• Title/Summary/Keyword: Microbiological assay

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Simultaneous Removal of Phenol and Hexavalent Chromium by Rhodococcus sp. CP01 (Rhodococcus sp. CP01에 의한 페놀과 6가 크롬이온의 동시 제거)

  • 최광현;오영숙;김병동;최성찬
    • Korean Journal of Microbiology
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    • v.36 no.4
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    • pp.279-284
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    • 2000
  • Simultaneous reduction of Cr(VI) and degradation of phenol was observed in batch and bench-scale continuous stirred tank reactors using Rhodococcus sp. CP01 isolated from leachate. The strain CP01, which was capable of utilizing phenol as a sole source of carbon and energy, completely reduced added hexavalent chromium (0.25 mM) to its trivalent form during 60 hr batch assay under optimal conditions (pH 7.0 and 1,000 mg/L of phenol concentration). The rates of Cr(VI) reduction and phenol degradation were estimated as 4.17 $\mu$M Cr(VI) and 38.4 mg phenol.$L^{-1}{\cdot}hr^{-1}$, respectively. The continuous culture experiment was conducted for 46 days using synthetic feed containing different levels of chromate (0.0625 to 0.25 mM) and phenol(1,000 to 4,000 mg/L). With a hydraulic retention time of 100 hr, Cr(VI) reduction efficiency was mostly 100% for influent Cr(VI) and phenol concentrations of 0.125 mM and 3,000 mg/L, respectively. During quasi-steady-state operation, specific rate of Cr(VI) reduction was calculated as 0.34 mg Cr(VI).g $protein^{-1}{\cdot}hr^{-1}$ which was comparable to reported values obtained by using glucose as growth substrate. The results suggest the potential application of biological treatment for detoxification of wastewater contaminated simultaneously with Cr(VI) and pheonol.

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Purification and Assay of Extracellular Autolysin from Moraxella sp. CK-l (Moraxella sp. CK-1의 세포외 Autolysin의 분리 정제 및 활성도 측정)

  • 오영상;이장현;한명수;윤문영
    • Korean Journal of Microbiology
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    • v.39 no.3
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    • pp.148-154
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    • 2003
  • Moraxella sp. CK-l is known to inhibits the growth of Anabaena cylindrica, a cyanobacterium. It has been documented that the ability of this growth inhibition of Anabaena cylindrica was attributed to extracellular autolysin from Moraxella sp. CK-l. However, it remains to be elucidated identification and characterization of autolysin have yet been elucidated. In this study, we tried to purify and identify autolysin secreted from Moraxella sp. CK-l. Cells were grown in a complex liquid medium (BGC-11) and culture supernatants were collected, followed by ammonium sulfate fractionation. Fractions were further separated with anion exchange column, Mono-Q, in FPLC system and analyzed by SDS/PAGE. The fraction containing high autolysin activity showed a single distinct protein peak in anion column and molecular mass of about 17 kDa in SDS/PAGE. Nterminal amino acid sequencing of the protein was analyzed, of which result showed the homology with some proteases, including extracellular serine protease, Dichelobacter nodosus.

Antifungal Activity of Rheum undulatum on Candida albicans by the Changes in Membrane Permeability (막투과성 변화로 인한 대황의 Candida albicans에 대한 항진균 활성)

  • Lee, Heung-Shick;Kim, Younhee
    • Korean Journal of Microbiology
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    • v.50 no.4
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    • pp.360-367
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    • 2014
  • Candida albicans is an opportunistic and the most prevalent fungal pathogen that can cause superficial and systemic infections in immunocompromised patients. C. albicans can promote the transition from budding yeast to filamentous form, generating biofilms. Infections associated with C. albicans biofilms are frequently resistant to conventional antifungal therapy. Therefore, the development of more effective antifungal drugs related with biofilm formation is required urgently. The roots of Rheum undulatum have been used for medicinal purposes in Korea and China traditionally. The aim of present study was to evaluate the effect of R. undulatum extract upon preformed biofilms of 12 clinical C. albicans isolates and the antifungal activities. Its effect on preformed biofilms was evaluated using XTT reduction assay, and metabolic activity of all tested strains was reduced significantly ($49.4{\pm}6.0%$) at 0.098 mg/ml R. undulatum. The R. undulatum extract blocked the adhesion of C. albicans biofilms to polystyrene surfaces, and damaged the cell membrane integrity of C. albicans which was analyzed by CFDA, AM, and propidium iodide double staining. It caused cell lysis which was observed by Confocal laser scanning and phase contrast microscope after propidium iodide and neutral red staining, respectively. Membrane permeability was changed as evidenced by crystal violet uptake. The data suggest that R. undulatum inhibits biofilm formation by C. albicans, which can be associated with the damage of the cell membrane integrity, the changes in the membrane permeability and the cell lysis of C. albicans.

