ tube elongation of microconidia of F. oxysporum f. sp. cucumerinum on Water Agar (WA). P. fluorescens was the strongest inhibitor. Several my co parasitisms were observed on dual culture of WA between antagonistic fungi and F. oxysporum f. sp. cucumerinum such as coiling, penetration, overgrowing, and lysis. Mycelial lysis of the pathogen was the most severe at pH 4.6, followed by 3.6, 5.6 and 6.6 of the medium in decreasing order. At pH 6.6, mycelia of the pathogen were not conspicuously damaged, however, the antagonistic fungi formed abundant chlamydospores especially Gliocladium sp. T. harzianum revealed the most excellent antagonism in vitro.

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Probiotic Characterization of Acid- and Bile-tolerant Lactobacillus salivarius subsp. salivarius from Korean Faeces

  • Bae, H.C.;Nam, M.S.;Lee, J.Y.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.12
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    • pp.1798-1807
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    • 2002
  • This study was conducted to investigate lactobacillus salivarius subsp. salivarius having probiotic properties to be used as the health adjuncts with fermented milk products. Acid- and bile-tolerant lactobacillus salivarius subsp. salivarius was isolated with lactobacilli MRS broth from faeces of 80 healthy persons (infants, children and adults). It was used as a probiotic strain in fermented milk products. The pH of fermented milk decreased from pH 6.7 to 5.0 and titratable acidity increased from 0.3% to 1.0% by L. salivarius subsp. salivarius (isolation strain 20, 35, and 37), when incubated for 36 h at 37$^{\circ}C$. The number of viable cell counts of fermented milk was maximized at this incubation condition. The SDS-PAGE evidenced no significant change of casein but distinct changes of whey protein were observed by isolated L. salivarius subsp. salivarius for titratable acidity being incubated by 0.9-1.0% at 37$^{\circ}C$. All of the strains produced 83.43 to 131.96 mM of lactic acid and 5.39 to 26.85 mM of isobutyric acid in fermented products. The in vitro culture experiment was performed to evaluate ability to reduce cholesterol levels and antimicrobial activity in the growth medium. The selected L. salivarius subsp. salivarius reduced 23-38% of cholesterol content in lactobacilli MRS broth during bacterial growth for 24 h at 37$^{\circ}C$. All of the isolated L. salivarius subsp. salivarius had an excellent antibacterial activity with 15-25 mm of inhibition zone to E. coli KCTC1039, S. enteritidis KCCM3313, S. typhimurium M-15, and S. typhimurium KCCM40253 when its pH had not been adjusted. Also, all of the isolated L. salivarius subsp. salivarius had partial inhibition zone to E. coli KCTC1039, E. coli KCTC0115 and S. enteritidis KCCM3313 when it had been adjusted to pH 5.7. The selected strains were determined to have resistances of twelve antibiotic. Strains 27 and 35 among the L. salivarius subsp. salivarius showed the highest resistance to the antibiotics. These results indicated that some of the L. salivarius subsp. salivarius (strain 27 and 35) are considered as effective probiotic strains with a potential for industrial applications, but the further study is needed to establish their use as probiotics in vivo.

Isolation and Characterization of Dextrans Produced by Leuconostoc sp. strain JYY4 from Fermented Kimchi

  • Gu, Ji-Joong;Ha, Yoo-Jin;Yoo, Sun-Kyun
    • Journal of the Korean Applied Science and Technology
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    • v.32 no.4
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    • pp.758-766
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    • 2015
  • Dextran is a generic term for a bacterial exopolysaccharide synthesized from sucrose and composed of chains of D-glucose units connected by ${\alpha}$-1,6-linkages by using dextransucrases. Dextran could be used as vicosifying, stabilizing, emulsifying, gelling, bulking, dietary fiber, prebiotics, and water holding agents. We isolated new strain capable of producing dextran from Korean traditional kimchi and identified as Leuconostoc sp. strain JYY4. Batch fermentation was conducted in bioreactor with a working volume of 3 L. The media was MMY and 15% (w/v) sucrose. Mineral medium consisted of $3.0g\;KH_2PO_4$, $0.01g\;FeSO_4$, $H_2O$, $0.01g\;MnSO_4$, $4H_2O$, $0.2g\;MgSO_4\;7H_2O$, 0.01 g NaCl, $0.05g\;CaCl_2$ per 1 liter deionized water. The pH of media was initially adjusted to 6.0. The inoculation rate was 1.0% (v/v) of the working volume. Temperature was maintained at $28^{\circ}C$. The agitation rate was 100 rpm. The production pattern of dextran was associated with the cell growth. After 24 hr dextran reached its highest concentration of 59.4 g/L. The sucrose was consumed completely after 40 hr. Growth reached stationery phase when sucrose became limiting, regardless of the presence of fructose or mannitol. When the specific growth rate was 0.54 hr-1, utilization averaged 5.8 g/L-hr. The yield and productivity of dextran were 80% and 2.0 g/L-hr, respectively. Dextrans produced by were separated to two different size by an alcohol fraction method. The size of high molecular weight dextran (45% alcohol, v/v), less soluble dextran, was between MW 500,000 and 2,000,000. Soluble dextran (55% alcohol, v/v) was between 70,000 and 150,000. The molecular weight average of total dextran (70% alcohol, v/v) was between 150,000 to 500,000. The enzymatic hydrolyzates of total dextran of ATCC 13146 showed branched dextrans by Penicillium dextranase contained of glucose, isomaltose, isomaltotriose, and isomaltooligosaccharides greater than DP4 (degree of polymerization) that had branch points. Compounds greater than DP4 were branched isomaltooligosaacharides. Hydrolysates by the Lipomyces dextranase produced the same composition of oligosaccharides as those by Penicillin dextranase.