Isolation and Phylogenetic Characterization of Chitinase Producing Oligotrophic Bacteria (Chitinase생산 저영양세균의 분리 및 계통분류학적 특성)

  • Kim Soo-Jin;Kim Min-Young;Koo Bon-Sung;Yoon San-Hong;Yeo Yun-Soo;Park In-Cheol;Kim Yoon-Ji;Lee Jong-Wha;Whang Kyung-Sook
    • Korean Journal of Microbiology
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    • v.41 no.4
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    • pp.293-299
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    • 2005
  • Many isolates from soil of Korean ginseng rhizosphere did not show remarkable growth on full strength of the conventional nutrient broth (NB medium) but grew on its 100-fold dilution (DNB medium). Six hundred-forty strains were isolated as oligotrophic bacteria. In the course of screening for new bioactive compounds from oligotrophic bacteria from soil, 8 strains which had appeared to form of clear zone on a medium containing colloidal chitin as a sole carbon source were selected for further studies. Strain CR42 hydrolyzed a fluorogenic analogue of chitin, 4-methylumbelliferyl-D-glucosaminide (MUF-NAG) . Mo st of the culture supernatant of these isolates hydrolyzed 4-methylumbelliferyl-D-N,N'-diacetylchitobioside (MUF-diNAG). The isolates were heterogeneous and categorized to gamma- and beta-proteobacteria, Bacillaceae, Actinobactepia, and Bacteroides by 16S rRNA analysis. Two strains, WR164 and CR18, had a 16S rRNA sequence of $95-96\%$ identical to uncultured bacteria. It was observed that CR2 and CR75 could inhibit the growth of Colletotrichum gloeosporioides with hyphal extention-inhibition assay on PDA plate supplemented with $1\%$ colloidal chitin.

Characterization and Antimicrobial Activity of Lactic Acid Bacteria Isolated from Vaginas of Women of Childbearing Age (가임기 여성의 질에서 분리한 젖산 세균인 Lactobacillus plantarum UK-3의 특성 및 항균활성)

  • Ahn, Hye-Ran;So, Jae-Seong;Oh, Kye-Heon
    • Korean Journal of Microbiology
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    • v.47 no.4
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    • pp.308-315
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    • 2011
  • The purpose of this work was to examine the antimicrobial activity derived from the lactic acid bacterium, UK-3 isolated from the vaginas of women of childbearing age. Various physiological and biochemical properties of this strain were characterized. Both the BIOLOG system and phylogenetic analysis using 16S rRNA sequencing were utilized for identification, and the strain was designated as Lactobacillus plantarum UK-3, and registered in GenBank as [JK266589]. Growth rate, production of organic acids (e.g., lactic acid and acetic acid), and pH during growth were monitored. The maximum concentrations of lactic acid and acetic acid were approximately 684.11 mM and 174.26 mM, respectively, and pH changed from 7.0 to 3.7 after 72 h of incubation. High performance liquid chromatography was used to confirm lactic acid and acetic acid production. Significant antimicrobial activity of the concentrated supernatant was demonstrated against various Gram-positive (e.g., Staphylococcus aureus, Staphylococcus epidermidis, Methicillin-resistant Staphylococcus aureus, Enterococcus faecalis, Neisseria species., Listeria monocytogenes), Gram-negative bacteria (e.g., Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis), and yeast (e.g., Candida albicans) by the plate diffusion method. As a result, the concentrated L. plantarum UK-3 cultures had lower acidity and inhibited the growth of all microorganisms tested, whereas the growth of L. acidophilus was not affected.

Analysis of diversity of hemolytic microbiome from aquafarm of arkshell, Scapharca broughtonii (피조개 양식장 내 용혈성 미생물의 다양성 분석)

  • Gwon, Byeong-Geun;Kim, Young-Ok;Nam, Bo-Hye;Kim, Woo-Jin;Kong, Hee Jeong;Kim, Bong-Seok;Jee, Young-Ju;Lee, Sang-Jun;An, Cheul Min;Kim, Dong-Gyun
    • Journal of fish pathology
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    • v.26 no.3
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    • pp.193-206
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    • 2013
  • The ark shell, Scapharca broughtonii is a marine bivalve mollusks belonging to the family Arcidae and important seafood for Korean and Japanese, and southern coast is brisk bays for the ark shell aquaculture. However, productivity of ark shell from these regions were rapidly reduced during the last decade due to mass mortality. The reason of this great damage has not yet been identified. To overcome this economic loss, diverse investigations were focused on environmental factors that affects in the physiology of S. broughtonii, but microbiological researches were performed insufficiently. Hemoglobin is one of the major blood component of ark shell and is damaged by some species of bacterial toxins. We concentrated on this red pigment because hemolysis could be the cause of ark shell mortality. In this study, we analyzed microbial diversity of underwater sediments in coastal regions and also existences in the body of S. broughtonii. We investigate about 4,200 isolates collected from June to September for microbial diversity of sediments and ark shell. We screened all of culturable microorganisms, and identified 25 genera 118 species, 24 genera 89 species, 30 genera 109 species and 39 genera 141 species, and selected 140 unique colonies for identification and challenge assay.

H-NS binding on dicA promoter DNA inhibits dicA gene expression (dicA promoter DNA에 붙는 H-NS 단백질에 의한 dicA 유전자의 발현 조절)

  • Yun, Sang Hoon;Lee, Yonho;Lim, Heon M.
    • Korean Journal of Microbiology
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    • v.55 no.3
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    • pp.191-198
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    • 2019
  • H-NS binds to promoter DNA and works as a general transcription silencer. DicA protein, by binding to the promoter DNA of dicA, activates dicA expression and at the same time inhibits expression of dicF and dicB, thus, exerting cell division control in Escherichia coli. H-NS complexed with a nucleoid protein Cnu was known to be involved in dicA expression. However, the exact nature of H-NS binding to dicA promoter DNA and the consequences of H-NS binding in expression of dicA is not clear. In this study, we explored the DNA binding activity of H-NS on the promoter DNA of dicA and found that H-NS binding occurs exclusively to the dicA promoter DNA. We never observed, however, H-NS binding at the vicinity of the dicA promoter. Temperature dependent oligomerization of H-NS was observed during DNA binding and the Cnu protein enhances the oligomerization process of H-NS binding. In vivo measurement of dicA expression in an hns deleted strain showed that dicA expression increased. These results demonstrated that H-NS binds specifically to dicA promoter DNA and functions as a transcription silencer.

Improving amber suppression activity of an orthogonal pair of Saccharomyces cerevisiae tyrosyl-tRNA synthetase and a variant of E. coli initiator tRNA, fMam tRNACUA, for the efficient incorporation of unnatural amino acids (효율적인 비천연 아민노산 도입을 위한 효모균 타이로신-tRNA 합성효소와 대장균 시작 tRNA 변이체의 엠버써프레션 활성증가)

  • Tekalign, Eyob;Oh, Ju-Eon;Park, Jungchan
    • Korean Journal of Microbiology
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    • v.54 no.4
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    • pp.420-427
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    • 2018
  • The orthogonal pair of Saccharomyces cerevisiae tyrosyl-tRNA synthetase (Sc YRS) and a variant of E. coli initiator tRNA, fMam $tRNA_{CUA}$ which recognizes the amber stop codon is an effective tool for site-specific incorporation of unnatural amino acids into the protein in E. coli. To evolve the amber suppression activity of the orthogonal pair, we generated a mutant library of Sc YRS by randomizing two amino acids at 320 and 321 which involve recognition of the first base of anticodon in fMam $tRNA_{CUA}$. Two positive clones are selected from the library screening with chloramphenicol resistance mediated by amber suppression. They showed growth resistance against high concentration of chloramphenicol and their $IC_{50}$ values were approximately 1.7~2.3 fold higher than the wild type YRS. In vivo amber suppression assay reveals that mutant YRS-3 (mYRS-3) clone containing amino acid substitutions of P320A and D321A showed 6.5-fold higher activity of amber suppression compared with the wild type. In addition, in vitro aminoacylation kinetics of mYRS-3 also showed approximately 7-fold higher activity than the wild type, and the enhancement was mainly due to the increase of tRNA binding affinity. These results demonstrate that optimization of anticodon recognition by engineered aminoacyl tRNA synthetase improves the efficiency of unnatural amino acid incorporation in response to nonsense codon.

Effect of rearing system (free-range vs cage) on gut and muscle histomorphology and microbial loads of Italian White breed rabbits

  • Caterina Losacco;Antonella Tinelli;Angela Dambrosio;Nicoletta C. Quaglia;Letizia Passantino;Michele Schiavitto;Giuseppe Passantino;Vito Laudadio;Nicola Zizzo;Vincenzo Tufarelli
    • Animal Bioscience
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    • v.37 no.1
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    • pp.151-160
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    • 2024
  • Objective: The growing consumers' interest on animal welfare has raised the request of products obtained by alternative rearing systems. The present study was conducted to assess the influence of housing system on gut and muscle morphology and on microbial load in rabbits reared under free-range (FR) and cage system (CS). Methods: A total of forty weaned (35 days of age) male Italian White breed rabbits were allotted according to the rearing system, and at 91 days of age were randomly selected and slaughtered for the morphological evaluation of tissue from duodenum and longissimus lumborum. Morphometric analysis of the villus height, villus width, crypt depth, villus height/crypt depth ratio, and villus surface was performed. The microbial loads on hind muscle was determined by total mesophilic aerobic count (TMAC), Escherichia coli and Enterobacteriaceae; whereas, total anaerobic bacteria count (TABC) and TMAC, E. coli and Enterobacteriaceae was determined on caecal content. Results: Rearing system did not interfere with the duodenum and muscle histomorphology in both rabbit groups. Similarly, microbial load of caecal content showed no significant differences on the TABC and TMAC. Conversely, significant difference was found for E. coli strains in caecal content, with the lower counts in FR compared to CS rabbits (p<0.01). Microbiological assay of muscle revealed significant lower TMAC in FR vs CS rabbits (p< 0.05). All rabbit meat samples were negative for E. Coli and Enterobacteriaceae. Conclusion: Free-range could be considered a possible alternative and sustainable rearing system in rabbits to preserve gut environment and muscle quality.

Total Folate Contents of 15 Edible Plants Consumed in Korea Using Trienzyme Extraction Method (국내 소비되는 봄나물의 Trienzyme 추출법을 적용한 엽산 함량 분석)

  • Kim, Bo Min;Kim, So-Min;Oh, Ji Yeon;Cho, Young-Sook;Kim, Se-Na;Choi, Youngmin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.11
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    • pp.1796-1800
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    • 2014
  • Trienzyme digestion (AOAC Official Method 2004.05) procedure using protease, ${\alpha}$-amylase, and chicken pancreas conjugase was evaluated to determine its usefulness in the microbiological quantitation of total folate in foods. Folate values obtained by alkali hydrolysis (Korean Food Standards Codex) were compared to those obtained by the trienzyme method for four certified reference materials (CRM) representing diverse matrixes. Trienzyme treatment increased measurable folate from most CRM compared to levels found after alkali hydrolysis. The largest increases were observed with CRM 487 (pig liver, 5.8-fold) and CRM 121 (whole meal flour, 3.1-fold) after trienzyme digestion. Using trienzyme digestion method, total folate contents of raw and blanched edible plants were determined. Eleutherococcus senticosus ($146.9{\mu}g/100g$) showed the highest total folate content, followed by Aster glehni F. Schmidt ($142.8{\mu}g/100g$) and Ledebouriella seseloides H. Wolff ($140.4{\mu}g/100g$) on a wet weight basis. Blanching of samples resulted in an insignificant decrease in folate content for five samples and 11~63% reduction for nine samples. Our finding suggests that trienzyme digestion method is accurate for the determination of food folate in leafy vegetables.